Pseudomonas aeruginosa

Biochem J, 1986 Nov 1, 239(3), 733 - 8
Isolation and identification of uridine(5')-diphospho(1)-2,3-diacetamido-2,3-dideoxy-alpha-D- glucopyra nuronic acid from Pseudomonas aeruginosa P1-III; Okuda S et al.; A new uridine nucleotide was isolated from Pseudomonas aeruginosa P1-III (Habs serotype 5) . On the basis of 13C-n.m.r . and p.m.r . spectroscopy, mass spectrometry, i.r.-absorption spectroscopy and circular dichrometry, the structure of the new compound was unequivocally identified as uridine(5')-diphospho(1)-2,3-diacetamido-2,3-dideoxy-alpha-D-gl ucopyranuronic acid.

Mol Gen Mikrobiol Virusol, 1986 Nov, (11), 18 - 23
{Genetic characteristics and physical organization of the R-plasmid pBS52 with a broad range of bacterial hosts}; Polevoda BV et al.; The genetic and physical data on Pseudomonas aeruginosa plasmid pBS52 coding for the resistances to ampicillin, streptomycin and sulfonamids have been obtained . This conjugative plasmid is transferable to a broad range of gram-negative bacterial hosts and compatible with the broad host-range plasmids from all known incompatibility groups . The plasmid size has been determined (38 Kb) and a physical map has been constructed using restriction endonucleases EcoRI, EcoRV, BamHI, BglII, PstI, PvuII, SalI, SlaI . The presence of a fragment, approximately 200 bp in size, which contains the sites for many of widely used restriction endonucleases is a characteristic feature of the plasmid pBS52.

Mol Biol (Mosk), 1986 Nov-Dec, 20(6), 1589 - 92
{Effect of ligand orientation on the redox potential of homologous proteins}; Denisenko AN; Using experimental g-values of homologous cytochromes isolated from the horse heart and bacterial cyt-c-551 from Pseudomonas aeruginosa, the electron energy difference of redox-orbital of Fe(III) ion of heme was calculated during reduction . Data are in a good accordance with experimental redox-potential values for these proteins . The model gives opportunity to predict redox-potential values for other homologous proteins.

Genetika, 1986 Nov, 22(11), 2721 - 7
{Cloning and characteristics of recA gene in Pseudomonas aeruginosa}; Zaitsev EN et al.; A recA-like gene from Pseudomonas aeruginosa was cloned and identified by means of interspecific complementation of gene recA repair defect in Escherichia coli . The gene was mapped in the PvuII-HindIII Ps . aeruginosa chromosome fragment of 1.5 kbp in length . Having been recloned in pUC18 or 19 plasmids in either of possible orientations, this fragment was shown to complement three different defects of E . coli recA mutants: in repair, recombination and SOS functions.

Genetika, 1986 Nov, 22(11), 2637 - 48
{2 types of molecular structure (composition) of a genome in one species of transposable bacteriophages of Pseudomonas aeruginosa}; Krylov VN et al.; Comparison of heteroduplexes (HD) between DNAs of different transposable phages of Pseudomonas aeruginosa belonging to two previously described subgroups (D3112 and B3) revealed two types of structure (composition) of the bacteriophages, designated "type A" and "type B" . The properties of genome structure of type A (phages of D3112 subgroup) are as follows: high level of conservation (up to 70% of genomes of different phages are represented as blocks of homologous DNA sequences); substitutions in genomes revealed as nonhomology regions in HD are, as a rule, small and located in certain sites; the distribution of the nonhomologous regions in HD of these phages is highly reproducible in independent experiments . Bacteriophages of subgroup B3 have genomes of type B: only a small part (approx . 30%) of genomes retain homology general for all of the phages; the nonhomologous regions are distributed in a large number of sites in HD; the sizes of nonhomologous regions are substantially larger than for the phages of subgroup D3112; distribution of the regions in HD is highly variable, which is characteristic of DNAs with partial homology . There is no difference between genomes of types A and B in G + C content (approx . 61-63%) . Viable recombinants can be formed in crosses between phages of different genome types not only in regions with earlier revealed large DNA/DNA homology (right ends of genomes), but also in central portions of the genomes . Nevertheless, functional incompatibility of some regions of phage genomes of types A and B was demonstrated.

Antimicrob Agents Chemother, 1986 Nov, 30(5), 802 - 5
Distribution of porin and lipopolysaccharide antigens on a Pseudomonas aeruginosa permeability mutant; Godfrey AJ et al.; Pseudomonas aeruginosa common surface antigens were compared in a permeability mutant (PCC118) and its parent (PAO503) . The distribution of lipopolysaccharide and porin antigens in the mutant supports the conclusion that beta-lactam permeability was affected by lipopolysaccharide-side chain presentation rather than by a change in porin number.

J Trauma, 1986 Nov, 26(11), 1013 - 23
Factor XIII as a modulator of plasma fibronectin alterations during experimental bacteremia; Kiener JL et al.; Fibronectin is found in plasma as well as in association with connective tissue and cell surfaces . Depletion of plasma fibronectin is often observed in septic trauma and burned patients, while experimental rats often manifest hyperfibronectinemia with sepsis . Since Factor XIII may influence the rate of clearance and deposition of plasma fibronectin into tissues, we evaluated the temporal changes in plasma fibronectin and plasma Factor XIII following bacteremia and RE blockade in rats in an attempt to understand the mechanism leading to elevation of fibronectin levels in bacteremic rats, which is distinct from that observed with RE blockade . Clearance of exogenously administered fibronectin after bacteremia was also determined . Rats received either saline, Pseudomonas aeruginosa (1 X 10(9) organisms), gelatinized RE test lipid emulsion (50 mg/100 gm B.W.), or emulsion followed by Pseudomonas . Plasma fibronectin and Factor XIII were determined at 0, 2, 24, and 48 hours post-blockade or bacteremia . At 24 and 48 hr following bacteremia alone or bacteremia after RE blockade, there was a significant elevation (p less than 0.05) of plasma fibronectin and a concomitant decrease (p less than 0.05) of plasma factor XIII activity . Extractable tissue fibronectin from liver and spleen was also increased at 24 and 48 hours following R.E . blockade plus bacteremia . In addition, the plasma clearance of human fibronectin was significantly prolonged (p less than 0.05) following bacterial challenge . Infusion of activated Factor XIII (20 units/rat) during a period of hyperfibronectinemia (908.0 +/- 55.1 micrograms/ml) resulted in a significant (p less than 0.05) decrease in plasma fibronectin (548.5 +/- 49.9 micrograms/ml) within 30 min . Thus Factor XIII deficiency in rats with bacteremia may contribute to the elevation in plasma fibronectin by altering kinetics associated with the clearance of fibronectin from the blood.

J Bacteriol, 1986 Nov, 168(2), 574 - 80
Expression of pili from Bacteroides nodosus in Pseudomonas aeruginosa; Elleman TC et al.; The pili of Bacteroides nodosus, the causative agent of ovine footrot, constitute the major host-protective immunogen against homologous serotypic challenge . The pilin gene from B . nodosus 198 has been cloned and morphologically expressed as extracellular pili in Pseudomonas aeruginosa by using a plasmid-borne, thermoregulated expression system . B . nodosus pilin could not be detected in cultures of P . aeruginosa grown at 32 degrees C, but after induction at 37 degrees C, B . nodosus pili were expressed on the cell surface of P . aeruginosa to the virtual exclusion of the host cell pili . Pili harvested from induced P . aeruginosa cultures were used to immunize sheep against footrot . The serum agglutinating antibody titers of vaccinated sheep were comparable to those of sheep receiving pili from B . nodosus . Subsequent challenge of the sheep with B . nodosus 198 indicated that the recombinant- DNA-derived pili vaccine and the B . nodosus pili vaccine provided similar levels of protection against footrot.

Arzneimittelforschung, 1986 Oct, 36(10), 1511 - 7
Antibacterial activity and ototoxicity in guinea pigs, and nephrotoxicity in rats of arbekacin; Kurebe M et al.; Arbekacin (HBK), 1-N{(S)-4-amino-2-hydroxybutyl}-3',4'-dideoxykanamycin B, showed broad antibacterial spectra against gram-positive and gram-negative bacteria including Pseudomonas aeruginosa . It was also effective against gentamycin- or tobramycin-resistant bacteria . HBK was resistant to various aminoglycoside-inactivating enzymes except for AAC (2') and AAC (6')-IV, both of which slowly inactivated it . Even at higher dosages (150 mg/kg i.m . or greater, which resulted in some deaths), HBK never decreased the pinna reflex in guinea pigs, while 150 mg/kg or more of dibekacin (DKB) or amikacin (AMK) caused loss of this reflex . HBK has less ototoxicity than do DKB and AMK . This was confirmed by histopathological examination of the inner ear . The degree of nephrotoxicity of HBK was suggested to be similar to that of DKB as judged from serum biochemical tests, urinalysis, and histopathological findings.

Biochem J, 1986 Oct 1, 239(1), 221 - 4
The determination of specificity constants in enzyme-catalysed reactions; Crompton IE et al.; A convenient and accurate procedure for determining the kinetic parameter Vmax./Km is described . This avoids the error in the usual method of taking the observed first-order rate constant of an enzymic reaction at low substrate concentration as Vmax./Km . A series of reactions is used in which the initial concentration of substrate is below Km (e.g . from 5% to 50% of Km) . Measurements are taken over the same extent of reaction (e.g . 70%) for each member of the series, and treated as if the kinetics were truly first-order . The reciprocal of the observed first-order rate constant is then plotted against the initial concentration of substrate: the reciprocal of the ordinate intercept is Vmax./Km . The procedure, as well as being applicable to simple reactions, is shown to be valid when there is competitive inhibition by the product, or when the reaction is reversible, or when there is competitive or mixed inhibition . The hydrolysis of cephalosporin C by a beta-lactamase from Pseudomonas aeruginosa is used to illustrate the method.

Antimicrob Agents Chemother, 1986 Oct, 30(4), 614 - 6
Pharmacokinetics and sputum penetration of ciprofloxacin in patients with cystic fibrosis; Smith MJ et al.; Levels of ciprofloxacin in serum and sputum were studied for eight patients with cystic fibrosis who were infected with Pseudomonas aeruginosa . Patients were studied in a steady state on a dosage of 500 mg every 8 h . Peak levels in serum ranged from 1.27 to 5.6 mg/liter (mean, 3.16 +/- 1.27), and absorption was rapid, the time to peak concentration ranging from 0.5 to 3.0 h (mean, 1.5 +/- 0.9) . The antibiotic penetrated sputum well, achieving areas under the curve of approximately 46% of those obtained in serum.

Scand J Clin Lab Invest, 1986 Oct, 46(6), 511 - 8
Increased mole fraction of arachidonic acid in bronchial phospholipids in patients with cystic fibrosis; Gilljam H et al.; The fatty acid pattern of the phospholipids in the bronchial secretion of patients with cystic fibrosis (CF) showed an increase of the mole fraction of arachidonic acid (AA) in most phospholipid classes compared to normals . Increase of AA in some classes was also found in patients with chronic bronchitis and in patients chronically colonized with Pseudomonas aeruginosa but in the diphosphatidylglycerol, lysophosphatidylethanolamine and sphingomyelin phospholipids, high fractions of AA was found exclusively in CF patients . Arachidonic acid was found to attain the highest ratios in CF in seven of the nine major bronchial phospholipids compared to the controls . There was no difference in the ratio saturated/unsaturated fatty acids between the CF patients and the control groups . A tendency towards unsaturation of the fatty acids in the bronchial secretion seems to be characteristic of infection and inflammation but AA appears to be more markedly increased in CF . Thus, the recorded changes may be characteristic for CF and not secondary to infection and/or inflammation in general, nor to P . aeruginosa colonization.

Exp Mol Pathol, 1986 Oct, 45(2), 193 - 206
Morphologic and microbiologic features of Pseudomonas aeruginosa pneumonia in normal hamsters; Coalson JJ et al.; A model of pneumonia due to Pseudomonas aeruginosa was produced in hamsters by an intratracheal bolus instillation of microorganisms . Sequential lung changes from 4 hr through 11 days were studied by morphologic and microbiologic methods . Hamsters inoculated with greater than 10(6) pseudomonads survived but consistently had histologic evidence of mild bronchopneumonia 24 hr postinoculation, whereas a severe bronchopneumonia and a 100% mortality were elicited with a 10(8) inoculum of organisms in 0.5 ml phosphate-buffered saline (PBS) . An inoculum of 10(7) pseudomonads/0.5 ml PBS was then used to define the changes in the bacterial population in Pseudomonas pneumonia and to obtain serial histopathologic observations . Quantitative lung cultures obtained within 1 hr postinoculation demonstrated a mean of 10(6) colony forming units per lung, and none of the hamsters were bacteremic . However, by 24 hr bacterial counts had increased and all animals were bacteremic . Bacterial proliferation continued through 48 hr; however, the number of bacteremic animals had decreased . By 72 hr, bacterial counts had decreased with total Pseudomonas clearance noted by 120 hr . A striking polymorphonuclear leukocyte-rich alveolar exudate was present by 12 hr . Pseudomonas "vasculitis" was evident by 24 hr . The evolution of this vascular lesion correlated with the bacteremic state of the hamsters . By 11 days, resolution of the pneumonic process was seen . The macroscopic and microscopic features of this hamster model of Pseudomonas pneumonia are very similar to those reported in infected patients.

Drug Intell Clin Pharm, 1986 Oct, 20(10), 767 - 9
Therapy of choice for the empiric treatment of the febrile neutropenic patient; Barriere SL; Patients who become neutropenic secondary to chemotherapy for malignancy are at high risk for bacterial infection . Since the most common organisms include enteric gram-negative bacilli and staphylococci, drug combinations have traditionally been used . In addition, available clinical evidence suggests that synergistic drug combinations are most effective in the treatment of infections due to Pseudomonas aeruginosa . Comparative clinical trials have not shown any particular drug combination to be superior to others tested . Therefore, the drug choice for the combination should be based on local susceptibility patterns, clinical experience, and cost . Newer beta-lactam compounds are broader in spectrum and more potent than older drugs, and have been tested as sole agents in this setting . Although results appear promising for drugs such as cefoperazone or ceftazidime, the development of resistance and lack of adequate antistaphylococcal activity preclude routine use of these drugs . Imipenem is currently undergoing clinical trials for this purpose and appears to have the appropriate breadth of spectrum . However, frequent development of resistance among isolates of P . aeruginosa is problematic . At the present time a combination of an aminoglycoside and an extended-spectrum penicillin seems to be the therapy of choice for the febrile, neutropenic patient.

Surgery, 1986 Oct, 100(4), 679 - 90
A critical comparison of the hematologic, cardiovascular, and pulmonary response to steroids and nonsteroidal anti-inflammatory drugs in a model of sepsis and adult respiratory distress syndrome; Kopolovic R et al.; Improved survival of patients receiving high-dose steroid therapy in sepsis and adult respiratory distress syndrome (ARDS) has been reported, but such therapy and its benefits remain controversial . Recently research has been directed toward manipulation of the arachidonic acid cascade . Improved survival and hemodynamics with administration of nonsteroidal anti-inflammatory drugs (NSAID) have been reported in animal models of sepsis and ARDS . The purpose of this study was to compare the effects of steroids (methylprednisolone) and NSAID (ibuprofen) in a porcine model of septic ARDS induced by a continuous infusion of live Pseudomonas aeruginosa . Cardiopulmonary parameters were monitored in animals intubated, paralyzed, and ventilated at a 250 ml tidal volume and 0.5 Fio2 . Pigs were randomly assigned to one of five groups: groups I and II received respective doses of 12.5 mg/kg ibuprofen and 30 mg/kg methylprednisolone at 20 and 210 minutes after baseline; group III had P . aeruginosa only; groups IV and V received respective doses of ibuprofen and methylprednisolone at 20 and 210 minutes of sepsis . Significant pulmonary edema, increased intrapulmonary shunting, hypoxemia, hemoconcentration, and systemic hypotension occurred with P . aeruginosa infusion . In septic animals treated with ibuprofen normal systemic arterial pressure was maintained, hemoconcentration was decreased, and oxygenation was improved with a significant decrease in shunting and pulmonary edema . Administration of methylprednisolone improved hemoconcentration and cardiac index, but no significant effect on pulmonary edema, intrapulmonary shunting, or oxygenation was observed . The results of this study demonstrated a significant beneficial effect of ibuprofen and we would encourage controlled clinical trials of this drug in the management of sepsis and ARDS . On the other hand, methylprednisolone was found to be relatively ineffective in treatment of circulatory collapse and ARDS associated with sepsis.

Invest Ophthalmol Vis Sci, 1986 Oct, 27(10), 1466 - 9
The role of the polymorphonuclear leukocyte in the induction of corneal edema; Chusid MJ et al.; Corneal inflammation is frequently associated with the development of corneal edema . It has been suggested the development of corneal edema might in some way be related to the presence of polymorphonuclear leukocytes (PMNLs) within inflamed corneas . In the present studies, it was found that corneal thickness markedly increased after experimental infection with Pseudomonas aeruginosa, but in guinea pigs made neutropenic by whole body irradiation, significantly less of an increase in corneal thickness occurred . Furthermore, corneas from non-neutropenic animals experimentally infected with P . aeruginosa consistently showed a greater increase in water content than did infected corneas from neutropenic animals . Over the first 48 hr of infection, the increase in corneal water was directly proportional to the corneal ingress of radiolabelled PMNLs . Corneal inflammation induced by intracorneal injection of the PMNL chemotactic agents phorbol myristate acetate (PMA) or endotoxin was also associated with a significant increase in corneal water compared with neutropenic animals . These data strongly suggest that activated PMNLs in the cornea are responsible for the induction of corneal edema in infected corneas.

Antimicrob Agents Chemother, 1986 Oct, 30(4), 528 - 31
Efficacy of ciprofloxacin in experimental aortic valve endocarditis caused by a multiply beta-lactam-resistant variant of Pseudomonas aeruginosa stably derepressed for beta-lactamase production; Bayer AS et al.; The emergence of multi-beta-lactam resistance is a limiting factor in treating invasive Pseudomonas infections with newer cephalosporins . The in vivo efficacy of ciprofloxacin, a new carboxy-quinolone, was evaluated in experimental aortic valve endocarditis caused by a strain of Pseudomonas aeruginosa which is stably derepressed for beta-lactamase production and is resistant to ceftazidime and multiple other beta-lactam agents . A total of 51 catheterized rabbits with aortic catheters in place were infected with this strain and then received no therapy (controls), ceftazidime (75 mg/kg per day), or ciprofloxacin (80 mg/kg per day) . Ciprofloxacin sterilized all blood cultures and significantly lowered vegetation densities of P . aeruginosa by day 2 of treatment versus controls (P less than 0.0005) and animals receiving ceftazidime (P less than 0.0005) . This beneficial effect of ciprofloxacin was also noted on therapy days 6 and 11 . Ciprofloxacin rendered most vegetations (85%) culture negative over the 11-day treatment period and achieved bacteriologic cure in 73% of animals (P less than 0.0005 versus other therapy groups) . Ciprofloxacin prevented bacteriologic relapse at 6 days posttherapy . No ciprofloxacin resistance was detected among Pseudomonas isolates from cardiac vegetations . Ciprofloxacin warrants further evaluation in vivo versus multi-drug-resistant gram-negative bacillary infections.

Microbiol Sci, 1986 Oct, 3(10), 302 - 8
Pseudomonas aeruginosa and cystic fibrosis: unusual bacterial adaptation and pathogenesis; Govan JR et al.; Pseudomonas aeruginosa is an adaptable, saprophytic bacterium with the potential to cause a variety of opportunistic infections in compromised hosts . In patients with cystic fibrosis, chronic pulmonary colonization with mucoid alginate-producing mutants of P . aeruginosa is a major cause of morbidity and mortality and is an interesting example of microbial adaptation and host-bacterium interaction.

Acta Pathol Microbiol Immunol Scand {C}, 1986 Oct, 94(5), 175 - 9
Effect of Pseudomonas aeruginosa proteases on human leukocyte phagocytosis and bactericidal activity; Kharazmi A et al.; The effect of Pseudomonas aeruginosa alkaline protease (AP) and elastase (Ela) on neutrophil phagocytosis and bactericidal activity was examined . It was found that both proteases reduced the phagocytic activity of the leukocytes against P . aeruginosa, whereas they had little effect on the phagocytosis of S . aureus . AP and Ela at concentration of up to 250 micrograms per ml (much higher than the levels detectable under in vivo conditions) did not interfere with the bactericidal activity of the leukocytes against both test organisms . Inhibition of phagocytosis by AP and Ela without effect on the bactericidal activity suggests that the P . aeruginosa proteases most probably exert their effect on the cell surface perhaps by proteolytic cleavage of the cell receptors which are necessary for phagocytosis.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1986 Oct, 182(5-6), 551 - 7
{Decontamination of drinking water taps colonized with Pseudomonas aeruginosa}; Schoenen D et al.; Water taps in hospitals are quite often contaminated with Pseudomonas aeruginosa . It is possible that Pseudomonas aeruginosa is disseminated with the water . It seems necessary to disinfect the water taps in order to prevent an endangering of the patients and a contamination of wetted areas . It has been looked for a method of decontamination which could be used under practical conditions . The experimentally contaminated taps could be disinfected in a short time when hot water was lead into the taps . The taps could not be disinfected by heating with a flexible electric heating ribbon.

J Inorg Biochem, 1986 Oct-Nov, 28(2-3), 329 - 36
Isolation and properties of the complex nonheme-iron-containing cytochrome b557 (bacterioferritin) from Pseudomonas aeruginosa; Moore GR et al.; A nonheme-iron-containing cytochrome b557, also known as bacterioferritin, was isolated from Pseudomonas aeruginosa . Gel electrophoresis showed that the protein subunits had a molecular weight of 18 kD and it is suggested that the intact molecule contained 24 subunits . The isolated protein contained 8.7% by weight Fe and 8% by weight phosphate . Most of the Fe was contained in the nonheme-iron core and could be readily removed by dialysis against 0.12 M thioglycollic acid . The resulting apobacterioferritin contained approximately one protoporphyrin IX group for every five subunits and, in addition, an unidentified fluorophor . The nonheme-iron core was found to be amorphous with a mean core size of 60-65 A.

Biochem Int, 1986 Oct, 13(4), 547 - 53
Genes encoding threonine tRNAs with the anticodon CGU from Escherichia coli and Pseudomonas aeruginosa; Dalrymple B et al.; Homologous genes for threonine tRNAs with the anticodon CGU have been identified in the region of the proBA operon of Escherichia coli and downstream from the fimbrial subunit gene of Pseudomonas aeruginosa . tRNAs with the anticodon CGU have not previously been identified from either of these bacterial species . Sequence analyses have shown that these genes are similar to other bacterial tRNA genes, and that the predicted structure conforms to the standard cloverleaf model, including retention of all invariant and semi-invariant bases . Analysis of upstream sequences suggests that these genes have associated promoters and are probably expressed in vivo.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Oct, (10), 72 - 5
{Development of a polyvalent erythrocyte diagnostic agent based on the antigens of Pseudomonas aeruginosa slime}; Aleksandrova IA et al.; The study has revealed the fundamental possibility, and established the concrete conditions of obtaining standard erythrocyte polyvalent diagnosticum for the detection of antibodies to the antigens of P . aeruginosa, belonging to 5 types most frequently occurring in clinical practice, in the passive hemagglutination test . The data on high type- and species-specificity of the new erythrocyte diagnosticum have been obtained, which indicates that slime antigens have been correctly chosen as sensitins and confirms the necessity of using the polyvalent preparation . Preliminary data on the clinical trial of the erythrocyte diagnosticum show the expediency of its use for the control of specific immune response in donors in the process of obtaining anti-P . aeruginosa hyperimmune plasma, as well as the possibility of using this method for the serological diagnosis of P . aeruginosa infection . The diagnostic preparation has been found to retain its activity for 10 months (the term of observation).

Antimicrob Agents Chemother, 1986 Oct, 30(4), 584 - 9
Effect of clavulanic acid on the activity of ticarcillin against Pseudomonas aeruginosa; Tausk F et al.; We studied the ability of clavulanic acid (CA) to induce beta-lactamase in Pseudomonas aeruginosa isolates and what effect this might have on the susceptibilities to beta-lactam agents . We first used a disk approximation method to test 4 laboratory and 16 clinical P . aeruginosa isolates against antipseudomonal beta-lactam agents for truncation by CA and found this to be very common . All antimicrobial compounds except imipenem demonstrated truncation in the vicinity of CA . We also evaluated the extent to which chromosomal beta-lactamase is induced by CA and found this to occur to some degree in most isolates and to be dependent on the concentration of CA . Finally, we performed time kill curves on these isolates to compare bacterial growth in ticarcillin alone with growth in ticarcillin-CA (the CA at 2 or 4 micrograms/ml) . We found that CA at this concentration has neither an antagonistic nor a synergistic antibacterial effect in combination with ticarcillin.

Antibiot Med Biotekhnol, 1986 Oct, 31(10), 778 - 81
{Treatment of an infection due to the intraperitoneal inoculation of mice with Pseudomonas aeruginosa with tobramycin and hyperimmune Pseudomonas aeruginosa plasma used per se and in combination}; Minukhin VV et al.; Schemes for treating infections caused by intraperitoneal administration of P . aeruginosa to mice were tested . The animals were treated with tobramycin and hyperimmune pyocyanic plasma (HPP) used per se or in combination . In a dose of 5 mg/kg body weight tobramycin protected 72.9 per cent of the animals from death . HPP with a titer of the antipyocyanic antibodies of 1:320-1:160 had a stable 100 per cent protective effect on the infected animals . However, no complete elimination of P . aeruginosa from the host was observed . The combined use of tobramycin and HPP LD50 protected 97.23 per cent of the mice from lethal infection, the drugs being titrated for their separate use . The combined administration of tobramycin and HPP in treatment of mice with acute infection due to P . aeruginosa was more efficient than their use alone.

J Clin Pathol, 1986 Oct, 39(10), 1124 - 9
Development of enzyme linked immunosorbent assay (ELISA) to detect antibodies to Pseudomonas aeruginosa cell surface antigens in sera of patients with cystic fibrosis; Brett MM et al.; An enzyme linked immunosorbent assay (ELISA) to measure free serum IgG antibodies to Pseudomonas aeruginosa in patients with cystic fibrosis was developed . Seven strains of P aeruginosa cells, treated with glutaraldehyde and representing the most commonly isolated serotypes in our cystic fibrosis unit, were used . The specificity of the test was confirmed by the absence of cross reacting antibodies to other Gram negative bacteria . The results showed differences in the titres of antibodies at different stages of P aeruginosa infection . Because of its reproducibility, specificity, and sensitivity these preliminary results suggest that this test may be of value in monitoring the progress of P aeruginosa infection in patients with cystic fibrosis.

J Antibiot (Tokyo), 1986 Oct, 39(10), 1419 - 29
Studies on 6 alpha-substituted penicillins . II . Synthesis and structure-activity relationships of 6 beta-(2-aryl-2-sulfoacetamido)-6 alpha-methoxy penicillanic acids; Burton G et al.; The synthesis and antibacterial activity of 6 alpha-methoxysulbenicillin analogues (2) are described . Structure-activity studies of these derivatives bearing hydrophilic substituents in the phenyl ring led to the identification of disodium 6 beta-{D-2-(3,4-dihydroxyphenyl)-2-sulfoacetamido}-6 alpha-methoxypenicillanate (2m) as a compound with potent activity against Pseudomonas aeruginosa including beta-lactamase producing strains . Additional substitution of 2m gave derivatives 2p, 2q, 2r, with a further improvement in activity against Gram-negative bacteria.

Microbiologica, 1986 Oct, 9(4), 455 - 60
Evaluation of Pseudomonas aeruginosa influence on the cytoskeleton of Hep-2 cells; Zanetti S et al.; The cell cytoskeleton (microtubules, microfilaments and intermediate filaments) plays an important role in many cell functions, such as maintenance of cell locomotion, movement and compartimentalization of intracellular organelles and cell-to-cell interaction . Therefore, cytoskeleton alterations may result in sever impairment of cell functions . The aim of this paper was to study the in vitro effects of Pseudomonas aeruginosa on the cytoskeleton of cultivated Hep-2 cells . Cytoskeleton modifications were evidenced by immunofluorescence using monoclonal and polyclonal antibodies against tubulin and vimentin, and rhodamine conjugated phalloidin, that specifically binds to actin microfilaments . We report here that P . aeruginosa has a strong cytopathic effect on monolayers within a few hours of contact with the cells, and influences the organization of microfilaments, but has no discernable effect on microtubules or intermediate filaments.

J Antimicrob Chemother, 1986 Oct, 18(4), 453 - 8
Incidence of strains producing plasmid determined beta-lactamases among carbenicillin resistant Pseudomonas aeruginosa; Tirado M et al.; A total of 1056 strains of Pseudomonas aeruginosa isolated from human clinical specimens from patients was collected from eight laboratories in order to study the frequency of plasmid-determined beta-lactamase producers and the different enzymes represented . The strains from each laboratory comprised consecutive, non-repeated, clinical isolates . In the 166 strains selected because they were carbenicillin resistant, the isoelectric points of the beta-lactamases were studied by means of analytical isoelectric focusing and the different types of plasmid-determined beta-lactamases identified . Seventy-five of the strains (45.18%) were plasmid-mediated beta-lactamase producers; the frequency varied among laboratories from 0% to 100% . Overall the most frequently identified beta-lactamase type was PSE-1 (49.34%) followed by TEM-1 (37.34%) . The remaining carbenicillin-resistant strains did not produce plasmid-determined beta-lactamases.

Arch Ophthalmol, 1986 Oct, 104(10), 1540 - 4
Quantitation of corneal inflammation by chemiluminescense; Chusid MJ et al.; Various inflammatory agents, including Pseudomonas aeruginosa, bacterial filtrates, endotoxin, and phorbol myristate acetate were found to induce significant increases in corneal chemiluminescense (CLM) . Disruption of polymorphonuclear leukocytes within corneas by sonication, freeze-thawing or cryotherapy, or reduction of corneal infiltration by induction of neutropenia resulted in marked decreases of CLM . Increased corneal CLM was associated with significant increases in corneal thickness and water content . Oxygen-free radical scavengers significantly inhibited CLM of experimentally infected corneas in vitro, as did the anti-inflammatory agents prednisolone acetate, indomethacin, and salicylic acid . In vivo therapy of infected corneas with prednisolone resulted in significant reductions in corneal CLM, thickness, and water content compared with saline-treated eyes . The CLM assay is a simple technique that allows quantitation of corneal inflammation and evaluation of the effect of therapeutic agents on corneal inflammation.

Infect Immun, 1986 Oct, 54(1), 239 - 44
Polyclonal and monoclonal antibody therapy for experimental Pseudomonas aeruginosa pneumonia; Pennington JE et al.; A human immunoglobulin G preparation, enriched in antibodies to lipopolysaccharide (LPS) Pseudomonas aeruginosa antigens (PA-IGIV) and murine monoclonal antibodies (MAb) to P . aeruginosa Fisher immunotype-1 (IT-1) LPS antigen and outer membrane protein F (porin), were evaluated for therapeutic efficacy in a guinea pig model of P . aeruginosa pneumonia . The concentration of antibodies to IT-1 LPS was 7.6 micrograms/ml in PA-IGIV and 478 micrograms/ml in the IT-1 MAb preparation . No antibody to IT-1 was detected in MAb to porin . For study, animals were infected by intratracheal instillation of IT-1 P . aeruginosa and then treated 2 h later with intravenous infusions of PA-IGIV, IT-1 MAb, or porin MAb . Control groups received intravenous albumin, and routinely died from pneumonia . Both PA-IGIV (500 mg/kg) and IT-1 MAb (greater than or equal to 2.5 mg/kg) treatment resulted in increased survival (P less than 0.01 to 0.001), and also improved intrapulmonary killing of bacteria . Porin MAb failed to protect from fatal pneumonia . IT-1 MAb treatment produced more survivals than did PA-IGIV treatment but only at dosages of MAb resulting in serum antibody concentrations greater than those achieved with PA-IGIV . PA-IGIV and IT-1 MAb demonstrated in vitro and in vivo (posttreatment guinea pig serum) opsonophagocytic activity for the IT-1 challenge strain . However, the polyclonal preparation required complement, whereas the MAb did not . We conclude that passive immunization with polyclonal hyperimmune P . aeruginosa globulin or with MAb to LPS antigens may be useful in the treatment of acute P . aeruginosa pneumonia . The relative efficacies of such preparations may be limited, however, by their type-specific LPS antibody concentrations.

Infect Immun, 1986 Oct, 54(1), 149 - 53
Degradation of soluble laminin and depletion of tissue-associated basement membrane laminin by Pseudomonas aeruginosa elastase and alkaline protease; Heck LW et al.; Purified Pseudomonas aeruginosa elastase and alkaline protease rapidly cleaved soluble laminin, with each enzyme yielding different cleavage products . These cleavage fragments were separated from the intact laminin A and B polypeptide chains by sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis and detected by their characteristic Coomassie blue staining patterns . Pseudomonas elastase produced rapid and extensive degradation of both A and B chains, including the disulfide-rich regions . Apparently complete degradation to limit digests was obtained after 30 min with a substrate/enzyme ratio of 30:0.5 . Under similar conditions, alkaline protease rapidly degraded the A chain while slowly degrading the B chain . In addition, immunoreactive laminin was released from authentic basement membranes after incubation with either enzyme as detected by an enzyme-linked immunoabsorption assay and by immunofluorescence . The results from these studies suggest a direct role for elastase and alkaline protease in both tissue invasion and hemorrhagic tissue necrosis in P . aeruginosa infections.

J Infect Dis, 1986 Oct, 154(4), 682 - 8
Pseudomonas aeruginosa polysaccharide-tetanus toxoid conjugate vaccine: safety and immunogenicity in humans; Cryz SJ Jr et al.; A Pseudomonas aeruginosa polysaccharide-tetanus toxoid (Ttxd) conjugate vaccine was produced . Polysaccharide was derived from lipopolysaccharide (LPS) and covalently linked to Ttxd by using carbodiimide with adipic acid dihydrazide as a spacer molecule . The conjugate possessed a relative molecular weight of greater than 350,000 and was nontoxic and nonpyrogenic . The vaccine bound serospecific monoclonal antibodies with an avidity similar to LPS and reacted with murine and human opsonic antibody . The vaccine was immunogenic in rabbits and mice and elicited IgG antibody to both LPS and Ttxd . The vaccine was safe when parenterally administered to humans and evoked only mild, transient reactions . Mean titers of IgG antibody to LPS rose 19-fold after immunization, with 82% of the volunteers responding with a fourfold or greater rise in titer . IgG antibody to LPS evoked after immunization was opsonic and highly effective at preventing fatal experimental burn wound sepsis due to P . aeruginosa.

J Gen Microbiol, 1986 Oct, 132 ( Pt 10), 2667 - 75
The arcABC operon required for fermentative growth of Pseudomonas aeruginosa on arginine: Tn5-751-assisted cloning and localization of structural genes; Luthi E et al.; Pseudomonas aeruginosa is able to utilize L-arginine as the energy source for growth under anaerobic, nitrate-free conditions . Mutations in the chromosomal arcABC gene cluster specifying the inducible arginine deiminase pathway enzymes abolish fermentative growth on arginine . From two different arc::Tn5-751 insertion mutants of P . aeruginosa recombinant plasmids have been derived which carry a resistance marker of transposon Tn5-751 plus flanking parts of the arc region . These recombinant plasmids served to reconstruct in vitro the functional arcABC cluster on a 5.6 kb fragment, which was inserted into the broad-host-range vector pKT240 . In P . aeruginosa this 5.6 kb segment complemented arcABC mutations in trans and contained the control region necessary in cis for arc enzyme induction by oxygen limitation and arginine . The results of subcloning experiments and transcriptional lacZ fusions, the polarity of transposon insertions and the effect of external promoters led to the conclusion that the structural genes arcA (for arginine deiminase), arcB (for catabolic ornithine carbamoyltransferase) and arcC (for carbamate kinase) are contiguous and transcribed in the same direction . Thus, the arcABC cluster appears to have the characteristics of an operon . In Escherichia coli the cloned arcABC genes were expressed at low, non-inducible levels; strong vector promoters enhanced arc expression up to 100-fold . This indicates that transcriptional initiation at the arc promoter(s) is poor in E . coli.

Eur J Biochem, 1986 Oct 1, 160(1), 149 - 53
Pseudomonas aeruginosa cytochrome oxidase . Product inhibition by low thermodynamic driving force; Blatt Y et al.; The steady-state kinetics of Pseudomonas aeruginosa cytochrome oxidase were studied . Reduced cytochrome c551 and azurin from the same bacteria were used as the electron-donating substrates, while dioxygen served as the electron acceptor . Oxidized cytochrome c551 and azurin exhibited product inhibition of the reaction . However, apo-azurin and azurin derivatives in which the copper was substituted by the redox-inert ions Ni2+, Co2+, Cd2+ and Zn2+, did not show any effect on the kinetics . These observations implied that complex formation between the substrates or the products and the enzyme is not a rate-limiting step and is not the cause for product inhibition . The integrated rate law for a reaction scheme in which we assumed that complex formation was not rate limiting was fitted to the complete reaction traces . The results suggested that it is the low thermodynamic driving force, expressed in the small differences in redox potential between the substrates and heme c of the enzyme, which cause the observed product inhibition.

Mol Gen Mikrobiol Virusol, 1986 Oct, (10), 13 - 6
{Physical map of Pseudomonas aeruginosa phage phi kF77 genome . Localization of sites sensitive to restriction endonucleases}; Kulakov LA et al.; A physical map of the P . aeruginosa bacteriophage phi kF77 has been constructed using the restriction endonucleases SalI, HindIII, EcoRI, EcoRV, MuI, XbaI, ClaI . The phi kF77 DNA is resistant to cleavage by the restriction endonucleases BamHI, BglII, HpaI, PstI, PvuII, SmaI, XhoI.

Bioorg Khim, 1986 Oct, 12(10), 1422 - 4
{Structure of O-specific polysaccharide chains of Pseudomonas aeruginosa II (Sandwick) and V (Verder-Evans) lipopolysaccharides containing N-acetyl-D-galactosaminuronic acid}; Knirel' IuA et al.; Acidic polysaccharides were isolated from the Pseudomonas aeruginosa II (Sandvik) and V (Verder-Evans) lipopolysaccharides on mild acid hydrolysis followed by gel filtration on Sephadex G-50 . The Sandvik II polysaccharide consists of 2-acetamido-2-deoxy-D-galacturonic acid, 2-acetamido-2,6-dideoxy-D-glucose, and L-rhamnose in the ratio 1:1:2 . The Verder-Evans V polysaccharide contained the same monosaccharides and, in addition, a D-glucose residue . On the basis of 13C NMR data, methylation analysis, Smith degradation and solvolysis with hydrogen fluoride, the following structures were determined for the repeating units of the polysaccharides: (Formula: see text).

Bioorg Khim, 1986 Oct, 12(10), 1384 - 90
{Antigenic polysaccharides of bacteria . 18 . Structure of O-specific polysaccharide chains of Pseudomonas aeruginosa 05 (Lányi) lipopolysaccharide}; Knirel' IuA et al.; Polysaccharide chains of P . aeruginosa O5a, b, c, O5a, b, d and O5a, d (Lanyi classification) lipopolysaccharides contain D-xylose, N-acetyl-D-fucosamine (FucNAc) and a derivative of 5,7-diamino-3,5,7,9-tetradeoxy-L-glycero-L-manno-nonulosonic acid (pseudaminic acid, PseN2) carrying acetyl or (R)-3-hydroxybutyryl (Hb) and formyl (Fm) groups as N-acyl substituents . Degradation of the lipopolysaccharides with dilute acetic acid caused depolymerisation of the polysaccharide chains as a result of cleavage of glycosidic linkage of pseudaminic acid to give trisaccharides representing chemical repeating units of the polysaccharides . Basing on analysis of the trisaccharides using 1H and 13C NMR spectroscopy and mass-spectrometry, the following structures of the polysaccharide chains were established: (Formula: see text) . O5a, d polysaccharide is identical to P . aeruginosa immunotype 6 O-specific polysaccharide.

Biochim Biophys Acta, 1986 Sep 26, 873(2), 214 - 27
The pH and redox-state dependence of the copper site in azurin from Pseudomonas aeruginosa as studied by EXAFS; Groeneveld CM et al.; The EXAFS of the K-edge of copper in azurin from Pseudomonas aeruginosa has been measured in solutions of the oxidized and reduced protein, at both low and high pH . Model compounds of known molecular structure, exhibiting Cu-N and Cu-S bonds of varying length, were studied as well . The major shell of the high-pH oxidized azurin EXAFS contains contributions of two N(His) at 1.95 +/- 0.03 A, and one S(Cys) at 2.23 +/- 0.03 A . Some minor contributions from the carbon atoms of the histidine residues and the distal sulfur atom are observed in the 3-4 A region . Upon reduction a decrease is seen in amplitude of the main peak in the Fourier transform, due to a lengthening of one of the Cu-N(His) bonds (2.05 +/- 0.03 A), and a shortening of the other (1.89 +/- 0.03 A), both by approx . 0.1 A . Indications for a Cu-S(Met) bond are found in the reduced azurin data (2.70 +/- 0.05 A) . However, in the oxidized protein, this bond could not be determined unambiguously, in line with results of a model compound featuring weak Cu-thioether coordination . The effect of pH is only slight for both the oxidized and the reduced protein, and no significant changes in bond lengths are found upon a change of pH from 4.1 to 9.1 . The relevance of these findings for the interpretation of the existing data on the redox activity of the protein is discussed.

Presse Med, 1986 Sep 20, 15(30), 1417 - 20
{Antibiotic therapy of Pseudomonas aeruginosa infections}; Modai J et al.; Antibiotics active against Pseudomonas aeruginosa are carboxy- and ureidopenicillins, some cephalosporins, aztreonam, imipenem, aminoglycosides, fluoroquinolones and, in some special circumstances, polymyxins and fosfomycin . The choice of the antibiotic should be based not only on minimal inhibitory concentrations in vitro, but also on the inoculum effect, on the speed with which bacteria are killed, on natural or acquired resistance and on data obtained from experimental infections in animals . Antibiotics used against P . aeruginosa infections must be rapidly bactericidal . Since the prognosis is severe, the initial treatment given before the sensitivity of the strain is known must combine a beta-lactam antibiotic and an aminoglycoside, both being selected among the most regularly active.

J Immunol, 1986 Sep 15, 137(6), 2025 - 30
In vitro T cell-mediated killing of Pseudomonas aeruginosa . IV . Nonresponsiveness in polysaccharide-immunized BALB/c mice is attributable to vinblastine-sensitive suppressor T cells; Powderly WG et al.; We reported previously that BALB/c mice immunized with a polysaccharide (PS) antigen isolated from immunotype 1 Pseudomonas aeruginosa and vinblastine sulfate develop T cell-mediated protective immunity, despite their failure to produce specific antibody . In vitro, Lyt-1-,2+, I-J+ T cells from vinblastine- and PS-immunized mice kill P . aeruginosa by secretion of a bactericidal lymphokine . BALB/c mice immunized with PS alone generate neither protective antibodies nor a protective T cell response . The current studies indicate that T cells from mice immunized with PS alone significantly suppress the bactericidal activity of T cells from mice immunized with vinblastine and PS . The suppressor T cells are of the same Lyt-1-,2+, I-J+ phenotype as the bactericidal T cells . Suppression is mediated by a soluble product of these suppressor T cells which both inhibits T cell proliferation and interferes with the production or release of the bactericidal lymphokine . Cyclophosphamide, used in other systems to remove suppressor T cells, fails to enhance bacterial killing and does not inhibit suppressor cell activity . These studies indicate that immunization with PS elicits responses in two functionally distinct subgroups of Lyt-1-,2+, I-J+ T cells, and that these cells are distinguishable by their sensitivity to vinblastine sulfate.

Biochim Biophys Acta, 1986 Sep 11, 860(3), 699 - 707
Demonstration and chemical modification of a specific phosphate binding site in the phosphate-starvation-inducible outer membrane porin protein P of Pseudomonas aeruginosa; Hancock RE et al.; The interaction of phosphate ions with the Pseudomonas aeruginosa anion-specific protein P channel was probed . The single-channel conductance of protein P incorporated into planar lipid bilayer membranes in the presence of 0.3 M H2PO-4 was shown to be 6.0 pS, demonstrating that protein P channels allowed the permeation of phosphate . When large numbers of protein P channels were incorporated into lipid bilayer membranes in the presence of 40 mM Cl-, addition of small concentrations of phosphate resulted in reduction of macroscopic Cl- conductance in a dose- (and pH-) dependent fashion . This allowed calculation of an I50 value of e.g . 0.46 mM at pH 7.0, suggesting that the affinity of protein P for its normal substrate phosphate was at least 60-100-fold greater than the affinity of the channel for other ions such as chloride . Pyrophosphate and the phosphate analogue, arsenate, also inhibited macroscopic Cl- conductance through protein P with I50 values at pH 7.0 of 4.9 mM and 1.3 mM, respectively . To probe the nature of the phosphate binding site, the epsilon-amino groups of available lysine residues of protein P were chemically modified . Acetylation and carbamylation which produced uncharged, modified lysines destroyed both the anion (e.g . Cl-) binding site and the phosphate binding site as determined by single-channel experiments and macroscopic conductance inhibition experiments respectively . Nevertheless, the modified proteins still retained their trimeric configuration and their ability to reconstitute single channels in lipid bilayer membranes . Methylation, which allowed retention of the charge on the modified lysine residues, increased the Kd of the channel for Cl- 33-fold and the I50 for phosphate inhibition of macroscopic Cl- conductance 2.5-4-fold . A molecular model for the phosphate binding site of the protein P channel is presented.

Klin Monatsbl Augenheilkd, 1986 Sep, 189(3), 240 - 2
{Studies of the penetrating ability of fosfomycin into the aqueous humor and vitreous body of the eye}; Philipp W et al.; Fosfomycin is an antibiotic with a new simple chemical structure and a broad spectrum of antibacterial activity, especially against staphylococci . In the study reported here, the penetration of fosfomycin into the aqueous humor and vitreous body was investigated after intravenous infusion of 4 and 8 g . The highest average concentration approximately 2 hours after administration was 28.3 micrograms/ml after the 4 g dose and 52 micrograms/ml after the 8 g dose . Both the 4 g and 8 g doses thus produced aqueous humor levels exceeding the minimum inhibitory concentrations required for most gram-positive and gram-negative organisms commonly responsible for bacterial endophthalmitis, including strains of pseudomonas aeruginosa . Extremely high aqueous humor levels of fosfomycin were observed in two patients with inflamed eyes because of a change in the blood-aqueous barrier.

J Antibiot (Tokyo), 1986 Sep, 39(9), 1243 - 56
Semisynthetic beta-lactam antibiotics . III . Synthesis and antibacterial activity of 7 beta-{2-(2-aminothiazol-4-yl)-2-(substituted carbamoylmethoxyimino)acetamido}cephalosporins; Arimoto M et al.; Syntheses of cephalosporins modified with a 7 beta-{2-(2-aminothiazol-4-yl)-2-(substituted carbamoylmethoxyimino)acetamido} group at the C-7 position and with various hetero aromatics at the C-3 position are described . The effects of substituents on the carbamoyl group in the 7-side chain were investigated in order to improve antibacterial activity . Some of these compounds exhibited high antibacterial activity against Gram-positive and Gram-negative bacteria, including Pseudomonas aeruginosa, as well as good resistance to beta-lactamase.

Am J Reprod Immunol Microbiol, 1986 Sep, 12(1), 21 - 4
Bacteriology of cervix in cases of infertility: effect on human sperm; Kaur M et al.; Microorganisms such as Escherichia coli, Pseudomonas aeruginosa, and Bacillus subtilis isolated from cervices of infertile females possessed spermicidal activity . They also agglutinated the human spermatozoa in vitro and showed tail-to-tail agglutination . Cell-free supernatant of these organisms was found to be spermicidal but did not agglutinate spermatozoa in vitro . Spermicidal activity was increased with increase in age of the culture.

Laryngoscope, 1986 Sep, 96(9 Pt 1), 1021 - 3
Malignant external otitis with optic neuritis; Holder CD et al.; Malignant external otitis (MEO) is a progressive necrotizing infection which spreads to the skull base . The causative organism is usually Pseudomonas aeruginosa and 90% of the patients are diabetic . The infection gains access to the skull base at the temporal bone . Cranial nerve involvement is common . We present a case of malignant external otitis causing blindness due to optic neuritis . Progressive vascular involvement along the skull base is the pathogenic mechanism that best explains spread from the temporal bone to the orbital apex.

Infection, 1986 Sep-Oct, 14(5), 246 - 9
Fosfomycin-ampicillin versus gentamicin-ampicillin in the treatment of critically ill patients with pneumonia; Nissen LR et al.; Thirty-two patients with severe pneumonia (22 on assisted ventilation) were entered into a prospective randomised trial, in which fosfomycin plus ampicillin (17 patients) was compared with gentamicin plus ampicillin (15 patients) . Treatment was either 4 g fosfomycin or 80 mg gentamicin every 8 h and 1 g ampicillin every 6 h . Complete or partial clinical success was attained in 94% (16/17) in the fosfomycin group and in 80% (12/15) in the gentamicin group . Bacteriological success was 87.5% with fosfomycin-ampicillin and 90% with gentamicin-ampicillin . An intermediary sensitive Klebsiella pneumoniae strain developed complete resistance in the fosfomycin group, and an in vitro sensitive Pseudomonas aeruginosa strain was resistant in vivo in the gentamicin group . Two of three patients in the fosfomycin group receiving the infusion through a peripheral vein developed thrombophlebitis . No other side-effects were observed . We conclude that fosfomycin is at least as effective as gentamicin . Since fosfomycin is widely atoxic and may be given in large doses, irrespective of kidney function, it is considered to have advantages over gentamicin in the combined therapy of pneumonia.

Antimicrob Agents Chemother, 1986 Sep, 30(3), 435 - 9
Antibiotic-resistant bacteria in surveillance stool cultures of patients with prolonged neutropenia; Wingard JR et al.; The value of stool surveillance for antibiotic-resistant gram-negative bacteria was analyzed in 86 neutropenic bone marrow transplant patients . Twice-weekly specimens were inoculated onto culture medium containing gentamicin plus carbenicillin . The recovered organisms were identified to the species level and tested for antibiotic susceptibility . Forty-eight resistant organisms were recovered from 35 patients . Thirteen isolates persistently colonized patients . Escherichia coli (29%) and Pseudomonas aeruginosa (19%) were the most frequently recovered organisms . Although most organisms were recovered while patients were on antibiotics, 15 isolates, including eight of nine resistant P . aeruginosa, were detected before antibiotics were initiated . The duration of antibiotic use was longer for patients persistently colonized than for those not colonized (P = 0.03) . Of the 15 resistant organisms which caused infection, 12 were detected in the surveillance cultures . Infections by antibiotic-resistant organisms occurred more frequently in patients colonized than in those not colonized (P = 0.006) and more frequently in patients persistently colonized than in those colonized only once (P = 0.01) . The absence of colonization or persistent colonization correlated well with the absence of infection (negative predictive values of 94 and 91%, respectively).

Pediatr Med Chir, 1986 Sep-Oct, 8(5), 715 - 20
{Occurrence of hospital infections in a department of pediatric heart surgery}; Mantero E et al.; The incidence of nosocomial infections (NI) and the related risk factors in a Department of Pediatric Cardiovascular Surgery were studied, during a 6 months period . 155 successive admissions were considered . Nosocomial infections were 17 (11%), nosocomial colonizations 18 (11.6%) . The most important risk factors for nosocomial infections were: age, cyanosis, duration of hospitalization, hospitalization in Intensive Care Unit and central venous catheter only as a risk factor for sepsis . The most important risk factors for nosocomial colonizations were: tracheal intubation and central venous catheter . In 4 cases the NI was related to nosocomial colonization (2 sepsis, 1 pneumonia, 1 wound infection) . The most frequently isolated microorganisms were Pseudomonas aeruginosa and Staphylococcus spp . The Authors found that a longer than 5 days period of antibiotic prophylaxis did not reduce the incidence of nosocomial infections.

Pathol Biol (Paris), 1986 Sep, 34(7), 855 - 8
{Choice of a rapidly bactericidal beta-lactamin-aminoglycoside combination in the treatment of Pseudomonas aeruginosa infections at a child intensive care unit}; Lambert-Zechovsky N et al.; Morbidity and mortality among children with Pseudomonas aeruginosa infection in Pediatric Intensive Care Unit remains high . Delays in bacterial killing may be responsible for the poor outcome . Antimicrobial sensitivity and timed-killing assays were determined for ticarcillin, azlocillin, piperacillin, cefsulodin, ceftazidime, gentamicin, tobramycin and amikacin alone and in combination against 40 strains of Pseudomonas aeruginosa isolated from blood cultures and tracheal aspirate . Antibiotic concentrations used were at clinically achievable level . None bactericidal effect was observed with each beta-lactamin alone . However with the combinations azlocillin or piperacillin or cefsulodin or ceftazidime plus amikacin a bactericidal effect was observed at 4.5 hours.

Mol Biol Evol, 1986 Sep, 3(5), 449 - 58
Structure and regulation of the anthranilate synthase genes in Pseudomonas aeruginosa: II . Cloning and expression in Escherichia coli; Crawford IP et al.; The genes for the large and small subunits of anthranilate synthase (trpE and trpG, respectively) have been cloned from Pseudomonas aeruginosa PAC174 into E . coli by R-prime formation with the broad-host-range plasmid R68.44 . Sequential subcloning into plasmid vectors reduced the active Pseudomonas DNA fragment to a length of 3.1 kb . We obtained evidence that this region contains the promoter for its own expression and retains a vestigial regulatory response to tryptophan scarcity or excess.

Mol Biol Evol, 1986 Sep, 3(5), 436 - 48
Structure and regulation of the anthranilate synthase genes in Pseudomonas aeruginosa: I . Sequence of trpG encoding the glutamine amidotransferase subunit; Crawford IP et al.; We have determined the DNA sequence of the distal 148 codons of trpE and all of trpG in Pseudomonas aeruginosa . These genes encode, respectively, the large and small (glutamine amidotransferase) subunits of anthranilate synthase, the first enzyme in the tryptophan synthetic pathway . The sequenced region of trpE is homologous with the distal portion of E . coli and Bacillus subtilis trpE, whereas the trpG sequence is homologous to the glutamine amidotransferase subunit genes of a number of bacterial and fungal anthranilate synthases . The two coding sequences overlap by 23 bp . Codon usage in these Pseudomonas genes shows a marked preference for codons ending in G or C, thereby resembling that of trpB, trpA, and several other chromosomal loci from this species and others with a high G + C content in their DNA . The deduced amino acid sequence for the P . aeruginosa trpG gene product differs to a surprising extent from the directly determined amino acid sequence of the glutamine amidotransferase subunit of P . putida anthranilate synthase (Kawamura et al . 1978) . This suggests that these two proteins are encoded by loci that duplicated much earlier in the phylogeny of these organisms but have recently assumed the same function . We have also determined 490 bp of DNA sequence distal to trpG but have not ascertained the function of this segment, though it is rich in dyad symmetries.

Quad Sclavo Diagn, 1986 Sep, 22(3), 273 - 82
{Frequency of isolation and drug resistance of bacterial strains isolated from respiratory material in patients with cystic fibrosis}; Rosselli P; During 1985, 251 respiratory samples from 61 patients were examined at the tuscan cystic fibrosis Center (Florence), and isolated strains were tested against various antimicrobial drugs . Pseudomonas aeruginosa and Staphylococcus aureus were the predominant pathogens isolated . Infections caused by St . aureus were the commonest during the first years of life, whereas those caused by Ps . aeruginosa, occasional at early age, became chronic and the main source of infection with growth . Sensitivity testings against antibiotics confirmed that Ps . aeruginosa isolated from cystic fibrosis patients has a high percentage of resistant strains . The most active drugs were ceftazidime and aztreonam . An increased resistance to aminoglycosides was observed.

Acta Paediatr Scand, 1986 Sep, 75(5), 840 - 5
Does centralized treatment of cystic fibrosis increase the risk of Pseudomonas aeruginosa infection?
Pedersen SS, Jensen T, Pressler T, Hoiby N, Rosendal K.
Two hundred and forty Danish patients with cystic fibrosis (97% of the total CF population in Denmark) participated in a point-prevalence study of Pseudomonas aeruginosa infection . One hundred and ninety-two patients were treated at the Danish CF centre and 48 patients were treated in other places . The age distribution was significantly different as no patients older than 19 years were found in the non-centre group . Pathogenic bacteria were isolated from the sputum of 96% of the patients . P . aeruginosa was more prevalent in patients from the centre, whereas Staphylococcus aureus was more prevalent in the non-centre group . No difference in serogroup and phage pattern of P . aeruginosa was found . There was a tendency that non-centre treated patients acquired chronic broncho-pulmonary P . aeruginosa infection later, but at the age of 16 years 90% of all patients will be chronically infected . Chronic P . aeruginosa infection was significantly more common in the age group 10-14 years at the centre than outside the centre . It is not possible to prevent chronic P . aeruginosa infection in CF patients treated in small groups and because of the better prognosis of centralized treatment the latter must be recommended.

Jpn J Antibiot, 1986 Sep, 39(9), 2355 - 66
{Cefsulodin concentration in exudates from drainage of patients with acute peritonitis following intravenous administration}; Nakamura T et al.; Cefsulodin (CFS), a new antipseudomonal cephalosporin, shows a potent antibacterial activity against Pseudomonas aeruginosa and some Gram-positive bacteria, whereas it shows low activity against many Gram-negative rods . Against clinical isolates of P . aeruginosa, CFS was about 10 times more active than sulbenicillin and carbenicillin, and had a similar activity to gentamicin and dibekacin . The CFS was administered by an intravenous bolus injection at a dose of 1 g to each of 14 patients operated for acute peritonitis with drainage or radical mastectomy with drainage to treat breast cancer . These cases included 3 of localized peritonitis due to perforative appendicitis, 3 of diffuse peritonitis due to perforative duodenal ulcer, 2 of panperitonitis due to intestinal obstruction and perforative sigmoid colon cancer, 4 of subacute cholangitis, localized peritonitis T-tube choledochal drainage due to choledocholithiasis, and 2 of breast cancer . Materials from drain exudate were taken at intervals with sterilized paper discs and CFS concentrations were determined by the paper disc bioassay method with P . aeruginosa NCTC 10490 as the test organism . Serum concentrations of CFS just after injection reached 135.4 +/- 66.1 micrograms/ml, and they were 2.7 +/- 1.5 micrograms/ml at 6 hours after injection . Concentrations in purulent exudates of patients with acute peritonitis increased quickly after intravenous bolus injections, and reached maximum levels relatively early after injection in cases 2 to 3 days after operation . In cases 10 to 13 days after operation, CFS levels were comparatively low and reached to peak levels at 4 to 5 hours after injection . Levels of CFS in purulent exudate tended to increase in proportion to the severity of symptoms, as did CFS levels in appendix wall . Pseudomonas spp . were not isolated in this study, but MICs of CFS were mostly around 1.56 to 3.13 micrograms/ml when clinically isolated Pseudomonas spp . were present at 10(6) cells/ml . Levels of CFS in infected exudate were higher than the above MIC values against Pseudomonas spp . Therefore, CFS were a useful drug for the chemotherapy against pseudomonal infections.

Eur J Epidemiol, 1986 Sep, 2(3), 182 - 5
Prevalence of protease and elastase production by clinical isolates of Pseudomonas aeruginosa in relation to aeruginocine typing patterns; Kapur R et al.; Sixty six consecutive P . aeruginosa isolates from heterogeneous clinical specimens were subjected to aeruginocine (pyocine) typing and assayed for in vitro protease and elastase production by a simple and reproducible qualitative test . The 45.4% of the clinical isolates were found to be both protease and elastase (P + E +) producers; 40.9% were only protease producers (P + E -) and 13.6% were non producers (P - E -) . Aeruginocine code 7777 strains were found to be predominant among P + E + and P + E - types, as 48.2% and 51.7% isolates belonged to the types, respectively, suggesting thereby the virulence of this aeruginocine type in P . aeruginosa infections and the possible association of protease and elastase production with aeruginocine production.

Arch Microbiol, 1986 Sep, 145(4), 408 - 10
Alginate biosynthesis by Pseudomonas aeruginosa: effect of arsenite and other metabolic inhibitors; Banerjee PC; Biosynthesis of alginic acid in presence of metabolic inhibitors by resting cells of mucoid Pseudomonas aeruginosa was studied . Among the inhibitors tested, arsenite exhibited very interesting results, while the others showed no remarkable effect . Firstly, arsenite stopped alginate production from all the substrates during initial hours of incubation; secondly, degradation of newly synthesized alginates to smaller molecular weight fragments took place if it was added after a few hours of incubation with the substrate; and thirdly, uncontrolled synthesis of alginate started after several hours of inhibition . Presence of arsenite was needed for the initial inhibitory phase of alginate synthesis; but once the cells were capable of synthesizing alginate after initial hours of inhibition, arsenite may be omitted from the medium.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Sep, (9), 90 - 2
{Dynamics of the contamination of mice during the treatment of burn sepsis due to Pseudomonas aeruginosa using tobramycin alone or combined with active and passive immunization}; Minukhin VV et al.; An experimental study was made with a view to finding out the possible bacteriological advantages of the combined use of tobramycin (Tb) and P . aeruginosa corpuscular polyvalent vaccine (PaCPV) or P . aeruginosa hyperimmune plasma (PaHIP) in burn sepsis caused by P . aeruginosa . The use of the median therapeutic dose of Tb (2.5 mg/kg body weight per day), alone or in combination with immunopreparations, ensured the survival rate of the animals equal to 100% . The contamination of the body with P . aeruginosa after treatment with Tb and PaHIP or PaCPV was lower than after the administration of Tb alone, this phenomenon becoming manifest starting from day 5 of observation in the first case and from day 10 in the second case . The combined use of Tb and immunopreparations (PaCPV or PaHIP) in acute P . aeruginosa infection proved to be more effective than treatment with Tb alone.

Can J Microbiol, 1986 Sep, 32(9), 751 - 5
{Bacterial interference with skin colonization in germfree hairless mice}; Barc MC et al.; Bacterial colonization of the digestive tract and the skin was studied over a 3-week period in a group of 10 germfree HRS mice using Staphylococcus epidermidis, Staphylococcus aureus, and Pseudomonas aeruginosa . Sequential utilization of two strains allowed us to carry out six assays and to show the presence of interference phenomena during colonization of the skin . When P . aeruginosa was given after challenge with S . aureus or S . epidermidis, it did not colonize the skin . If the first challenge was done with P . aeruginosa, this bacteria was eliminated within 10 days by S . aureus and S . epidermidis on the skin, but it succeeded in colonizing the digestive tract . When the first challenge was done with S . aureus, colonization of the skin and the digestive tract with S . epidermidis was prevented, whereas these two species were found in association when S . aureus was given in second place . None of the in vitro assays (mixed culture, bacteriocin production, adherence inhibition, antimicrobial activity) could explain the in vivo observations.

Antimicrob Agents Chemother, 1986 Sep, 30(3), 453 - 7
Evidence for multiple forms of type I chromosomal beta-lactamase in Pseudomonas aeruginosa; Gates ML et al.; The multiple stages of derepression of the type I chromosomal beta-lactamase in Pseudomonas aeruginosa were examined . Mutants partially and fully derepressed for beta-lactamase were selected from a wild-type clinical isolate . An analysis of the beta-lactamase produced by these mutants and the induced wild type revealed significant differences in the products of derepression at each stage . Beta-lactamase produced by the fully derepressed mutant showed a lower affinity (Km, 0.113 mM) for cephalothin than that produced by the partially derepressed mutant (Km, 0.049 mM) . However, due to a very large Vmax, the former possessed a much greater hydrolytic efficiency . Differences in substrate profile were also noted . Only beta-lactamase from the fully derepressed mutant hydrolyzed cefamandole, cefoperazone, and cefonicid . The partially derepressed mutant possessed a single beta-lactamase band with a pI of 8.4 . The fully derepressed mutant possessed this band and an additional major band with a pI of 7.5 . Induction of the wild type with cefoxitin produced both bands . The changes in physiologic parameters of the enzymes produced in the different stages of derepression suggest a complex system for beta-lactamase expression in P . aeruginosa . This may involve at least two distinct structural regions, each of which is under control of the same repressor.

Tohoku J Exp Med, 1986 Sep, 150(1), 69 - 75
Electron microscopic study of the cell surface of dibekacin-treated Pseudomonas aeruginosa; Aonuma S et al.; Electron microscopy of thin sections of Pseudomonas aeruginosa IAM 1007 treated with dibekacin revealed blebs, disintegration of the outer membrane of the cell wall and degenerative features of the cytoplasm . In the next experiment, the cell wall fraction was isolated from the mechanically disrupted cells, incubated with dibekacin and was subjected to electron microscopy, in order to find a clue to the action mechanism of dibekacin on the cell wall of Pseudomonas aeruginosa IAM 1007 . As a result, it was found that unidentified substances were released from the surface of the cell wall and that the outer membrane of the cell wall disappeared . The degree of changes of the cell wall ultrastructure was almost proportional to the length of incubation with dibekacin . These findings strongly suggest that dibekacin directly disintegrates the outer membrane of the cell wall of Pseudomonas aeruginosa IAM 1007.

Mol Gen Genet, 1986 Sep, 204(3), 519 - 23
The Hfr status of Pseudomonas aeruginosa is stabilized by integrative suppression; Herrmann H et al.; A temperature-sensitive mutant (dna-11) with the phenotype of a mutant defective in the initiation of DNA replication, was isolated from an Hfr-like FP2 donor of Pseudomonas aeruginosa . Reversion of its temperature-sensitive character was achieved by integrative suppression rather than by backmutation or an additional suppressor mutation . The dna-11 mutant proved to be helpful in stabilizing the Hfr status of the original host.

J Infect, 1986 Sep, 13(2), 125 - 31
Ceftazidime alone for treating Pseudomonas aeruginosa septicaemia in neutropenic patients; Verhagen C et al.; Twenty-nine patients with primary or secondary hypoplastic bone-marrow were treated successfully with ceftazidime alone for established septicaemia caused by Pseudomonas aeruginosa of which 38% strains were gentamicin-resistant . All the patients were neutropenic at the start of therapy; most were cured with microbiological confirmation before the bone marrow had regenerated . One patient died of cerebral haemorrhage due to profound thrombocytopenia without evidence of infection at autopsy . Significant toxicity was not observed . Ceftazidime alone is, therefore, a safe and effective treatment for infections caused by this organism even in the neutropenic patient.

J Clin Microbiol, 1986 Sep, 24(3), 372 - 6
Antibiograms, serotypes, and plasmid profiles of Pseudomonas aeruginosa associated with corneal ulcers and contact lens wear; Mayo MS et al.; Pseudomonas aeruginosa was isolated from the corneal scrapings of 11 of 14 patients with gram-negative corneal ulcers and from salt tablet-prepared saline solutions from 6 of these patients wearing soft contact lenses . Comparison of physiological properties, antibiograms, serotypes, and plasmid profiles for five of the patients indicated that the isolates from the ulcer and the saline solution of a given patients were of the same strain . Improper hygienic practices of contact lens wearers appeared to be a major factor in the epidemiology of pseudomonad corneal ulcers.

J Clin Microbiol, 1986 Sep, 24(3), 317 - 23
Variation and adaptation of Pseudomonas aeruginosa toxicity to HeLa cells and fibroblasts; Muller H et al.; The toxic components of supernatants from Pseudomonas aeruginosa cultures directed against HeLa cells and Staphylococcus aureus were evaluated with the aim of discovering interactions . Supernatants of eight different strains of P . aeruginosa were assayed for cytotoxic activity . All were active against HeLa cells; seven were toxic for S . aureus . On repeated suspension of P . aeruginosa in 0.9% sodium chloride solution, a shift from HeLa cell toxicity to staphylococcal lytic activity occurred along with a change of toxic activity from a high (50,000 +/- 5,000) to a low (8,000 +/- 400) molecular weight (MW) range on gel filtration . Addition of protein to the minimal medium of cultures producing material toxic only for S . aureus reactivated the generation of HeLa cell-toxic material . Cultivation of P . aeruginosa in the presence of HeLa cells and a chloramphenicol supplement produced suppression of the generation of material toxic for S . aureus but facilitated that of HeLa-toxic material of high MW . Adaptation of toxicity against fibroblasts developed only on cocultivation of P . aeruginosa together with S . aureus and in the presence of fibroblasts . Under these conditions a strong lytic activity for S . aureus appeared, even in the presence of chloramphenicol . Chloramphenicol caused the material toxic for fibroblasts to elute at a low MW well separated from that toxic for HeLa cells . In contrast to the high-MW toxic substances, the low-MW material did not induce antibodies after injection into rabbits . This may explain failures of vaccination against P . aeruginosa infection and of serum therapy of homologous sepsis in humans.

Eur J Biochem, 1986 Sep 1, 159(2), 309 - 13
Purification and structural analysis of pyocyanin and 1-hydroxyphenazine; Watson D et al.; Pyocyanin and related members of the phenazine family are produced by Pseudomonas aeruginosa and have been associated with events of pathophysiological importance . Pyocyanin and its base hydrolysis product 1-hydroxyphenazine were purified to homogeneity by reverse-phase high-pressure liquid chromatography . Their mass spectrometric behaviour was examined with a view to evaluating the use of high-resolution chromatography/mass spectrometry in studying phenazine-mediated effects in man . The molecular mass of naturally derived pyocyanin was determined as 210 Da by thermospray liquid chromatography/mass spectrometry and confirmed by desorption electron-impact mass spectrometry . Mass spectrometric data could not be obtained by fast-atom bombardment or desorption chemical ionisation, techniques commonly used to determine molecular mass of polar or thermally labile species . The thermal lability of underivatised pyocyanin precluded analysis by gas chromatography/mass spectrometry . In contrast to pyocyanin, mass spectrometric data were readily obtained for 1-hydroxyphenazine, using direct probe analysis as well as with gas and liquid chromatography inlet systems.

Infect Immun, 1986 Sep, 53(3), 621 - 7
Monoclonal antibodies to Pseudomonas aeruginosa ferripyochelin-binding protein; Sokol PA et al.; Hybridomas secreting specific monoclonal antibodies against the Pseudomonas aeruginosa ferripyochelin-binding protein (FBP) were isolated . These monoclonal antibodies reacted with FBP in immunoblots of outer membrane preparations from all serotypes of P . aeruginosa . Two of the monoclonal antibodies also reacted with FBP in strains of P . putida, P . fluorescens, and P . stutzeri . These antibodies did not react with outer membranes of P . cepacia, "P . multivorans," P . maltophilia, or other gram-negative organisms . The monoclonal antibodies were opsonophagocytic and blocked the binding of {59Fe}ferripyochelin to isolated outer membranes of strain PAO . By indirect immunofluorescence techniques, the monoclonal antibodies were used to demonstrate that FBP is present on the cell surface of P . aeruginosa cells grown in low-iron but not high-iron medium . These observations were confirmed by using 125I in surface-labeling techniques.

Infect Immun, 1986 Sep, 53(3), 611 - 5
T-cell-independent macrophage activation in mice induced with rRNA from Listeria monocytogenes and dimethyldioctadecylammonium bromide; van den Bosch JF et al.; Purified rRNA from Listeria monocytogenes or Pseudomonas aeruginosa injected in combination with dimethyldioctadecylammonium bromide (DDA), protects mice nonspecifically against a lethal challenge of various extra- and intracellular bacteria . In the present study vaccination of BALB/c as well as C57BL/Ka mice with listerial RNA-DDA resulted in activation of fixed-tissue macrophages, as measured by an enhanced in vivo L . monocytogenes killing in spleen and liver . Evidence was found that macrophage activation by vaccination with rRNA-DDA occurred by a T-cell-independent mechanism . Treatment of mice with cyclosporin A had no effect on the enhanced L . monocytogenes killing induced with RNA-DDA; in vitro exposure of RNA-DDA to spleen cell cultures did not give rise to any lymphocyte proliferation . No evidence could be found for a possible adjuvant activity for RNA-DDA in cellular responses; in fact, RNA-DDA had an inhibitory effect on lymphocyte proliferative responses to Listeria antigen and to concanavalin A.

Bioorg Khim, 1986 Sep, 12(9), 1268 - 73
{Antigenic polysaccharides of bacteria . 17 . The structure of O-specific polysaccharide chain of Pseudomonas aeruginosa X (Meitert) lipopolysaccharide}; Knirel' IU et al.; O-Specific polysaccharide composed of L-rhamnose and 2-acetamido-2-deoxy-D-mannose was obtained on mild acid degradation of P . aeruginosa X (Meitert classification) lipopolysaccharide . On the basis of non-destructive analis using 1H, 13C NMR spectroscopy and Klyne's rule calculation, as well as chemical methods (acid hydrolysis, methylation, Smith degradation), it was established that the polysaccharide is built up of disaccharide repeating units of the following structure: ----4)-alpha-L-Rha-(1----3)-beta-D-ManNAc-(1----.

Bioorg Khim, 1986 Sep, 12(9), 1263 - 7
{Antigenic polysaccharides of bacteria . 16 . The structure of O-specific polysaccharide chain of Pseudomonas aeruginosa 025 (Wokatsch) lipopolysaccharide}; Knirel' IuA et al.; O-Specific polysaccharide built up of trisaccharide repeating units containing 3-acetamidino-2-acetamido-2,3-dideoxy-D-mannuronic acid (ManNAcAmA), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid (Man(NAc)2A), N-acetyl-D-fucosamine (FucNAc), and O-acetyl group was obtained on mild acid hydrolysis of P . aeruginosa O25 (Wokatsch classification) lipopolysaccharide . Basing on de-O-acetylation of polysaccharide with aqueous triethylamine accompanied by hydrolysis of acetamidino group to acetamido group, as well as on the 1H and 13C NMR data, the following structure of the repeating unit of the polysaccharide was established: (Formula: see text) P . aeruginosa O25 polysaccharide has the same carbohydrate skeleton as that of P . aeruginosa O3a,b (Lanyi classification) and differs from the latter only by the presence of the O-acetyl group at position 4 of N-acetylfucosamine.

Infect Immun, 1986 Sep, 53(3), 656 - 62
Functionally active monoclonal antibody that recognizes an epitope on the O side chain of Pseudomonas aeruginosa immunotype-1 lipopolysaccharide; Stoll BJ et al.; A murine monoclonal antibody (MAb) was prepared against Pseudomonas aeruginosa immunotype-1 (It-1) lipopolysaccharide (LPS) . The MAb bound It-1 LPS in the enzyme-linked immunosorbent assay and in the immunodiffusion and immunoblotting assays, agglutinated and opsonized It-1 bacteria, and protected against challenge with live It-1 organisms in a murine burn infection model . All of these activities were immunotype specific . Correlation of the opsonic and protective properties of the MAb with its recognition site on the LPS O side chain confirmed that such immunotype-specific determinants are important targets for protective antibodies in Pseudomonas disease . The functional equivalence of this MAb and polyclonal antibodies from hyperimmune plasma underscores the therapeutic potential of single MAbs which recognize critical determinants in the LPS O side chain.

J Biol Chem, 1986 Aug 25, 261(24), 11404 - 8
Pseudomonas exotoxin A . Membrane binding, insertion, and traversal; Farahbakhsh ZT et al.; Using vesicle targets composed of phosphatidylcholine and cholesterol (1:1 molar ratio), we found that Pseudomonas aeruginosa exotoxin A (PTx) binding and insertion are not only dependent on pH (Zalman, L.S., and Wisnieski, B.J . (1985) Infect . Immun . 50, 630-635) but also on ionic strength, reaching a maximum in pH 4 buffer that contains 150-200 mM NaCl . Insertion was monitored by photolabeling with an intramembranous probe . Higher levels of binding and insertion were attained with vesicles that contained 2.5 mol% dicetylphosphate than with neutral vesicles . Positively charged vesicles (2.7 mol% stearylamine) were the least effective targets . At pH 7.4, all binding levels were depressed . While PTx binding increased with increasing temperature, the relative proportion of the vesicle-associated toxin that was photolabeled decreased . The most likely explanation for the decrease is that the bilayer translocation rates increased with increasing temperature, and hence fewer PTx molecules were accessible at the time of photolabeling . At 37 degrees C, binding and insertion both plateaued within 10 min of lowering the pH to 4 . After 10 min, the amount of bound toxin decreased slightly with time but there was a dramatic decrease in photolabeling, indicating that inserted PTx had begun to cross the bilayer . This was verified by the finding that when PTx was incubated with vesicles that contained trypsin, cleavage occurred only in those samples in which the pH was shifted down to pH 4 . Entry is triggered by an acid-induced conformational change that promotes productive binding and insertion . After insertion, the kinetics of membrane traversal appear to be regulated by the physical properties of the bilayer.

J Biol Chem, 1986 Aug 25, 261(24), 11259 - 65
Isolation and characterization of a lysine-specific protease from Pseudomonas aeruginosa; Elliott BW Jr et al.; We report here a procedure which results in the purification of an extracellular protease (designated Ps-1) from Pseudomonas aeruginosa . This enzyme cleaves fibrinogen so that the modified molecules form microcrystals and large single crystals . Precise knowledge of the Ps-1 cleavage sites is essential for the interpretation of the structural information provided by these crystals (Weisel, J . W., Stauffacher, C . V., Bullitt, E., and Cohen, C . (1985) Science 230, 1388-1391) . Ps-1 is a single-chain polypeptide of Mr 30,000 which appears to function as a monomer . The pH optimum is 8-9 . The activity of the protease is not decreased by inhibitors of thiol, carboxyl, or metallo proteases; the abolishment of activity by N alpha-p-tosyl-L-lysine chloromethyl ketone and the partial inhibition obtained with serine-reactive inhibitors suggests that Ps-1 may be a serine protease with an unusual active-site conformation . Studies with synthetic peptide substrates show that Ps-1 exhibits one of the most restricted specificities known for an endoproteinase: only peptide, ester, and amide bonds containing the carbonyl group of lysine are hydrolyzed . The limited specificity of Ps-1 should make it useful for other applications requiring the selective cleavage of proteins, such as sequence analysis and the isolation of domains.

Biochim Biophys Acta, 1986 Aug 21, 860(2), 263 - 7
Role of lysines in ion selectivity of bacterial outer membrane porins; Hancock RE et al.; The epsilon-amino groups of available lysine residues of the OmpC, OmpF and PhoE porin proteins of Escherichia coli and of the protein P porin of Pseudomonas aeruginosa, were modified by the bulky reagent trinitrobenzenesulphonic acid . Approximately 78% of the lysines of the anion-selective protein P and PhoE porins were modified whereas only 40-50% of the lysines of the cation selective OmpF and OmpC porins were altered . After modification, the three E . coli porins had very similar high selectivities for cations over anions, in contrast to the native porins which varied 86-fold in ion selectivity . Despite the large size of the trinitrophenyl group attached to modified lysines (i.e., a disc of approx . 0.86 nm diameter X 0.36 nm high) relative to the reported size of the constrictions of the E . coli porins (1.0-1.2 nm diameter), only the anion-selective PhoE porin was substantially blocked after trinitrophenylation . The protein P porin channel was relatively unaffected by trinitrophenylation, in contrast to previous data showing dramatic effects of acetylation of lysines on protein P conductance and selectivity . This favoured a model in which the critical lysines involved in anion binding by protein P were present in a constriction of the channel that was too small for trinitrobenzenesulphonic acid to enter . Overall, the data suggest that both the number and relative position of charged lysines are major determinants of ion selectivity.

J Antibiot (Tokyo), 1986 Aug, 39(8), 1092 - 107
Synthesis and structure-activity relationships of a new series of cephalosporins, BMY-28142 and related compounds; Naito T et al.; The synthesis of a series of 7-{(Z)-2-(2-aminothiazol-4-yl)-2-oxyiminoacetamido} -3-ammoniomethyl-3-cephems is described . Variations of an oxyimino moiety in the 7-side chain and a quaternary ammonium moiety in the 3-side chain were examined and structure-activity relationships studied . BMY-28142, the 3-(N-methylpyrrolidinio)methyl derivative of the 7-alpha-methoxyimino series of cephalosporins, exhibited broad antimicrobial activity against both Gram-positive and Gram-negative bacteria including Pseudomonas aeruginosa.

Ophthalmic Surg, 1986 Aug, 17(8), 499 - 501
Eyelid necrosis in an anophthalmic patient; Woog JJ et al.; A 72-year-old woman with chronic lymphocytic leukemia developed periorbital swelling, erythema, and discharge involving her anophthalmic left socket in the course of pseudomonas aeruginosa septicemia . Histopathologic examination of debrided eyelid tissues revealed ischemic necrosis of the eyelid margin and deeper eyelid tissues . Underlying malignancy, disseminated systemic infection, and alteration in the vascular supply to the eyelids following enucleation may have contributed to the development of ischemic eyelid necrosis in this patient.

Antimicrob Agents Chemother, 1986 Aug, 30(2), 260 - 6
Pharmacokinetics and pharmacodynamics of ciprofloxacin in cystic fibrosis patients; LeBel M et al.; The pharmacokinetics and blister fluid penetration of oral ciprofloxacin were compared in 11 cystic fibrosis (CF) patients who had sputum colonization but were asymptomatic and in 12 healthy volunteers after a single dose (500 mg) and at steady state (500 mg every 8 h) . The antibacterial effect of ciprofloxacin therapy was also evaluated by bacterial counts of colonizing pathogens in the respiratory secretions of CF patients . The CF patients were 15.9% lighter in weight than the controls (P less than 0.05) . After a single dose, the elimination half-life of ciprofloxacin was decreased by a third in the CF patients as compared with the controls (2.62 versus 3.93 h, respectively; P less than 0.01) . This was the result of a diminished apparent volume of distribution in CF subjects . Interestingly, we observed no statistically significant difference in total apparent and renal clearances between the groups . Suction-induced blister fluid penetration was not different between CF patients and healthy volunteers . In CF patients, ciprofloxacin exhibited levels in respiratory secretions above the reported MIC for Pseudomonas aeruginosa: 1.36 and 1.86 micrograms/ml at 2 h after a single dose and at steady state, respectively . An important fall (mean, 3.9 log10/ml) in the log titer in 10 patients with P . aeruginosa in their respiratory secretions was observed after 5 days of treatment . However, this improvement was short-lived; the secondary increase in bacterial counts observed in five patients and the development of five resistant strains were causes for concern . The pharmacokinetic results presented here showed that ciprofloxacin should be administered every 8 or even every 6 h in CF patients.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Aug, (8), 42 - 5
{Experimental basis for the combined use of tobramycin and immune preparations for treating acute Pseudomonas aeruginosa infections}; Minukhin VV et al.; The possibility of using tobramycin (Tb) in combination with P . aeruginosa polyvalent corpuscular vaccine (PaPCV) or pyocyanosis hyperimmune plasma (PHP) for the treatment of P . aeruginosa sepsis was experimentally studied . The combined use of Tb and PHP, administered in amounts corresponding to ED50 of each preparation used separately, ensured the survival of 90% of the infected mice, and the injected of PaPCV with ED50 of the antibiotic ensured the survival of 73% of the experimental animals . The combined use of Tb and the two immuno-preparations (PaPCV and PHP) for the treatment of P . aeruginosa infection proved to be more effective than their separate administrations.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Aug, (8), 18 - 21
{Tetrathionate reductase as a diagnostic trait in identifying Pseudomonas aeruginosa}; Kalina GP et al.; The tetrathionate reductase test may be used for the identification of P . aeruginosa . The most reliable results have been obtained with the use of a medium containing sodium tetrathionate for this purpose, and replacing bromthymol blue used as an indicator for phenol red excludes the possibility of false negative reactions.

Am Rev Respir Dis, 1986 Aug, 134(2), 214 - 6
Lack of bacterial aerosols associated with heat and moisture exchangers; Saravolatz LD et al.; Contaminated condensate might serve as a source for cross infection . Heat and moisture exchangers (HME) are devices that humidify inspired gases, which pass through a hygroscopic felt pad surrounded by a cellulose sponge housed in a plastic case . In our study, we used a Servo 150 HME in place of a cascade humidifier in mechanical ventilator circuits . We performed 2 studies to evaluate the microbiologic safety of the HME . First, 42 HMEs used by patients for 24 h were tested in the laboratory for contamination . To simulate patient/air exchange, the HMEs were connected to the Andersen Sampler (flow at 35 L/min x 20 min) . Although the inner felt pad of the HMEs was contaminated in 74% of the units (31 of 42), only 4.8% (2 of 42) generated 1 to 2 bacteria/702 L of air . In a second study, HMEs contaminated with either Staphylococcus aureus or Pseudomonas aeruginosa (at 10(3), 10(5), or 10(8) organisms/ml) were connected to an Andersen Air Sampler to simulate a ventilator circuit . Bacterial aerosols were not generated, with the exception of 2 to 4 bacteria recovered after contamination with 10(8) bacteria . The HME can provide humidification for mechanically ventilated patients with little risk of generating respirable bacterial aerosols.

J Infect Dis, 1986 Aug, 154(2), 289 - 94
Emergence of resistance to imipenem during therapy for Pseudomonas aeruginosa infections; Quinn JP et al.; We studied the mechanism of resistance to imipenem in three clinical isolates of Pseudomonas aeruginosa . Two of these isolates arose from imipenem-susceptible strains isolated during therapy with imipenem and were associated with treatment failure . One of these two strains had previously been broadly resistant to beta-lactams; the second acquired resistance to imipenem alone . One isolate of the third strain was resistant to imipenem but susceptible to other antipseudomonal beta-lactams . No isolate contained beta-lactamase activity capable of hydrolyzing imipenem at a detectable rate . Studies of the penicillin-binding proteins of all isolates revealed no differences in the number of proteins, molecular weight of, affinity for penicillin, or affinity for imipenem in any isolate . In each case the resistant isolate lacked one or more outer membrane proteins that were present in a susceptible isolate of the same strain . The observed alterations in outer membrane proteins may be associated with diminished permeability of the bacterial outer membrane to imipenem and may be the major factor responsible for resistance in these isolates.

Genetika, 1986 Aug, 22(8), 2048 - 54
{Effective, plasmid RP4-dependent replication-transposition of DNA of the transposable phage D3112 of Pseudomonas aeruginosa in a heterologous Escherichia coli system}; Akhverdian VZ et al.; The processes of replication and transposition of Pseudomonas aeruginosa transposable phage D3112 in cells of Escherichia coli (D3112) and E . coli (RP4::D3112) were studied . D3112 genome is a "silent cassette" ("conex-phage"--conditionally expressible) in E . coli cells incubated at 42 degrees C . Two compulsory conditions for D3112 genome expression are incubation at 30 degrees C and the presence in cells of RP4 plasmid . Processes of replication and transposition in E . coli are coupled . RP4 plasmid stimulates D3112 DNA synthesis in E . coli at least by two order of magnitude . In correspondence with this observation is the fact that when Mg2+ is present in high concentration (0.1 M) in a cultural medium, the production of mature phage is enhanced by two order of magnitude in E . coli (RP4::D3112) or in E . coli (D3112, RP4) cells, and is approx . 10(-1)-10(-2) phage per cell . No influence of Mg on phage production is observed in E . coli (D3112) cells.

J Clin Microbiol, 1986 Aug, 24(2), 210 - 3
Oral succession of gram-negative bacilli in myelosuppressed cancer patients; Minah GE et al.; Aerobic and facultative gram-negative bacilli (GNB) have been reported to increase on various body surfaces in the seriously ill and debilitated patient . This study examined quantitative aspects of GNB succession at five oral sites in cancer patients before and during myelosuppressive chemotherapy . GNB concentrations increased sharply during chemotherapy at 25 to 50% of the oral sites in both acute nonlymphocytic leukemia and small-cell lung carcinoma patients . Most sites did not exhibit shifts of GNB to levels higher than 0.1% of the cultivable flora . When shifts occurred, all sites sampled in the mouth were usually affected and GNB usually represented more than 10% of the cultivable flora . Low levels of indigenous microflora were observed in most sites exhibiting GNB shifts . None of the subjects harboring high levels of GNB developed the symptoms of acute infection which are commonly observed in myelosuppressed patients . Although Pseudomonas aeruginosa and Klebsiella pneumoniae were recovered from some sites, most GNB were nonpathogenic species of Pseudomonas; Pseudomonas pickettii was the most frequently recovered.

J Bacteriol, 1986 Aug, 167(2), 473 - 9
Expression in Escherichia coli and function of Pseudomonas aeruginosa outer membrane porin protein F; Woodruff WA et al.; The gene encoding porin protein F of Pseudomonas aeruginosa was cloned onto a cosmid vector into Escherichia coli . Protein F was expressed as the predominant outer membrane protein in a porin-deficient E . coli background and was clearly visible on one-dimensional sodium dodecyl sulfate-polyacrylamide gels in a porin-sufficient background . The identity of the protein F from the E . coli clone and native P . aeruginosa protein F was demonstrated by their identical mobilities on sodium dodecyl sulfate-polyacrylamide gel electrophoretograms, 2-mercaptoethanol modifiabilities, and reactivities with monoclonal antibodies specific of two separate epitopes of protein F . In the course of gene subcloning, a 2-kilobase DNA fragment was isolated, with an apparent truncation of the part of the gene encoding the carboxy terminus of protein F . This subclone produced a 24,000-molecular-weight, outer membrane-associated, truncated protein F derivative which was not 2-mercaptoethanol modifiable and which reacted with only one of the two classes of protein F-specific monoclonal antibodies . The 2-kilobase fragment was used in Southern blot hybridizations to construct a restriction map of the cloned and subcloned fragments and to demonstrate with restriction digests of whole P . aeruginosa DNA that only one copy of the protein F gene was present in the P . aeruginosa chromosome . The protein F produced by the original cosmid clone in a porin-deficient E . coli background was purified . To demonstrate retention of porin function after cloning, the protein F from the E . coli clone was incorporated into black lipid bilayer membranes . Two major classes of channels were revealed . The predominant class of channels had an average conductance of 0.36 nS in 1 M KCl, whereas larger channels (4 to 7 nS) were seen at a lower frequency . Similar channel sizes were observed for porin protein F purified by the same method from P . aeruginosa outer membranes.

Endocrinology, 1986 Aug, 119(2), 515 - 21
Effects of an acute bacterial infection on serum thyroid hormones and nuclear triiodothyronine receptors in mice; Burgi U et al.; Serum T3, T4, and rT3 levels as well as liver nuclear T3 receptors (NT3R) were measured in mice with a bacterial infection . Pseudomonas aeruginosa were injected into one thigh of ICR mice, resulting in a severe infection at sacrifice 15 h later . Since food intake, which influences serum thyroid hormone levels and NT3R, was 75% lower in infected than in control mice, infected mice were either fed and compared with pair-fed controls or fasted and compared with fasted and fed controls . Fasting induced a fall in serum T3 and T4 levels, which was even more pronounced in infected fasted animals . However, while fasting caused an approximately 80% increase in serum rT3 concentrations, serum rT3 levels in infected fasted animals were not different from those in fed controls . The combination of infection and fasting thus prevented the rise in serum rT3 otherwise invariably associated with fasting . NT3R measurements on isolated nuclei revealed the presence of NT3R in mouse liver similar to those reported in rat liver . The NT3R Kd (approximately 2 X 10(-10) M) was not affected by decreased food intake, infection, or a combination thereof . The NT3R maximum binding capacity (MBC) was decreased in fasted animals (460 vs . 306 pg/mg DNA) . However, the MBC of infected fasted mice was not different from that of fasted mice . Similarly, no difference in MBC was found between infected fed and pair-fed control mice . In mice injected with heat-killed P . aeruginosa to evaluate potential effects of endotoxins, neither serum thyroid hormone levels nor hepatic NT3R were different from those of controls . These data show that in mice, a severe bacterial infection with P . aeruginosa has effects on serum hormone levels not explained by the disease-associated diminished food intake, whereas it has no effects on liver NT3R beyond those due to the disease-related decreased food intake.

Acta Pathol Microbiol Immunol Scand {B}, 1986 Aug, 94(4), 273 - 6
Population analyses of the susceptibility to ciprofloxacin of eight clinical strains of Pseudomonas aeruginosa; Gerner-Smidt P et al.; Population analyses of the susceptibility to ciprofloxacin of eight strains of Pseudomonas aeruginosa, including mucoid strains and strains resistant to aminoglycosides or anti-Pseudomonas beta-lactams, were carried out . All strains were sensitive as judged by the broth-dilution technique, but four strains were found to yield resistant mutants with a frequency of less than 10(-6) . Two strains yielded mutants homogeneously sensitive to ciprofloxacin at a level 8-16 times the MIC of the parent strains . Two other strains yielded mutants resistant to different higher concentrations of ciprofloxacin . One of these mutants was examined for production of ciprofloxacin-inactivating enzyme; no enzyme production could be detected . Cross-resistance was found with another quinolone antibiotic, ofloxacin, but not with aminoglycosides or beta-lactam antibiotics.

J Clin Microbiol, 1986 Aug, 24(2), 189 - 96
Polysaccharide surface antigens expressed by nonmucoid isolates of Pseudomonas aeruginosa from cystic fibrosis patients; Pier GB et al.; We tested nonmucoid Pseudomonas aeruginosa isolates obtained from cystic fibrosis (CF) patients for the expression of lipopolysaccharide (LPS) serotype antigens, serum sensitivity, and production of mucoid exopolysaccharide (MEP) . When all nonmucoid isolates were compared with a set of random mucoid isolates, 20 of 52 (38%) nonmucoid isolates were typable and serum resistant, compared with 13 of 51 (24%) mucoid isolates (P = 0.16 by chi-square analysis) . However, nonmucoid strains from CF patients colonized only with nonmucoid strains were more frequently typable and serum resistant (67%) than were nonmucoid isolates from patients cocolonized with mucoid strains (31%) (P = 0.012, Fisher exact test) . An inhibition enzyme-linked immunosorbent assay done with bacterial extracts, a direct-whole-cell enzyme-linked immunosorbent assay done with affinity-purified antibody to MEP, and immune electron microscopy all demonstrated production of MEP by all nonmucoid P . aeruginosa isolates tested, including nonmucoid revertants of mucoid strains . No other bacterial species tested positive in these assays . These findings suggest that MEP is produced by all P . aeruginosa isolates obtained from CF patients, that the initial colonizing nonmucoid strains produce a smooth LPS, and that once LPS-rough, mucoid strains appear in the sputum, the predominant LPS phenotype is rough regardless of colony morphology.

J Clin Invest, 1986 Aug, 78(2), 375 - 80
T lymphocyte-mediated protection against Pseudomonas aeruginosa infection in granulocytopenic mice; Powderly WG et al.; BALB/c mice immunized with Pseudomonas aeruginosa immunotype 1 polysaccharide develop protective T cell immunity to bacterial challenge . In vitro, T cells from immunized mice kill P . aeruginosa by production of a bactericidal lymphokine . The present study demonstrates that adoptive transfer of T cells from immunized BALB/c mice to granulocytopenic mice resulted in 97% survival on challenge with P . aeruginosa, compared with 17% survival with adoptive transfer of T cells from nonimmune BALB/c mice . This protection is specifically elicited by reexposure to the original immunizing antigen; adoptive recipients cannot withstand challenge with immunotype 3 P . aeruginosa . However, the adoptive recipients do survive simultaneous infection with both P . aeruginosa immunotypes 1 and 3 . Adoptive transfer of T cells from the congenic CB.20 mice, which are unable to kill P . aeruginosa in vitro, provides only 20% protection to granulocytopenic mice . These studies indicate that transfer of specific immune T lymphocytes can significantly enhance the resistance to P . aeruginosa infection in granulocytopenic mice.

J Bacteriol, 1986 Aug, 167(2), 611 - 5
Overproduction and assay of Pseudomonas aeruginosa phosphomannose isomerase; Gill JF et al.; Phosphomannose isomerase activity was undetectable in extracts of mucoid (alginate-producing) Pseudomonas aeruginosa . When a P . aeruginosa gene previously shown to complement an alginate-negative mutant was overexpressed under the control of the tac promoter in the broad-host-range controlled-expression vector pMMB22, phosphomannose isomerase activity could be measured in extracts of P . aeruginosa and in a manA (phosphomannose isomerase-negative) mutant of Escherichia coli . P . aeruginosa extracts containing induced levels of enzyme were shown to interconvert fructose 6-phosphate and mannose 6-phosphate . A 56,000-dalton polypeptide was visualized on sodium dodecyl sulfate-polyacrylamide gels after induction in both hosts . When RNA-DNA dot- blot hybridization analysis was used, transcription of algA, the gene coding for P . aeruginosa phosphomannose isomerase, was not measurable from the chromosomes of either mucoid or nonmucoid P . aeruginosa . However, a high level of algA transcription was detected after expression of algA under tac promoter control in pMMB22.

J Immunol, 1986 Aug 1, 137(3), 988 - 94
Antigenic specificities of Pseudomonas aeruginosa alkaline protease and elastase defined by human T cell clones; Parmely MJ et al.; Virulent strains of Pseudomonas aeruginosa derive their pathogenicity, in part, from their secretion of two proteolytic enzymes, alkaline protease (AP) and elastase (E) . Human T lymphocytes specific for AP and E can be detected in the blood of immune donors and have afforded the opportunity to characterize the antigenicity of these proteins . To accomplish this goal, we have recently selected 68 human T cell clones from five different Pseudomonas-immune donors and determined their fine specificities . Fifty-five (81%) were found to be protease specific, demonstrating the immunogenicity of the exoenzymes in humans . These clones defined five AP and three E specificities and suggested the existence of at least five allomorphic determinants expressed on the proteases of various Pseudomonas strains . Limiting dilution analysis confirmed a number of antigenic relationships suggested by the long-term T cell clones and revealed that T cells specific for allomorphic protease determinants were at least as frequent in the blood of immune donors as were T cells committed to conserved determinants . Thus, both primary and long-term human T cell clones showed specificity patterns that distinguished proteases from different Pseudomonas strains . These observations describe a heretofore unknown antigenic system of Pseudomonas aeruginosa that should assist in defining the nature and specificity of Pseudomonas immunity in humans.

Am J Med, 1986 Jul 28, 81(1A), 73 - 7
Infection in patients with cystic fibrosis; Rubio TT; Cystic fibrosis is the most common lethal genetic disease of Caucasians . The disease affects the exocrine gland secretions throughout the body, and as a result, major pathologic changes develop in the pancreas and in the bronchi . Obstruction of the respiratory airways results in chronic infection, and in time, this leads to progressive deterioration of lung function . In the initial stages of the disease, usually during infancy, infection with Staphylococcus aureus plays an important role . Hemophilus influenzae infections are also common . As the disease progresses, infection with Pseudomonas aeruginosa develops . Exacerbation of bronchopulmonary infection is often initiated by respiratory viral or mycoplasmal infection, with superimposed S . aureus and P . aeruginosa infections contributing to the severity of the infection . Frequent courses of antibiotic therapy are usually required, and some patients may have to receive antibiotics continuously . Oral cephalosporins, ampicillin, and the combination of trimethoprim/sulfamethoxazole are commonly used for relatively mild infections . In the treatment of exacerbation of infection, intravenous penicillinase-resistant penicillins and anti-Pseudomonas antibiotics are the drugs of choice . For Pseudomonas infections, ticarcillin, carbenicillin, the ureidopenicillins, and the aminoglycosides are indicated . The combination of an anti-Pseudomonas penicillin and an aminoglycoside are most commonly used . Of the third-generation cephalosporins, ceftazidime appears to be the most efficacious . The quinolones (such as ciprofloxacin) are also active against P . aeruginosa, and preliminary studies of these drugs in patients with cystic fibrosis appear to indicate that they are as efficacious as the already available antibiotics . In many centers, Pseudomonas cepacia has emerged as a serious problem in patients with cystic fibrosis . This organism tends to develop resistance to multiple antibiotics . In some centers, infection with P . cepacia has been associated with a severe, frequently fatal, pneumonia.

FEBS Lett, 1986 Jul 28, 203(2), 131 - 4
A copper-containing protein that inhibits nitrite reductase from Pseudomonas aeruginosa; Karapetian AV et al.; A non-blue copper-containing glycoprotein was isolated from Pseudomonas aeruginosa . The protein has a molecular mass of 10 kDa and contains 1 atom of EPR-detectable type II copper . The protein inhibits oxidation of both azurin and cytochrome c-551 catalyzed by nitrite reductase from Ps . aeruginosa . Thus, it may be considered as an endogenous inhibitor of nitrite reductase.

Am Rev Respir Dis, 1986 Jul, 134(1), 22 - 6
Bacterial infection and acute lung injury in hamsters; Seidenfeld JJ et al.; Bacterial pneumonia is a common complication of lung injury that can be an important determinant of outcome . We studied experimental lung injury produced in hamsters by injecting 20 mg/kg paraquat (PQ) intraperitoneally; control animals received saline vehicle . Three days later, Pseudomonas aeruginosa (PAO1), 10(8) organisms in 0.25 ml, or saline, 0.25 ml, was inoculated intratracheally . Lung and systemic antibacterial defenses were studied at death 24 h later . Paraquat alone produced focal interstitial pneumonitis and neutrophilic alveolitis, and resulted in a 12% (3 of 26) mortality . PAO1 alone caused focal pneumonias and no deaths . Animals receiving both agents (PAO1/PQ) had extensive diffuse alveolar damage characterized by alveolar hemorrhage, edema, influx of neutrophils, and vasculitis; 50% (16 of 32) died within 96 h of PQ injection . Mean lung counts of PAO1 at death were 7.6 X 10(4) colony forming units/g in PAO1 and 2.8 X 10(7) in PAO1/PQ animals (p less than 0.05) . PAO1 colony counts in liver were increased nearly 100-fold in PAO1/PQ animals (p less than 0.05) . Half-time of clearance of P . aeruginosa from the blood was prolonged in PAO1 and in PAO1/PQ animals (p less than 0.05) but not in PQ animals . Phagocytosis of Staphylococcus aureus by leukocytes lavaged from the lung was not impaired in any group compared with that in control animals, but intracellular killing was impaired in PAO1 and PAO1/PQ but not in PQ animals . Paraquat injury impairs lung antibacterial defenses by uncertain mechanisms . Superinfection of PQ-injured lungs by PAO1 appears responsible for defects in intrapulmonary and systemic antibacterial defenses.

Pediatr Infect Dis, 1986 Jul-Aug, 5(4), 440 - 3
Comparison of piperacillin vs . ticarcillin plus tobramycin in the treatment of acute pulmonary exacerbations of cystic fibrosis; Jackson MA et al.; During a 22-month period 35 children with cystic fibrosis received 52 courses of antibiotic therapy for acute pulmonary exacerbations, including 26 cases of therapy with piperacillin and 26 courses with ticarcillin plus tobramycin . Groups were similar in age (5 vs . 5.4 years), disease severity based on Schwachman scores and presenting symptoms . Pseudomonas aeruginosa was the most common organism isolated in 90% of sputum cultures . Mean minimal inhibitory concentrations for piperacillin, ticarcillin and tobramycin were 8, 64 and 1 microgram/ml, respectively . Piperacillin pharmacokinetic data revealed an average half-life in serum of 36 minutes . Peak serum concentrations averaged 144 micrograms/ml, and after 4 hours serum concentrations continued to exceed the P . aeruginosa 90% minimal inhibitory concentration in 50% of children . The dosage requirement for tobramycin was quite variable, necessitated monitoring of aminoglycoside serum concentrations and in most cases resulted in at least one dosage adjustment . Emergence of resistant bacteria was not seen in 26 courses of piperacillin therapy . Both regimens were effective and well-tolerated . Single agent therapy has the advantage of providing reliable serum concentrations and, in contrast to the standard therapy, does not necessitate monitoring of serum drug concentrations.

J Biomed Mater Res, 1986 Jul-Aug, 20(6), 731 - 8
Omiderm, a new synthetic wound covering: physical properties and drug permeability studies; Behar D et al.; Omiderm (Omikron Scientific Ltd., Rehovot, Israel), a new synthetic wound covering based on hydrophilized polyurethane, was found to be highly permeable to water . Values in the region of 5000 g/m2 24 h were found for the water permeability of Omiderm in comparison to 1400 and 500 g/m2 24 h for Biobrane (Hall, Woodroof Inc., Santa Ana, CA) and OP site (Smith and Nephew Ltd.), respectively . Permeabilities of antibacterial agents through Omiderm were found to be two to three orders of magnitude greater than those through Biobrane . The in vitro effectiveness of various antibacterial agents in lowering bacterial growth of different bacterial strains were applied to seeded agar plates through Omiderm membrane was investigated . NBH ointment (1% Neomycin, 1% Bacitracin, and 0.5% Hydrocortisone) was found to be the most effective material in inhibiting bacterial growth, except for Pseudomonas aeruginosa where silver sulfadiazine was superior . In in vivo experiments bacterial counts of infected wounds covered with Omiderm and topically treated with NBH were lowered to less than 10(3) organism/g tissue after one day of treatment.

J Infect Dis, 1986 Jul, 154(1), 64 - 8
Failure of therapy in pseudomonas endocarditis: selection of resistant mutants; Jimenez-Lucho VE et al.; Despite optimal use of available antibacterial agents, endocarditis due to Pseudomonas aeruginosa is commonly associated with poor response to medical treatment . Two patients are described in whom emergence of resistance to beta-lactam antibiotics was associated with clinical failure . A subpopulation of resistant mutants (10(-7)) was found within the initial, apparently sensitive population of bacteria . These resistant mutants were similar to posttherapy isolates in their increased production of beta-lactamase and in their identical pattern of resistance to beta-lactam antibiotics . Moreover, the only beta-lactamase produced was type Id, and this enhanced production proved to be constitutive . A relatively large inoculum (10(6) colony-forming units/g of tissue) of bacteria was found postoperatively in the heart valves of both patients . The failure to respond is postulated to be due to the selection of these producers of high levels of beta-lactamase in a large bacterial inoculum.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1986 Jul, 261(4), 400 - 6
O-serotypes of Pseudomonas aeruginosa from animal and inanimate sources in Saudi Arabia; Barbour EK et al.; A total of 1012 samples were examined for Pseudomonas aeruginosa and 257 (25.4%) were positive . The incidence of Ps . aeruginosa in samples collected from animal sources (N = 730) was significantly higher (28.7%) than that in 282 samples of inanimate sources (16.7%) . The percentage of samples infected with these organisms was lowest in poultry feed (2.8%) and highest in sewage effluent (57.1%) . Nine serotypes were defined from all sources . P5 was the common predominant individual O type in infected chicken navels and in the nasal cavities of Najdi sheep (a Saudi Arabian sheep breed), while P3 and P6 were predominate in the nasal cavities of Somali sheep (a breed imported from Somalia) . No Ps . aeruginosa serotype was predominant in sheep faeces . In inanimate sources, P4 was predominant in water and sewage effluent . The isolate from the animal feed was untypeable . In using the slide agglutination technique for serotyping, most of the unusual agglutination reaction types of Ps . aeruginosa (70%) were of strains isolated from Somali sheep.

J Antimicrob Chemother, 1986 Jul, 18(1), 45 - 9
Susceptibility of Pseudomonas aeruginosa strains isolated from hospitalized children; Patzer J et al.; Pseudomonas aeruginosa strains, 1285 in total, were isolated from hospitalized children in the period 1981-1984 . Serotypes 011 and 06 and nontypable strains were the most frequently identified . 44.6% of serotype 011 strains and between 12.4 and 37.7% of other serotypes were resistant to gentamicin, or carbenicillin, or both agents . The aminoglycosides with the highest activity against P . aeruginosa isolates were amikacin and netilmicin . Among the seven beta-lactam antibiotics tested the most active were cefsulodin, piperacillin and azlocillin.

Antimicrob Agents Chemother, 1986 Jul, 30(1), 35 - 8
Discrepancies between disk diffusion and broth susceptibility studies of the activity of ticarcillin plus clavulanic acid against ticarcillin-resistant Pseudomonas aeruginosa; Manian FA et al.; Ticarcillin and clavulanic acid in combination were tested against 40 Pseudomonas aeruginosa isolates resistant to ticarcillin by disk diffusion . A total of 21 isolates (53%) were susceptible to ticarcillin-clavulanate by disk diffusion, under currently recommended criteria for ticarcillin susceptibility . Macro-broth dilution tests (ticarcillin plus clavulanic acid, 2 micrograms/ml) confirmed susceptibility (MIC less than or equal to 64 micrograms/ml) of only 8 (38%) of 21 isolates . Time-kill studies of disk diffusion susceptible isolates indicated 2 log10 or greater killing of most isolates at 6 h in broth containing ticarcillin (64 micrograms/ml) combined with clavulanic acid (1, 2, 5, or 10 micrograms/ml) . After 6 h, regrowth was common in all concentrations of clavulanic acid except 10 micrograms/ml . Regrowth populations were resistant to ticarcillin-clavulanate by MIC determination . Poor bactericidal activity of ticarcillin-clavulanate against ticarcillin-resistant P . aeruginosa was confirmed, as most isolates did not undergo 99.9% or greater killing at 24 h in all concentrations of clavulanic acid . Serotype O-11 was our most common serotype and was associated with disk diffusion "pseudosusceptibility." Concomitant disk diffusion testing of ticarcillin-clavulanate and ticarcillin is recommended for testing the susceptibility of P . aeruginosa to ticarcillin-clavulanate by disk diffusion . P . aeruginosa isolates resistant to ticarcillin should as a rule be considered also resistant to ticarcillin-clavulanate, despite apparent susceptibility by disk diffusion.

Antimicrob Agents Chemother, 1986 Jul, 30(1), 25 - 30
In vitro activity of piperacillin, ticarcillin, and mezlocillin alone and in combination with aminoglycosides against Pseudomonas aeruginosa; Lyon MD et al.; A total of 103 isolates of Pseudomonas aeruginosa were studied to compare the in vitro effectiveness of three beta-lactam antibiotics (piperacillin, ticarcillin, and mezlocillin) when used alone and in combination with four aminoglycosides (tobramycin, gentamicin, amikacin, and netilmicin) . All drugs were tested as single agents against a standard inoculum (5 X 10(5) CFU/ml) . The three antipseudomonal penicillins were also tested against the isolates at a higher inoculum concentration (10(7) CFU/ml) . Synergy testing was performed by the two-dimensional checkerboard method and was defined by a fractional bactericidal index of less than or equal to 0.5 and bacterial killing accomplished at antibiotic concentrations no greater than those achievable in serum . All combinations were assessed for synergy . The degree of synergy was further analyzed by dividing the isolates into groups based on their susceptibility and resistance to the individual agents in the combination . The overall effectiveness of the various aminoglycoside-antipseudomonal penicillin combinations was assessed regarding their ability to kill the isolates either as single agents or through synergy . Piperacillin was the most active antipseudomonal penicillin, and tobramycin and amikacin were the most active aminoglycosides when used as single agents . When tested against isolates at a higher inoculum concentration, ticarcillin was significantly more active than the other beta-lactams . The highest degree of overall synergy was noted with gentamicin-ticarcillin (78.2% of strains) and amikacin-piperacillin (77% of strains) . When assessed for overall effectiveness, all combinations containing amikacin were the most active . The combination of amikacin-piperacillin was the most effective, with activity against 96% of all isolates.

Antimicrob Agents Chemother, 1986 Jul, 30(1), 122 - 6
Serum bactericidal activity and killing rate for volunteers receiving imipenem, imipenem plus amikacin, and ceftazidime plus amikacin against Pseudomonas aeruginosa; Van der Auwera P et al.; Serum bactericidal activity against 20 strains of Pseudomonas aeruginosa was studied in 10 volunteers after administration of imipenem (25 mg/kg), imipenem (25 mg/kg) plus amikacin (7.5 mg/kg), and ceftazidime (25 mg/kg) plus amikacin (7.5 mg/kg) . Eight strains were susceptible and 12 were resistant to ticarcillin . Serum levels were measured microbiologically after 30 and 60 min and were, respectively, 97 and 46 micrograms/ml for imipenem given alone and 79 and 45 micrograms/ml for imipenem given with amikacin . Despite the very large dose of imipenem used, imipenem and imipenem plus amikacin appeared slightly less active than ceftazidime plus amikacin (P less than or equal to 0.1; Wilcoxon matched-pairs test), with respective median titers at 30 min of 1:128, 1:128, and 1:256 against ticarcillin-susceptible strains and 1:32, 1:32, and 1:64 against ticarcillin-resistant strains; however, more than 90% of the serum determinations, regardless of the regimen, had a serum bactericidal activity greater than or equal to 1:8 . Amikacin significantly increased the rate of killing in serum of P . aeruginosa by imipenem . Imipenem plus amikacin appeared as effective as ceftazidime plus amikacin in reducing the viable counts of P . aeruginosa after 24 h of incubation.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Jul, (7), 44 - 8
{Effect of Pseudomonas aeruginosa exotoxin on blood biochemical indices in an experiment}; Dziubak ST; Some problems connected with the pathogenic action of P . aeruginosa exotoxin under experimental conditions have been studied . The study has revealed that in the process of P . aeruginosa intoxication the development of hypoproteinemia, an increase in the activity of aminotransferases, lactate dehydrogenase and its isoforms occurs in the body . The characteristic features of the process are phasic changes in the activity of ceruloplasmin and in the amount of sulfhydryl groups.

Rev Infect Dis, 1986 Jul-Aug, 8 Suppl 4, S420 - 5
Immunoglobulin G: potentiation of tobramycin and azlocillin in the treatment of Pseudomonas aeruginosa sepsis in neutropenic mice and neutralization of exotoxin A in vivo; Collins MS et al.; Mice with cyclophosphamide-induced neutropenia were challenged with four immunotypes of Pseudomonas aeruginosa by contamination of a small dorsal surface wound . The infections were lethal; 100% of control animals (n = 80) treated only with albumin died . Administration of an immunoglobulin G intravenous preparation (IGIV) and/or therapy with tobramycin or azlocillin was begun 16 hr after challenge . Mortality among mice (n = 120) treated only with an antibiotic was 75.0%, while that among mice (n = 80) treated only with IGIV was 78.8% . Combination therapy with IGIV and an antibiotic (n = 120) resulted in mortality of 38.3% . The protection afforded by IGIV may have resulted in part from neutralization of exotoxin A, as mice treated with IGIV before challenge with exotoxin A were subject to lower mortality and had lower levels of serum aspartate and alanine aminotransferases than controls.

Genetika, 1986 Jul, 22(7), 1093 - 8
{Differences in the allele state of genes controlling the specificity of adsorption in transposable phages of Pseudomonas aeruginosa}; Krylov VN et al.; To reveal possible differences in adsorptional specificities of transposable phages of Pseudomonas aeruginosa and to study the genetical control of this character, we isolated a group of phage-resistant P . aeruginosa mutants using some temperate and virulent phages . The study of resistance of the mutants to all the phages permitted us to find some types of mutants and to build a formal scheme of distribution of adsorptional receptors on the surface of P . aeurginosa cell . According to the results obtained, there are two main "receptor chains", where the receptors for all phages under study are grouped . For the majority of phages, just a single adsorptional receptor is obligatory, and at least two essential receptors are needed for adsorption of virulent phage E79 . Two receptors were found also for another virulent phage, phi 11, one of them only being essential . Transposable phages can be grouped into three types, according to their adsorptional specificities . No correlations of adsorptional specificity types and all other characteristics of transposable phages studied (including the sub-groups of transposable phages belonging to different DNA homology types) were found . Genes of natural transposable phages controlling the differences in adsorptional specificities revealed can recombine in phage crosses.

Drug Intell Clin Pharm, 1986 Jul-Aug, 20(7-8), 575 - 81
Pseudomonas aeruginosa susceptibility in a university hospital: recognition and treatment; Rotschafer JC et al.; Pseudomonas aeruginosa continues to be a leading cause of nosocomial bacteremia and other serious, often life-threatening infections . The incidence of P . aeruginosa infection appears to be increasing . The resilience of Pseudomonas in the hospital environment, its endogenous virulence factors, and its current level of resistance to antimicrobials make it a formidable pathogen, particularly in a compromised host . Despite the availability of several effective antipseudomonal antibiotics, infections caused by this pathogen are still associated with significant morbidity and mortality . Early recognition and prompt intervention with appropriate antimicrobial agents are vital to successful management . Combination therapy with an aminoglycoside and an extended-spectrum penicillin or cephalosporin is recommended in the initial management of suspected or documented P . aeruginosa infections.

Chirurg, 1986 Jul, 57(7), 452 - 6
{Long-term intubation following tracheotomy}; Kramer W et al.; Between 1981 and 1983 tracheotomy was performed on 61 patients in the Surgical University Clinic of Tubingen . The dominant factors in indication of tracheotomy was for 49% of patients the persistingly necessary artificial respiration, for 26% a better bronchial toilet and other reasons for 25% . With 40.5% of all cases pneumonia was the most frequent complication encountered with our patients . The most frequent bacteria was Pseudomonas aeruginosa . Because of the decisive advantage constituted by the possibility of unproved bronchial toilet an earlier realisation of tracheotomy is to be recommended.

Ann Surg, 1986 Jul, 204(1), 48 - 52
Effects of infection on oxygen consumption and core temperature in experimental thermal injury; Aulick LH et al.; Oxygen consumption (VO2) and colonic temperature (Tc) were measured in groups of rats before and after 30% total body surface, full thickness burns . Some wounds were seeded with Pseudomonas aeruginosa or Staphylococcus epidermidis, and some seeded wounds were treated with Sulfamylon or Silvadene . Three groups became bacteremic (B) during the 2-3 week period of observation . At an ambient temperature (Ta) of 32 C, VO2 of the B group rose from 0.83 +/- 0.01 to 1.20 +/- 0.01 ml/hr/g (mean +/- S.E., p less than 0.001) versus 0.81 +/- 0.01 to 0.99 +/- 0.02 for nine nonbacteremic (NB) groups (p less than 0.001) . Tc increased only in the B groups--from 36.8 +/- 0.1 to 37.7 +/- 0.1 C (p less than 0.001) . In the second or third week postinjury, VO2 of the NB rats was reduced when Ta was increased to 34 C; Tc followed changes in Ta . Sulfamylon lowered VO2 of P . aeruginosa seeded, NB rats . The metabolic cost of wound contamination appeared to vary with bacterial strain . The metabolic effects of infection appear to be a continuum, beginning with a modest rise in VO2 and progressing to greater increases in VO2 and Tc with wound invasion and systemic infection.

J Clin Invest, 1986 Jul, 78(1), 196 - 204
Gentamicin and gram-negative bacteremia . A synergism for the development of experimental nephrotoxic acute renal failure; Zager RA et al.; To explore whether bacteremia potentiates gentamicin nephrotoxicity, we injected rats with either 1 X 10(9) Escherichia coli (E . coli), Pseudomonas aeruginosa, or Staphylococcus aureus, and then gave them gentamicin, 100 mg/kg . Renal injury was assessed over the next 24-48 h . Staphylococcus/gentamicin or gentamicin alone induced no renal injury . However, E . coli/gentamicin and Pseudomonas/gentamicin caused acute renal failure (severe azotemia; tubular necrosis; cast formation) . This effect was not due to acute reductions in arterial blood pressure or renal blood flow, it could be reproduced by substituting nonviable for viable gram-negative organisms, and it was associated with increased renal gentamicin uptake . E . coli without gentamicin induced only mild azotemia and no tubular necrosis . Endotoxin-tolerant rats were significantly protected against the E . coli/gentamicin nephrotoxic interaction . We conclude that gram-negative bacteremia and gentamicin exert synergistic nephrotoxicities; and that this effect is mediated, at least in part, by endotoxin and in part by increased renal gentamicin uptake.

J Bacteriol, 1986 Jul, 167(1), 7 - 11
Mapping of mutations in Pseudomonas aeruginosa defective in pyoverdin production; Ankenbauer R et al.; Twelve mutants of Pseudomonas aeruginosa PAO defective in pyoverdin production were isolated (after chemical and transposon mutagenesis) that were nonfluorescent and unable to grow on medium containing 400 microM ethylenediaminedi(o-hydroxyphenylacetic acid) . Four mutants were unable to produce hydroxamate, six were hydroxamate positive, one was temperature sensitive for pyoverdin production, and another was unable to synthesize pyoverdin on succinate minimal medium but was capable of synthesizing pyoverdin when grown on Casamino Acids medium (Difco Laboratories, Detroit, Mich.) . The mutations were mapped on the PAO chromosome . All the mutations affecting pyoverdin production were located at 65 to 70 min, between catA1 and mtu-9002.

J Bacteriol, 1986 Jul, 167(1), 291 - 8
Nucleotide sequence and expression of a phosphate-regulated gene encoding a secreted hemolysin of Pseudomonas aeruginosa; Pritchard AE et al.; A 3.3-kilobase-pair fragment of Pseudomonas aeruginosa DNA containing the phospholipase C (heat-labile hemolysin) gene was sequenced, and the location of the gene was determined . The gene product contains at its NH2 terminus a 38-amino acid sequence which structurally resembles the signal peptides of other secreted proteins but is unusually long and positively charged (6+) . The location of the translation start codon was determined by constructing a series of plasmids in which the promoter of a transcription vector was ligated to Pseudomonas DNA containing deletions at the 5' end of the gene . The plasmids were used to transform Escherichia coli, and the resulting clones were assayed for hemolysin activity . In addition, sizes of truncated proteins produced by mutants with translation terminators introduced at specific sites were analyzed in E . coli maxicells . The gene is transcribed, starting just upstream of the hemolysin gene, as an mRNA of approximately 2,800 bases . Analysis of the nucleotide sequence, analysis of mutants in maxicells, and transcriptional studies indicate that the hemolysin is part of an operon composed of two genes . Phosphate regulation of the operon is at the transcriptional level . The location of the 5' end of the transcript was determined by S1 mapping.

Br J Ophthalmol, 1986 Jul, 70(7), 507 - 9
Experimental contamination of Minims of fluorescein by Pseudomonas aeruginosa; Claoue C; Contamination of fluorescein solutions by Pseudomonas aeruginosa has been a concern of ophthalmologists for many years because of the severity of pseudomonas keratitis . Attempts to prevent contamination have been directed at stringent sterility control during manufacture and the introduction of single-dose disposable containers such as Minims . Deliberate contamination of Minims fluorescein with Pseudomonas aeruginosa was attempted . Under conditions likely to be met with in clinical practice the contents remained sterile . However, under extreme conditions of immersion in pure broth culture of Pseudomonas aeruginosa contamination could be achieved . The relevance of these results to clinical practice is discussed.

Rev Infect Dis, 1986 Jul-Aug, 8 Suppl 4, S426 - 33
Active and passive immunization strategies for Pseudomonas aeruginosa pneumonia; Pennington JE et al.; The unusually high mortality associated with Pseudomonas aeruginosa pneumonia has provided an incentive for the development of immunologic strategies for preventing or treating this infection . A guinea pig model of experimental P . aeruginosa pneumonia was employed to determine prophylactic efficacy of active immunization with a detoxified lipopolysaccharide vaccine; efficacy of passive immune therapy utilizing a new hyperimmune immunoglobulin G preparation enriched for antibodies to P . aeruginosa immunotypes 1, 2, 4, and 6; and efficacy of active and passive immunization against the mucoid exopolysaccharide antigen associated with mucoid strains of P . aeruginosa . Each of these immunologic methods provided an element of protection against P . aeruginosa pneumonia.

J Bacteriol, 1986 Jul, 167(1), 243 - 50
Nucleotide sequence of the gene encoding the two-subunit pilin of Bacteroides nodosus 265; Elleman TC et al.; The nucleotide sequence of the gene encoding pilin from Bacteroides nodosus 265 has been determined . The pilin is encoded by a single-copy gene, from which can be predicted a prepilin comprising a single protein chain of Mr 16,637 . The prepilin sequence differs in several respects from the mature protein sequence . Seven additional N-terminal amino acid residues are present in prepilin, whereas residue 8, phenylalanine, undergoes posttranslational modification to become the N-methylated amino-terminal residue of mature pilin . In addition, further processing occurs through internal cleavage to produce two noncovalently linked subunits characteristic of pilins from serogroup H of B . nodosus, of which strain 265 is a member . The position of cleavage has been identified between alanine residues at positions 72 and 73 of the mature 149-residue pilin protein . The predicted pilin sequence of B . nodosus 265 shows extensive N-terminal amino acid sequence homology with other pilins of the N-methylphenylalanine type . In addition this sequence also shows homology with these N-methylphenylalanine-type pilins in the C-terminal region of the molecule, especially with pilin from Pseudomonas aeruginosa PAK.

Infect Immun, 1986 Jul, 53(1), 207 - 12
Nonopsonic phagocytosis of nonmucoid Pseudomonas aeruginosa by human neutrophils and monocyte-derived macrophages is correlated with bacterial piliation and hydrophobicity; Speert DP et al.; We have shown previously that some strains of Pseudomonas aeruginosa from patients with cystic fibrosis are phagocytized by human polymorphonuclear leukocytes in the absence of serum opsonins . The purpose of this study was to identify the bacterial features which render certain strains susceptible to nonopsonic phagocytosis . Three strains were phagocytized by human neutrophils and monocyte-derived macrophages, and two were not, as determined by luminol-enhanced chemiluminescence, visual inspection of stained smears, and bactericidal assay . Strains that were phagocytized formed pellicles when grown in static broth, but the phagocytosis-resistant strains did not . The phagocytosis-susceptible strains were more heavily piliated and more hydrophobic than the resistant strains . Bacteria exposed to heat (60 degrees C) or UV irradiation were depiliated, as assessed by electron microscopy, and rendered resistant to phagocytosis . When P . aeruginosa was grown on agar, it was piliated, hydrophobic, and susceptible to nonopsonic phagocytosis, but when grown to stationary phase in shaken broth, it was nonpiliated, less hydrophobic, and resistant to phagocytosis . It appears that nonopsonic phagocytosis of certain P . aeruginosa strains by human polymorphonuclear leukocytes and macrophages is facilitated by hydrophobic interactions which may be determined in part by pili.

Bioorg Khim, 1986 Jul, 12(7), 995 - 7
{The structure of O-specific polysaccharide of Pseudomonas aeruginosa immunotype 3; revision of the structure of acetoamidine derivative of 2,3-diamino-2,3-dideoxy-D-mannuronic acid}; Knirel' IuA et al.; O-Specific side chain of P . aeruginosa immunotype 3 lipopolysaccharide is composed of N-acetyl-D-fucosamine (FucNAc), 2,3-diacetamido-2,3-dideoxy-L-guluronic acid (GulN2Ac2A) and 3-acetamidino = 2-acetamido = 2,3 = dideoxy = D-mannuronic acid (ManNAcAmA) . The latter sugar is identified on the basis of solvolysis with anhydrous hydrogen fluoride, 13C NMR spectroscopy and fast-atom bombardment mass spectrometry analysis, as well as of reactions of acetamidino function (alkaline hydrolysis to acetamido group and reductive deamination to ethylamino group) . Earlier, in the course of investigation of P . aeruginosa O3 lipopolysaccharides, the structure of 1-methyl-2-imidazoline was erroneously ascribed to the acetamidino group . The following structure was established for the repeating unit of immunotype 3 polysaccharide which is identical to P . aeruginosa O3(a),3c polysaccharide: ----4)-beta-D-ManNAcAmA-(1----4)-alpha-L-GulN2Ac2A-(1----3)- beta-D-FucNac-(1----.

Bioorg Khim, 1986 Jul, 12(7), 992 - 4
{Identification of 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid in lipopolysaccharide from Pseudomonas aeruginosa immunotype 7}; Knirel' IuA et al.; O-Specific polysaccharide chain of Pseudomonas aeruginosa immunotype 7 lipopolysaccharide is composed of 3-acetamidino-2-acetamido-2,3-dideoxy-L-guluronic acid (GulNAcAmA), 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid (ManN2Ac2A), and N-acetyl-D-fucosamine (FucNAc) . On solvolysis with anhydrous hydrogen fluoride, the polysaccharide afforded a trisaccharide containing all its components . Borohydride reduction of the trisaccharide in boric acid solution resulted in conversion of reducing fucosamine into fucosaminitol, whereas in water the reduction was accompanied by reductive deamination of acetamidino function into ethylamino group . On hydrolysis with aqueous triethylamine, acetamidino group gave acetamido group . Analysis of the trisaccharides thus obtained by 1H NMR spectroscopy (including nuclear Overhauser effect), 13C NMR spectroscopy, and fast-atom bombardment mass spectrometry allowed the determination of the structure of the unusual uronic acid derivative and the following structure of the polysaccharide repeating unit: -4)-alpha-L-GulNAcAmA-(1-4)-beta-D-ManN2Ac2A-(1-3)-alpha-D-+ ++FucNAc-(1-.

Am Rev Respir Dis, 1986 Jul, 134(1), 49 - 56
Lysosomal enzymes from polymorphonuclear leukocytes and proteinase inhibitors in patients with cystic fibrosis; Goldstein W et al.; In serum and sputum samples from 15 patients with cystic fibrosis (CF) suffering from chronic Pseudomonas aeruginosa lung infections, concentrations and/or activities of elastase derived from polymorphonuclear leukocytes (PMN), cathepsin G, myeloperoxidase (MPO), and superoxide dismutase (SOD), as well as concentrations of the proteinase inhibitors alpha 1-proteinase inhibitor (alpha 1-PI) and alpha 2-macroglobulin (alpha 2-M), were determined . High enzyme concentrations compared with those in normal control subjects were found for PMN elastase (mean, 96.1 +/- 91.7 micrograms/ml), cathepsin G (mean, 5.9 +/- 6.0 micrograms/ml), and MPO (mean, 138.0 +/- 177 micrograms/ml) in patients' sputum samples . Superoxide dismutase was not detectable in any of the sputum specimens (below 1 ng/ml) . Proteinase inhibitor concentrations were elevated in serum samples (alpha 1-PI: mean, 3,457 +/- 1,084 micrograms/ml; alpha 2-M: mean, 4,835 +/- 1,334 micrograms/ml) . Means of 61 +/- 38 micrograms/ml alpha 1-PI and 29 +/- 31 micrograms/ml alpha 2-M were present in the sputum specimens . Both proteinase inhibitors were functional in the serum samples . However, sputum alpha 1-PI was proteolytically degraded, as shown by western blot technique, and was not able to bind 125I-labeled PMN elastase, as shown by autoradiography . Only 10.9 +/- 8.5% of the total alpha 1-PI in the sputum samples was complexed to PMN elastase and 3.6 +/- 3.2% to cathepsin G . On the other hand, 96.2 +/- 96.8% of the total PMN elastase and 78.0 +/- 100% of cathepsin G were unbound in the sputum samples . The study suggests that the imbalance between PMN proteinases and their inhibitors is due to inactivation of alpha 1-PI in the sputum caused by proteolytic or oxidative attack from PMN enzymes.

Am Rev Respir Dis, 1986 Jul, 134(1), 17 - 21
Effects of Pseudomonas aeruginosa on bronchial epithelial ion transport; Stutts MJ et al.; During chronic lung infections the effects of bacterial exoproducts on the function of the airways epithelium could play a role in the overall pathophysiology of infection . We measured the effects of Pseudomonas aeruginosa culture on ion transport and solute permeability of dog bronchial epithelium to determine if this organism produces toxins that affect these epithelial functions . Thirty-six-hour trypticase soy broth cultures of P . aeruginosa at 1:40 dilutions decreased short-circuit current and active sodium absorption and lowered conductance and chloride unidirectional fluxes . At 1:4 dilutions, P . aeruginosa increased conductance and the permeability coefficient of 14C-mannitol, suggesting that solute flow around cells was increased . These effects appear to be caused by a heat-stable glycolipid hemolysin produced by P . aeruginosa.

Am J Med, 1986 Jun 30, 80(6B), 190 - 4
Future trends in aminoglycoside therapy; Levin S et al.; The aminoglycosidic aminocyclitols have been utilized extensively for three decades . Nonetheless, the future use of this class of agents has been questioned of late . Recognized inadequacies of the aminoglycosides and the development of new antibiotics with significant activity against gram-negative bacilli are commonly cited reasons for the theorized decline of these compounds . However, resistance to newly developed antibiotics already has become evident . This insures a continuing role for the aminoglycosides in the treatment of nosocomial infections . Aminoglycosides will have continued use as empiric, potentially synergistic therapies for hospital-acquired infections in neutropenic patients with bacteremia, in enterococcal endovascular infections, and in patients with serious infections associated with Pseudomonas aeruginosa . Those factors that will influence the future role of aminoglycosides in these settings will include economic, administrative, and space pressures to restrict the number of antibiotics available in hospitals, the discovery of novel antibiotics, the utility of combination therapies employing an aminoglycoside and newly available drugs, the comparative toxicities of new antimicrobial regimens, and considerations of cost containment.

Am J Med, 1986 Jun 30, 80(6B), 156 - 60
In vitro models for the study of combination antibiotic therapy in neutropenic patients; Zinner SH et al.; Neutropenic patients are at risk of serious infection caused by gram-negative bacilli and staphylococci . The mortality rate associated with gram-negative bacteremia in these patients is extremely high, especially in those with persistent and profound granulocytopenia . In these latter patients, the best results have been obtained by administering combinations of antibiotics in which both agents are active and/or show in vitro synergism against the infecting organism . Most combinations include an aminoglycoside such as amikacin and a broad-spectrum beta-lactam antibiotic, such as azlocillin, mezlocillin, piperacillin, or ceftazidime . The International Antimicrobial Therapy Project Group of the European Organization for Research and Treatment of Cancer has completed several studies evaluating various antibiotic combinations in the empiric treatment of febrile neutropenic patients . These trials have evaluated cephalothin plus gentamicin, carbenicillin plus gentamicin, and cephalothin plus carbenicillin; carbenicillin plus amikacin and carbenicillin plus amikacin plus cefazolin; azlocillin plus amikacin, ticarcillin plus amikacin, and cefotaxime plus amikacin; and azlocillin plus amikacin versus ceftazidime plus long- or short-course amikacin . The preclinical evaluation of antibiotic combinations usually involves the in vitro testing of antibiotics alone and in combination by the checkerboard method or with the use of time-kill curves . However, these methods expose the bacterial culture to a static or constant concentration of the drugs . During the in vivo treatment of infections, bacteria are exposed to changing concentrations of antibiotics, which are contingent on the individual pharmacokinetics of these drugs . We have designed a two-compartment in vitro pharmacokinetic model that allows the simultaneous study of the activity of two antibiotics with similar or different half-lives against a number of bacteria . Amikacin and azlocillin have been studied alone and in combination in this model against Pseudomonas aeruginosa, a frequent cause of bacteremia in neutropenic patients . In pharmacologically relevant doses, amikacin alone produced rapid bacterial killing, followed by regrowth of resistant subpopulations . Azlocillin alone produced a more gradual reduction of the bacterial inoculum, with ultimate bacteriostasis . Amikacin plus azlocillin produced rapid and complete eradication of the organism . In vitro pharmacokinetic models may prove to be more predictive of clinical outcome than are traditional static in vitro methods used to study antibiotic combinations.

Am J Med, 1986 Jun 30, 80(6B), 76 - 81
Surveillance of aminoglycoside resistance . European data; Van Landuyt HW et al.; The susceptibility patterns of gram-negative aerobic organisms to aminoglycosides differ widely from one European health care center to another and depend upon local antibiotic prescribing policies . Reports of the susceptibility of Pseudomonas aeruginosa to gentamicin and tobramycin have ranged from as low as 49.8 percent and 77.7 percent, respectively, in Greece, to as high as 96.6 percent and 99.2 percent, respectively, in the United Kingdom . The susceptibility of P . aeruginosa to gentamicin, tobramycin, and amikacin decreased in our hospital from 73.1 percent, 94.8 percent, and 95.6 percent, respectively, in 1982, to 43.1 percent, 70.6 percent, and 74.3 percent, respectively, in 1984 . A prospective surveillance study of the susceptibility of gram-negative aerobic bacilli to four aminoglycosides (gentamicin, tobramycin, amikacin, and netilmicin) was performed over a period of 17 months . Gentamicin and tobramycin were freely used, while the use of amikacin was restricted throughout the hospital during a four-month baseline period (May through August 1984) . Gentamicin and tobramycin accounted for 94 percent of the aminoglycoside use . During the following 13 months (September 1984 through September 1985), amikacin was used as the first-line aminoglycoside and accounted for more than 97 percent of the aminoglycoside usage . A total of 1,866 organisms were analyzed during the baseline period; 5,429 were analyzed during the amikacin-usage period . The overall susceptibility to gentamicin, tobramycin, amikacin, and netilmicin increased from 86.9 percent, 90.4 percent, 94.2 percent, and 88.3 percent, respectively, to 92.3 percent, 94.0 percent, 97.3 percent, and 92.3 percent, respectively . P . aeruginosa isolates had the most striking changes, with the susceptibility to gentamicin, tobramycin, amikacin, and netilmicin increasing from 43.1 percent, 70.6 percent, 74.3 percent, and 50.6 percent, respectively, during the baseline period, to 64.5 percent, 81.6 percent, 90.8 percent, and 65.1 percent, respectively, during the amikacin-usage period . The use of amikacin as a first-line aminoglycoside, while use of the other aminoglycosides was restricted, seemed to have a favorable influence on the susceptibility pattern of gram-negative aerobic isolates in our hospital.

Am J Med, 1986 Jun 30, 80(6B), 182 - 9
The potential for discovery and development of improved aminoglycosides; Price KE; Following the development of amikacin, pharmaceutical companies made intensive efforts to find even more potent and broader-spectrum aminoglycosides . This effort was justifiable in view of the fact that over the preceding decade, these agents, because of their unique properties, had proven to be the primary weapons in the therapeutic armamentarium for the treatment of seriously ill patients . Since the toxicities associated with the aminoglycosides were beginning to limit their use in general medicine, researchers ultimately shifted their emphasis from probing for higher-potency, broader-spectrum agents to finding those with a reduced potential for toxicity . This article addresses the issue of whether superior aminoglycoside derivatives will reach the marketplace in the future . A comparison is made of several key properties of virtually all aminoglycosides that have reached an advanced preclinical development stage, gone into the clinic, or been registered for commercial use over the past 10 years . The following parameters are used for comparisons with already marketed aminoglycosides: antibacterial potency, as measured by relative minimum inhibitory concentrations for 50 percent of the strains tested, against wild-type Pseudomonas aeruginosa; degree of resistance to inactivation by microbial enzymes; and potential for toxicity utilizing comparative acute intravenous lethal doses for 50 percent of the population in mice, values that appear to predict the maximum recommended daily doses in man . An assessment of a number of compounds, including three structurally related to gentamicin, two to sisomicin, two to kanamycin A, three to kanamycin B, and two to fortimicin, revealed that none had overall properties superior to those already being utilized commercially . In no case did a compound prove to be less toxic, and in many instances, the antibacterial potency of the newer agents was lower than that exhibited by the older aminoglycosides . Some increase in resistance to inactivating enzymes was seen, but only BB-K 311 proved refractory to more enzymes than did amikacin . In view of this and the fact that no new agents of promise have moved into the development stage during the past five years, it seems safe to say that the current armamentarium of aminoglycosides is all that will be available for use in the foreseeable future.

Cancer, 1986 Jun 15, 57(12), 2291 - 4
Fournier's gangrene complicating aggressive therapy for hematologic malignancy; Berg A et al.; The authors describe two cases of Fournier's gangrene complicating aggressive therapy for hematologic malignancies . Fournier's gangrene is a fulminant necrotizing fasciitis of the scrotum and penis, often with an infectious etiology . Only one prior case has been reported in a patient receiving aggressive chemotherapy for a hematologic malignancy . Unique to these three cases was profound granulocytopenia and the culturing of Pseudomonas aeruginosa in both the blood and necrotic perineal-scrotal tissue . Maintaining a high index of suspicion with early recognition and aggressive therapy may decrease the morbidity and mortality of this devastating and life-threatening infection in the compromised host.

Antimicrob Agents Chemother, 1986 Jun, 29(6), 972 - 6
In vitro activity of BRL 36650, a new semisynthetic penicillin; Hoy JF et al.; BRL 36650 {sodium 6 beta-(D-2-{(4-ethyl-2, 3-dioxopiperazin-1-yl)carbonylamino}-2-(3,4-dihydroxyphenyl) acetamido)-6 alpha-formamido-penicillinate} is a new semisynthetic penicillin . It was tested in vitro for activity against 884 organisms cultured from blood specimens of cancer patients . BRL 36650 had broad-spectrum activity against the gram-negative bacilli tested but had no gram-positive activity . The MIC against 90% of the Pseudomonas aeruginosa isolates was 3.12 micrograms/ml . The activity of BRL 36650 was superior to that of piperacillin, comparable or slightly inferior to that of aztreonam and ceftazidime, and lower than that of imipenem and amifloxacin . BRL 36650 should prove useful for the management of gram-negative bacillary infections, including those caused by P . aeruginosa.

Invest Ophthalmol Vis Sci, 1986 Jun, 27(6), 958 - 65
Effect of inflammation on antibiotic penetration into the anterior segment of the rat eye; Badenoch PR et al.; Bacterial infections were established in the right cornea of rats . Animals infected with Staphylococcus aureus were given cephradine intravenously (IV) (40 mg/kg) or topically (50 mg/ml) to both eyes . Animals infected with Pseudomonas aeruginosa were given gentamicin sulfate IV (40 mg/kg) or topically (10 mg/ml) . Antibiotic concentrations in cornea and aqueous humor were measured for 4 hrs following dosing using bioassay and radioimmunoassay . In general, infection significantly increased the concentrations obtained soon after dosing . Topically applied cephradine passed through infected eyes more quickly than through normal eyes . Of the pharmacokinetic parameters derived, the permeability of the corneal epithelium to gentamicin in the rat more closely agrees with reported human values than does the rabbit, while the coefficient of elimination from aqueous in the rat is considerably greater than that for either humans or rabbits . This suggests that there are both advantages and disadvantages in using the rat for therapeutic studies of ocular disease.

Infect Immun, 1986 Jun, 52(3), 846 - 52
Pulmonary microvascular injury induced by Pseudomonas aeruginosa cytotoxin in isolated rabbit lungs; Seeger W et al.; The effects of Pseudomonas aeruginosa cytotoxin on the pulmonary microvasculature were studied in blood-free, perfused, isolated rabbit lungs . Cytotoxin was administered to the recirculating Krebs Henseleit albumin (1%) buffer during two consecutive 30-min-perfusion phases (phases 1 and 2) at a concentration of 13 micrograms/ml, followed by a third perfusion phase (phase 3) without toxin . After perfusion phases 2 and 3, the capillary filtration coefficient (Kf,c) and vascular compliance were determined gravimetrically from two-step microvascular pressure increments under zero-flow conditions . Cytotoxin caused a continuous release of K+ and lactate dehydrogenase, which started within the first 5 min and amounted to about 50% of the total lung cellular K+ and 5 to 7% of the total lactate dehydrogenase by the end of the experiment . The toxin caused the continuous generation of prostaglandin I2, which was detectable in the perfusates of all perfusion phases at maximum values five times above the control values and which was measured in the bronchoalveolar lavage fluid at the end of the experiment . Thromboxane generation in toxin-treated lungs did not significantly exceed that of control lungs or of lungs with mechanically induced edema . Cytotoxin caused a gradual increase in pulmonary vascular resistance, to maximum values 2.5 times above the control, starting within 1 min; the increase was partially reversible after washout of the toxin . After a lag period of 20 to 30 min, the lungs gained weight, amounting to a mean gain of 9.1 g at the end of the experiments . After perfusion phases 2 and 3, an almost fourfold increase in Kf,c, which was not reversible after washout of the toxin, was measured, whereas the values of vascular compliance were not altered . We conclude that pseudomonal cytotoxin may be an important factor in the pathogenesis of prolonged microvascular injury, encountered in states of P . aeruginosa sepsis or acute lung failure with secondarily acquired P . aeruginosa pneumonia.

Pathol Biol (Paris), 1986 Jun, 34(5 Pt 2), 621 - 4
{Present status of the sensitivity of Pseudomonas aeruginosa to aminoglycosides in France}; Thabaut A et al.; 9,119 strains of P . aeruginosa were collected from 10 French hospitals from 1981 to 1984 . The MIC of gentamicin (GM), sisomicin (SIS), tobramycin (TOB), dibekacin (DBK), netilmicin (NET), amikacin (AMK), and habekacin (HBK) was determined by the two fold microdilution method . 81% of the strains were susceptible to the 4 aminoglycosides . The MICs (geometric mean, mg/l) for these susceptible strains were: GM: 1.69, SIS: 1.58, TOB: 1.06, DBK: 1.12, NET: 2.73, AMK: 4.16, HBK: 2.70 . 19% of the 9 119 strains were resistant to SIS, 18% to GM, 16% to TOB and DBK, 11% to NET, 3% to AMK and HBK . Among the 1 758 strains resistant to one or several aminoglycosides, the frequency of the resistance phenotypes was GM-SIS: 11%, GM-SIS-NET: 4.95%, GM-SIS-TOB-DBK: 32.7%, GM-SIS-TOB-DBK-NET: 34.45%, SIS-TOB-DBK-NET: 2.5%, SIS-TOB-DBK-NET-AMK-HBK: 2.39%, GM-SIS-TOB-DBK-NET-AMK-HBK: 11.61% . The mechanism of resistance of the phenotypes 1 to 6 is enzymatic . The mechanism of resistance of the phenotype 7 (resistance to the six aminoglycosides) is either enzymatic or due to decreased uptake or penetration of antibiotic . HBK and AMK are the aminoglycosides most frequently active against P . aeruginosa . The patterns of resistance to both antibiotics are the same . On a weight-for-weight basis, HBK is more active than AMK.

Jpn J Antibiot, 1986 Jun, 39(6), 1487 - 93
{In vivo combination effects of astromicin and beta-lactam antibiotics against Pseudomonas aeruginosa}; Sato K et al.; Astromicin (ASTM, Fortimicin) is a pseudodisaccharide aminoglycoside antibiotic . The ASTM exhibited excellent activity against Gram-positive and Gram-negative bacteria but was only weakly active against Pseudomonas aeruginosa . In vitro synergistical activities of ASTM combined with beta-lactam antibiotics have been reported against P . aeruginosa previously . In this paper, we investigated the in vivo combination efficacies of ASTM and beta-lactam antibiotics (latamoxef (LMOX), cefoperazone (CPZ), piperacillin (PIPC) and cefsulodin (CFS) against experimental infection with P . aeruginosa in both normal and immunosuppressed mice . In normal mice, the combination of ASTM with these beta-lactam antibiotics produced significantly greater protective effects than the single use of individual antibiotics against both strains of P . aeruginosa BMH No . 1 and E-2 . In mice immunosuppressed with cyclophosphamide, the combination of ASTM with LMOX or CFS also exhibited synergistic protective effects against P . aeruginosa BMH No . 1, but PIPC and CPZ did not . From the above results, the combination therapy of ASTM with beta-lactam antibiotics appeared to be effective against experimental infections with P . aeruginosa in mice.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Jun, (6), 67 - 9
{Antigenic properties of Pseudomonas aeruginosa anatoxin and the protective action of antitoxic anti-Pseudomonas aeruginosa serum}; Podgornaia LG et al.; The antigenic properties of P . aeruginosa toxoid, prepared with the use of casein culture medium, were not inferior to those of the toxoid obtained in Martin broth . In experiments on white mice antisera obtained by the immunization of rabbits with the toxoid prepared on the basis of casein culture medium showed sharply pronounced protective properties against P . aeruginosa homologous and heterologous strains, as well as toxigenic reference strain PA-103.

Eur J Clin Microbiol, 1986 Jun, 5(3), 292 - 6
In vitro activity of carumonam (RO 17-2301) and twelve other antimicrobials against clinical isolates of Pseudomonas aeruginosa; Vurma-Rapp U et al.; Minimal inhibitory concentrations of the monobactam carumonam (RO 17-2301) and twelve other antimicrobials were determined using agar dilution against 140 recent non-replicate clinical isolates of Pseudomonas aeruginosa . The most active drugs were ciprofloxacin, amikacin, imipenem and ceftazidime, inhibiting 96, 91, 90 and 86 percent of the strains, respectively, at or below the susceptibility threshold . The monobactams carumonam and aztreonam were active against 78 and 65 percent of the strains, respectively . Tobramycin inhibited 68 percent of the strains, and gentamicin and netilmicin 50 and 21 percent, respectively . Analysis of correlation coefficients revealed a low correlation between imipenem and the other beta-lactams and a remarkably good correlation between the beta-lactams (excepting imipenem) and the aminoglycosides.

Arzneimittelforschung, 1986 Jun, 36(6), 899 - 904
Bacteriological and ultrastructural studies on the effect of subinhibitory beta-lactam concentrations on intraphagocytic killing of Pseudomonas aeruginosa by human polymorphonuclear leukocytes; Stubner G et al.; The effect of azlocillin, ticarcillin and cefsulodin, respectively, on the susceptibility of Pseudomonas aeruginosa to the antimicrobial action of human polymorphonuclear leukocytes (PMN) was investigated under two different experimental conditions . Firstly, phagocytic capacity as well as bactericidal activity of PMN were assessed in a homologous system, i.e . the clinical isolate as well as the PMN and serum were obtained from the same patient . Secondly, ultrastructural studies were performed by electron microscopy . Preincubation of bacteria with subinhibitory beta-lactam concentrations augmented the phagocytic capacity as well as the antibacterial activity of PMN; azlocillin tended to be the most effective drug in this respect . The enhanced susceptibility to leukocyte killing is not due to an increased antibacterial action of the beta-lactams in the presence of PMN . These findings suggest that a non-immunological linkage between bacteria and PMN may exist which may be based on the interaction between bacterial- and eukaryotic surface structures, respectively . It may be assumed that the antipseudomonal beta-lactam antibiotics may cause changes in the surface structures of P . aeruginosa, thus rendering them more susceptible to phagocytosis . Preliminary data indicate that the lectins on the outer membrane of P . aeruginosa are not mannose sensitive . Electron microscopic studies revealed that azlocillin pretreatment of bacteria brought about a high undulation and a disruption of the outer membrane . These morphological changes may render bacteria more vulnerable to the antimicrobial action of PMN . It may be speculated that an interference with surface adhesins and induction of morphological changes may affect engulfment and intracellular killing of bacteria.

Antibiot Med Biotekhnol, 1986 Jun, 31(6), 456 - 61
{Antibiotic resistance of clinical strains of Pseudomonas aeruginosa from various O-serogroups}; Iskhakova KhI et al.; The results of the study on antibiotic resistance of 745 strains of P . aeruginosa isolated in hospitals from surgical patients, environment and carriers are presented . 89.7 per cent of the strains were typed by the commercial O-sera . The isolates were most sensitive to tobramycin (99 per cent), amikacin (95.1 per cent), cefsulodin (95.1 per cent), polymyxin (89.7 per cent) and gentamicin (73.5 per cent) . Comparison of the antibiotic resistance of the typed and nontyped cultures revealed that the former were more resistant to tetracycline, carbenicillin, rifampicin, kanamycin and cefotaxim, while the latter were more resistant to gentamicin and polymyxin . It was also noted that the proportion of the strains resistant to all of the tested antibiotics was higher among the nontyped cultures . Antibiotic sensitivity of P . aeruginosa was in many respects defined by the strain origin: the strains isolated from patients were more resistant to tetracycline, carbenicillin, rifampicin, cefotaxim and kanamycin and more sensitive to gentamicin and polymyxin than the strains isolated from the environment . The cultures belonging to different O-serogroups (O-11 and O-2) markedly differed by their antibioticograms.

Acta Pathol Microbiol Immunol Scand {B}, 1986 Jun, 94(3), 135 - 8
A comparison of different methods for determining elastase activity of Pseudomonas aeruginosa strains from mink; Elsheikh LE et al.; We have characterized 20 Pseudomonas aeruginosa strains isolated from pneumonic mink lungs with regard to elastase production and serotype . P . aeruginosa PAO1, a well-characterized elastase-producing strain, and two elastase-deficient mutants of PAO1 were used for comparative purposes . Elastase activity was assayed on elastin agar and by using 14C-elastin coated microtiter plates . Elastase antigen was measured using a double antibody sandwich ELISA (enzyme-linked immunosorbent assay) . Total proteolytic activity was determined on skim milk agar plates . The results from ELISA showed that all strains produced antigenically similar elastase, although the amounts produced varied considerably between strains . There was a good correlation regarding elastase assays between ELISA and 14C-elastin, elastin agar and total proteolytic activity on skim milk agar . No correlation was found between serotype and elastase activity . The results showed that the 14C-elastin assay is a simple and sensitive method of determining elastolytic activity of P . aeruginosa strains.

Methods Find Exp Clin Pharmacol, 1986 Jun, 8(6), 391 - 2
Comparison of antimicrobial susceptibilities of Pseudomonas aeruginosa isolates, with and without plaques, from two hospitals; Flournoy DJ et al.; Disc-agar diffusion plates, used for antimicrobial susceptibility testing, of 368 Pseudomonas aeruginosa isolates with and without plaques were compared . Seventeen percent of all isolates from the Veterans Administration Medical Center and eleven percent from Oklahoma Memorial Hospital produced plaques . When percentages of susceptibility for plaque and non-plaque containing isolates were compared, there were some statistically significant differences which are discussed in the text.

J Antimicrob Chemother, 1986 Jun, 17(6), 755 - 62
Resistance to cefsulodin and gentamicin in Pseudomonas aeruginosa strains in five areas of Japan between 1980 and 1983; Furusawa T et al.; The activities of cefsulodin and gentamicin against Pseudomonas aeruginosa isolated from clinical specimens from five hospitals in different geographical areas of Japan, from 1980 to 1983, were compared in vitro . The incidence of resistant strains was higher for gentamicin . In 1982 the sensitivity to both drugs decreased from 1980 and 1981 levels, largely because of the isolation of numbers of cefsulodin-gentamicin cross-resistant bacteria from three of the five hospitals . In 1983, the incidence of resistant strains was similar to that in 1982 . This linked cefsulodin-gentamicin resistance may have been selected by the use of the cephalosporins, cefmenoxime, cefoperazone, cefotaxime, ceftizoxime and latamoxef, which had been prescribed extensively in the hospitals where cross-resistance was encountered . Although cefsulodin-resistant strains of P . aeruginosa have increased since 1982, the in-vitro activity of cefsulodin in 1983 remained greater than that of these third-generation cephalosporins.

J Antimicrob Chemother, 1986 Jun, 17(6), 717 - 23
In-vitro emergence of beta-lactam-resistant variants of Pseudomonas aeruginosa; Eng RH et al.; The frequency with which resistant variants could be found in Pseudomonas aeruginosa ATCC 27853 during growth in media with and without antibiotics was determined for ticarcillin, piperacillin, cefotaxime, latamoxef, cefoperazone, ceftriaxone, ceftazidime, aztreonam and imipenem . The resistance frequency was highest for ceftriaxone, cefotaxime, cefoperazone and piperacillin (up to 2 X 10(-4)) and lowest for ticarcillin, latamoxef, ceftazidime, aztreonam and imipenem (less than 10(-8)) . The resistant variants showed cross-resistance to all of the beta-lactam antibiotics tested with the exception of imipenem . The MIC profiles of all variants were similar regardless of which antibiotic the cultures had been exposed to previously . Although all resistant variants produced and excreted cephalosporinases, increased resistance was demonstrated to both cephalosporinase-susceptible and cephalosporinase-stable beta-lactam antibiotics.

Can J Microbiol, 1986 Jun, 32(6), 531 - 3
Opsonic activity of antisera to ribosomal vaccine fractions with live and formalinized Pseudomonas aeruginosa; Lieberman MM et al.; The opsonic capacity of antisera to Pseudomonas aeruginosa ribosomal vaccine fractions was determined by a chemiluminescent technique . Antiserum to a vaccine fraction ("peak A") containing lipopolysaccharide (antiserum A), and antiserum to a vaccine fraction ("peak B"), which did not contain detectable amounts of lipopolysaccharide (antiserum B), were used to opsonify live or formalin-treated bacteria . Polymorphonuclear leukocytes were then stimulated by the opsonified bacteria in the presence of the chemiluminigenic probe, luminol, resulting in the observed chemiluminescence . The data obtained indicated that the antisera had comparable opsonic activity with live (untreated) bacteria . However, antiserum B had far less opsonic activity than did antiserum A when formalinized bacteria were used . Owing to the effects of formaldehyde on protein, these results were interpreted as evidence to suggest that the opsonic activities of the two antisera are dependent on different antigens on the bacterial cell surface . Antiserum A activity is probably dependent on lipopolysaccharide to a great extent, whereas antiserum B activity is most likely dependent primarily on a protein(s).

Can J Microbiol, 1986 Jun, 32(6), 513 - 5
C-390 as sole selective agent for isolation of Pseudomonas aeruginosa from hospital waste water; Havelaar AH et al.; Pseudomonas aeruginosa was recovered (in numbers ranging from 10(2) to 10(5) colony-forming units per millilitre) from heavily contaminated hospital waste water when grown at 41.5 degrees C on a differential medium agar containing 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan (C-390) at a final concentration of 30 micrograms/mL . The medium appeared to be highly selective for P . aeruginosa with 95-100% of all colonies isolated from four different hospital waste waters being identified as P . aeruginosa . Many strains of P . aeruginosa isolated from hospital waste waters failed to hydrolyse casein when grown on skim milk agar and this medium appeared to restrict pigment production to only pyoverdin (detectable only under ultraviolet light) . However, most strains were capable of casein hydrolysis when grown on a modified skim milk medium.

J Trauma, 1986 Jun, 26(6), 525 - 33
Comparison of the effect of three adrenal corticosteroids on human granulocyte function against Pseudomonas aeruginosa; Baltch AL et al.; The effect of hydrocortisone, methylprednisolone, and dexamethasone on the phagocytic and bactericidal capabilities of normal human granulocytes (PMN) was studied under previously described optimal conditions for Pseudomonas aeruginosa, PA 1348A . At hydrocortisone and methylprednisolone concentrations of 1,000 micrograms/ml, delayed phagocytosis was clearly observed, whereas dexamethasone 400 micrograms/ml had no effect on phagocytosis . The bactericidal effect of PMN on PA 1348A was significantly reduced by all three corticosteroids at highest concentrations (p less than 0.05) . However, the effect of methylprednisolone was greatest and that of dexamethasone was least evident, 25% and 10% reduction in PMN bactericidal activity, respectively . Following exposure to the highest concentrations of corticosteroids, TEM observations correlated well with the PMN functional assays . While the observations of PMN and bacteria in controls, hydrocortisone, and dexamethasone preparations were similar, evidence for incomplete phagocytosis, lack of vacuole coalescence, minimal disruption of bacterial cell walls, and dividing bacteria in phagosomes were evident in methylprednisolone preparations . These PMN functional and TEM observations suggest that of the three corticosteroids studied, methylprednisolone appears most deleterious to the PMN phagocytic and bactericidal activity.

J Med Microbiol, 1986 Jun, 21(4), 331 - 6
Transfer of a chromosomal locus responsible for mucoid colony morphology in Pseudomonas aeruginosa isolated from cystic fibrosis patients to P . aeruginosa PAO; MacGeorge J et al.; The locus responsible for mucoid colony morphology in five independent clinical isolates of Pseudomonas aeruginosa from cystic fibrosis patients have been transferred by means of pM060-mediated conjugation to the genetically characterised strain P . aeruginosa PAO . Genetic mapping has shown that in all five strains the locus is on the chromosome between 89' and 94', although it is not possible to say that the same locus is involved in each case . The way is now open for a more detailed genetic analysis of the loci responsible for mucoid colony morphology.

J Clin Microbiol, 1986 Jun, 23(6), 1134 - 5
Imipenem susceptibility testing with a commercially prepared dry-format microdilution tray; Staneck JL; MICs of imipenem, concurrently generated in commercially prepared microdilution trays containing predried antibiotic dilutions (Sensititre), and in a standard agar dilution assay (as recommended by the National Committee for Clinical Laboratory Standards, Villanova, Pa.), were within +/- 1 twofold dilution for 94% of 226 bacterial isolates . Imipenem biological activity remained stable over 5 months of tray storage at room temperature against Pseudomonas aeruginosa ATCC 27853 . Activity of imipenem was shown by microdilution testing with 890 clinical isolates to be high, with only 4% of isolates having MICs of greater than or equal to 16 micrograms/ml (in vitro resistance).

J Clin Microbiol, 1986 Jun, 23(6), 1096 - 8
In vitro study of bacterial growth in continuous ambulatory peritoneal dialysis fluids; Sheth NK et al.; We examined the in vitro survival of bacteria in continuous ambulatory peritoneal dialysis effluents of patients with clinical peritonitis and those without peritonitis . Standard strains of coagulase-negative staphylococci (CNS), Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa were inoculated into the fluids, and portions were plated for bacterial counts at 0.5, 4, 24, 48, 72, and 96 h . Unused dialysate fluid was also inoculated simultaneously . Our results show that CNS increased minimally up to 48 h in the noninfected continuous ambulatory peritoneal dialysis effluents and decreased by 96 h, whereas survival was only minimal in the infected effluent . S . aureus showed trends similar to those of CNS, but differences in survival in infected and noninfected effluents were less marked . By contrast, E . coli and P . aeruginosa increased by greater than 1,000-fold in all solutions tested . Based on the above findings, it is likely that a proportionate number of culture-negative cases of peritonitis are due to gram-positive cocci, especially CNS, which are not retrievable by standard culture techniques because of poor survival rate.

Infect Immun, 1986 Jun, 52(3), 853 - 7
Lung defenses against Pseudomonas aeruginosa in C5-deficient mice with different genetic backgrounds; Cerquetti MC et al.; Lung defenses against Pseudomonas aeruginosa were investigated in C5-deficient strains of mice with different genetic backgrounds . We studied pulmonary clearance and cell responses after aerosol exposure to P . aeruginosa in C5-deficient B10.D2/oSnJ and DBA/2J mice and their closest C5-sufficient counterparts, B10.D2/nSnJ and DBA/1J mice . Different patterns of lung clearance and pulmonary cell responses were found for the two C5-deficient strains . C5-deficient B10.D2/oSnJ mice showed defective lung clearance of P . aeruginosa 4 h after challenge compared with C5-sufficient B10.D2/nSnJ animals . This finding was associated with a decreased number of polymorphonuclear leukocytes recruited into the airways during the same time . Interestingly, C5-deficient DBA/2J mice recruited higher numbers of polymorphonuclear leukocytes than did C5-sufficient DBA/1J mice by 4 h after aerosolization . Nevertheless, lung clearance of P . aeruginosa in DBA/2J mice was not as effective as in C5-sufficient DBA/1J mice, suggesting that other functions of C5 besides chemotaxism could be involved . Lung clearance of P . aeruginosa was also investigated in C5-deficient and -sufficient hybrids sharing the same genetic background (DBA/2J X B10.D2) . The results suggested that murine lung clearance of P . aeruginosa is markedly affected by lack of C5 in a specific genetic background (B10.D2).

J Infect Dis, 1986 Jun, 153(6), 1098 - 107
Temporal relationships among immunologic alterations in a guinea pig model of thermal injury; Bjornson AB et al.; Temporal relationships among various humoral and cellular alterations of host defense mechanisms were investigated in a guinea pig model of thermal injury during three weeks after burning . Reduction in serum concentration of C3 and fixation of C3 on Pseudomonas aeruginosa, presence of activated C3 in plasma, and elevations in levels of 6-ketoprostaglandin F1 alpha and thromboxane B2 in wound fluid were observed at 3-6 hr after burning . These alterations were accompanied by reduction in intrinsic bactericidal activity of polymorphonuclear neutrophils (PMNs) against P . aeruginosa, suppression of bactericidal activity of PMNs by serum, and decreased blood clearance of P . aeruginosa . All parameters returned to normal values by seven to nine days after burning . Proliferative responses of splenic lymphocytes to T cell mitogens were depressed at four days after burning and were maximally reduced at eight days . These data support the concept that there is a continuum of immunologic alterations resulting from thermal injury and that consumption of complement and increase in arachidonic acid metabolism are early events.

Mol Gen Genet, 1986 Jun, 203(3), 511 - 9
IS21 insertion in the trfA replication control gene of chromosomally integrated plasmid RP1: a property of stable Pseudomonas aeruginosa Hfr strains; Reimmann C et al.; Broad host range IncP-1 plasmids are able to integrate into the chromosome of gram-negative bacteria . Strains carrying an integrated plasmid can be obtained when the markers of a temperature-sensitive (ts) plasmid derivative are selected at non-permissive temperature; in this way Hfr (high frequency) donor strains can be formed . The integrated plasmids, however, tend to be unstable in the absence of continuous selective pressure . In order to obtain stable Hfr donor strains of Pseudomonas aeruginosa PAO, we constructed a derivative of an RP1 (ts) plasmid, pME134, which was defective in the resolvase gene (tnpR) of transposon Tn801 . Chromosomal integration of pME134 was selected in a recombination-deficient (rec-102) PAO strain at 43 degrees C . Plasmid integration occurred at different sites resulting in a useful set of Hfr strains that transferred chromosomal markers unidirectionally . The tnpR and rec-102 mutations prevented plasmid excision from the chromosome . In several (but not all) Hfr strains that grew well and retained the integrated plasmid at temperatures below 43 degrees C, the insertion element IS21 of RP1 was found to be inserted into the trfA locus (specifying an essential trans-acting replication function) of the integrated plasmid . One such Hfr strain was rendered rec+; from its chromosome the pME134::IS21 plasmid (= pME14) was excised and transferred by conjugation to Escherichia coli where pME14 could replicate autonomously only when a helper plasmid provided the trfA+ function in trans . Thus, it appears that trfA inactivation favours the stability of chromosomally integrated RP1 in P . aeruginosa.

Mol Gen Genet, 1986 Jun, 203(3), 430 - 4
Heterologous expression and regulation of the lysA genes of Pseudomonas aeruginosa and Escherichia coli; Martin C et al.; The Pseudomonas aeruginosa lysA gene encoding diaminopimelate decarboxylase (DAP-decarboxylase) was cloned into a broad host range vector . This gene complemented a lys mutation at the lys-12 locus of P . aeruginosa and a lysA defect in Escherichia coli . The P . aeruginosa DAP-decarboxylase was synthesized constitutively in P . aeruginosa as well as in E . coli, where the Pseudomonas lysA gene was poorly expressed . By contrast, the E . coli lysA gene was expressed well in P . aeruginosa and subject to lysine regulation when the E . coli LysR activator protein was provided . This indicates that the mechanism of transcriptional activation for the E . coli lysA gene is effective in the heterologous host.

Mol Gen Genet, 1986 Jun, 203(3), 421 - 9
Expression of biosynthetic genes from Pseudomonas aeruginosa and Escherichia coli in the heterologous host; Jeenes DJ et al.; We examine the expression of constitutive or repressible, monocistronic genes from Pseudomonas aeruginosa and Escherichia coli after their transfer to the heterologous host . To this end, chromosomal DNA from P . aeruginosa was cloned into the mobilizable broad-host-range vector pKT240; recombinant plasmids carrying the argA, argF, or proC genes were identified by complementation of the corresponding auxotrophic mutations . The isofunctional E . coli genes and the E . coli proB gene were subcloned into pKT240 from existing recombinant plasmids . The enzyme expression specified by the Pseudomonas genes in E . coli, calculated per gene copy, ranged from 0.3%-5% of the levels observed in Pseudomonas . Fusion of the P . aeruginosa proC gene to the E . coli consensus tac promoter resulted in very high proC enzyme production in E . coli, indicating that, at least in this case, the expression barrier is essentially at the level of transcriptional initiation . The E . coli argA and argF enzymes, which are controlled by repression in their native host, were synthesized constitutively in P . aeruginosa at 5% of the levels measured in E . coli under derepressed conditions . The constitutive E . coli proB and proC genes were expressed at high levels (ca . 50%) in the heterologous host . These results support the idea that P . aeruginosa may be a more permissive host than E . coli for the heterologous expression of genes from gram-negative bacteria.

Genetika, 1986 Jun, 22(6), 929 - 38
{Coupling and rec-independence of the processes of replication and transposition of Pseudomonas aeruginosa phage D3112 . The effect of the phage genes controlling the replication of DNA of D3112}; Akhverdian VZ et al.; D3112 phage was shown to replicate via the process of coupled replication--transposition: the phage DNA is not excised from the chromosome after prophage induction and new phage copies insert into many different sites . The transposition is controlled by two D3112 early genes--A (mapped in the 1.5-3 kbp region) and B (3-4.5 kbp), and requires intact attL site (involvement of the phage right end attR not studied) . D3112 is capable to transpose RP4 plasmid into the chromosome; both the D3112 and RP4 transpositions are rec-independent . The product of the early C gene which is not required for D3112 transposition has pleiotropic effect on the development of D3112 and is necessary for the process of D3112 DNA excision from the chromosome, for cell lysis as well as for mature phage production . We suggest that this gene is responsible for positive regulation of D3112 late genes expression, similar to the C gene of Mu phage or Q gene of lambda . Mutations in four D3112 late genes ts25, ts35, ts73 and ts110 do not affect transposition or excision processes . No detectable (less than 0.02 copies per cell) amount of linear or circular D3112 DNA is formed during the replication--transposition . Hence, in the course of replication and transposition processes D3112 genome has its ends permanently bound covalently to the chromosome . The excision of the D3112 DNA takes place at late stages.

Appl Environ Microbiol, 1986 Jun, 51(6), 1239 - 46
Growth of Pseudomonas aeruginosa on nitrous oxide; Bazylinski DA et al.; Three strains of Pseudomonas aeruginosa were grown anaerobically on exogenous N2O in a defined medium under conditions that assured the maintenance of highly anaerobic conditions for periods of 1 week or more . The bacteria were observed reproducibly to increase their cell density by factors of 3 to 9, but not more, depending on the initial amount of N2O . Growth on N2O was cleanly blocked by acetylene . Cell yields, CO2 production, and N2O uptake all increased with initial PN2O at PN2O less than or equal to 0.1 atm . Growth curves were atypical in the sense that growth rates decreased with time . This is the first observation of growth of P . aeruginosa on N2O as the sole oxidant . N2O was shown to be an obligatory, freely diffusible intermediate during growth of strains PAO1 and P1 on nitrate . All three strains used this endogenous N2O efficiently for growth . For strains PAO1 and P1, it was confirmed that exogenous N2O had little effect on the cell yields of cultures growing with nitrate; thus, for these strains exogenous N2O neither directly inhibited growth nor was used significantly for growth . On the other hand, strain P2 grew abundantly on exogenous N2O when small and growth-limiting concentrations of nitrate or nitrate (2 to 10 mM) were included in the medium . The dramatic effect of these N-anions was realized in large part even when the exogenous N2O was introduced immediately after the quantitative conversion of anion-nitrogen to N2 . No evidence was found for a factor in filter-sterilized spent medium that stimulated fresh inocula to grow abundantly on N2O.(ABSTRACT TRUNCATED AT 250 WORDS)

Transplantation, 1986 Jun, 41(6), 725 - 9
Infections in heart-lung transplant recipients; Dummer JS et al.; Infectious episodes were analyzed in 14 heart-lung transplant recipients who survived more than one week after transplantation . These patients had higher rates of infection than heart transplant recipients at our institution (P less than 0.01) and greater than 90% of all infections were potentially life-threatening . A total of 67% of all infections involved the lung or thoracic cavity as a primary site, and most of the rest were disseminated viral or fungal infections . Pneumocystis carinii infections occurred in six patients and were more common in this group than in patients who received heart transplants in the same period (P less than 0.005) . Two patients followed more than one year developed a syndrome of chronic sputum production and bronchial colonization with Pseudomonas aeruginosa, which required recurrent treatment with i.v . antibiotics for symptomatic relief . The high rate of pulmonary infections in these patients presents a challenge to clinical management, and suggests that intensive and invasive monitoring for pulmonary infection is desirable.

J Bacteriol, 1986 Jun, 166(3), 1134 - 6
IS222, a new insertion element associated with the genome of Pseudomonas aeruginosa; Gertman E et al.; A new insertion element, IS222, was identified to be associated with the DNA of a mutant strain of the converting Pseudomonas aeruginosa bacteriophage D3 . The insertion sequence was 1,350 base pairs in size and possessed terminal inverted repeats . The nucleotide sequence contained single cleavage sites for EcoRI and PvuI but none for BamHI, PstI, HindIII, SmaI, or SalI . By Southern hybridization analysis, no homology was found with genomic DNA from P . aeruginosa PAT or Escherichia coli . Genomic DNA from the phage host, P . aeruginosa PAO, contained two sequences homologous to IS222.

Clin Exp Immunol, 1986 Jun, 64(3), 597 - 605
Elastase from polymorphonuclear leucocytes: a regulatory enzyme in immune complex disease; Doring G et al.; In sputa of 21 cystic fibrosis patients suffering from chronic Pseudomonas aeruginosa lung infections, activities of lysosomal proteases released from polymorphonuclear leukocytes (PMN) and immune complexes were quantitatively determined . The results showed that low immune complex values correlated with high protease activities and vice versa . In a longitudinal study fluctuating, reciprocal changes of the parameters were observed . Therefore, the hypothesis was investigated as to whether PMN elastase cleaves immune complexes and regulates inflammation . It was shown that treatment of immune complex-positive sputa with PMN elastase decreased IgG and IgA immune complex values and lead to cleavage of the immunoglobulins in the hinge region . PMN elastase-treated samples were not able to stimulate the oxidative burst of PMN . Cleavage of in vitro built immune complexes did not lead to liberation of bound antigen . Western blot analysis of one sputum sample revealed split products of the IgG heavy chain . The results suggest that PMN elastase splits off the Fc portion of immunoglobulins in immune complexes and thus regulates inflammation by a feedback mechanism leading to cyclic inflammatory states.

Infect Immun, 1986 Jun, 52(3), 885 - 91
Siderophore-mediated iron acquisition from transferrin by Pseudomonas aeruginosa; Sriyosachati S et al.; Pseudomonas aeruginosa placed across a dialysis membrane from {55Fe}transferrin caused the mobilization of the iron from the transferrin side to the bacterial or dialysate side of the membrane . Although the bacteria were capable of obtaining iron from transferrin for growth, the siderophores of P . aeruginosa failed to convert iron bound to transferrin into dialyzable, low-molecular-weight chelates . The crucial factor produced by the bacteria which was not present when the siderophores were added alone was the acid produced from the glucose minimal medium . The siderophores mobilized considerable iron from transferrin when used in the dialysis assay at pH values between 5.0 and 6.0, values which were commonly found during incubation of bacteria in the assays . When the siderophores were tested individually, pyoverdin was more effective than pyochelin in mobilizing iron across dialysis membranes at pH values of 5.0 and 6.0, but neither had appreciable activity at pH 7.4 . The amounts of iron mobilized from conalbumin were comparable to the amounts from transferrin, but there was negligible release from lactoferrin at the three pH values . When the two siderophores were combined, the level of iron mobilization was identical to that demonstrated by pyoverdin alone . When the dialysis membrane was removed and the bacteria were mixed with the siderophores and transferrin, pyoverdin was again more active than pyochelin in mediating iron transport . Although no pyochelin-mediated iron mobilization could be detected at pH 7.4, there was transport . Therefore, the bacteria appeared to be aiding the siderophores in iron mobilization from transferrin.

Bioorg Khim, 1986 Jun, 12(6), 848 - 51
{The structure of O-specific polysaccharide from Pseudomonas aeruginosa 013 containing 5,7-diacetoamido-3,5,7,9-tetradeoxy-D- glycero-L-galacto-nonulosonic acid and 2-acetamidino-2,6-dideoxy-L-galactose}; Knirel' IuA et al.; O-Specific polysaccharide chain of P . aeruginosa 013 (Lanyi) lipopolysaccharide is composed of N-acetyl-D-quinovosamine (QuiNAc), acetamidino derivative of L-fucosamine (FucNAm), and 5,7-diacetamido-3,5,7,9-tetradeoxy-D-glycero-L-galacto-nonuloso nic acid (Sug) . On solvolysis with HF in methanol, the polysaccharide afforded methylglycosides of a disaccharide and a trisaccharide both containing fucosamine and ulosonic acid derivatives . Chemical transformations (alkaline hydrolysis, reductive deamination, acetylation accompanied by intramolecular acylation of acetamidino group by ulosonic acid), 1H and 13C NMR analysis and mass spectral data proved the following structure of the trisaccharide unit of the polysaccharide: -8)-beta-Sug-(1-3)-alpha-L-FucNAm-(1-3)-alpha-D-QuiNAc -(1-

J Biochem (Tokyo), 1986 Jun, 99(6), 1551 - 61
The structure of the O-specific chain of lipopolysaccharide from Pseudomonas aeruginosa IID 1008 (ATCC 27584); Yokota S et al.; Structural studies were carried out on the O-polysaccharide fraction obtained from the lipopolysaccharide of Pseudomonas aeruginosa IID 1008 (ATCC 27584) . The O-polysaccharide comprises L-rhamnose, N-acetyl-D-quinovosamine, N-acetyl-D-galactosaminuronic acid, and N-formyl-D-galactosaminuronic acid . The characterization of oligosaccharide fragments resulting from acid hydrolysis, Smith degradation and alkaline degradation of the O-polysaccharide, together with 1H-NMR and 13C-NMR spectroscopic data of the polysaccharide, led to the following structure for the repeating units: ----3)Rha(alpha 1----4)GalNAcA(alpha 1----4 GalNFoA(alpha 1----3)QuiNAc(alpha 1---- . Almost all of the carboxyl groups of the N-acetylgalactosaminuronic acid residues and about half of the same groups of the N-formylgalactosaminuronic acid residues were in an amide form.

Antimicrob Agents Chemother, 1986 Jun, 29(6), 1079 - 87
Gentamicin interaction with Pseudomonas aeruginosa cell envelope; Martin NL et al.; Gentamicin, an aminoglycoside antibiotic known to inhibit protein synthesis, had a detrimental effect on the integrity of the cell wall of Pseudomonas aeruginosa ATCC 9027 (a susceptible strain) as shown by electron microscopy using negative-staining, thin-sectioning, and freeze-fracture techniques . The disruption occurred in a sequential manner, moving from the outer membrane to the inner membrane, and could result in lysis of the cell . During this process the outer membrane lost 34% of its total protein and 30% of its lipopolysaccharide (measured as 2-keto-3-deoxyoctonate) upon exposure to 25 micrograms of gentamicin per ml for 15 min . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the outer membrane proteins showed altered banding patterns after exposure to gentamicin . Atomic absorption spectrophotometry revealed a decrease in magnesium and calcium content (18 and 38%, respectively) in the cell envelopes after gentamicin treatment . It is proposed that gentamicin displaces essential metal cations within the outer membrane, consequently destabilizing and extracting organic constituents . Small transient holes are thereby produced which make the outer membrane more permeable to the antibiotic and which expose the protoplast to high concentrations of gentamicin . This membrane effect may contribute to the effects of protein synthesis inhibition during the killing process.

Infect Immun, 1986 Jun, 52(3), 756 - 62
Functionally distinct monoclonal antibodies reactive with enzymatically active and binding domains of Pseudomonas aeruginosa toxin A; Chia JK et al.; Monoclonal antibodies (MAbs) are described which react with two discrete structural domains of Pseudomonas aeruginosa toxin A and which have two distinct functional profiles . The MAbs designated T3-1C7 and T4-1F2 reacted with a 46,000-dalton peptide similar to the putative B or binding fragment of toxin A . These antibodies neutralized the cytotoxic and lethal properties of toxin but had no effect on its ADP-ribosyl transferase activity . T4-1F2 interfered with the binding of toxin A to membrane receptors on mouse fibroblasts (L cells), although the epitope for the antibody appears to be distinct from the actual receptor binding site . The MAb designated T2-1H2 reacted with intact toxin A and with a cloned, enzymatically active carboxy-terminal polypeptide similar to the toxin A fragment . This MAb neutralized the ADP-ribosyl transferase activity of activated holotoxin and of the cloned peptide, but inhibited neither binding of toxin to membrane receptors nor its cytotoxic and lethal actions . The complementary specificity and function of these MAbs confirm the functional specialization of discrete structural domains within the toxin A molecule . Our findings suggest the greater antitoxic potential of antibodies that block binding, compared with those which inhibit the enzymatic activity of toxin A.

Am J Med, 1986 May 30, 80(5C), 64 - 9
Toxicologic and pharmacologic considerations in the choice of empiric parenteral antibiotics; Glauser MP et al.; The last five years have produced an explosive development of new beta-lactam compounds with extended bacterial spectra to include most of the gram-positive cocci as well as the gram-negative bacilli . With the exception of activity against enterococci, there are now third-generation cephalosporins with a spectrum of activity similar to that of the aminoglycosides . In addition, beta-lactams retain a high level of activity against anaerobes and against aerobes surviving in anaerobic conditions, a capacity that is lacking with the aminoglycoside group of antibiotics . The beta-lactams in general and some specific compounds in particular (moxalactam, ceftriazone) have a good penetration into the cerebrospinal fluid, a capacity that is also lacking with the aminoglycosides . Since there is a wide range of different spectra of activity among penicillins and cephalosporins, it is also possible to choose an antibiotic with a spectrum restricted to the offending microorganisms . Finally, because the potential toxicity of beta-lactams is less than that of the aminoglycosides (with the possible exception of myelotoxicity), the first choice of antibiotic in a patient with suspected severe infection should be a beta-lactam . The addition of an aminoglycoside should be reserved for those situations in which a synergistic action between the beta-lactam and the aminoglycoside antibiotics is expected to increase the efficacy of the beta-lactam alone, e.g., enterococcal infections, Pseudomonas aeruginosa infections, infections in severely neutropenic patients, or bacterial endocarditis.

Am J Med, 1986 May 30, 80(5C), 40 - 4
In vitro models in the study of antibiotic therapy of infections in neutropenic patients; Zinner SH et al.; Most conventional methods for in vitro testing of antibiotics involve exposure of a bacterial inoculum to a constant, static concentration of drug . The in vivo concentrations of antibiotics change continually according to their pharmacokinetics . When two drugs are used, the ratios of their concentrations also change with time . The usual checkerboard tests for combined activity of two or more antibiotics do not consider the pharmacokinetic properties . An in vitro two-compartment pharmacokinetic model has been developed that presents changing concentrations of one or two antibiotics to isolated bacterial inocula . This model simulates the treatment of a bacterial infection in the absence of host defenses and thus mimics infection in a neutropenic patient . This model has been used to study the synergistic activity of beta-lactam/aminoglycoside combinations compared with conventional checkerboard and time-kill methods . Also, in this model, the addition of azlocillin or ceftazidime to netilmicin prevented the selection of resistant subpopulations of Pseudomonas aeruginosa that occurred with the aminoglycoside alone . In vitro pharmacokinetic models add kinetic parameters to conventional susceptibility testing and may prove useful in the design of trials of the optimal dosing and administration of antibiotics for infected neutropenic patients.

Am J Med, 1986 May 30, 80(5C), 53 - 8
Efficacy of single-agent therapy with azlocillin, ticarcillin, and amikacin and beta-lactam/amikacin combinations for treatment of Pseudomonas aeruginosa bacteremia in granulocytopenic rats; Johnson DE et al.; The efficacy of azlocillin, ticarcillin, and amikacin as single agents and the penicillin/amikacin combinations for treatment of Pseudomonas aeruginosa bacteremia during cyclophosphamide-induced severe neutropenia in a rat model were assessed . Equivalent antibiotic dosing was based on the time rat serum antibiotic levels were above the minimal bactericidal concentration for the challenge organism . Antibiotic therapy was administered for 62 hours after bacterial challenge . Antimicrobial efficacy was based on the rate of bacteremia, the emergence of resistant organisms during therapy, life-table survival analysis, and rat survival seventy-two hours after bacterial challenge . For infection with a P . aeruginosa strain susceptible to all study antibiotics, therapy with azlocillin and ticarcillin (given so as to be equipotent) were equivalent, as judged by bacteremia rates or rat survival . However, combination therapy prevented the emergence of organisms resistant to azlocillin, but not to ticarcillin . Amikacin-containing combinations were more effective than single-agent regimens.

Am J Med, 1986 May 30, 80(5C), 13 - 20
Empiric antibiotic therapy for granulocytopenic cancer patients; Schimpff SC; Many cancer patients become granulocytopenic as a result of therapy and, as such, are likely to have fever during neutropenic episodes . Approximately 20 percent of these episodes have an associated gram-negative rod bacteremia; these infections occur among the most profoundly granulocytopenic patients and are associated with the highest mortality . Most infections are caused by one of three organisms: Escherichia coli, Pseudomonas aeruginosa, or Klebsiella pneumoniae . The standard approach to therapy has been the empiric utilization of an antibiotic combination, most often an aminoglycoside with either an anti-Pseudomonas penicillin or a cephalosporin . In patients for whom concern about aminoglycoside-associated nephro- or ototoxicity is high, a double beta-lactam combination has been considered . Also, with the introduction of increasingly active, exceptionally broad-spectrum antimicrobials, empiric therapy with single agents has been considered . Beta-lactam/aminoglycoside combinations more often than not are synergistic, although antagonism can be detected on occasion . Some double beta-lactam combinations demonstrate antagonism, whereas in other cases, synergism, or at least partial synergism, can be observed . Antibiotic combinations can be evaluated through in vitro models, such as the capillary model system, or through animal models designed to mimic the neutropenic state with gram-negative bacteremia, to determine potential agents or combinations of agents for this patient population . These preclinical approaches have suggested that some agents may prove effective as monotherapy and, indeed, have been comparable in activity to some of the standard antibiotic combinations . However, clinical trials have had insufficient numbers of particularly high-risk patients with profound, persistent granulocytopenia and gram-negative rod bacteremia to be able to assess their usefulness in such patients . In general, it still appears to be advantageous to use combinations such as those used in the most recent European Organization for Research on Treatment of Cancer antimicrobial trial, which compared azlocillin/amikacin with ceftazidime/amikacin . In order to reduce aminoglycoside toxicity, patients were randomly assigned to receive amikacin either for a short period or for the entire length of therapy . The study should help to determine whether it is possible to maintain the advantages of two-drug combinations while reducing the disadvantages of prolonged aminoglycoside therapy.

Eur J Biochem, 1986 May 15, 157(1), 33 - 8
A small diffusion pore in the outer membrane of Pseudomonas aeruginosa; Yoneyama H et al.; The permeability properties of the outer membrane of Pseudomonas aeruginosa were re-examined, since the reported conclusions are conflicting {Decad, M . G . and Nikaido, H . (1976) J . Bacteriol . 128, 325-336; Caulcott, C . A., Brown, M . R . W . and Gonda, I . (1984) FEMS Microbiol . Lett . 21, 119-123} . On the basis of the experimental evidence to be described below we conclude that the exclusion limit of the outer membrane of P . aeruginosa is smaller than the size of uncharged disaccharides but larger than the size of hexose . This conclusion is based on the following evidence . Penetration of monosaccharides into the expanded periplasm was large and that of disaccharides was small, after the cells were plasmolyzed with 600 mosM NaCl . A significant amount of protein was released after osmotic down-shock of cells treated with the hypertonic monosaccharides but not of cells treated with the hypertonic saccharides larger than disaccharides . Centrifuged pellets of cells treated with hypertonic di, tri and tetrasaccharides weighed about 15-20% less than that of cells treated with the isotonic monosaccharide, suggesting that the osmotic pressure was exerted on the outer membrane causing dehydration and shrinking of the cells . By contrast, cells treated with the hypertonic pentose and hexoses weighed about 0.1% and 6% less, respectively, than cells treated with the isotonic saccharide, suggesting that pentose diffused through the outer membrane freely.

Eur J Biochem, 1986 May 15, 157(1), 129 - 38
Somatic antigens of Pseudomonas aeruginosa . The structure of O-specific polysaccharide chains of P . aeruginosa O10 (Lányi) lipopolysaccharides; Knirel YA et al.; Mild acid degradation of lipopolysaccharides from Pseudomonas aeruginosa O10a and O10a,b (Lanyi classification) resulted in O-specific polysaccharides built up of trisaccharide repeating units containing 2-acetamido-2,6-dideoxy-D-glucose (N-acetylquinovosamine, DQuiNAc), 2-acetamido-2,6-dideoxy-D-galactose (N-acetylfucosamine, DFucNAc), and 5-acetamido-3,5,7,9-tetradeoxy-7-{(R)-3-hydroxybutyramido} -L-glycero-L-manno-nonulosonic acid . The latter is a di-N-acyl derivative of a new sialic-acid-like sugar which was called by us pseudaminic acid (PseN2) . A 3-hydroxybutyric acid residue was also found in natural carbohydrates for the first time . In the O10a,b polysaccharide pseudaminic acid carried an O-acetyl group at position 4 . For selective cleavage of the O10a polysaccharide, solvolysis with hydrogen fluoride was employed which, owing to the relatively high stability of the glycosidic linkage of pseudaminic acid, led to the disaccharide with this sugar on the non-reducing terminus . Performing the solvolysis in methanol afforded the methyl glycoside of this disaccharide which proved to be more advantageous for further analysis . Carboxyl-reduction made the glycosidic linkage of pseudaminic acid extremely labile, and mild acid hydrolysis of the carboxyl-reduced 010a polysaccharide afforded the trisaccharide with a ketose derivative on the reducing terminus . Establishing the structure of the oligosaccharide fragments obtained and interpreting the 13C nuclear resonance spectra of the polysaccharides allowed to determine the following structure for their repeating units: (formula: see text) In the polysaccharides the N-acetylquinovosamine residue is attached not to pseudaminic acid itself, but to its N-acyl substituent, 3-hydroxybutyryl group, and thus the monomers are linked via both glycosidic and amidic linkages.

Biochemistry, 1986 May 6, 25(9), 2448 - 55
Studies of thermally induced denaturation of azurin and azurin derivatives by differential scanning calorimetry: evidence for copper selectivity; Engeseth HR et al.; Azurin, a blue copper protein from Pseudomonas aeruginosa, and several derivatives of azurin have been studied by differential scanning calorimetry . Two well-separated, irreversible transitions are observed in a scan of apoazurin under a variety of conditions, and they are assigned to distinct steps in the denaturation process . No specific structural component can be assigned to the lower temperature transition, but a "flap" structure which is found near the metal binding site may be involved . Circular dichroic spectra suggest that melting of the beta-sheet structure, the main structural motif in the native protein, occurs during the second transition . With the exceptions of the Ni(II) and p-(hydroxymercuri)benzoate derivatives, the transitions are superposed in the metalated forms, and the enthalpies of denaturation are more endothermic . By comparison with other first-row divalent transition ions and especially Zn(II), the Cu(II) derivative exhibits the most endothermic denaturation process . Along with other data, this suggests that the binding energy is greater for Cu(II) . It is postulated that the selectivity for copper over zinc arises because of the irregular binding geometry offered by the folded protein . Denaturation of the Hg(II) derivative is even more endothermic, confirming that the type 1 binding site has a very great affinity for Hg(II) . Finally, when substoichiometric amounts of Hg(II) are added to the apoprotein, there is evidence that a novel mercury-bridged dimer of azurin forms.

Jpn J Antibiot, 1986 May, 39(5), 1241 - 9
{The prophylactic effects of latamoxef against the postoperative infections to open heart surgery . Studies on the penetration of latamoxef into the pericardial fluid and the auricle of heart}; Matsuura Y et al.; Latamoxef (LMOX, Siomarin) at a dose of 2 g was intravenously administered to each of 23 patients undergoing the open heart surgery and the concentrations in serum, pericardial fluid and auricle of heart were measured . Pharmacokinetic observations are summarized below . The peak serum concentration (t = 0) was 227.3 micrograms/ml and the serum half-life (T1/2 beta) was 1.74 hours . In pericardial fluid, LMOX reached the peak concentration of 28.44 micrograms/ml at 4.9 hours and the half-life was 9.99 hours . In auricle of heart, LMOX reached the peak concentration of 42.78 micrograms/g at 6.9 minutes and the half-life was 1.74 hours . It was shown that LMOX penetrates well into the pericardial fluid and the auricle of heart, and it is considered that their levels exceed the minimal inhibitory concentration against a majority of clinical isolates except Pseudomonas aeruginosa.

Am J Vet Res, 1986 May, 47(5), 1101 - 4
Comparison of antipseudomonad activity of chlorine dioxide/chlorous acid-containing gel with commercially available antiseptics; Kenyon AJ et al.; A chlorine dioxide-containing gel was compared with 3 commercially available antimicrobials and 1 antibiotic intended for topical use . This gel was tested at 0.5 X and 4 X and was found to be more effective as a 4 X gel in treating Pseudomonas aeruginosa-infected excised wounds on mice . To further compare this gel with other antiseptics, a murine bioassay was developed . This wound model consisted of an excised cutaneous wound on the dorsum of mice which were irradiated (800 rad) and inoculated with P aeruginosa at 10-fold dilutions, from 10(-2) to 10(-10) . The wounds were observed for latency of infection or mice survival time as a function of concentration of viable organisms remaining after treatment . The advantage of this model was demonstrated where a standard curve based on latency did not consume as many test subjects and yet provided an estimate of viable organisms in each wound . In this model, the chlorine dioxide-containing gel was more active than were preparations of providone-iodine, chlorhexidene, or silver sulfadiazine and was similar to polymyxin-bacitracin-neomycin ointment as a topical antiseptic . The effectiveness of the tested gel was reduced if delays in treatment were longer than 1 hour.

Pediatr Res, 1986 May, 20(5), 453 - 9
Altered antibody isotype in cystic fibrosis: possible role in opsonic deficiency; Moss RB et al.; Patients with cystic fibrosis (CF) whose respiratory tracts are colonized with Pseudomonas aeruginosa (PA) may develop a specific opsonic deficiency for alveolar macrophage phagocytosis of PA . We examined the possible role of altered antibody (Ab) isotype in this phenomenon by measuring serum levels and distribution of IgG and IgG subclass Ab (IgG1, IgG2, IgG3, and IgG4) to the major opsonic immunodeterminant, serotype-specific lipopolysaccharide (LPS), by means of enzyme-linked immunosorbent assays employing monoclonal secondary antibodies, and comparing these results to the serum opsonic capacity in an in vitro murine alveolar macrophage phagocytic assay . Twenty-one patients with CF who were colonized with PA had approximately a 30-fold elevation of PA LPS IgG Ab levels and higher IgG subclass 1-4 Ab compared to 10 uncolonized patients with CF and 11 healthy controls (p less than 0.05-0.0005 depending on the isotype) . Colonized patients with CF had a shift in PA LPS Ab distribution toward IgG3 compared to uncolonized patients with CF (p less than 0.02) . A surprising finding was that uncolonized patients with CF had lower levels (p less than 0.05) and proportion (p less than 0.002) of PA LPS IgG2 Ab than controls, with an apparent shift to higher levels and proportion of PA LPS IgG4 (p less than 0.01) . Serum from colonized patients with CF showed diminished opsonic capacity for phagocytosis of PA compared to uncolonized patients and controls (p less than 0.005), with 42% showing inhibitory activity . Functional Ab was also found to be inhibitory at high (greater than 500 ng/ml) concentrations . Serum opsonic capacity appeared to include a noncomplement cofactor for optimal activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Am Rev Respir Dis, 1986 May, 133(5), 784 - 8
Bacterial adherence to respiratory tract cells . Relationships between in vivo and in vitro pH and bacterial attachment; Palmer LB et al.; Alterations in in vitro pH have been shown to have a significant effect on the bacterial binding capacity of epithelial cells for certain organisms . We investigated the effect of in vivo pH and in vitro changes in pH on the adherence of Pseudomonas aeruginosa to buccal and tracheal cells of 19 chronic tracheostomy patients . In addition, airway pH was measured in 5 normal volunteers . Oropharyngeal and endobronchial pH were measured with a flexible electrode that could be passed through a bronchoscope under direct visualization . In vitro adherence of Pseudomonas to respiratory epithelial cells was determined at pH 6.5 and 7.2 . Colonization status of the patients was determined by culture of oropharyngeal and tracheal secretions . The pH value of each site in the respiratory tract and its secretions were different in the tracheostomy patients (buccal pH 6.3; tracheal, 6.9; sputum, 7.5) . No difference was noted in pH of the 2 sites in control subjects (buccal, 6.7; tracheal, 6.7) . Changes in in vitro pH had a significant effect (p less than 0.004) on bacterial binding to epithelial cells from both sites . The pH had its greatest effect on tracheal cells of patients colonized with Pseudomonas . Increased in vitro binding of these organisms was noted at the more alkaline pH . The magnitude of the effect of in vitro pH alteration on bacterial binding correlated directly with the degree of binding . These results demonstrated that pH has a significant effect on in vitro Pseudomonas adherence and the effect is most marked on cells obtained from the lower respiratory tract of patients colonized with this organism.(ABSTRACT TRUNCATED AT 250 WORDS)

J Pediatr, 1986 May, 108(5 Pt 2), 830 - 4
Antibiotic resistance of Pseudomonas species; Prince A; Pseudomonas species are highly versatile organisms with genetic and physiologic capabilities that allow them to flourish in environments hostile to most pathogenic bacteria . Within the lung of the patient with cystic fibrosis, exposed to a number of antimicrobial agents, highly resistant clones of Pseudomonas are selected . These may have acquired plasmid-mediated genes encoding a variety of beta-lactamases or aminoglycoside modifying enzymes . Frequently these resistance determinants are on transposable elements, facilitating their dissemination among the population of bacteria . Mutations in chromosomal genes can also occur, resulting in constitutive expression of normally repressed enzymes, such as the chromosomal cephalosporinase of Pseudomonas aeruginosa or Pseudomonas cepacia . These enzymes may confer resistance to the expanded-spectrum beta-lactam drugs . Decreased cellular permeability to the beta-lactams and the aminoglycosides also results in clinically significant antibiotic resistance . The development of new drugs with anti-Pseudomonas activity, beta-lactam agents and the quinolones, has improved the potential for effective chemotherapy but has not surpassed the potential of the organisms to develop resistance.

J Antimicrob Chemother, 1986 May, 17 Suppl C, 123 - 6
Timentin in the treatment of invasive burn wound infection with sepsis; Diem E et al.; Twenty-one patients with infected burns were treated with 5.2 g Timentin (ticarcillin 5 g + potassium clavulanate 200 mg) three times a day for an average of 8.3 days . Staphylococcus aureus was the commonest pathogen, there being ten cases of septicaemia with this organism and all were cured . Three patients with Pseudomonas aeruginosa septicaemia and infections with methicillin-resistant S . aureus failed . No drug related side effects were noted.

Antimicrob Agents Chemother, 1986 May, 29(5), 838 - 44
Bactericidal effects of ticarcillin-clavulanic acid against beta-lactamase-producing bacteria in vivo; Boon RJ et al.; The comparative efficacies of ticarcillin and ticarcillin plus clavulanic acid have been determined in the mouse against experimental infections caused by ticarcillin-resistant bacteria . The infections studied comprised an intraperitoneal infection, local tissue infections, pyelonephritis, and pneumonia . Both ticarcillin and clavulanic acid penetrated readily to the sites of infection studied and at the doses employed were present at concentrations of the same order as those obtained in humans after the administration of ticarcillin-clavulanic acid formulations (Timentin; Beecham) . At these concentrations, the ticarcillin-clavulanic acid combination caused significant bactericidal effects at the sites of infection against the ticarcillin-resistant strains of Bacteroides fragilis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus investigated . The efficacy of ticarcillin plus clavulanic acid against the infections resistant to therapy with ticarcillin demonstrated the beta-lactamase-inhibitory activity of clavulanic acid in vivo.

J Infect Dis, 1986 May, 153(5), 902 - 9
Levels of free granulocyte elastase in bronchial secretions from patients with cystic fibrosis: effect of antimicrobial treatment against Pseudomonas aeruginosa; Suter S et al.; Large amounts of free granulocyte elastase (GE), an enzyme capable of mediating airway damage, have been found in bronchial secretions of patients with cystic fibrosis who are infected with Pseudomonas aeruginosa . This finding indicates an imbalance between GE and its antiproteases, alpha 1-proteinase inhibitor (alpha 1-PI) and bronchial mucosal inhibitor (BMI), in the airways of these individuals . The effect of intravenous antimicrobial treatment against P . aeruginosa on activity and concentration of GE, BMI, and alpha 1-PI was evaluated in 30 treatment courses of 20 patients with cystic fibrosis . Although sputum volume and level of immunoreactive GE decreased and concentrations of alpha 1-PI and BMI increased significantly (P less than .05), a high level of free GE persisted . No active alpha 1-PI and BMI were detectable after treatment . High levels of GE correlated with a poor pulmonary condition (rs = .98, P less than .001) . In vitro, elastolytic activity of bronchial secretions from patients with cystic fibrosis was significantly inhibited by eglin C and an oxidation-resistant variant of alpha 1-PI, both compounds currently produced by recombinant DNA technology.

Mol Biol Evol, 1986 May, 3(3), 191 - 204
Nucleotide sequence of the genes for tryptophan synthase in Pseudomonas aeruginosa; Hadero A et al.; We have determined the DNA sequence of the two adjacent genes for the alpha and beta chains of tryptophan synthase in Pseudomonas aeruginosa, along with 34 5'-flanking and 799 3'-flanking base pairs . The gene order is trpBA as predicted from earlier genetic studies, and the two cistrons overlap by 4 bp; a ribosome binding site for the second gene is evident in the coding sequence of the first gene . We have also determined the location of three large deletions eliminating portions of each gene . A detailed comparison of the deduced P . aeruginosa amino acid sequence with those published for E . coli, Bacillus subtilis, and Saccharomyces cerevisiae shows much similarity throughout the beta and most of the alpha subunit . Most of the residues implicated by chemical modification or mutation as being critical for enzymatic activity are conserved, along with many others, suggesting that three-dimensional structure has remained largely constant during evolution . We also report the construction of a recombinant plasmid that overproduces a slightly modified alpha subunit from P . aeruginosa that can form a functionally effective multimer with normal E . coli beta 2 subunit in vivo.

Ann Inst Pasteur Microbiol, 1986 May-Jun, 137A(3), 253 - 66
Possibility of using purified pyocins for typing Pseudomonas aeruginosa: purification of pyocins and sensitivity of P . aeruginosa in different tests; Jurado Chacon D et al.; Two types of pyocins were simultaneously found in Pseudomonas aeruginosa strains HCP2 . According to their structures, they belonged to the types classified as R and F, respectively, and were named HCP2-R and HCP2-F . The sensitivity of 87 strains of P . aeruginosa of clinical origin to pyocins of strain HCP2 was compared in different types of tests . Results indicated that the use of the purified pyocins for the typing of P . aeruginosa provides data which are easier to control and interpret.

Pathol Biol (Paris), 1986 May, 34(5), 431 - 5
{Bactericidal activity of cefpiramide on P . aeruginosa using an in vitro pharmacokinetic simulation model}; Drigues P et al.; Cefpiramide (CPM) is a new cephalosporin with good activity against Pseudomonas . Sustained high serum concentrations are observed . We studied CPM killing kinetics using an in vitro model that simulates the pharmacokinetic profile observed in humans following a single intramuscular injection . The strain tested was Pseudomonas aeruginosa NCTC 8203 . CPM was compared to cefoperazone (CPZ), cefsulodin (CFS) and ceftazidime (CTZ) . These drugs differed only for the time-interval to bacterial regrowth that was in the following ascending order: CFS, CPZ, CTZ and CPM . This finding corroborates the fact that cefpiramide's pharmacokinetic properties allow wider space intervals between doses than for other drugs.

Pathol Biol (Paris), 1986 May, 34(5), 419 - 23
{Beta-lactamase induction in Pseudomonas aeruginosa by cefpiramide and 3 other antipyocyanic cephalosporins}; Drigues P et al.; Cefpiramide (SR 95445) (CPM) is a new cephalosporin with activity against Pseudomonas and a good bioavailability following parenteral administration . This drug is a first rather than second choice treatment in Pseudomonas infections . For this reason, investigation into cefpiramide's capacity to induce beta-lactamase production is especially interesting . A heavy inoculum of P . aeruginosa NCTC 8203, a strain that produces and inducible cephalosporinase (pI = 8.7) was incubated for 4 hours with CPM, cefsulodin (CFS), cefoperazone (CPZ) and ceftazidime (CTZ) in various concentrations . After collection and sonic treatment of the bacteria, the beta-lactamase activity was assayed using an acidimetric method and expressed as units of enzyme activity per mg proteins in the cell-free extract . The smallest increase in beta-lactamase production was recorded with CPM . The strongest inductor was CTZ . CFS and CPZ had an intermediate effect.

Pathol Biol (Paris), 1986 May, 34(5), 415 - 8
{Bacteriostatic activity of azlocillin, gentamycin, amikacin singly or in combination on 200 strains of Pseudomonas aeruginosa}; Texier-Maugein J et al.; In vitro activity of azlocillin, gentamicin, and amikacin, alone or in combination was evaluated against 200 clinical isolates of Pseudomonas aeruginosa . The minimum inhibitory concentrations (MICs) were determined by a standard technique . For the evaluation of synergistic activities, one antibiotic was added in a concentration equivalent to one-fourth its MIC to increasing concentrations of the other antibiotic . The MIC for 50% of the strains was 3.25 micrograms/ml for gentamicin, 3 micrograms/ml for amikacin and 7 micrograms/ml for azlocillin . No significant difference could be seen between the two combinations, the percentage of synergy was 37% for azlocillin-gentamicin and 36% for azlocillin-amikacin . No antagonism was observed.

Infection, 1986 May-Jun, 14(3), 105 - 7
Antibody determinations by counterimmunoelectrophoresis in the diagnosis and management of Pseudomonas aeruginosa bone and joint infections; Wagner DK et al.; Antibody titers were measured by counterimmunoelectrophoresis in eight patients with Pseudomonas aeruginosa and in eight patients with Staphylococcus aureus bone and joint infections . Titers were obtained at the beginning of therapy in all patients with pseudomonas infections and at various intervals during and after completion of therapy in seven patients . These patients were followed clinically, with 99mTc-MDP bone and 67Ga scans, and with serial erythrocyte sedimentation rates . Six of eight of these patients had detectable pseudomonas antibodies . A fall in antibody titer occurred in all six successfully treated antibody-positive patients . By contrast, none of the control patients with staphylococcal infections had antibodies to pseudomonas heptavalent antigen . The predictive value of a positive test was 100% and of a negative test, 80% . Counterimmunoelectrophoresis is a helpful adjunctive test in supporting the diagnosis and documenting a successful treatment of chronic pseudomonas bone or joint infections.

Appl Environ Microbiol, 1986 May, 51(5), 985 - 9
Pilot plant production of rhamnolipid biosurfactant by Pseudomonas aeruginosa; Reiling HE et al.; Rhamnolipid biosurfactants were continuously produced with Pseudomonas aeruginosa on the pilot plant scale . Production and downstream processing elaborated on the laboratory scale were adapted to the larger scale . Differences in performance resulting from the scale-up are discussed . A biosurfactant concentration of approximately 2.25 g liter-1 was achieved . The biosurfactant yield on glucose was 77 mg g-1 h-1, and the productivity was 147 mg liter-1 h-1, corresponding to a daily production of 80 g of biosurfactant . The first enrichment step consisted of an adsorption chromatography which was followed by an anion-exchange chromatography . The resulting product was 90% pure, and the overall recovery of active material was above 60% with the downstream processing used.

Antibiot Med Biotekhnol, 1986 May, 31(5), 366 - 9
{Significance of cellular pharmacokinetic data in predicting antibiotic activity in body cells: comparative research on preparations acting on Pseudomonas aeruginosa}; Kivman GIa et al.; Penetration of carbenicillin, cefotaxime, gentamicin and sisomicin into leukocytes of peripheral blood in man and their intracellular distribution were studied . By its capacity for penetration into leukocytes cefotaxime was superior to the other antibiotics . The levels of sisomicin and carbenicillin absorption by the cells were equal . The level of gentamicin penetration into the cells was the lowest . After penetration into the cells the main part of carbenicillin was preserved in its active form in the cytoplasm . Absorption of the antibiotic by the nuclei, granules, mitochondria and microsomes was insignificant . Cefotaxime was bound to the organoids and not more than 20 per cent of the antibiotic were preserved in its active form . The aminoglycosides were mainly absorbed by the cell granule fraction and the level of the active form of sisomicin in the cytoplasm was twice as higher as that of gentamicin . On the basis of the data on the cell pharmacokinetics of the above antibiotics, their mean therapeutic serum levels and mean geometric values of their MICs for P . aeruginosa it was suggested that in case of intracellular localization of P . aeruginosa the increase in the antibiotic efficacy was of the following order: gentamicin less than cefotaxime less than or equal to sisomicin less than carbenicillin.

Zh Mikrobiol Epidemiol Immunobiol, 1986 May, (5), 7 - 12
{Hospital infection due to Pseudomonas aeruginosa in resuscitation departments}; Svirskaia LM et al.; The bacteriological survey of resuscitation and intensive care units has revealed the presence of P . aeruginosa strains in the microflora in 69.4% of cases . The circulation of 1-2 P . aeruginosa strains, identical in their serovar and pyocin type, is indicative of the presence of hospital infection and, therefore, the endogenous character of the contamination of patients . P . aeruginosa hospital strains are the main causative agents of infectious complications in the patients treated in resuscitation units.

J Antimicrob Chemother, 1986 May, 17 Suppl C, 41 - 6
Antibacterial effects of ticarcillin/clavulanic acid in animal models of infection; Boon RJ et al.; The therapeutic effects produced by ticarcillin plus clavulanic acid were compared with those of ticarcillin and clavulanic acid separately against infections in the mouse caused by beta-lactamase-producing bacteria . The infections studied included a pneumonia model, a local tissue infection and pyelonephritis . The distribution of ticarcillin and clavulanic acid in infected animals was evaluated by measurement of the concentrations of the substances present at sites of infection . The results showed that both ticarcillin and clavulanic acid were well-distributed in the mouse and at the doses employed were present at the sites of infection at concentrations of the same order as those obtained in man after administration of ticarcillin/clavulanic acid formulations (Timentin) . The protection of ticarcillin by clavulanic acid from inactivation by the beta-lactamases produced in vivo by Bacteroides fragilis, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa was demonstrated by the pronounced bactericidal effects produced by the ticarcillin/clavulanic acid combination against the ticarcillin-refractory infections studied.

Can J Microbiol, 1986 May, 32(5), 436 - 8
Effect of lipopolysaccharide mutations and temperature on plasmid transformation efficiency in Pseudomonas aeruginosa; Berry D et al.; We have isolated, and characterized electrophoretically, two new lipopolysaccharide-defective (rough) mutants of Pseudomonas aeruginosa strain PAO . These strains, AK1401 and AK1414, together with two previously characterized isolates, AK1012 and AK1282, were used as recipients in transformation experiments with plasmid pR01614 DNA . The roughest mutant, AK1282, was not transformable, while the transformation efficiency of AK1012, and to a lesser extent the wild-type strain, was dependent upon the growth temperature . The two new isolates which are less rough than AK1012 were transformed at a frequency equivalent to that of the wild type-strain.

J Clin Microbiol, 1986 May, 23(5), 967 - 9
Production of elastase and other exoproducts by environmental isolates of Pseudomonas aeruginosa; Nicas TI et al.; Pseudomonas aeruginosa isolates from environmental sources and bacteremic patients were compared for their levels of elastolytic activity . No significant differences were found . The incidence of production of toxin A, phospholipase C, alkaline protease, and elastase among the environmental strains was also as high as that previously reported for clinical isolates.

Diagn Microbiol Infect Dis, 1986 May, 5(1), 17 - 23
In vitro inhibitory and bactericidal activity of cefpiramide and seven antipseudomonal agents against Pseudomonas aeruginosa; Pfaller M et al.; Cefpiramide was tested against 493 clinical isolates of Pseudomonas aeruginosa and the results of minimal inhibitory concentration, minimal bactericidal concentration, bactericidal rate, and time-kill synergy studies were compared with those obtained with seven other antipseudomonal agents . The minimal inhibitory concentrations of cefpiramide for P . aeruginosa were comparable to all of the agents tested . Minimal bactericidal concentration results were generally within one twofold dilution of the minimal inhibitory concentration values for all agents tested . Bactericidal rate studies showed that at concentrations of four times the minimal inhibitory concentration, all of the agents produced rapid killing . Results of time-kill synergy studies showed a marked synergistic interaction between cefpiramide and each of three aminoglycosides, gentamicin, tobramycin, and amikacin . These results suggest that cefpiramide may be useful in the therapy of infections due to P . aeruginosa.

Medicine (Baltimore), 1986 May, 65(3), 180 - 9
Left-sided endocarditis due to Pseudomonas aeruginosa . A report of 10 cases and review of the literature; Wieland M et al.; Ten confirmed cases of left-sided endocarditis due to Pseudomonas aeruginosa were reported in detail and the English literature was reviewed . In recent years, venous access (usually illicit) has been the major predisposing factor to this infection and abuse of pentazocine and tripelennamine has been particularly associated with endocarditis due to this organism . This infection involves previously damaged as well as normal valves . The development of congestive heart failure did not adversely affect the prognosis of this infection . However, the development of azotemia was associated with a greater likelihood of a fatal outcome . In the current series, deaths were due to uncontrolled infection . This often occurred despite inhibitory and bactericidal activity in serum generally considered adequate for treatment of endocarditis . Medical treatment alone rarely produced cure of infection . Our experience with a high frequency of major vessel embolization (4/10) and the improved survival after medical/surgical treatment suggests that prompt valve replacement combined with high doses of an aminoglycoside plus carbenicillin or ticarcillin provide the best opportunity for successful outcome in patients with left-sided endocarditis due to P . aeruginosa.

J Med Microbiol, 1986 May, 21(3), 199 - 202
Chemical composition of the extracellular slime glycolipoprotein of Pseudomonas aeruginosa and its relation to gentamicin resistance; Arsenis G et al.; The slime glycolipoproteins (GLPs) extracted from Pseudomonas aeruginosa strain C2 and its laboratory-induced gentamicin-resistant variant were analysed for gross chemical composition . The GLP of the wild-type strain contained significantly greater amounts of neutral sugars, uronic acid and thiobarbituric-reactive material (p less than 0.001) than the GLP of the gentamicin-resistant variant . Also significantly higher (p less than 0.01) was the amino-sugar content of the GLP from the wild-type strain . Paper chromatographic analyses of the hydrolysates of the GLPs revealed that two neutral sugars, rhamnose and mannose, were absent from the GLP of the resistant variant . The GLP of strain C2 contained significantly less protein than the GLP of the gentamicin-resistant variant.

J Pediatr, 1986 May, 108(5 Pt 2), 817 - 23
Immunologic aspects of surface infections in the lung; Piedra P et al.; The hallmark of cystic fibrosis is progressive bronchopulmonary damage associated with chronic infection with Pseudomonas aeruginosa, leading to respiratory failure and, ultimately, death . P . aeruginosa is an essentially nonvirulent organism in an immunocompetent host, but it has been shown to colonize the respiratory tract progressively in more than 80% of patients with CF . Patients with CF do not exhibit any evidence of immunologic compromise, and the infection is limited to the mucosal surfaces of the respiratory tract . Thus, P . aeruginosa in bronchopulmonary disease is a unique chronic mucosal infection resulting in a progressive pathologic process . It is therefore conceivable that locally induced alteration in the pulmonary mucosal defense is a major mechanism underlying the development of lung disease in patients with CF . An understanding of immunologic homeostasis in the mucous membranes and in the bronchopulmonary epithelium should provide a better perspective on the factors contributing to the acquisition and chronicity of P . aeruginosa infection in the lower respiratory tract and the development of progressive pulmonary damage unique to CF.

Mol Gen Mikrobiol Virusol, 1986 May, (5), 24 - 6
{New specific endonucleases PaeI and PaeII from Pseudomonas aeruginosa}; Sokolov NN et al.; Specific endonuclease activities have been found it two Pseudomonas aeruginosa strains . Isolation and purification of enzymes and determining their specific activities have permitted one to find out that PaeI is an isoshizomer of SphI and digests the sequence 5'-GCATG C-3' . Another isolated enzyme PaeII is an isoshizomer of SmaI and cleaves DNA in a fragment 5'-CCC GGG-3' . The use of PaeI and PaeII enzymes in genetical engineering and their advantages are discussed.

Arch Microbiol, 1986 May, 144(4), 405 - 7
Accumulation of fructose 1,6-bisphosphate in mutant cells of mucoid Pseudomonas aeruginosa as an evidence of phosphofructokinase activity; Banerjee PC; Phosphoglucose isomerase negative mutant of mucoid Pseudomonas aeruginosa accumulated relatively higher concentration of fructose 1,6-bisphosphate (Fru-1,6-P2) when mannitol induced cells were incubated with this sugar alcohol . Also the toluene-treated cells of fructose 1,6-bisphosphate aldolase negative mutant of this organism produced Fru-1,6-P2 from fructose 6-phosphate in presence of ATP, but not from 6-phosphogluconate . The results together suggested the presence of an ATP-dependent fructose 6-phosphate kinase (EC 2.7.1.11) in mucoid P . aeruginosa.

Curr Eye Res, 1986 May, 5(5), 343 - 55
Age alters ADPase positive dendritic (Langerhans) cell response to P . aeruginosa ocular challenge; Hazlett LD et al.; The morphology, distribution and quantitation of dendritic (Langerhans) cells (LC) was determined by analysis of ADPase stained epithelial flat mounts from 6-8 week young adult (resistant) and 24 month old (susceptible) aged mice before and after experimental infection with P . aeruginosa topically applied to the scarified cornea . The contralateral eye (controls) was also scarified and phosphate buffered saline applied similarly . This study has examined the changes in ADPase positive cell populations of the conjunctival limbal epithelium and corneal epithelium of naturally resistant mice (Swiss-Webster and CD2F1) following corneal infection with Pseudomonas aeruginosa at two different ages, young adult (8 week old) and aged (24 month old) . The young adult mice recover from their infection and restore corneal clarity while the aged mice have extensive ocular destruction and corneal scarring . Conjunctival limbal dendritic cell numbers in young adult mice were found to be significantly increased at day seven post infection and then returned to baseline levels . In contrast, conjunctival limbal dendritic cell numbers in aged mice were found to increase slowly and to peak at fourteen days after infection . Other differences between the two ages (young adult and aged) included an initial increase in dendritic cells five hours post infection in the young adult groups and an initial decrease at five hours in the aged groups of mice.

J Pediatr, 1986 May, 108(5 Pt 2), 800 - 5
Pseudomonas aeruginosa: biology, mechanisms of virulence, epidemiology; Vasil ML; Pseudomonas aeruginosa is a gram-negative pathogen, versatile and opportunistic in terms of its genetics, metabolic potential, and mechanisms of virulence . This versatility enables it to respond to variable and frequently adverse environmental conditions . Considered by many to be an aerobic organism, it is capable of growing anaerobically if certain substrates are available, for example, nitrates or arginine . Diversity of mechanisms of genetic exchange, including transformation, transduction, and conjugation, help P . aeruginosa adapt to changing conditions by acquiring new genetic information . Genetic manipulations have been exploited in recent years to study the basic biology of this bacterial species and the roles of its numerous virulence factors . Recently, transposon mutagenesis techniques and recombinant DNA methods (cloning) have been used to study some of the virulence factors of P . aeruginosa . The pathogenesis of P . aeruginosa infections is multifactorial, as manifested by the numerous toxins, or virulence factors, it produces and the variety of diseases it causes . P . aeruginosa is invasive and toxigenic . Infections appear to occur in stages: bacterial adherence, colonization, invasion and dissemination, and systemic or toxemic disease . Virulence factors can contribute to one or several stages of pathogenesis . Surface factors, including pili, lipopolysaccharide, and polysaccharide slime (alginate), probably contribute to the first two stages . Polysaccharide slime and lipopolysaccharide may also contribute to other processes later in the course of infection . Toxins, including exotoxin A and phospholipase C (hemolysin), and proteases of P . aeruginosa may contribute to tissue damage and dissemination . They may also aid in the procurement of nutrients required by the bacteria in the early stages of infection . The significance of the different virulence factors probably depends on the infection . Alginate production and phospholipase C are likely to have special significance in respiratory infections, particularly in cystic fibrosis.

J Bacteriol, 1986 May, 166(2), 392 - 8
Cloning of a gene cluster encoding biphenyl and chlorobiphenyl degradation in Pseudomonas pseudoalcaligenes; Furukawa K et al.; A gene cluster encoding biphenyl- and chlorobiphenyl-degrading enzymes was cloned from a soil pseudomonad into Pseudomonas aeruginosa PAO1161 . Chromosomal DNA from polychlorinated biphenyl-degrading Pseudomonas pseudoalcaligenes KF707 was digested with restriction endonuclease XhoI and cloned into the unique XhoI site of broad-host-range plasmid pKF330 . Of 8,000 transformants tested, only 1, containing the chimeric plasmid pMFB1, rendered the host cell able to convert biphenyls and chlorobiphenyls to ring meta cleavage compounds via dihydrodiols and dihydroxy compounds . The chimeric plasmid contained a 7.9-kilobase XhoI insert . Subcloning experiments revealed that the genes bphA (encoding biphenyl dioxygenase), bphB (encoding dihydrodiol dehydrogenase), and bphC (encoding 2,3-dihydroxybiphenyl dioxygenase) were coded for by the 7.9-kilobase fragment . The gene order was bphA-bphB-bphC . The hydrolase activity, which converted the intermediate meta cleavage compounds to the final product, chlorobenzoic acids, and was encoded by a putative bphD gene, was missing from the cloned 7.9-kilobase fragment.

Infect Immun, 1986 May, 52(2), 538 - 48
Molecular studies of Pseudomonas exotoxin A gene; Vasil ML et al.; A 2.7-kilobase DNA fragment carrying the entire exotoxin A (ETA) structural gene was divided into three nonoverlapping probes . Two probes covering the ETA structural gene were used in colony hybridization experiments to determine whether sequences homologous to the ETA gene could be detected in genera other than Pseudomonas or in Pseudomonas species other than Pseudomonas aeruginosa . The majority of strains examined other than the P . aeruginosa strains failed to react in the colony hybridization assays . Some Pseudomonas spp . other than P . aeruginosa and some Bordetella spp . did react in colony hybridization assays with the probes . However, additional studies in which we used Southern hybridization methods indicated that these reactions were apparently nonspecific and that the ETA gene is limited to P . aeruginosa . Studies in which we used all three ETA-related probes in Southern hybridization experiments to analyze the ETA gene and surrounding sequences in P . aeruginosa strains isolated from diverse sources revealed the following: (i) the incidence of the ETA gene in P . aeruginosa is approximately equal to 95%; (ii) there are strains which have been isolated from human infections that do not carry the ETA structural gene; (iii) there is a maximum of one copy of the ETA gene per genome in any given strain; (iv) sequences within and 4 to 5 kilobases downstream of the ETA structural gene appear to be well conserved in different strains of P . aeruginosa; and (v) in contrast, sequences immediately upstream of the ETA structural gene are considerably rearranged from strain to strain . A multicopy plasmid carrying the entire cloned ETA gene was transferred to a tox- P . aeruginosa strain . This strain synthesized and secreted mature, full-length ETA, but the amount produced was small considering the multicopy nature of the plasmid . Synthesis of toxin in this strain was only minimally affected by iron . Our data suggest that the synthesis of ETA is positively regulated . Finally, we found that the presence of the ETA gene is independent of the ability of P . aeruginosa to produce several other recognized virulence factors, supporting the concept of the multifactorial nature of P . aeruginosa virulence.

Z Naturforsch {C}, 1986 May-Jun, 41(5-6), 497 - 506
{Pyoverdin-type siderophores from Pseudomonas aeruginosa}; Briskot G et al.; From Pseudomonas aeruginosa three siderophores belonging to the pyoverdine group have been isolated which differ only in the acid side chains bound to the quinoline chromophore.

Antimicrob Agents Chemother, 1986 May, 29(5), 927 - 9
Influence of medium and method on the in vitro susceptibility of Pseudomonas aeruginosa and other bacteria to ciprofloxacin and enoxacin; Blaser J et al.; Ciprofloxacin and enoxacin were two- to fourfold less active against Pseudomonas aeruginosa in calcium- and magnesium-supplemented broth compared with unsupplemented broth regardless of inoculum size, presence of serum, or use of inhibitory or bactericidal endpoints (P less than 0.01) . The effect of cation supplementation was less pronounced and less consistent for Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus.

J Antimicrob Chemother, 1986 May, 17(5), 641 - 9
Ciprofloxacin in experimental aortic valve endocarditis due to Pseudomonas aeruginosa; Bayer AS et al.; Left-sided endocarditis caused by Pseudomonas aeruginosa is frequently associated with failure of medical therapy in man . The efficacy of ciprofloxacin and netilmicin + azlocillin has been studied in 79 rabbits with aortic valve endocarditis caused by a serum-resistant strain of P . aeruginosa . Infected animals received either: no therapy; ciprofloxacin (80 mg/kg/day); or netilmicin (6.5 mg/kg/day) + azlocillin (400 mg/kg/day) . Ciprofloxacin significantly lowered vegetation titers of P . aeruginosa at days 6 and 10 of therapy compared with netilmicin + azlocillin (P less than 0.001) . Similarly, ciprofloxacin was significantly more effective in sterilizing vegetations (P less than 0.005), curing P . aeruginosa endocarditis (P less than 0.001), and preventing bacteriological relapse after discontinuing antibiotic therapy (P less than 0.005) . Both antibiotic regimens were equally effective in sterilizing renal abscesses . Resistance to azlocillin was occasionally observed in vivo among P . aeruginosa isolates within cardiac vegetations during the second week of therapy, but not to ciprofloxacin or netilmicin.

J Biol Chem, 1986 Apr 25, 261(12), 5450 - 4
Effect of phospholipid composition on activity of sodium-dependent leucine transport system in Pseudomonas aeruginosa; Uratani Y et al.; The sodium-dependent leucine transport system of Pseudomonas aeruginosa was reconstituted into liposomes of specific polar head group composition . Na+-dependent counterflow and Na+ gradient-driven transport were measured as reconstituted transport activities . Proteoliposomes containing phosphatidylethanolamine exhibited increased transport activities . Phosphatidylglycerol, second to phosphatidylethanolamine, also enhanced the reconstituted transport activities . Proteoliposomes composed of phosphatidylcholine did not accumulate leucine . The enhanced transport activity by phosphatidylethanolamine was significantly influenced by its fatty acid composition . Dioleoylphosphatidylethanolamine was more effective in stimulating counterflow activity than dilauroylphosphatidylethanolamine . These results show that the leucine transport system of P . aeruginosa is sensitive to not only the polar head group composition but also the acyl group composition of phospholipids.

CMAJ, 1986 Apr 15, 134(8), 909 - 13
Nosocomial outbreak of Pseudomonas aeruginosa folliculitis associated with a physiotherapy pool; Schlech WF 3rd et al.; Outbreaks of community-acquired Pseudomonas aeruginosa folliculitis have recently been described in association with health spa whirlpools . In February 1984 we detected an outbreak of Pseudomonas folliculitis among hospital staff and patients using a swimming pool in a newly constructed physiotherapy unit . A rash developed in 5 (45%) of the 11 physiotherapists who had used the pool, as compared with 0 of the 17 who had not (p less than 0 005) . Pseudomonas folliculitis also developed in 6 (21%) of 29 outpatients and 4 (33%) of 12 inpatients who had used the facility; Pseudomonas infection of a surgical wound also developed in 1 of the 4 inpatients . The epidemic curve was consistent with a continuing common-source outbreak . P . aeruginosa, serotype O:10, was isolated from three physiotherapists, the patient with an infected surgical wound and the pool . A case-control study of pool users did not identify risk factors for infection, although the physiotherapists had spent longer in the pool than had the patients . After hyperchlorination and structural repairs to the pool, no further cases were identified among pool users . This outbreak is the first reported nosocomial outbreak of Pseudomonas folliculitis . Further investigation is needed to determine the risk of serious Pseudomonas infections in hospitalized patients using physiotherapy pools.

J Bacteriol, 1986 Apr, 166(1), 66 - 71
Exopolysaccharides of the phytopathogen Pseudomonas syringae pv . glycinea; Osman SF et al.; Exopolysaccharides (EPS) of the soybean pathogen Pseudomonas syringae pv . glycinea were isolated from culture filtrates and infected soybean leaves . Levan (a polyfructan with a C-2----C-6 backbone and C-2----C-1 branching) or acetylated alginate (a linear polyuronide of C-1----C-4-linked mannuronic and guluronic acids) was isolated from culture filtrates when bacterial strains were grown in a semisynthetic medium containing sucrose or glucose, respectively, as the primary carbon source . Acetylated alginate was the only EPS isolated from soybean {Glycine max (L.) Merr.} leaves inoculated with compatible (disease-inducing) strains of P . syringae pv . glycinea . The acetyl content of the P . syringae pv . glycinea alginates varied from 3 to 14%, and the amount of guluronic acid varied from less than 1 to 20% . The P . syringae pv . glycinea alginates from in vitro batch cultures were of lower molecular weight and polydispersity than those from in planta cultures, and both were of lower molecular weights than alginates produced by Pseudomonas aeruginosa.

J Antimicrob Chemother, 1986 Apr, 17(4), 499 - 503
Synergistic activity of apalcillin and gentamicin in a combination therapy in experimental Pseudomonas bacteraemia of neutropenic mice; Fu KP et al.; The therapeutic activity of a combination of apalcillin and gentamicin was evaluated in experimental Pseudomonas aeruginosa infection in neutropenic mice . Mice made neutropenic by administration of cyclophosphamide were more susceptible to P . aeruginosa infection than normal mice . At both challenge levels of 2LD50 and 20LD50, therapy with gentamicin alone was more effective than that with apalcillin or piperacillin . However, therapy with apalcillin-gentamicin combinations was significantly more effective than that by either component alone, and was as active as that with piperacillin-gentamicin . These in-vivo findings correlated with those of in-vitro studies, thus establishing a synergistic effect when apalcillin and gentamicin were combined . The results show that apalcillin when combined with gentamicin is effective in treating serious P . aeruginosa bacteraemia in neutropenic mice.

Eur J Respir Dis, 1986 Apr, 68(4), 263 - 6
Cystic fibrosis: protease activity in saliva evaluated with chromogenic substrates; Kittang E et al.; The protease activities in saliva from individuals with cystic fibrosis (CF) were studied using four different chromogenic substrates . In the CF-group a significantly decreased protease activity in the range 50-70% was found, compared to an age- and sex-matched control group, but with considerable overlap between the CF-patients and the control patients . The trypsin-like activity found in CF-patients without chronic colonisation with Pseudomonas aeruginosa was significantly decreased and without overlap compared to the control patients . The results indicate that determination of salivary protease activity using chromogenic substrates may give additional information in patients with suspected cystic fibrosis, and indicate the possibility of an additional diagnostic test.

Eur J Clin Invest, 1986 Apr, 16(2), 143 - 8
Demonstration of neutrophil chemotactic activity in the sputum of cystic fibrosis patients with Pseudomonas aeruginosa infection; Kharazmi A et al.; The pathogenesis of tissue damage in the lungs of cystic fibrosis patients with chronic P . aeruginosa infection is quite complex and not well understood . Inflammatory cells, particularly neutrophils, are accumulated in the damaged tissues and in the sputum . This study demonstrates the presence of a heat-stable neutrophil chemotactic factor(s) in the sputum of CF patients . The chemotactic activity seems to be associated with the endotoxin present in the sputum . Chemotaxis of sol phase sputum was determined in a modified Boyden chamber assay . It was found that the sputum obtained from the patients showed strong chemotactic activity for peripheral blood neutrophils of normal individuals . The activity was greater than twice that of casein, a strong neutrophil chemo-attractant . Sputum at about dilution of 1:50 exhibited chemotactic activity equal to that of casein . Heat treatment of the sputum at 56 degrees C for 30 min, to inactivate complement, and at 100 degrees C for 15 min, to denature other proteins, somewhat reduced the chemotactic activity, but there was still strong chemotactic activity . The presence of endotoxin was demonstrated in both the non-heated and the heated samples by LAL and rocket immunoelectrophoresis . It is concluded that the sputum of CF patients contain chemotactic activity of heat-stable nature and most likely of bacterial origin endotoxin . These factors are involved in attraction of neutrophils to the lungs and sputum of these patients and contribute to the tissue damage of cystic fibrosis.

Acta Pathol Microbiol Immunol Scand {B}, 1986 Apr, 94(2), 63 - 8
Pseudomonas aeruginosa virulence factors: modifications by sub-inhibitory concentrations of carbenicillin or gentamicin; Ogaard AR et al.; A virulent strain of Pseudomonas aeruginosa was assayed for adhesion to HEp-2 cells, production of toxin A, and production of elastase, in the presence of sub-inhibitory concentrations of carbenicillin and gentamicin . Both antibiotics, assayed in a concentration of 1:12 of their minimum bactericidal concentration (MBC), inhibited the production of toxin A . Gentamicin at this concentration totally abolished the production of elastase, whereas carbenicillin had little or no effect on this factor . Both antibiotics inhibited the bacterial adhesion, but in different ways . While gentamicin had a strong activity of slow onset, carbenicillin had a transitory activity of rapid onset, with return towards normal values after 90 min incubation.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Apr, (4), 14 - 7
{Immunoprophylaxis and immunotherapy of experimental Pseudomonas aeruginosa sepsis}; Grishina IA et al.; The protective activity of pyoimmunogen II, lot 9, was studied on P . aeruginosa experimental sepsis used as a model . In experiments on rats the preparation was shown to be nontoxic according to the results of the determination of acute and chronic toxicity . The preparation under test produced a high protective effect in experiments on animals infected with various P . aeruginosa strains, irrespective of their virulence and immunotype . Anti-P . aeruginosa plasma, obtained by plasmapheresis from donors immunized with pyoimmunogen II, showed a curative effect when injected into experimental animals in a dose of 0.02 ml/g body weight at early stages of the infection.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Apr, (4), 10 - 4
{Development of a latex agglutination method for the diagnosis of Pseudomonas aeruginosa infection}; Aleksandrova IA et al.; The present investigation has revealed the possibility of using different kinds of monodispersed polystyrene latex, produced in the USSR, as carriers in the process of the preparation of antibody diagnosticums intended for the detection of water-soluble slime antigens of Pseudomonas aeruginosa strains belonging to the most widespread serological types . The optimum conditions for the preparation of latex reagents and for making the latex-agglutination test have been experimentally established . The new diagnosticums+ have been shown to be highly species- and type-specific, which permits making judgment on the presence or absence of slime antigens of P . aeruginosa strains belonging to definite serovars in the clinical material under study . The preparations thus obtained have been found to retain their sensitivity for 16 months (the term of observation).

Microbiologica, 1986 Apr, 9(2), 253 - 8
Serological and pyocin typing and antibiotic sensitivity of Pseudomonas aeruginosa strains; Del Piano M et al.; 94 strains of Pseudomonas aeruginosa, isolated from hospitalized patients, were typed with serological and pyocinic methods, also testing their sensitivity to 6 antibiotics . The serological method allowed for the typing of almost all the strains (98.9%), (the 0-11 serotype was the most frequent -23.4%-) vs . 80.6% by the pyocinic method . Among those tested, ceftazidime was the most active antibiotic, against the P . aeruginosa strains examined.

J Antimicrob Chemother, 1986 Apr, 17(4), 505 - 16
An epidemic spread of multiresistant Pseudomonas aeruginosa in a cystic fibrosis centre; Pedersen SS et al.; Early in 1983 an epidemic of a Pseudomonas aeruginosa resistant to aminoglycosides, carbenicillin, ureidopenicillins, ceftazidime, cefsulodin and imipenem occurred in a cystic fibrosis centre . Most of the epidemic could be attributed to a specific nosocomial strain by means of O-grouping and phage-typing . This strain was present in the centre at a low frequency in 1973 and developed resistance during courses of chemotherapy . The epidemic was stopped by isolating patients with the resistant strains . Restrictive and selective use of antibiotics have not been sufficient to eradicate the resistant strains, which persist in 42% of the patients . The extensive use of the third generation cephalosporins in the clinic is probably responsible for inducing and selecting for the resistant strains . Clustering of patients in the centre has facilitated the spread . First-line use of older beta-lactam antibiotics, close bacteriological monitoring and prompt isolation of patients with resistant strains are recommended.

J Antimicrob Chemother, 1986 Apr, 17(4), 415 - 22
New plasmid-mediated oxacillin-hydrolyzing beta-lactamase in Pseudomonas aeruginosa; Philippon AM et al.; A novel type of oxacillin-hydrolyzing beta-lactamase, termed OXA-4, has been detected in three Pseudomonas aeruginosa strains isolated in Paris between 1977 and 1981 . The strains contained similar plasmids that determined resistance to carbenicillin, chloramphenicol, gentamicin, kanamycin, streptomycin, sulphonamide, tetracycline, tobramycin, sodium borate, and mercuric chloride, had a size of approximately 150 megadaltons, and belonged to the P-5 incompatibility group . Compared to reference OXA-1 beta-lactamase produced by plasmid RGN238, OXA-4 beta-lactamase produced by these plasmids had a similar substrate profile, similar response to inhibitors, and identical immunological reactions but differed in isoelectric point . In a more recent survey of 10 French hospitals plasmid-determined OXA-4 beta-lactamase production was found in P . aeruginosa isolates from four hospitals in the Paris area.

Am Rev Respir Dis, 1986 Apr, 133(4), 648 - 52
Association of systemic immune complexes, complement activation, and antibodies to Pseudomonas aeruginosa lipopolysaccharide and exotoxin A with mortality in cystic fibrosis; Moss RB et al.; The relevance of circulating immune complexes, plasma complement activation, and serum antibodies against discrete antigens of Pseudomonas aeruginosa, to the clinical course in patients with cystic fibrosis (CF) is unknown . We related these factors to outcome in 49 patients with CF colonized by P . aeruginosa, comparing 14 who died of lung disease with 35 survivors of similar age and duration of colonization, as well as 9 uncolonized patients with CF, 24 patients with other bronchorrheic lung disease, and 10 healthy control subjects . The patients with CF colonized by P . aeruginosa who died had a higher incidence of immune complexes than did survivors (71 versus 40%, p less than 0.05) . Moreover, C4 activation was highly associated with immune complexes and mortality (p less than 0.001 for each) . Those who died also had much higher levels of IgG antibodies to P . aeruginosa lipopolysaccharide (LPS) and exotoxin A than did survivors colonized by P . aeruginosa (p less than 0.005 and p = 0.01, respectively), whereas both groups had similar levels of P . aeruginosa sonicate, elastase, alkaline protease, and endotoxin core antibodies . We conclude that increasing levels of serum IgG antibodies to P . aeruginosa LPS and exotoxin A and the presence of systemic immune complexes and complement activation are associated with poor prognosis in CF, and may provide useful noninvasive markers for studying the possible immunopathogenesis of CF lung disease.

Invest Ophthalmol Vis Sci, 1986 Apr, 27(4), 507 - 15
The immune system in experimental Pseudomonas keratitis . Model and early effects; Twining SS et al.; A model to study the immune system in Pseudomonas keratitis was developed using defined flora rats (WAG/RijMCW) that have not been exposed to Pseudomonas aeruginosa . One group of rats was made immunocompetent towards P . aeruginosa by intraperitoneal injection of phenol-killed P . aeruginosa while a second group remained naive to this organism . Corneas of both groups were scratched centrally with a 21-g needle, before inoculation with 2 X 10(8) P . aeruginosa organisms . Corneas of control animals were either only scratched or only inoculated with the bacterium . At 18 hr, the naive animals were killed . In naive rat corneas, light and electron microscopy showed bacteria throughout the cornea, polymorphonuclear leukocytes (PMNs) distributed from the limbus towards the center, and little stromal degradation . In contrast, massive corneal degradation was observed in the immunocompetent rats; PMNs were present, but no bacteria were observed free in the stroma . The total acid protease content was higher in the immunocompetent than in the naive rat corneas, a possible reason for the observed difference in corneal degradation . This difference was not due to increased numbers of PMNs since nearly equal numbers of PMNs were counted after enzymatic disaggregation of both types of corneas . Glycogen-induced peritoneal PMNs from both types of rats migrated equally well towards P . aeruginosa culture media and media of corneas incubated with this bacterium . The authors conclude that immune recognition is (1) involved in the corneal host response to P . aeruginosa and (2) required for efficient phagocytosis by PMNs but not their recruitment.

Infect Immun, 1986 Apr, 52(1), 90 - 5
Physiological effects of a bactericidal protein from human polymorphonuclear leukocytes on Pseudomonas aeruginosa; Hovde CJ et al.; The physiological changes seen in Pseudomonas aeruginosa after exposure to a bactericidal protein (BP) from the granules of human polymorphonuclear leukocytes were studied . It was demonstrated, using radiolabeled proline or leucine, that both the rate of cellular uptake and amino acid incorporation into trichloroacetic acid-insoluble material were markedly decreased immediately after exposure to BP . The rate of O2 consumption by P . aeruginosa was decreased immediately after exposure to BP and continued to decline exponentially until it ceased completely 30 min after exposure to BP . In the presence of 30 mM CaCl2 or MgCl2, bacteria were protected from death due to BP and respiration rates were unaffected . The cellular ATP pool of P . aeruginosa remained constant for up to 2 h after exposure to BP . Membrane depolarization was measured by the influx of the lipophilic anion thiocyanate . It was shown that the cytoplasmic membrane of P . aeruginosa was partially depolarized after exposure to BP . Purified BP killed 95% of 5 X 10(6) CFU of P . aeruginosa at a concentration of 60 to 100 ng of protein per ml . Although the concentration of bacteria and BP varied with each type of experiment, the BP/bacteria ratio required to cause a 95 to 99% loss in viability remained constant . We propose that cytoplasmic membrane depolarization is the biochemical lesion responsible for the other physiological changes seen and ultimately for the death of P . aeruginosa induced by BP.

Infect Immun, 1986 Apr, 52(1), 76 - 84
Smooth lipopolysaccharide is the major protective antigen for mice in the surface extract from IATS serotype 6 contributing to the polyvalent Pseudomonas aeruginosa vaccine PEV; MacIntyre S et al.; The nature of the protective antigen in one of the sixteen monovalent extracts (viz., extract-6) contributing to the pseudomonas polyvalent extract vaccine (PEV) was studied in a mouse challenge assay . Selective removal, by filtration through Sep-Pak C18 cartridges, of two major protein antigens with molecular weights of 16,200 and 21,000 had no effect on the protection afforded by extract-6 . When analyzed on the basis of 2-keto-3-deoxyoctonate, lipopolysaccharide (LPS) purified by hot phenol extraction (LPS-A) from Pseudomonas aeruginosa (International Antigenic Typing System serotype 6) could account in full for the protective capacity of extract-6 . Comparative analysis of LPS heterogeneity by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by silver staining indicated that both extract-6 and LPS-A possessed similar spectra of smooth LPS molecules, containing between 10 and approximately equal to 50 O-antigen repeating units . Differences in the profiles of heterogeneity displayed by LPS in LPS-A and extract-6 were restricted to molecular species with short O-antigen chains . Subfractionation of LPS molecules on the basis of number of O-antigen repeating units was achieved by gel filtration in the presence of deoxycholate . Protection experiments performed on the subfractionated species of LPS-A revealed a relationship between O-antigen chain length and protective capacity; molecules with over 18 O-antigen repeating units being 50 to 100 times more protective than those with zero-two repeating units . The results indicate that most of the protection afforded by LPS-A and extract-6 can be accounted for by LPS molecules possessing extended (10 or more) O-antigen repeating units.

Infect Immun, 1986 Apr, 52(1), 161 - 5
Pseudomonas aeruginosa immunotype 5 polysaccharide-toxin A conjugate vaccine; Cryz SJ Jr et al.; Polysaccharide (PS) derived from Pseudomonas aeruginosa immunotype 5 lipopolysaccharide was covalently coupled to toxin A by reductive amination with adipic acid dihydrazide as a spacer molecule . The resulting PS-toxin A conjugate was composed of 27.5% PS and 72.5% toxin A . The conjugate was composed of heterogeneous high-molecular-weight species, all of which possessed an Mr greater than 670,000 . The conjugate was nontoxic for mice and nonpyrogenic at a dose of 50 micrograms/kg of body weight when intravenously administered to rabbits . Immunization of rabbits with the conjugate evoked both an antilipopolysaccharide immunoglobulin G (IgG) and an anti-toxin A IgG response . Anticonjugate IgG was capable of neutralizing the cytotoxic effect of toxin A . Immunization of mice with the conjugate increased the mean lethal dose from 4.5 X 10(1) P . aeruginosa for control mice to 9.6 X 10(5) P . aeruginosa for vaccinated mice . Similarly, immunization raised the mean lethal dose for toxin A from 0.2 to 4.67 micrograms per mouse.

J Infect Dis, 1986 Apr, 153(4), 676 - 81
The effects of hyperoxia on pulmonary clearance of Pseudomonas aeruginosa; Dunn MM et al.; Among the toxic effects of hyperoxia may be impaired pulmonary clearance of gram-negative bacteria . To better define this effect, we exposed BALB/c mice to 100% O2 for 24, 48, or 72 hr and intrabronchially inoculated them with 10(6) Pseudomonas aeruginosa . Clearance was assessed 4 hr later by quantitative lung cultures of air- and O2-exposed mice . Clearance was first reduced at 48 and 72 hr in mice exposed to O2 . To determine the mechanism responsible, we measured bronchoalveolar lavage neutrophil (PMN) counts and neutrophil chemotactic activity at 0, 2, and 4 hr after instillation of P . aeruginosa into mice first exposed to air or O2 for 48 hr . Air-exposed mice had more PMNs than did O2-exposed mice after challenge (13.3 +/- 2.1 X 10(5) vs . 4.4 +/- 0.6 X 10(5)) . There was no difference in neutrophil chemotactic activity between air- and O2-exposed mice at any time, although chemotactic activity increased in both groups after challenge . Our data suggest that hyperoxia impairs pulmonary clearance of P . aeruginosa by decreasing the influx of PMNs and that this effect is not due to diminished chemotactic activity in bronchoalveolar lavage fluid.

Genetika, 1986 Apr, 22(4), 566 - 75
{Two loci in the genome of the Pseudomonas aeruginosa transposable phage B39 which affect the integration process . II . Mapping of the pdeX and pdeY loci by restriction and heteroduplex analysis}; Akhverdian VZ et al.; The coordinate function of two loci - pdeX and pdeY - in the genome of a transposable phage (TP) provides the phage function pde+ (good growth on bacteria with Rms163 plasmid) . When these two loci in hybrid phages originate from different TP, some of the hybrid phages have Pde- phenotype . To localize pdeX and pdeY, the structure of hybrid TP genomes with Pde+ and Pde- phenotype obtained in crosses between B39ts+ and PH132 were studied using restriction and heteroduplex analysis . On the basis of data obtained, pdeX and pdeY were mapped in 2.85-6.4 and 6.4-16 kbp regions, respectively.

Arch Microbiol, 1986 Apr, 144(3), 228 - 32
Isolation and characterization of TMPD-oxidase mutants of Pseudomonas aeruginosa; Yang TY; Mutants showing negative oxidase-reaction have been isolated from Pseudomonas aeruginosa following mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine . These mutants were compared to the wild type cells with respect to their respiratory activities and cytochrome contents . They exhibit lower respiration rates and contain much less cytochrome c's which are responsible for their weak or negative oxidase-reaction in these mutants . This is supported in part from an initial linear relationship observed between the measured oxidase activities and the lower cytochrome c contents in these mutants . Further evidence comes from analyzing oxidase-negative cells of P . syringae, in which low cytochrome c contents similar to these oxidase mutants account for negative oxidase activities . Cytochrome o was the sole oxidase found among these mutants as well as in the wild type cell, suggesting that cytochrome c + o complex is responsible for the tetramethyl-p-phenylenediamine-oxidase activity in these mutants as the case in the wild-type cells . From the spectral characteristics it seems that all mutants contain about the same amount and type of terminal oxidase as that of the wild-type cells . The mutation occurred which altered the oxidase activities in these mutants appears to affect cytochrome c gene(s), but not the terminal oxidase gene(s).

Infect Immun, 1986 Apr, 52(1), 96 - 100
Variables which affect suppression of the immune response induced by Pseudomonas aeruginosa exotoxin A; Holt PS et al.; Pseudomonas exotoxin A has been shown previously to induce suppression of the murine immune response . In the present study, various parameters were examined which may have an effect on immunosuppression . The addition of 10(-4) ng of exotoxin A induced suppression of the immune response to trinitrophenylated Ficoll from days 3 to 10, while 10 ng of toxin exerted no suppressive effect over the same examination periods . When the toxin was administered 1 or 2 days before antigen stimulation, suppression of the response was observed with both 10 and 10(-4) ng . Priming splenocytes with toxin either in vivo or in vitro for 1 or 2 days suppressed the response of fresh cultured splenocytes to antigenic stimulation . Heated toxin, photoaffinity-labeled toxin, or preincubation of the toxin with rabbit anti-exotoxin A antiserum eliminated the toxin-induced suppression . These results suggest that Pseudomonas exotoxin A can generate multiple biological effects.

Eur J Clin Microbiol, 1986 Apr, 5(2), 197 - 200
Penetration of ciprofloxacin into bronchial secretions; Bergogne-Berezin E et al.; The penetration of ciprofloxacin into bronchial secretions was evaluated in 21 patients after a single oral dose of 500 mg of ciprofloxacin . Ten successive serum samples were collected in the interval 0-12 h after administration, and bronchial samples were taken 2, 3, 4 and 6 h after administration . Concentrations were measured in all samples using a standard microbiological assay . The results showed that the kinetics in serum did not differ from those determined in previous studies, a peak level of 2.2 +/- 1.3 mg/l being achieved at 2 h followed by a slow decrease of the levels to 0.6 +/- 0.4 mg/l at 6 h . The bronchial concentrations reached about 0.5 mg/l at 2 h and remained stable until 6 h, ranging between 0.5 and 0.8 mg/l . The ratio between simultaneous bronchial and serum levels ranged from 0.19 at 2 h to 0.95 at 6 h . These data indicate that ciprofloxacin might be suitable for treatment of severe respiratory infections, especially pneumonia caused by Pseudomonas aeruginosa, since the MICs of ciprofloxacin for most bacteria involved in bronchopulmonary infections are very low and tissue diffusion of the drug in the respiratory tract is good.

Infect Immun, 1986 Apr, 52(1), 263 - 70
Role of pyocyanin in the acquisition of iron from transferrin; Cox CD; Pseudomonas aeruginosa produces a blue pigment called pyocyanin . In the presence of oxidizable substrates, bacteria reduce this pigment to a colorless product, leukopyocyanin . Pyocyanin can also be nonenzymatically reduced by NADH . Leukopyocyanin formed by cell- or NADH-mediated reduction nonenzymatically reduces oxygen or Fe(III) . Pyocyanin-dependent iron reduction by whole bacterial cells was measured by the formation of the ferrous-ferrozine complex . In addition, leukopyocyanin reduced chelated Fe(III) including ferric iron in complex with transferrin, the serum iron-binding protein . High-pressure liquid chromatography was used to display the reductive removal of iron from transferrin and the accumulation of iron in the ferrous-ferrozine complex . Pyocyanin stimulated the accumulation of 55Fe from {55Fe}transferrin when it was added to bacteria incubated under low-oxygen conditions . Although bacteria grown in the presence of 100 microM FeCl3 reduced pyocyanin just as rapidly as iron-limited bacteria, these cells did not accumulate iron in the presence or absence of pyocyanin . Therefore, P . aeruginosa participates indiscriminantly in the reduction of pyocyanin, but soluble or available iron generated by the pyocyanin is taken up specifically by iron-limited bacteria.

J Infect Dis, 1986 Apr, 153(4), 707 - 14
The importance of pharmacodynamics in determining the dosing interval in therapy for experimental pseudomonas endocarditis in the rat; Ingerman MJ et al.; The efficacy of ciprofloxacin, BMY-28142, and ceftazidime was compared in vitro and in experimental left-sided endocarditis due to Pseudomonas aeruginosa in the rat . The dose, dosing interval, and duration of therapy were varied, and the resulting antibiotic levels in serum and vegetations were correlated with bacterial clearance from vegetations . These studies demonstrated that beta-lactams such as BMY exhibited a slow rate of bactericidal action and had no postantibiotic effect against P . aeruginosa in vitro or in vivo . As a consequence, BMY had to be given in multiple doses at relatively short intervals during which concentrations of antibiotics in vegetations were continuously in excess of the MBC for the pathogen . The earlier onset of rapid bactericidal action and the prolonged postantibiotic effect of ciprofloxacin (demonstrated in vivo and in vitro) were, in all likelihood, the factors that allowed the successful use of fewer doses of this antimicrobial agent at relatively longer dosing intervals.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Apr, (4), 68 - 71
{Comparative study of the antibody concentration and protective activity against Pseudomonas aeruginosa of gamma-globulin and IgM-enriched immunoglobulin preparations}; Kiseleva IA et al.; The protective effect of IgM-enriched immunoglobulin isolated by the caprylate-alcohol technique has been found to be more pronounced in comparison with that of the commercial preparation of gamma globulin in experiments on mice infected with P . aeruginosa live culture . The effectiveness of this protective action correlates with a higher content of antibodies to P . aeruginosa O-antigen in IgM-enriched immunoglobulin . In mice injected with P . aeruginosa toxin both preparations have shown equal intensity of their protective action.

Eur J Immunol, 1986 Apr, 16(4), 434 - 40
Effects of pyocyanine, a blue pigment from Pseudomonas aeruginosa, on separate steps of T cell activation: interleukin 2 (IL 2) production, IL 2 receptor formation, proliferation and induction of cytolytic activity; Muhlradt PF et al.; Pyocyanine was isolated by chloroform extraction of cultures of Pseudomonas aeruginosa, and purified by thin layer chromatography . The effects of pyocyanine on the various stages of T cell activation were studied with concanavalin A-stimulated CBA/J mouse splenocytes . At 12.5 microM concentration pyocyanine totally inhibited Con A-dependent proliferation and development of cytotoxic effector cells . Protein and RNA synthesis was only 50% inhibited at this concentration . Inhibitory doses of pyocyanine were nontoxic, in that cell viability was maintained, and the inhibitory effects were reversible after removal of the drug . Pyocyanine did not interfere with interleukin 2 synthesis, nor did it affect the lytic stage of cytotoxic effector T cells . However, T blasts generated by Con A in the presence of pyocyanine did not grow in response to IL2 even in the absence of pyocyanine, and IL2 receptors, detected by indirect immunofluorescence with the receptor-specific monoclonal antibody AMT-13, were diminished in pyocyanine-treated cells . Pyocyanine also inhibited IL2-dependent proliferation of T blasts with fully developed IL2 receptors . The substance thus interferes with several discrete stages of T cell activation.

J Mol Biol, 1986 Mar 20, 188(2), 225 - 32
Structure and composition of ferritin cores isolated from human spleen, limpet (Patella vulgata) hemolymph and bacterial (Pseudomonas aeruginosa) cells; Mann S et al.; Ferritin cores isolated from human spleen, limpet (Patella vulgata) hemolymph and bacterial (Pseudomonas aeruginosa) cells have been investigated by high resolution transmission electron microscopy, electron diffraction and chemical analysis . Hemosiderin particles isolated from thalassemic spleens also have been studied . The results show that there is a marked difference in structure and composition of the biomineral phases . Human ferritin and hemosiderin particles are single domain crystals of hydrated iron (III) oxide (ferrihydrite) . Lattice fringes were low in contrast and often discontinuous within the central regions of the core . Heat treatment of human ferritins results in a 5 A shrinkage in particle size and an increase in the single crystalline nature of the core . In contrast, lattice images and electron diffraction of limpet and bacterial cores show no evidence of long-range crystallographic order . Chemical analysis indicates a high inorganic phosphate (Pi) (Fe/Pi = 1.71) content in bacterial ferritin compared with human ferritin (thalassemic) (Fe/Pi = 21.0) . The high Pi content of bacterial ferritin suggests a hydrated amorphous iron (III) phosphate mineral core . Structural disorder within the limpet and bacterial cores may be associated with increased Pi content and increased oxidation in Fe(II), resulting in rapid mineral deposition . Growth of the iron (III) oxide cores in human ferritin is discussed on the basis of high resolution electron microscopy results.

Eur J Biochem, 1986 Mar 17, 155(3), 659 - 69
Somatic antigens of Pseudomonas aeruginosa . The structure of O-specific polysaccharide chains of the lipopolysaccharides from P . aeruginosa O1 (Lányi), O3 (Habs), O13 and O14 (Wokatsch), and the serologically related strain NCTC 8505; Knirel YA et al.; Lipopolysaccharides from Pseudomonas aeruginosa O1 (Lanyi classification), O3 (Habs classification), O13 and O14 (Wokatsch classification), and strain NCTC 8505, which is also related to serogroup O3 (Habs), have structurally similar O-specific polysaccharide chains built up of tetrasaccharide repeating units involving L-rhamnose (Rha), 2-acetamido-2-deoxy-D-glucose (GlcNAc), 2-acetamido-2-deoxy-L-galacturonic acid (GalNAcA), and a di-N-acyl derivative of bacillosamine (BacN): 2,4-diacetamido-2,4,6-trideoxy-D-glucose or 2-acetamido-2,4,6-trideoxy-4-{(S)-3-hydroxybutyramido}-D-glucose . The latter derivative was obtained free by solvolysis with hydrogen fluoride of carboxyl-reduced Habs O3 polysaccharide, and was identified by 1H-nuclear magnetic resonance spectroscopy and by mass spectrometry of the corresponding methylated alditol . Habs O3, Lanyi O1, and Wokatsch O14 polysaccharides contained O-acetyl groups . Solvolysis with hydrogen fluoride of the native Habs O3 polysaccharide resulted in selective cleavage of the glycosidic linkages of 6-deoxy sugars to give the trisaccharide fragment involving all three N-acylated amino sugars . Similar solvolysis of NCTC 8505 polysaccharide afforded a mixture of disaccharide and trisaccharide with N,N'-diacetylbacillosamine at the reducing end . Smith degradation of Habs O3 polysaccharide resulted in selective oxidation of rhamnose to give a glycoside of a trisaccharide with glyceraldehyde as the aglycone . Smith degradation of NCTC 8505 polysaccharide was complicated by the formation of the glycoside of a trisaccharide with an aglycone of unknown structure . A trisaccharide with rhamnose at the reducing end was also isolated after Smith degradation of the latter polysaccharide . Analysis of the composition and structure of all oligosaccharides obtained, and detailed examination of the 13C-nuclear magnetic resonance spectra of these oligosaccharides, and of both intact and modified polysaccharides, revealed the following structures of the repeating units . The structure for the NCTC 8505 polysaccharide differs from that proposed previously {Tahara, Y . and Wilkinson, S.G . (1983) Eur . J . Biochem . 134, 299-304} in the configurations assigned to the glycosidic linkages of rhamnose and bacillosamine . The results obtained show the P . aeruginosa strains studied to represent three different O-serotypes in a single O-serogroup (Formula: see text).

Biochim Biophys Acta, 1986 Mar 13, 855(3), 325 - 36
Bacterial cytotoxins, amphotericin B and local anesthetics enhance transbilayer mobility of phospholipids in erythrocyte membranes . Consequences for phospholipid asymmetry; Schneider E et al.; Incorporation of the channel-forming polyene antibiotic amphotericin B and of cytotoxins from Staphylococcus aureus (alpha-toxin) or Pseudomonas aeruginosa into erythrocyte membranes results in a concentration-dependent enhancement of the flip rates of exogenous lysophosphatidylcholine . The flip rate is also enhanced by incorporation of tetracaine and dibucaine . Removal of tetracaine and amphotericin B from the cells normalizes the flip rates . In parallel to the enhancement of flip rates, alpha-toxin produces a loss of transmembrane asymmetry of both phosphatidylethanolamine and phosphatidylserine . Pretreatment of cells with amphotericin or high concentrations (over 2.5 mmol . l-1) of tetracaine, followed by removal of the perturbing agent by washing, produces a selective loss of the asymmetric orientation of phosphatidylethanolamine to the inner membrane layer, as evaluated by the accessibility of the lipid towards cleavage by phospholipase A2 . The extent to which asymmetry is lost depends on the time of pretreatment with amphotericin or tetracaine, indicating a limitation by the rate of reorientation of phosphatidylethanolamine to the outer membrane surface . Evaluation of the accessibility of phosphatidylethanolamine towards cleavage by phospholipase A2 in the presence of local anesthetics indicates accessible fractions much higher than those obtained after removal of the perturbant . In the presence of tetracaine, endofacial phosphatidylethanolamine seems somehow to become accessible to phospholipase A2 . Phosphatidylserine does not exhibit this peculiarity . The results indicate that various types of perturbation of the lipid domain of the erythrocyte membrane may enhance the transbilayer mobility of phospholipids as well as destabilize the asymmetric distribution of aminophospholipids . However, as in other instances reported previously (Haest, C.W.M., Erusalimsky, J., Dressler, V., Kunze, I . and Deuticke B . (1983) Biomed . Biochim . Acta 42, 17-21), there is no tight coupling between transbilayer mobility and destabilization of asymmetry of the transbilayer distribution of phospholipids.

Biochemistry, 1986 Mar 11, 25(5), 1089 - 93
1H NMR investigation of cytochrome cd1: complexes with electron-donor proteins; Timkovich R; Mixtures of the dissimilatory nitrite reductase cytochrome cd1 from Pseudomonas aeruginosa and potential electron-donating proteins were prepared in both fully oxidized and fully reduced states and examined by 1H NMR spectroscopy . The relatively narrower lines of the donor proteins enabled them to be clearly observed in spectra in the presence of significant amounts of the high molecular weight cd1 . Mixtures of the physiological donor (Pseudomonas ferrocytochrome c-551) and ferrocytochrome cd1 showed specific line-broadening effects on the resonances of c-551 that depended on the mole ratio of c-551 to cd1 . The experimental broadening fit a model in which c-551 is in intermediate or fast exchange between free solution and a complex with cd1, with an association constant for the complex in excess of 10(4) M-1 . The model yields a minimum estimate for the forward bimolecular rate constant of 5 X 10(7) M-1 s-1 and suggests that the actual value may be much larger . The complexation was independent of pH in the range of 6-8, was independent of ionic strength over a salt concentration range of 20-1000 mM, and possessed a low thermal activation barrier . Mixtures of ferricytochrome c-551 and ferricytochrome cd1 showed no observable NMR perturbations, indicating that any hypothetical complex involving the oxidized forms must follow different dynamical and/or equilibrium conditions . No observable NMR perturbations existed in spectra of mixtures of cd1 and mammalian cytochrome c or Pseudomonas azurin in either oxidation state.

Biochim Biophys Acta, 1986 Mar 7, 870(1), 127 - 34
Mössbauer spectroscopic studies of the cores of human, limpet and bacterial ferritins; St Pierre TG et al.; Ferritin cores from human spleen, limpet (Patella vulgata) haemolymph and bacterial (Pseudomonas aeruginosa) cells have been investigated using 57Fe Mossbauer spectroscopy . The Mossbauer spectra were recorded over a range of temperatures from 1.3 to 78 K, all the spectra are quadrupole-split doublets with similar quadrupole splittings and isomer shifts, characteristic of iron(III), while at sufficiently low temperatures the spectra of all the samples show well-resolved magnetic splitting . At intermediate temperatures, the spectra from the human ferritin exhibit typical superparamagnetic behaviour, while those from the bacterial ferritin show behaviour corresponding to a transition from a magnetically ordered to a paramagnetic state . The spectra from the limpet ferritin show a complex combination of the two effects . The results are discussed in terms of the magnetic behaviour of small particles . The data are consistent with magnetic ordering temperatures of about 3 and 30 K for the bacterial and limpet ferritin cores, respectively, while the data indicate that the magnetic ordering temperature for the human ferritin cores must be above 50 K . These differences are interpreted as being related to different densities of iron in the cores and to variations in the composition of the cores . The human ferritin cores are observed to have a mean superparamagnetic blocking temperature of about 40 K, while that of the limpet ferritin cores is about 25 K . This difference is interpreted as being due not only to different mean numbers of iron atoms in the two types of core but also to the higher degree of crystallinity in the cores of the human ferritin.

JPEN J Parenter Enteral Nutr, 1986 Mar-Apr, 10(2), 146 - 50
Improvements in host immunity by partially purified interleukin 1 in rats with portacaval anastomosis and splenectomy; Hamawy KJ et al.; Despite the high incidence and severity of bacterial infections in individuals with chronic liver disease, the relative role of host immunity and the effects of immune stimulants have not been fully investigated . To study the role of the liver and spleen in reticuloendothelial system (RES) function and the host response to infection following portacaval anastomosis, 107 Sprague Dawley CD rats received a portacaval anastomosis either with or without an additional splenectomy, or a sham procedure . Animals that had undergone portacaval anastomosis and splenectomy were also administered a nonspecific host immune stimulant, interleukin 1, or saline and the effect on blood bacterial clearance and organ uptake examined . Three to four weeks following surgery, animals that received a portacaval anastomosis had a decreased ability to clear 59Fe-labeled Pseudomonas aeruginosa from the blood and an increased uptake of bacteria in the spleen (p less than 0.01) when compared to sham-treated animals . Rats that received a portacaval anastomosis and splenectomy showed further decreases in blood clearance and increased sequestration of bacteria in the liver (p less than 0.01) . Rats with a portacaval anastomosis and splenectomy that received interleukin 1 treatment prior to Pseudomonas bacteremia showed significantly improved blood bacteria clearance (p less than 0.05) and a reduced bacterial sequestration in the liver (p less than 0.001) when compared to similar animals receiving saline . These findings suggest that portacaval anastomosis and splenectomy result in impaired immune function as reflected by blood bacteria clearance and changes in organ sequestration of bacteria . Secondly, pretreatment of those immunocompromised animals with interleukin 1 improves immune function.

Laryngoscope, 1986 Mar, 96(3), 245 - 51
Osteomyelitis of the base of the skull; Chandler JR et al.; Infection in the marrow of the temporal, occipital, and sphenoid bones is an uncommon, but increasing occurrence . It is usually secondary to infections beginning in the external auditory canal and is caused almost uniformly by the gram negative Pseudomonas aeruginosa bacteria . Technetium and gallium scintigraphy help in the early detection of such infections while CT scans demonstrate dissolution of bone in well-developed cases . Headache is the predominant symptom . Dysphagia, hoarseness, and aspiration herald the inevitable march of cranial nerves . We have diagnosed and treated 17 cases of osteomyelitis of the skull base . Although the total mortality rate is 53%, it is now a curable disease . Six of our last 8 patients remain alive, although 1 is still under treatment . Treatment is medical and requires the long-term concomitant intravenous administration of an aminoglycoside and a broad spectrum semisynthetic penicillin effective against the causative organism.

Antimicrob Agents Chemother, 1986 Mar, 29(3), 519 - 20
Properties of a novel carbenicillin-hydrolyzing beta-lactamase (CARB-4) specified by an IncP-2 plasmid from Pseudomonas aeruginosa; Philippon AM et al.; CARB-4, a novel carbenicillin-hydrolyzing beta-lactamase with an isoelectric point of 4.3, was discovered in a strain of Pseudomonas aeruginosa from France . It was determined by a multiresistant transmissible plasmid belonging to the P-2 incompatibility group.

J Antimicrob Chemother, 1986 Mar, 17 Suppl A, 55 - 66
Ureidopenicillins, aztreonam, and thienamycin: efficacy as single-drug therapy of severe infections and potential as components of combined therapy; Winston DJ et al.; The ureidopenicillins (piperacillin, mezlocillin, and azlocillin), aztreonam, and thienamycin are new broad-spectrum beta-lactams with more potent antibacterial activity than the older cephalosporins and penicillins . However, therapy of severe infections with the ureidopenicillins alone is limited by the potential for emergence of resistant organisms, while monotherapy with aztreonam does not provide adequate coverage for Gram-positive aerobic organisms and anaerobes . In combination therapy with the aminoglycosides, the ureidopenicillins may be preferred to carbenicillin or ticarcillin in the treatment of Pseudomonas aeruginosa infections and in institutions with a high prevalence of carbenicillin or ticarcillin-resistant organisms . Double beta-lactam therapy with piperacillin plus either latamoxef (moxalactam) or ceftazidime may be advantageous in patients for whom aminoglycoside toxicity is a concern . Thienamycin is the most promising of the new beta-lactams for single-agent therapy of severe infections but may also be associated with the emergence of resistant P . aeruginosa during monotherapy . In febrile granulocytopenic patients or other severely ill patients at risk of severe P . aeruginosa infections, thienamycin or another antipseudomonal beta-lactam should be combined with an aminoglycoside.

J Antimicrob Chemother, 1986 Mar, 17 Suppl A, 1 - 5
In-vitro studies of antibiotic combinations with special emphasis on the evaluation of newly developed methods; Zinner SH et al.; We have described an in-vitro pharmacokinetic model that mimics the serum and tissue concentrations of antibiotics during therapy of human patients, and thus presents a changing concentration of antibiotics to the bacterial inoculum . This pharmacokinetic model has been used to study antibiotic combinations that are used in the treatment of infections in granulocytopenic patients . In this model the addition of piperacillin to amikacin or of ceftazidime or azlocillin to another aminoglycoside prevented the regrowth of resistant subpopulations of Pseudomonas aeruginosa . The activities of several antibiotic combinations were studied in the in-vitro model, the conventional checker-board method and the time-kill method . Discrepant results were found with the model and the conventional tests in one-third of the combinations . The model may be useful in the preclinical study of antibiotic combinations and may be proved more predictive of clinical outcome than conventional tests.

Can J Microbiol, 1986 Mar, 32(3), 248 - 53
Lipopolysaccharide changes and cytoplasmic polyphosphate granule accumulation in Pseudomonas aeruginosa during growth on hexadecane; Miguez CB et al.; Pseudomonas aeruginosa ATCC 9027 grew on 0.5% (v/v) hexadecane as a sole carbon source in a chemically defined medium which required the addition of Fe3+ and Ca2+ . There was a variable and extended lag period before an active growth rate was attained . Visible light microscopic evidence revealed that the bacteria did not adhere to hexadecane droplets suggesting the absence of a bioemulsifier . When compared with glucose-grown cells, hexadecane-grown cells produced 75% less lipopolysaccharide (on a total protein basis); this lipopolysaccharide contained 30-40% less carbohydrate, yet 50-75% more 2-keto-3-deoxyoctonate . These chemical changes made the cell surface appear more hydrophobic when tested in a biphasic hydrophobicity index system . Electron microscopy of thin sections and freeze etchings revealed hexadecane-grown cells contained granules which were judged to be polyphosphate by energy dispersive X-ray analysis . There was no apparent major morphological envelope alteration within the two cell types.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Mar, (3), 89 - 92
{Chemiluminescence of human neutrophils as affected by opportunistic microbes}; Belotskii SM et al.; Chemoluminescence of neutrophils obtained from 24 healthy donors in response to Staphylococcus aureus Cowan, Pseudomonas aeruginosa and Escherichia coli preopsonized with 5% fresh autologous serum or with pooled normal sera was studied . Chemoluminescent response to S . aureus was most pronounced in comparison with that to the other microbes . Neutrophils from most of the donors showed chemiluminescent response of medium intensity, their stimulation index (SI) being 10-12; neutrophils from some donors showed low response (their SI not exceeding 10), and some donors provided highly responsive neutrophils (their SI exceeding 20) . Neutrophils from the latter group of donors retained their high SI over the longest period of time (60 minutes and more) . Experiments made under the conditions of preopsonization with pooled normal sera indicated that differences in the response of neutrophils were linked with the individual features of these cells . Low response to P . aeruginosa and E . coli was, possibly, due to the antiphagocytic activity of these microorganisms . Differences in the response of neutrophils to antigens of opportunistic microbes, as well as in the dependence of this response from serum factors, may finally determine the result of the interaction between host defence factors and microorganisms at the infection atrium.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Mar, (3), 21 - 5
{Effect of a controlled abacterial environment on the Pseudomonas aeruginosa infection of suppurative wounds}; Samykina TD; The present work deals with the data on the isolation rate of P . aeruginosa from suppurative wounds of different origin during their treatment by the commonly used methods under dressings and by the open method under the conditions of controlled germ-free environment . The results of the immunotyping of P . aeruginosa strains isolated from patients treated by different methods are presented . The dynamics of changes in the isolation rate of P . aeruginosa at different periods of treatment, both by the open method and with the use of dressings, is shown . Among P . aeruginosa strains isolated from suppurative wounds, those belonging to immunotypes 6, 7 and 2, as well as nontyping strains, occurred most frequently . Treatment in the controlled germ-free environment permits the protection of the wound surface from hospital infection . During treatment with the use of dressings the cases of hospital infection were revealed (31.3%) . Such infection occurred, as a rule, at a later period of treatment.

Zh Mikrobiol Epidemiol Immunobiol, 1986 Mar, (3), 14 - 8
{Protective effect of a corpuscular polyvalent Pseudomonas aeruginosa vaccine in generalized chronic Pseudomonas aeruginosa infection in mice with cyclophosphamide-induced leukopenia}; Vetkova LG et al.; The prophylactic effect of immunization with P . aeruginosa polyvalent corpuscular vaccine has been shown on the model of P . aeruginosa generalized chronic infection in mice with leukopenia induced by the intraperitoneal injection of cyclophosphamids . This effect is manifested by the increased resistance of the animals to sublethal doses of P . aeruginosa strain, as well as by more intense general and specific immunological responses in the infected animals (the increase of specific antibody titers, the number of leukocytes in the blood serum and the phagocytic activity of the cells of peritoneal exudate).

J Clin Microbiol, 1986 Mar, 23(3), 655 - 9
Whirlpool-associated folliculitis caused by Pseudomonas aeruginosa: report of an outbreak and review; Ratnam S et al.; An outbreak of folliculitis caused by Pseudomonas aeruginosa serotype O:7 occurred among the guests of a hotel in St . John's, Newfoundland, Canada, and the source of the infection was traced to the hotel whirlpool . Of 36 persons who used the whirlpool, 26 (72%) developed folliculitis within 1 to 5 days after exposure; the attack rate was significantly higher for children (90%) than for adults (50%) . The rash characteristics were consistent with those of Pseudomonas folliculitis previously described (T . L . Gustafson, J . D . Band, R . H . Hutcheson, Jr., and W . Schaffner, Rev . Infect . Dis . 5:1-8, 1983) . This is considered to be the first outbreak in which P . aeruginosa serotype O:7 has been incriminated . Published reports to date of outbreaks of Pseudomonas folliculitis associated with the use of whirlpools, hot tubs, swimming pools, etc., were reviewed.

J Clin Microbiol, 1986 Mar, 23(3), 513 - 6
Association with phagocytic inhibition of anti-Pseudomonas aeruginosa immunoglobulin G antibody subclass levels in serum from patients with cystic fibrosis; Shryock TR et al.; Serum from cystic fibrosis patients colonized with Pseudomonas aeruginosa specifically inhibits phagocytosis of P . aeruginosa by alveolar macrophages . Serum was examined for P . aeruginosa lipopolysaccharide-specific immunoglobulin G (IgG) subclass levels (by enzyme-linked immunosorbent assay) and for the effect on macrophage phagocytosis (by radiolabeled P . aeruginosa uptake) . Sera from cystic fibrosis patients with no known P . aeruginosa colonization history had negligible amounts of lipopolysaccharide-specific IgG and a mean phagocytic enhancement of 5% . The sera of normal volunteers also had negligible amounts of lipopolysaccharide-specific IgG . Serum from cystic fibrosis patients with P . aeruginosa respiratory tract infections had substantial titers (range, 1:20 to 1:1,280) of lipopolysaccharide-specific IgG2, IgG3, and IgG4 and a mean phagocytic inhibition of 56% . However, these patients had low or absent titers of lipopolysaccharide-specific IgG1 . No consistent variation in the level of individual IgG subclasses in the sera of colonized patients was observed, as determined by radial immunodiffusion . The results suggest that during P . aeruginosa infection phagocytosis-inhibitory activity develops coincident with production of lipopolysaccharide-specific IgG subclasses.

Am Rev Respir Dis, 1986 Mar, 133(3), 418 - 22
Immunoglobulin-G subclasses in cystic fibrosis . IgG2 response to Pseudomonas aeruginosa lipopolysaccharide; Fick RB Jr et al.; Pulmonary macrophage phagocytosis of Pseudomonas aeruginosa is defective when this pathogen is opsonized with IgG antibodies isolated from serum samples from patients with cystic fibrosis (CF) . To evaluate this defect further, IgG subclasses in the serum and lung fluids of patients with CF were quantitated . The pattern of IgG subclasses in serum specimens from patients with CF (n = 15) and in patients without CF but with chronic obstructive airway disease and recurrent P . aeruginosa infection (n = 4) was significantly altered from that found in normal subjects (n = 31) . Immunoglobulin-G2 and IgG3 expressed as percentages of total IgG subclasses or in micrograms per milliliter of serum were significantly elevated in the serum specimens of these patients (p less than 0.05), and IgG1 was significantly decreased (p less than 0.01) . It appears that the increase in IgG2 in the serum of patients with CF and those without CF but with chronic P . aeruginosa infection may be in response to chronic antigenic stimulation by P . aeruginosa lipopolysaccharide . Evidence presented to support this includes: (1) IgG2 is not increased in CF serum if a history of P . aeruginosa infection is absent, (2) IgG2 levels expressed as percentages of total IgG subclasses in CF lung fluids were positively correlated (r = 0.73) with the number of colony-forming units of P . aeruginosa present in CF sputum specimens, and (3) IgG antibodies specifically eluted from P . aeruginosa lipopolysaccharide ligands on affinity gels were largely restricted to IgG2 . The opsonic index, ({IgG3} + {IgG1}) divided by ({IgG2} + {IgG4}), is inverted in CF lung fluids (0.73:1; normal, 2:1) . Because pulmonary macrophages show surface receptors binding primarily with IgG3 and IgG1, it may be that such an alteration in IgG subclasses in the respiratory secretions of patients with CF further inhibits opsonin-mediated clearance of P . aeruginosa.

Am J Med, 1986 Mar, 80(3), 528 - 9
Subcutaneous nodules in Pseudomonas sepsis; Bagel J et al.; Approximately 60 nontender, nonfluctuant, red, hot subcutaneous nodules developed on the trunk, extremities, and face due to Pseudomonas aeruginosa septicemia in a 56-year-old woman with stage III ovarian adenocarcinoma . Two years later, these lesions appeared atrophic with central scarring . Complete eradication with systemic antibiotics and without incision and drainage was accomplished.

Pediatr Infect Dis, 1986 Mar-Apr, 5(2), 223 - 5
Microbiology of chronic suppurative otitis media in children; Kenna MA et al.; The aerobic microorganisms present in 51 middle ears of 36 children with chronic suppurative otitis media without cholesteatoma were evaluated . Specimens were taken directly from the middle ear through a patent perforation or tympanostomy tube and cultured using standard techniques . Twenty-three microbiologic species were identified, with Pseudomonas aeruginosa being the most prevalent (67% of ears) and the only organism in 31% of ears . The initial treatment of all patients was intravenous administration of an antimicrobial selected on the basis of culture and susceptibility reports; this treatment was successful in all but four children, who subsequently required tympanomastoid surgery . These results indicate that medical therapy, based on the results of culture and susceptibility studies, provides a viable alternative to major mastoid surgery in the management of chronic suppurative otitis media without cholesteatoma.

Infect Immun, 1986 Mar, 51(3), 896 - 900
Characterization of antibody to the ferripyochelin-binding protein of Pseudomonas aeruginosa; Sokol PA et al.; An outer membrane protein of Pseudomonas aeruginosa was previously shown to bind 59Fe-labeled pyochelin . Antibodies to the purified ferripyochelin-binding protein (FBP) were characterized by using a variety of assays . Anti-FBP cross-reacted with several P . aeruginosa isolates in an enzyme-linked immunosorbent assay . Anti-FBP significantly enhanced phagocytosis of P . aeruginosa by human polymorphonuclear leukocytes . In a serum bactericidal assay we observed no difference in viability between cells incubated with antiserum to FBP and cells incubated with preimmune serum . Anti-FBP immunoglobulin G inhibited both binding and uptake of 59Fe-labeled pyochelin by whole cells . Passive protection by anti-FBP was examined in experimental P . aeruginosa burn infections in mice . The protection provided by this antibody was strain dependent but lipopolysaccharide serotype independent.

Afr J Med Med Sci, 1986 Mar-Jun, 15(1-2), 35 - 40
Cefoxitin: single agent treatment of septic abortion; Abudu O et al.; PIP: An uncontrolled clinical trail was carried out to assess the efficacy of cefoxitin as the sole antimicrobial agent in the treatment of 22 cases of septic abortion in Lagos, Nigeria . The mean age of patients studied was 21 years; 90% were from social classes IV and V . All study subjects satisfied the following diagnostic criteria: pyrexia of equal to or greater than 38 C, tachycardia equal to or greater than 90/min, foul smelling vaginal discharge, uterine tenderness, adnexal tenderness, peritonitis, or features of septicemia . All patients received initial doses of 2 gm of cefoxitin sodium by slow intravenous bolus and then 2 gm every 6 hours for the next 5 days . 17 (77%) of the patients improved on cefoxitin therapy . There was no mortality and no serious side effects . The mean duration of hospital stay was 9 days for the patients who responded to cefoxitin and 21 days for patients whose conditions was not improved by this treatment . Anaerobes, especially Bacteriodes bivius, were isolated from all 22 patients . 4 of the 5 patients who did not respond to 72 hours of treatment with cefoxitin harbored organisms such as Pseudomonas aeruginosa that are resistant to this drug . None of the patients treated with cefoxitin progressed to abscess formation . These findings suggest that cefoxitin can be used as a single antimicrobial agent in the treatment of septic abortion and may offer advantages over the use of a combination of antimicrobials .

Antimicrob Agents Chemother, 1986 Mar, 29(3), 496 - 500
Interaction of polycationic antibiotics with Pseudomonas aeruginosa lipopolysaccharide and lipid A studied by using dansyl-polymyxin; Moore RA et al.; A fluorescent derivative of polymyxin B (dansyl-polymyxin) was used to study the interaction of polycations with lipopolysaccharide (LPS) and lipid A from Pseudomonas aeruginosa . Dansyl-polymyxin became bound to LPS and lipid A sites, including Mg2+-binding sites, resulting in a 20-fold enhancement of fluorescence . A Hill plot of the binding data showed that the binding of dansyl-polymyxin to LPS was cooperative (n = 1.98) and of high affinity (S0.5 = 0.38 microM) . The maximal binding capacity of LPS was approximately four molecules of dansyl-polymyxin per mol of LPS . The dansyl-polymyxin interaction with lipid A displayed similar kinetics (n = 2.26; S0.5 = 0.38 microM), and the maximal binding capacity was approximately 2 mol of dansyl-polymyxin per mol of lipid A . A variety of polycationic compounds, including gentamicin, streptomycin, and polymyxin B, as well as Mg2+, were able to displace dansyl-polymyxin bound to LPS or to lipid A . Marked differences both in terms of the degree of displacement and in terms of the amount of competing polycation required to displace a given amount of dansyl-polymyxin were observed . Addition of excess polymyxin B resulted in displacement of all of the dansyl-polymyxin, demonstrating that only polymyxin-binding sites were being probed . Our data demonstrate that polymyxin B binds to multiple sites on LPS, including sites which bind aminoglycoside antibiotics and other polycationic compounds.

Proc Natl Acad Sci U S A, 1986 Mar, 83(5), 1320 - 4
Structure of exotoxin A of Pseudomonas aeruginosa at 3.0-Angstrom resolution; Allured VS et al.; Exotoxin A of Pseudomonas aeruginosa is a secreted bacterial toxin capable of translocating a catalytic domain into mammalian cells and inhibiting protein synthesis by the ADP-ribosylation of cellular elongation factor 2 . The protein is a single polypeptide chain of 613 amino acids . The x-ray crystallographic structure of exotoxin A, determined to 3.0-A resolution, shows the following: an amino-terminal domain, composed primarily of antiparallel beta-structure and comprising approximately half of the molecule; a middle domain composed of alpha-helices; and a carboxyl-terminal domain comprising approximately one-third of the molecule . The carboxyl-terminal domain is the ADP-ribosyltransferase of the toxin . The other two domains are presumably involved in cell receptor binding and membrane translocation.

Antimicrob Agents Chemother, 1986 Mar, 29(3), 501 - 5
Tissue penetration of ciprofloxacin after single and multiple doses; LeBel M et al.; The pharmacokinetics and the suction-induced blister fluid penetration of ciprofloxacin were compared after a single dose (500 mg) and after multiple dosing (500 mg q8h for 13 doses) . Significantly higher peak levels of ciprofloxacin in serum were observed after multiple dosing (3.51 versus 2.26 micrograms/ml; P less than 0.01) . Increased elimination half-life occurred after multiple dosing; this seems to be mostly related to decreased systemic clearance secondary to a diminished nonrenal clearance (240.0 versus 125.0 ml/min) . Ciprofloxacin appeared rapidly in the blister fluid, and the percentage of penetration (AUC0-tBF/AUC0-t serum) yielded values of 88.8 versus 84.7% after single and multiple doses, respectively . Ciprofloxacin levels in serum and blister fluid at the end of the dosing interval (8 h) were superior or almost superior to MICs for sensitive organisms including Pseudomonas aeruginosa . Comparative studies of ciprofloxacin metabolite excretion after multiple doses in healthy subjects and in renal insufficiency patients are needed.

Antimicrob Agents Chemother, 1986 Mar, 29(3), 395 - 9
Comparative evaluation of ciprofloxacin, enoxacin, and ofloxacin in experimental Pseudomonas aeruginosa pneumonia; Kemmerich B et al.; The therapeutic activity of ciprofloxacin, enoxacin, and ofloxacin was evaluated in guinea pigs with acute and chronic experimental Pseudomonas aeruginosa pneumonia . Intratracheal instillations of P . aeruginosa resulted in fatal pneumonia in all untreated animals within 36 h . Among treatment groups (80 mg/kg {body weight} per day), cumulative survival rates were: 47%, ciprofloxacin; 55%, enoxacin; and 42%, ofloxacin . These rates were not significantly different . Intrapulmonary killing of P . aeruginosa was equivalent 3 h after a single dose of ciprofloxacin or ofloxacin (20 mg/kg) or enoxacin (40 mg/kg) . The combination of ciprofloxacin with azlocillin, ceftazidime, or tobramycin did not increase the efficacy of intrapulmonary killing of P . aeruginosa over that of ciprofloxacin alone . A chronic, nonfatal bronchopneumonia was induced in guinea pigs by intratracheal instillation of microscopic agar beads impregnated with a mucoid strain of P . aeruginosa . Compared with no treatment, ciprofloxacin and enoxacin produced greater than or equal to 99.9% intrapulmonary killing, and ofloxacin sterilized the lungs completely, after 4 days of treatment . In no quinolone-treated animal did resistant strains of P . aeruginosa emerge during 4-day treatment periods . In further studies with the chronic model, oral and parenteral ciprofloxacin treatment were found to be equivalent in efficacy . We conclude that several quinolone derivatives may be effective for the treatment of P . aeruginosa pneumonia and that combinations of quinolones with beta-lactams or aminoglycosides may not increase efficacy against P . aeruginosa pneumonia.

Infection, 1986 Mar-Apr, 14(2), 82 - 5
{Synergism of ciprofloxacin with beta-lactam antibiotics, gentamicin, minocycline and pipemidic acid}; Dickgiesser N et al.; Studies on the Synergism of Ciprofloxacin with beta-Lactam Antibiotics, Gentamicin, Minocycline and Pipemidic Acid . Using Escherichia coli, Klebsiella pneumoniae/Klebsiella oxytoca, Pseudomonas aeruginosa and Staphylococcus aureus strains, we examined a possible influence of pipemidic acid, minocycline, gentamicin, cefazolin, mezlocillin and ampicillin on the antibiotic activity of ciprofloxacin . We found in all bacterial species synergistic influence by pipemidic acid . When ciprofloxacin was combined with gentamicin, synergism was observed in E . coli and in K . pneumoniae/oxytoca, additive effects in P . aeruginosa and S . aureus . In combination with minocycline we demonstrated synergism in S . aureus only . In all other bacterial species and antibiotic combinations we found neither synergism nor antagonism.

Mol Gen Genet, 1986 Mar, 202(3), 403 - 9
Isolation of a Tn501 insertion mutant lacking porin protein P of Pseudomonas aeruginosa; Poole K et al.; In order to demonstrate a role for anion-specific protein P channels in phosphate transport in Pseudomonas aeruginosa PAO, we wished to isolate a transposon insertion mutant deficient in protein P . A number of transposon delivery systems were tested which yielded, for the most part, whole plasmid inserts . Plasmid pMT1000 (Tsuda et al . 1984), a temperature-sensitive R68 plasmid carrying the transposon Tn501, was successfully employed in the isolation of a Tn501 insertion mutant lacking protein P under normally inducing conditions . To identify the mutant deficient in protein P, a protein P-specific polyclonal antiserum was used . This mutant, strain H576, was deficient in high-affinity phosphate transport exhibiting a Km for uptake (3.60 +/- 0.64 microM) almost ten times greater than that of the wild type strain (Km = 0.39 microM) . There was, however, no change in the Vmax for high-affinity phosphate transport as a result of the loss of protein P in this mutant . The protein P-deficiency of the mutant correlated with a growth defect in a phosphate-limited medium, resulting in an 18%-35% decrease in growth when compared with the wild type.

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