Infect Immun, 1989 Dec, 57(12), 3793 - 7 Pseudomonas aeruginosa elastase does not inactivate alpha 1-proteinase inhibitor in the presence of leukocyte elastase; Padrines M et al.; Pseudomonas aeruginosa elastase rapidly inactivates alpha 1-proteinase inhibitor by splitting its Pro-357-Met-358 peptide bond . The present study was aimed at testing whether this reaction takes place in the presence of leukocyte elastase . To this end was added alpha 1-proteinase inhibitor to a mixture of the two elastases, and we performed the following assays: (i) measurement of the residual leukocyte elastase activity, (ii) sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and (iii) immunoassay of the leukocyte elastase-alpha 1-proteinase inhibitor complex . These experiments were done with various concentrations of the three proteins . All experiments gave the same result: leukocyte elastase was fully inhibited by alpha 1-proteinase inhibitor in the presence of P . aeruginosa elastase even when the bacterial enzyme was 10-fold more concentrated than the neutrophil enzyme . We also measured the initial rate of the P . aeruginosa elastase-catalyzed inactivation of alpha 1-proteinase inhibitor as a function of the inhibitor concentration . The kcat/Km value derived from this experiment was 9 x 10(4) M-1 s-1, a value much lower than the rate constant for the leukocyte elastase-inhibitor association (kass, 1.7 x 10(7) M-1 s-1) . This rationalizes the above results . In conclusion, when alpha 1-proteinase inhibitor is faced with its target enzyme, leukocyte elastase, it will perform its physiologic antielastase function even if the bacterial elastase is present in excess. Infect Immun, 1989 Dec, 57(12), 3783 - 7 Degradation of basement membranes by Pseudomonas aeruginosa elastase; Bejarano PA et al.; Pseudomonas aeruginosa virulence has been attributed in part to extracellular proteinases . We found that one of these proteinases, elastase, extensively degrades intact basement membranes from bovine anterior-lens capsules, bovine glomeruli, and bovine lung, producing about 9, 14, and 9 fragments, respectively, with Mrs in the range of 15,000 to greater than 200,000, as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (nonreducing conditions) . Release of hydroxyproline showed that collagen IV was degraded by elastase . Degradation of the newly discovered alpha 3(IV) collagen chain was shown by immunoblotting of digests with Goodpasture's syndrome serum, which contains antibodies that react with an epitope located in the carboxyl-terminal globular (NCl) domain of alpha 3(IV) . Comparison of total protein release with collagen IV release showed that noncollagenous protein components were solubilized to the same extent as collagen IV . The extensive degradation of the basement membranes described here suggests a role for elastase in the pathogenic mechanism at the local level when P . aeruginosa infection is present. Zh Mikrobiol Epidemiol Immunobiol, 1989 Dec, (12), 68 - 74 {Immunosuppression in experimental staphylococcal infection and its effect on antimicrobial resistance}; Barsukov VS; Experiments on CBA and (CBA X C57BL)F1 mice have revealed that a prolonged period of antigen-nonspecific immunosuppression of humoral immunity develops in experimental staphylococcal infection; this period of suppression may be preceded by a short phase of antigen-nonspecific immunostimulation . Immunosuppression is linked with the accumulation of antigen-nonspecific T-suppressors in the spleen, these T-suppressors being capable of the manifestation of their activity both in vitro and in vivo in cases of their transplantation to semi-syngeneic recipients . Immunosuppression does not aggravate the course of staphylococcal infection and is accompanied by an increase in resistance to Pseudomonas aeruginosa superinfection, which is due to the stimulation of inflammatory reaction at the site of the injection of the superinfecting agent. APMIS, 1989 Dec, 97(12), 1146 - 8 Increased levels of IgG subclasses specific for Pseudomonas aeruginosa exoenzyme and polysaccharide antigens in chronically infected patients with cystic fibrosis; Albus A et al.; IgG subclass levels to Pseudomonas aeruginosa alginate, alkaline proteinase, elastase and exotoxin A in sera of healthy adults, non-infected and infected cystic fibrosis patients were investigated by enzyme linked immunosorbent assay . Whereas healthy adults and non-infected cystic fibrosis patients revealed mostly negative IgG subclass levels to the four antigens, infected cystic fibrosis patients had significantly elevated IgG1, IgG2, IgG3 and IgG4 levels to both the protein antigens as well as the polysaccharide antigen . The study does not support previous findings of an impaired natural IgG2 response to polysaccharide antigen . The study does not support previous findings of an impaired natural IgG2 response to polysaccharide in chronically infected cystic fibrosis patients. Am J Med, 1989 Nov 30, 87(5A), 70S - 75S Ciprofloxacin pharmacokinetics in critically ill trauma patients; Yuen GJ et al.; The steady-state pharmacokinetics of ciprofloxacin 200 mg intravenously every 12 hours was examined in 10 critically ill trauma patients . The mean parameter estimates for total clearance, renal clearance, non-renal clearance, and volume of distribution were 30.08 liters/hour/1.73 m2, 16.62 liters/hour/1.73 m2, 13.46 liters/hour/1.73 m2, and 2.10 liters/kg . Although the mean values were similar to those previously reported, significant individual differences were observed, with the coefficient of variation ranging from 41 to 61 percent . Non-renal clearance appeared to have a bimodal distribution . The dosage studied appeared to provide adequate serum concentration profiles to treat most pathogens found in infected trauma patients . However, the use of higher doses and more frequent dosing may be required to treat patients with Staphylococcus aureus and Pseudomonas aeruginosa infections. Am J Med, 1989 Nov 30, 87(5A), 23S - 27S Effect of the abscess environment on the antimicrobial activity of ciprofloxacin; Bryant RE et al.; The present studies were conducted to identify factors in human purulent material that might limit or enhance the activity of ciprofloxacin against bacteria causing suppurative infection . Ciprofloxacin, imipenem, and ampicillin were tested with regard to binding or inactivation by pus . The bactericidal activity of ciprofloxacin and imipenem were tested against Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, or Staphylococcus aureus in human pus with a pH of 6.0 incubated at 37 degrees C under aerobic or anaerobic conditions . The effect of single or combination drug therapy with 20 mg/kg of ciprofloxacin, imipenem, or rifampin given every 12 hours was tested against E . coli or P . aeruginosa in polymicrobic murine abscesses that had been produced by subcutaneous injection of either of those organisms mixed with Bacteroides fragilis and autoclaved human stool . Antibiotic levels and the number of bacteria surviving in pus were quantitated . Therapy of subcutaneous abscesses was delayed 72 hours to test drug efficacy against organisms in well-established infections . Levels of ampicillin, imipenem, or ciprofloxacin were reduced from 10 micrograms/ml to 3.1 +/- 4.0, 2.7 +/- 3, or 5.8 +/- 2 micrograms/ml, respectively, after incubation in eight pus specimens for 24 hours at 37 degrees C . Ampicillin levels were reduced to less than 1 microgram/ml in four pus specimens containing beta-lactamase . Imipenem levels were undetectable in two specimens and were 0.2 micrograms/ml in one specimen . Ciprofloxacin binding to pus supernate or sediment appeared to be explained by its binding to the deoxyribonucleic acid (DNA) present in pus . Activity of 5 micrograms/ml of ciprofloxacin against four E . coli or K . pneumoniae strains in pus in vitro was greater than that of twofold higher concentrations of imipenem . The bactericidal activity of ciprofloxacin and imipenem were comparable but substantially reduced against S . aureus and P . aeruginosa in pus . Ciprofloxacin alone or regimens combining ciprofloxacin with rifampin or rifampin plus imipenem reduced the number of E . coli in polymicrobic subcutaneous abscesses but had little effect on P . aeruginosa in polymicrobic abscesses . The anaerobic abscess milieu appeared to inhibit the growth of P . aeruginosa . Ciprofloxacin activity in abscess fluid did not appear to be adversely affected by acid pH, aerobic or anaerobic conditions of incubation, the abscess constituents, or the binding of ciprofloxacin to the DNA in pus . Ciprofloxacin was bound to DNA of bacterial or human origin . Binding by pus was reversible, and binding to DNA extracts of pus was blocked by pretreatment of extracts with deoxyribonuclease but not by pretreatment with ribonuclease.(ABSTRACT TRUNCATED AT 400 WORDS) Am J Med, 1989 Nov 30, 87(5A), 138S - 141S Ciprofloxacin treatment of malignant external otitis; Sade J et al.; The Ear, Nose, and Throat department of the Meir Hospital treated 91 patients with malignant external otitis during the past 16 years . The last 23 patients with malignant external otitis were treated with ciprofloxacin 750 mg twice daily, combined with local excision of the aural lesion . The records of 61 of our previous 68 patients who underwent surgery and were hospitalized and treated with an intravenous extended-spectrum penicillin and gentamicin for six to eight weeks, were analyzed . Twenty-one of 23 patients treated with ciprofloxacin were cured; therapy failed in two patients . Treatment averaged 16.8 days of hospitalization, and bacteriologic eradication was achieved after an average of 7.04 days, as compared with 49 and 15.3 days, respectively, in the group of patients with the intravenous treatment . The mean peak concentrations of ciprofloxacin in serum varied between 2.5 and 3.7 micrograms/ml, and the drug concentrations in different ear tissues were 0.2 to 13 micrograms/g . The treatment with ciprofloxacin was well tolerated with no significant side effects, whereas serious side effects were noted in 45.9 percent of the previous intravenously treated group . The concentrations of the drug in serum and ear tissues were higher than the average minimal inhibitory concentration for Pseudomonas aeruginosa . Use of ciprofloxacin treatment, combined with local excision of the aural lesion, will bring about healing of malignant external otitis in the majority of cases . Ciprofloxacin can be given on an ambulatory basis after a relatively short period of hospitalization. Am J Med, 1989 Nov 30, 87(5A), 123S - 127S Use of ciprofloxacin in cystic fibrosis patients; Bosso JA; In order to evaluate the clinical efficacy and safety of oral ciprofloxacin in the treatment of acute pulmonary exacerbations of cystic fibrosis and trace the possible development of resistance over time, three trials were conducted . In an open-label, uncontrolled trial, 25 courses of ciprofloxacin were administered to 16 patients . Efficacy and safety were assessed based on changes in short-term clinical scores, white blood cell counts, Pseudomonas aeruginosa counts in sputum, pulmonary function tests, and standard serum chemistries and urinalysis that were performed before therapy, weekly during therapy, at the end of therapy, and at a seven-day follow-up visit after therapy . In an open-label, randomized, controlled study, the efficacy and tolerance of oral ciprofloxacin were compared with those of intravenous tobramycin and azlocillin . In another study, the rate of susceptibility of P . aeruginosa isolated from cystic fibrosis patients during more than two years of clinical use was determined . In the uncontrolled trial, ciprofloxacin therapy was associated with clinical improvement in most cases with changes in short-term clinical score and forced expiratory volume in one second being statistically significant (p less than 0.05) . Twenty-five patients were entered in the controlled trial with 12 patients in each treatment group being evaluable . The groups were comparable based on admitting demographic and disease characteristics, and no differences in therapeutic response or side effects were noted between the two treatments (p greater than 0.5) . Bacterial susceptibility to ciprofloxacin has remained relatively stable over time . Based on these results as well as those from similar evaluations, ciprofloxacin appears to be efficacious in the treatment of acute pulmonary exacerbations in adults with cystic fibrosis, producing responses similar to those observed with standard intravenous antibiotic therapy. Am J Med, 1989 Nov 30, 87(5A), 28S - 31S Resistance to ciprofloxacin appearing during therapy; Chin NX et al.; The development of resistance to ciprofloxacin in nine clinical isolates of Pseudomonas aeruginosa was investigated . Isolates had increases in minimal inhibitory concentrations (MICs) from 0.25 to 16 micrograms/ml . The isolates also became resistant to ofloxacin and norfloxacin, but did not show increases in MICs to aminoglycosides, antipseudomonas penicillins, or cephalosporins . One isolate from a patient with endocarditis showed a reduction in a 43-kD outer membrane protein and simultaneous increase in the imipenem MIC . This isolate also showed impaired uptake of ciprofloxacin . Respiratory isolates from cystic fibrosis patients did not show loss of outer membrane protein . MICs were lowered by ethylene diaminetetra-acetic acid, suggesting changes in lipopolysaccharide . Resistant isolates were synergistically inhibited by combinations of ciprofloxacin plus tobramycin or ceftazidime, but MICs remained beyond the achievable serum level. Am J Med, 1989 Nov 30, 87(5A), 209S - 212S Intravenous/oral ciprofloxacin therapy of infections caused by multiresistant bacteria; Neu HC et al.; Sixty patients were treated with ciprofloxacin: 19 received only intravenous ciprofloxacin, 41 received intravenous followed by oral ciprofloxacin . The mean duration of therapy was 28 days in the intravenous only group and 10 days intravenously and 80 days orally in the intravenous/oral group . Ten (17 percent) patients received 200 mg intravenously every 12 hours and 49 (82 percent) 300 mg every 12 hours . The overall clinical response was 85 percent, with a bacteriologic response of 70 percent . The lowest bacteriologic response (38 percent) occurred in the 13 patients treated for Pseudomonas respiratory infection . Clinical response occurred in 24 of 26 patients with soft-tissue infection, and 10 of 13 patients with respiratory infection . Of three patients with endocarditis, therapy failed in two with resistance developing in Pseudomonas aeruginosa and Staphylococcus aureus . Overall, 19 percent of 26 P . aeruginosa isolates developed resistance to ciprofloxacin . Toxicity was minor, with phlebitis and nausea most commonly reported . Intravenously administered ciprofloxacin or intravenous followed by oral ciprofloxacin is a safe, effective therapy for serious infections due to multiply resistant gram-negative bacteria, including P . aeruginosa and S . aureus. S Afr Med J, 1989 Nov 18, 76(10), 543 - 5 The significance of Pseudomonas aeruginosa in chronic otitis media; Stolp DL et al.; Fifteen patients with chronic otitis media, where the organism cultured was Pseudomonas aeruginosa, had previously been treated with a variety of antibiotics, ear drops and surgical procedures . In 8 patients the discharge persisted . Between October 1986 and August 1987 these patients were treated with a combination of antibiotics (including piperacillin, ceftriaxone, amikacin and tobramycin) specific for Pseudomonas administered intravenously . In 7 patients the infection cleared completely . Follow-up ranged from 1 month to 1 year (average 4.5 months) . The regimen described could be of value in patients who do not respond to standard methods of treatment. J Biol Chem, 1989 Nov 15, 264(32), 19022 - 7 Ligand binding to cytochrome c peroxidase from Pseudomonas aeruginosa; Greenwood C et al.; The high potential heme site of Pseudomonas cytochrome c peroxidase has His and Met as ligands . On reduction, the Fe-met bond becomes photosensitive . Following photolysis, the bond reforms with a half-time of 35 ps . The low potential heme peroxidatic site of the fully reduced enzyme has been shown to bind to a range of ligands . The compounds with carbon monoxide, methyl, ethyl, n-butyl, and t-butyl isonitriles have been investigated by laser flash photolysis . All are photosensitive and show different degrees of geminate recombination of ligand in the picosecond and nanosecond time ranges . Carbon monoxide shows the least effect . The three straight-chain isonitriles show about 50% geminate recombination with half-times of the order of 10 ns . t-Butyl isonitrile shows more and faster recombination . These results imply considerable freedom of movement within the active site for the smaller ligands. J Biol Chem, 1989 Nov 15, 264(32), 18944 - 50 Difference in sodium requirement of branched chain amino acid carrier between Pseudomonas aeruginosa PAO and PML strains is due to substitution of an amino acid at position 292; Uratani Y et al.; Sodium dependence of leucine transport, a measure of the Na+-coupled leucine-isoleucine-valine II (LIV-II) transport system in Pseudomonas aeruginosa, was compared between two wild-type strains, PAO and PML . The leucine transport activity was saturated at 0.1 mM NaCl for PAO and at 5.0 mM for PML . From kinetics experiments, the apparent Km value for Na+ with respect to leucine transport was estimated to be 3 microM for PAO and 95 microM for PML . The Km value for leucine was 6 microM for PAO and 13 microM for PML . The LIV-II carrier gene (braB) of PML was isolated for comparison of its amino acid sequence with that of the PAO carrier cloned previously . The Km values for Na+ and leucine of the cloned LIV-II carriers of PAO and PML expressed in LIV-II defective mutants were similar to those in wild-type strains . Determination of the nucleotide and deduced amino acid sequences of the LIV-II carrier gene of PML showed an amino acid difference at position 292 between the PAO and PML carriers . The amino acid was threonine for PAO and alanine for PML . These results indicate that the substitution of the amino acid at position 292 of the LIV-II carrier causes a difference in the sodium requirement of the carriers of the PAO and PML strains. Infect Immun, 1989 Nov, 57(11), 3549 - 54 Photolabeling of Glu-129 of the S-1 subunit of pertussis toxin with NAD; Barbieri JT et al.; UV irradiation was shown to induce efficient transfer of radiolabel from nicotinamide-labeled NAD to a recombinant protein (C180 peptide) containing the catalytic region of the S-1 subunit of pertussis toxin . Incorporation of label from {3H-nicotinamide}NAD was efficient (0.5 to 0.6 mol/mol of protein) relative to incorporation from {32P-adenylate}NAD (0.2 mol/mol of protein) . Label from {3H-nicotinamide}NAD was specifically associated with Glu-129 . Replacement of Glu-129 with glycine or aspartic acid made the protein refractory to photolabeling with {3H-nicotinamide}NAD, whereas replacement of a nearby glutamic acid, Glu-139, with serine did not . Photolabeling of the C180 peptide with NAD is similar to that observed with diphtheria toxin and exotoxin A of Pseudomonas aeruginosa, in which the nicotinamide portion of NAD is transferred to Glu-148 and Glu-553, respectively, in the two toxins . These results implicate Glu-129 of the S-1 subunit as an active-site residue and a potentially important site for genetic modification of pertussis toxin for development of an acellular vaccine against Bordetella pertussis. J Foot Surg, 1989 Nov-Dec, 28(6), 542 - 6 Use of ceftazidime-impregnated polymethyl methacrylate beads in the treatment of Pseudomonas osteomyelitis; Tomczak RL et al.; Antibiotic-impregnated polymethyl methacrylate beads have been used in other countries to treat osteomyelitis . The drug of choice for this has historically been gentamicin . The authors have chosen ceftazidime to treat iatrogenic Pseudomonas aeruginosa osteomyelitis in rabbit femurs . After implanting the beads, the rabbits were killed at various times in the treatment period . Rabbits were killed at 15 days and later showed no signs of current osteomyelitis, including cultures and histologic examination . Blood antibiotic levels were measured at euthanasia and were minimal for all animals . It appears that ceftazidime may be an effective alternative to gentamicin, especially in treating gentamicin-resistant P . aeruginosa. Arch Intern Med, 1989 Nov, 149(11), 2579 - 83 Oral ciprofloxacin vs parenteral cefotaxime in the treatment of difficult skin and skin structure infections . A multicenter trial; Gentry LO et al.; A prospective, randomized, double-blind, multicenter study was conducted of hospitalized patients to compare the efficacy and safety of oral ciprofloxacin (dosage, 750 mg every 12 hours) with intravenous cefotaxime (dosage, 2.0 g every 8 hours) as monotherapy for difficult skin and skin structure infections requiring hospitalization . Five hundred seventy patients were assessed for an analysis of safety and 461 patients were assessed for an analysis of efficacy . The most common infections were infected ulcers and abscesses . At the end of therapy, there was a higher incidence of recurrent or persistent organisms in the cefotaxime group compared with ciprofloxacin . Adverse reactions related to either therapy were rare . By pathogens, there were no differences in activity, except the higher rate of recurrent or persistent Pseudomonas aeruginosa infection in the cefotaxime group . By diagnosis, the two drugs had comparable efficacy, except for the higher incidence of bacteriologic failure in patients with polymicrobial infected ulcers in the cefotaxime group . Larger studies are needed to evaluate emergence of resistance to ciprofloxacin . Oral ciprofloxacin therapy is as safe and effective as parenteral cefotaxime in the treatment of difficult infections of the skin and skin structure, and affords the prospect of early discharge from the hospital and significant cost savings. Infection, 1989 Nov-Dec, 17(6), 407 - 10 Adoptive transfer of resistance to Pseudomonas aeruginosa infection by splenocytes and bone marrow cells from BALB/c mice immunized by Pseudomonas aeruginosa lectin preparations; Avichezer D et al.; BALB/c mice immunized by intraperitoneal injection of purified Pseudomonas aeruginosa lectin preparations are fully protected against a lethal dose of the live bacteria . Intraperitoneal inoculation of splenocytes or bone marrow cells obtained from actively immunized mice into naive syngeneic mice was shown to significantly increase their resistance to P . aeruginosa infection . A similar transfer of splenocytes or bone marrow cells from untreated control mice did not provide protection against a lethal Pseudomonas challenge . Administration of immunocompetent cells from immunized animals to naive mice by the i.v . route was less efficient than the i.p . inoculation, probably due to different homing of the cells . Only the i.p . injection concentrated the immune cells to the site of infection. Ther Drug Monit, 1989 Nov, 11(6), 692 - 5 The use of aerosolized tobramycin in the treatment of a resistant pseudomonal pneumonitis; McCall CY et al.; Aerosolized tobramycin was given to a 68-year-old man with resistant Pseudomonas aeruginosa pneumonitis at a dose of 100 mg every 8 h via a tracheostomy, after the patient failed to respond adequately to parenteral aminoglycoside and ticarcillin therapy . Minimum inhibitory concentration and minimum bactericidal concentration for tobramycin and gentamicin were 16 micrograms/ml and greater than 16 micrograms/ml, which necessitated aerosol administration . Tracheal concentrations 15 min and 4 h after a dose were 1,560 and 930 micrograms/ml . The patient responded and eventually was discharged from the hospital . Thus, monotherapy with an aerosolized aminoglycoside may be effective in some patients with resistant Pseudomonas aeruginosa pneumonitis. J Hosp Infect, 1989 Nov, 14(4), 285 - 92 Two sources of contamination of a hydrotherapy pool by environmental organisms; Aspinall ST et al.; As a result of occasional water discolouration, the hydrotherapy pool of a large teaching hospital was monitored for free and combined chlorine, alkalinity, calcium hardness, total dissolved solids and cyanuric acid levels together with bacteriological analysis . The hose pipe supplying the pool and the dual water pumps were also examined as potential sources of bacterial contamination . The pool water yielded high counts of Pseudomonas vesicularis, Pseudomonas aeruginosa, and CDC Group IV C2, even in the presence of adequate levels of free chlorine . This was found to be due to high concentrations of cyanuric acid which resulted in a 'chlorine lock' . The source of the P . vesicularis and CDC Group IV C2 was found to be the pool hose and this problem was alleviated by flushing it with water each day before use . The source of the P . aeruginosa was the pool pumps, and was eradicated by regularly shock dosing them with 6-8 ppm of free chlorine. J Clin Microbiol, 1989 Nov, 27(11), 2589 - 93 Use of a pilin gene probe to study molecular epidemiology of Pseudomonas aeruginosa; Speert DP et al.; Strains of Pseudomonas aeruginosa from patients with cystic fibrosis (CF) are unusual . The majority have a rough lipopolysaccharide (LPS) which renders them nontypeable by conventional typing systems based on a serological reaction with the O polysaccharide of smooth LPS . We developed a new typing scheme using a pilin gene probe as a marker for hybridization with endonuclease-digested genomic DNA from P . aeruginosa . Twenty-one different restriction fragment length polymorphism (RFLP) types were found among 249 isolates . RFLP type 7 was recovered only from patients with thermal burns (9 of 14 isolates) in both Vancouver, British Columbia, and Edmonton, Alberta, Canada . None of the other RFLP types showed a clear predilection for disease state or environmental niche . Multiple morphologically different isolates from individual patients with CF were studied; each isolate in 33 of 40 sputum samples had an identical RFLP type, despite considerable LPS serotype heterogeneity . Sequential isolates from 23 patients were studied; in 10 isolates there was a clear change in both the RFLP and the LPS serotype . We conclude that patients with CF usually harbor a single P . aeruginosa RFLP type in their sputa, but that one strain can replace another as the predominant colonizing type. J Gen Microbiol, 1989 Nov, 135 ( Pt 11), 2819 - 27 Bacterial ethylene synthesis from 2-oxo-4-thiobutyric acid and from methionine; Mansouri S et al.; The ability of selected bacterial cultures to synthesize ethylene during growth in nutrient broth supplemented with methionine or 2-oxo-4-methylthiobutyric acid (KMBA) was examined . Although most cultures transformed KMBA into ethylene, only those of Escherichia coli SPAO and Chromobacterium violaceum were able to convert exogenously added methionine to ethylene . In chemically defined media, E . coli SPAO produced the highest amounts of ethylene from methionine and KMBA . This capability was affected by the nature of the carbon source and the type and amount of nitrogen source used for growth . When glutamate was used as sole source of carbon and nitrogen for growth, the activity of the ethylenogenic enzymes was reduced to 25% of that observed with cultures grown with glucose and NH4Cl . Neither methionine nor KMBA significantly affected the ethylenogenic capacity of E . coli SPAO . Menadione and paraquat, compounds that generate superoxide radicals, stimulated ethylene synthesis by harvested cells, but not by cell-free extracts of E . coli SPAO . In addition, cells of Pseudomonas aeruginosa, which produced no ethylene in culture in the presence of exogenously added KMBA, yet possessed the necessary enzymes in an active form, were able to synthesize ethylene from KMBA when incubated with menadione or paraquat. J Antimicrob Chemother, 1989 Nov, 24(5), 731 - 40 Synergistic interaction of josamycin with human neutrophils bactericidal function in vitro; Labro MT et al.; Josamycin, a 16-membered ring macrolide is concentrated up to 20-fold in phagocytic cells compared with serum . We have studied the in-vitro interaction of this drug with human neutrophils (PMN) bactericidal function by using two strains resistant to this antibiotic, Pseudomonas aeruginosa and Klebsiella pneumoniae, and a sensitive one, Staphylococcus aureus 209P . It was shown that josamycin-pretreated adherent PMN displayed an increased phagocytic activity (about 30 to 40%) for S . aureus or K . pneumoniae, mainly due to the recruitment of an additional phagocytizing subset of PMN . Furthermore, the bacterial killing was enhanced in josamycin-treated PMN in a dose-dependent manner for K . pneumoniae (60-130% increase in the range of concentration 0.1-25 mg/l) and independently of the dose for S . aureus (about 425-460% increase for josamycin 0.1-10 mg/l) . P . aeruginosa killing by whole blood was also significantly increased in the presence of 10 and 1 mg/l of josamycin . Other PMN functions were not much altered by josamycin except an enhancement of the formyl-methionyl-leucyl-phenylalanine-induced oxidative response . Chemotaxis was only increased by the presence of a high concentration (100 mg/l) of josamycin . These data suggest that the bactericidal synergy between PMN and josamycin could be related, partly at least, to a direct enhancing effect of josamycin on some PMN functions such as phagocytosis, chemotaxis and FMLP-induced chemiluminescence . On the other hand, alterations of bacteria, either inside the phagolysosome or in the extracellular medium, could lead to an enhanced susceptibility to the phagocytes' microbial mechanisms. Biochimie, 1989 Nov-Dec, 71(11-12), 1179 - 84 One-step affinity purification of amidase from mutant strains of Pseudomonas aeruginosa; Domingos A et al.; Amidases (acylamide amidohydrolase EC 3.5.1.4) from mutant strains (i.e., B6, AI3, AIU1N, OUCH 4 and L10) of Pseudomonas aeruginosa were purified in one-step by ligand affinity chromatography using Epoxy-activated Sepharose 4B-acetamide . The yields of the purified enzymes were about 90% for all mutant strains with purification factors of about 10 and were apparently homogeneous when analysed by SDS-PAGE and native PAGE . The protein bands on native PAGE coincided with the stained band of enzyme activity for all amidase preparations . Affinity columns had a maximum binding capacity of 0.5 mg amidase protein/ml of sedimented gel and could be regenerated and reused several times without any loss of binding capacity and resolution . Affinity gels containing either semicarbazide or urea were also found useful for the isolation of amidase . The differences in substrate specificity of these amidases reported previously were also observed in the elution behaviour of these enzymes from the affinity columns. Appl Environ Microbiol, 1989 Nov, 55(11), 3016 - 9 Correlation of nitrogen metabolism with biosurfactant production by Pseudomonas aeruginosa; Mulligan CN et al.; A direct relationship between increased glutamine synthetase activity and enhanced biosurfactant production was found in Pseudomonas aeruginosa grown in nitrate and Proteose Peptone media . A chloramphenicol-tolerant strain showed a twofold increase in biosurfactant production and glutamine synthetase activity . Increased ammonium and glutamine concentrations repressed both phenomena. Zh Mikrobiol Epidemiol Immunobiol, 1989 Nov, (11), 67 - 71 {The effect of staphylococci on the manifestation of opsonic cooperation in the complement system}; Konyshkina TM; The influence of the cultures of 14 Staphylococcus aureus and Staphylococcus epidermidis strains on the capacity of factor C3b of the complement for mediating the adhesive reaction of human neutrophils was studied . In experiments with 5 out of 11 S . aureus strains the essential weakening of reactions was registered, while one of the strains considerably enhanced reactions . Out of 3 S . epidermidis strains, the weakening of C3b-dependent reaction was noted in one case . The activity of the cultures did not correlate with the gelatinase properties of staphylococci and was absent in all gelatinase-positive Pseudomonas aeruginosa strains . The model worked out by the authors may be used for studying the influence of bacterial metabolites and biologically active substances on the effector properties of factor C3b of the complement. Radiobiologiia, 1989 Nov-Dec, 29(6), 765 - 8 {Possibilities of using bacterial cultures for the biological indication of relatively small doses of gamma radiation}; Torosian MV et al.; Some approaches to using bacterial cultures as test objects in biological indication of relatively low gamma-radiation doses have been considered . The survival rate, mutability and induction of prophages in the lysogenic strains have been investigate . A model for the prophage induction in Pseudomonas aeruginosa is proposed to estimate the biological effect of gamma-irradiation with doses of 0.25 to 10 Gy. Mol Microbiol, 1989 Nov, 3(11), 1493 - 9 Mapping the surface regions of Pseudomonas aeruginosa PAK pilin: the importance of the C-terminal region for adherence to human buccal epithelial cells; Lee KK et al.; The adherence of non-mucoid Pseudomonas aeruginosa strains is believed to be mediated by the pilus, which consists of a single protein subunit of 15,000 Daltons called pilin . Ten antipeptide antisera were raised to map the surface regions of pilin from P . aeruginosa strain K (PAK) . Only one of the antipeptide antisera to the eight predicted surface regions failed to react with PAK pili in direct ELISA . Five out of eight synthetic peptides representing the eight predicted surface regions reacted with anti-PAK pilus antiserum, indicating their surface exposure . Combining the antipeptide and antipilus antisera results, all eight predicted surface regions were demonstrated to be surface-exposed . The PAK 128-144-OH peptide produced the best binding antiserum to PAK pili . Only antipeptide Fab fragments directed against the disulphide bridged C-terminal region of PAK pilin blocked the adherence of pili to human buccal epithelial cells, which suggests that this region contains the receptor-binding domain of the PAK pilus. J Gen Microbiol, 1989 Nov, 135 ( Pt 11), 3023 - 34 Early cell envelope alterations by tobramycin associated with its lethal action on Pseudomonas aeruginosa; Raulston JE et al.; The immediate activities of the aminoglycoside antibiotic tobramycin were investigated in Pseudomonas aeruginosa PAO1 . The lethal action of a low concentration of tobramycin (8 micrograms ml-1) occurred rapidly (1-3 min) and was associated with leakage of certain cellular components into the supernatant . The presence of magnesium at the time of initial exposure protected cells by preventing uptake of tobramycin; however, magnesium addition following a brief exposure did not restore viability . Analyses of supernatant material revealed a rapid 2-fold increase in protein released following tobramycin treatment . A prominent 29 kDa protein, observed by SDS-PAGE in the released material was identified as the periplasmic beta-lactamase . Brief exposure to tobramycin did not result in major morphological damage or cell lysis as observed by transmission electron microscopy, and release of LPS was not a primary event . Although activity at the ribosomal level was observed by 2-3 min, leakage was detected after only 1 min . These data indicate that leakage of cellular components, particularly beta-lactamase, occurs simultaneously, if not prior to inhibition of protein synthesis by tobramycin. Curr Eye Res, 1989 Nov, 8(11), 1163 - 9 Tobramycin iontophoresis into corneas infected with drug-resistant Pseudomonas aeruginosa; Hobden JA et al.; Iontophoretic application of tobramycin was used to deliver drug to the cornea of rabbit eyes infected with a tobramycin-resistant strain of Pseudomonas aeruginosa (MIC = 31.25 micrograms/ml) . Corneas infected with P . aeruginosa 27853/pMG6 were treated 22 hours after infection with tobramycin delivered by either iontophoresis, mock iontophoresis (eye cup without current), or application of fortified topical drops . Corneal iontophoresis of tobramycin at 25 mg/ml caused more than a three log reduction in bacteria; yielding a significantly lower number of bacteria compared to untreated controls and all other treatments (P less than or equal to 0.0001) . Corneal iontophoresis of tobramycin at 10 mg/ml showed a one log reduction in the number of bacteria per cornea, yielding a significantly lower number of bacteria compared to untreated controls (P less than or equal to 0.0001), corneas treated with nine topical applications of 1.36% tobramycin drops (P less than or equal to 0.0001), and corneas treated with mock iontophoresis (P less than or equal to 0.02) . These results suggest that iontophoresis is a powerful ocular delivery system for tobramycin and may be suitable for use with other chemotherapeutic agents. Antimicrob Agents Chemother, 1989 Nov, 33(11), 1936 - 8 Phosphanilic acid inhibits dihydropteroate synthase; Eagon RG et al.; Intact cells of Pseudomonas aeruginosa were more susceptible to phosphanilic acid (PA) than cells of Escherichia coli . In cell extracts, the dihydropteroate synthases of P . aeruginosa and E . coli were about equally susceptible to inhibition by PA . These results suggest that cells of P . aeruginosa are more permeable to PA than cells of E . coli . Although a weak inhibitor, PA acted on dihydropteroate synthase in the same manner as the sulfonamides with which PA is structurally related . Inhibition of E . coli by PA in a basal salts-glucose medium was prevented by p-aminobenzoic acid (pABA) . However, pABA did not protect P . aeruginosa from PA under these conditions, possibly because pABA itself exhibited an inhibitory effect . PA also appeared to have a second mode of action . The mechanism was not elucidated. Antimicrob Agents Chemother, 1989 Nov, 33(11), 1921 - 6 Factors influencing the accumulation of ciprofloxacin in Pseudomonas aeruginosa; Celesk RA et al.; Ciprofloxacin accumulation in Pseudomonas aeruginosa was measured by a bioassay . Drug accumulation in strain PAO2 was compared with that of three spontaneous ciprofloxacin-resistant mutants selected with 0.5 micrograms of ciprofloxacin per ml . PAO4701 cfxA2 contains a mutation in the gyrA gene, PAO4742 cfxB5 may represent a permeability mutant based on pleiotropic drug resistance, and PAO4700 cfxA1 cfxB1 contains both types of mutations . In all strains, drug accumulation was similar, reaching steady state during the first minute of exposure . Drug accumulation was unsaturable over a range of 5 to 80 micrograms/ml, suggesting that ciprofloxacin accumulates by diffusion in P . aeruginosa . Although all four strains accumulated two- to sevenfold more ciprofloxacin in the presence of the inhibitor carbonyl cyanide m-chlorophenylhydrazone, the cfxB mutants accumulated two- to fourfold less drug than either PAO2 or the cfxA2 mutant . Polyacrylamide gel analysis revealed a protein common to cfxB mutants only, while all strains had similar lipopolysaccharide profiles . The results suggest that ciprofloxacin accumulation in P . aeruginosa is a complex phenomenon that may be affected by both an energy-dependent drug efflux process and outer envelope composition. Arch Dis Child, 1989 Nov, 64(11), 1599 - 603 Pseudomonas aeruginosa antibodies in blood spots from patients with cystic fibrosis; Thanasekaraan V et al.; The formation of antibodies to Pseudomonas aeruginosa may be the earliest indicator of pulmonary infection in patients with cystic fibrosis . To enable easy sampling in babies and young children an enzyme linked immunosorbent assay (ELISA) based on a blood spot sample taken on to blotting paper was developed . A sample of approximately 20 microliters of blood was required . A high correlation and level of absolute agreement was shown between paired finger prick and venepuncture blood spots, and between blood spot, serum spot, and serum samples . Healthy controls and non-infected patients with cystic fibrosis had low titres of antibody compared with patients with intermittent and chronic infection . The latter groups had significantly greater antibody titres than normal controls . This assay permits serial measurement of antibodies to P aeruginosa in patients of all ages with cystic fibrosis and may provide a means of assessing the value of such measurements in the detection and management of early infection. Circ Shock, 1989 Nov, 29(3), 245 - 56 Cardiopulmonary changes occurring with pulmonary intravascular clearance of live bacteria in sheep; Dehring DJ et al.; Sheep were infused with live bacteria to determine if the bacteria are phagocytized in the pulmonary circulation and to study the associated cardiopulmonary changes . Unanesthetized animals (n = 9) with chronic hemodynamic and pulmonary lymph catheters received a 1 hr central venous infusion of live Pseudomonas aeruginosa (5 x 10(7) Ps./minute) and were compared to a sham group (n = 7) . The pulmonary arterial levels of bacteria were five to 100 times higher than the aortic levels . Pulmonary intravascular clearance rates were 79-91% . Electron microscopy of the lungs 24 hr after the bacterial infusion showed that pulmonary intravascular macrophages and neutrophils phagocytosed the bacteria . Severe initial and mild persistent pulmonary hypertension occurred . The pulmonary lymph flow was elevated, initially from hydrostatic pressure and later from increased permeability . A hyperdynamic circulation occurred from 6 to 18 hr, with elevated cardiac index and lowered systemic vascular resistance and mean arterial pressure, mimicking cardiopulmonary changes seen in clinical sepsis . The removal of bacteria in the lungs may contribute to the injury in sheep. Arch Ophthalmol, 1989 Nov, 107(11), 1667 - 70 Effect of lid closure on contact lens-associated Pseudomonas keratitis; Aswad MI et al.; New or used extended-wear soft contact lenses, preincubated in suspensions of Pseudomonas aeruginosa, were placed on the corneas of rabbits . The lids were then sutured shut for either 1 or 2 weeks . Bacterial keratitis occurred in 9 of 9 eyes fitted with the used contaminated lenses but in none of 12 eyes fitted with new contaminated or new noncontaminated lenses . Similar experiments were carried out with other lenses specifically designed to fit the cornea of rabbits . Some of these lenses were preworn by rabbits for 1 week (used), whereas others were new . A significantly greater incidence of bacterial keratitis was found in eyes that had undergone lid closure after the placement of used contaminated lenses (4 of 5) than in closed eyes with new contaminated lenses (1 of 8) and in open eyes with used contaminated lenses (0 of 13) . These findings suggest that extended eyelid closure is a risk factor in the experimental model and may be a factor in clinical Pseudomonas keratitis associated with the wearing of extended-wear soft contact lenses. J Clin Microbiol, 1989 Nov, 27(11), 2616 - 8 Pseudomonas aeruginosa ATCC 49189, a new quality control strain for testing P . aeruginosa susceptibility to the aminoglycosides; Lally RT et al.; Quality control for in vitro antimicrobial susceptibility testing of Pseudomonas aeruginosa versus aminoglycosides is improved by P . aeruginosa ATCC 49189, which was developed in the Clinical Microbiology Laboratory at St . Paul-Ramsey Medical Center . This strain, used by us for daily testing for the past 6 years, requires MICs that approximate therapeutic concentrations, are midrange in most dilution schemes, and are stable and reproducible. J Bacteriol, 1989 Nov, 171(11), 6357 - 62 Cloning and expression of Pseudomonas aeruginosa flagellin in Escherichia coli; Kelly-Wintenberg K et al.; The flagellin gene was isolated from a Pseudomonas aeruginosa PAO1 genomic bank by conjugation into a PA103 Fla- strain . Flagellin DNA was transferred from motile recipient PA103 Fla+ cells by transformation into Escherichia coli . We show that transformed E . coli expresses flagellin protein . Export of flagellin to the E . coli cell surface was suggested by positive colony blots of unlysed cells and by isolation of flagellin protein from E . coli supernatants. J Bacteriol, 1989 Nov, 171(11), 6300 - 6 Cloning and nucleotide sequence of braC, the structural gene for the leucine-, isoleucine-, and valine-binding protein of Pseudomonas aeruginosa PAO; Hoshino T et al.; The gene for the leucine-, isoleucine-, and valine-binding protein (LIVAT-BP) in Pseudomonas aeruginosa PAO was isolated, and its nucleotide sequence was determined . The gene consisted of 1,119 nucleotides specifying a protein of 373 amino acid residues . Determination of the N-terminal amino acid sequence of the LIVAT-BP purified from P . aeruginosa shock fluid suggested that the N-terminal 26 residues of the gene product are cleaved off posttranslationally, showing the characteristic features of procaryotic signal peptides . The amino acid composition of the mature product predicted from the nucleotide sequence was in good agreement with that of the purified LIVAT-BP . The plasmid carrying the LIVAT-BP gene restored the activity of the high-affinity branched-chain amino acid transport system (the leucine, isoleucine, valine {LIV-I} transport system) in the braC310 mutant of P . aeruginosa, confirming that braC is the structural gene for LIVAT-BP . The mutant LIVAT-BP lacking a 16-amino-acid peptide in the middle was found to be functional in the LIV-I transport system . LIVAT-BP showed extensive homology (51% identical) to the LIV- and leucine-specific-binding proteins of Escherichia coli K-12, which are coded for by the livJ and livK genes, respectively, suggesting that the role of the proteins in the LIV-I transport systems is analogous in both organisms. Infect Immun, 1989 Nov, 57(11), 3491 - 7 Evidence for different pyoverdine-mediated iron uptake systems among Pseudomonas aeruginosa strains; Cornelis P et al.; Fourteen strains of Pseudomonas aeruginosa (P . aeruginosa ATCC 15692, P . aeruginosa ATCC 27853, and 12 clinical isolates) were checked for the production of pyoverdine and for pyoverdine-mediated iron uptake . Under iron restriction, two isolates produced undetectable amounts of pyoverdine, but all the other strains produced a compound with physicochemical properties identical or close to those of the pyoverdine of P . aeruginosa ATCC 15692 (strain PAO1) . The pyoverdines were purified and tested for their growth-promoting activity and for their ability to facilitate 59Fe uptake in homologous experiments involving each pyoverdine and its producing strain, as well as in heterologous systems involving all the other strains . The results of both types of experiments suggested the existence of three specificity groups . This was confirmed by analysis of the amino acid composition of the pyoverdines, which differed for each group . A partially purified polyclonal antiserum raised against a major 80-kilodalton (kDa) iron-regulated outer membrane protein (IROMP) of P . aeruginosa PAO1 recognized the 80-kDa IROMPs from P . aeruginosa PAO1 and the clinical isolates belonging to the same group, whereas the IROMPs from the strains belonging to the two other groups were not detected . A second antiserum raised against the P . aeruginosa ATCC 27853 80-kDa IROMP gave similar results by reacting specifically with the 80-kDa IROMP from the strains belonging to this group . Thus, together with the already known pyoverdine from P . aeruginosa PAO1, two new types of pyoverdines produced by strains belonging to this species were characterized. Carbohydr Res, 1989 Oct 31, 193, 75 - 90 Synthesis of a common polysaccharide antigen of Pseudomonas aeruginosa as the 6-aminohexyl glycoside; Tsvetkov YE et al.; The synthesis is described of a tritylated 1,2-O-cyanoethylidene derivative (3) of the trisaccharide alpha-D-Rha-(1----2)-alpha-D-Rha-(1----3)-D-Rha . Triphenylmethylium perchlorate-catalysed polycondensation of 3 in the presence of 6-phthalimidohexyl 2,4-di-O-benzoyl-3-O-trityl-alpha-D-rhamnopyranoside followed by deprotection afforded the 6-aminohexyl glycoside of a D-rhamnan corresponding to a common polysaccharide antigen of Pseudomonas aeruginosa. Biochem Biophys Res Commun, 1989 Oct 31, 164(2), 601 - 8 Control of alginate synthesis in Pseudomonas aeruginosa: regulation of the algR1 gene; Kimbara K et al.; The triggering of mucoidy (formation of the exopolysaccharide alginate) in Pseudomonas aeruginosa is accomplished primarily in Cystic Fibrosis lung environment through activation of the promoter of a gene algD, which encodes GDP-mannose dehydrogenase, by the product of a regulatory gene algR1 . Both algD and algR1 promoter regions have significant homology, including the presence of sequences recognized by the RNA polymerase sigma 54 . We demonstrate that the algR1 promoter is partly constitutive and its activation, similar to that of algD, is dependent on high osmolarity as well as the presence of its own gene product and is repressed by high concentrations of AlgR1 . The RpoN sigma factor also plays a critical role in the transcription of both algD and algR1 genes. J Immunol, 1989 Oct 15, 143(8), 2609 - 16 Bactericidal defect of neutrophils in a guinea pig model of thermal injury is related to elevation of intracellular cyclic-3',5'-adenosine monophosphate; Bjornson AB et al.; PG of the E series inhibit major effector functions of polymorphonuclear leukocytes (PMN) by elevating intracellular cAMP . The present study investigated the involvement of this mechanism in the bactericidal defect of PMN induced by thermal injury in a guinea pig model . Peripheral and peritoneal exudate PMN harvested from thermally injured guinea pigs at 1 or 2 days postburn had decreased bactericidal activity against Pseudomonas aeruginosa and a marked increase in cAMP content . Production of PGE1 by these cells in the absence of exogenous PMN activators was also increased . Treatment of PMN in vitro or in vivo with nonsteroidal anti-inflammatory drugs (indomethacin, ibuprofen, and piroxicam) restored bactericidal activity to normal and concomitantly reduced cAMP content and PGE1 production . A concomitant reduction in cAMP content and PGE1 production was also observed as bactericidal activity of PMN returned to normal under natural conditions during 4 to 7 days postburn . The enhancement of PMN bactericidal activity mediated by NSAID was fully counteracted by purified PGE1, theophylline, and by cAMP itself . These results suggest that the bactericidal defect of PMN induced by thermal injury is related to elevation of cAMP and that PGE1 plays a significant role in this phenomenon. Pol Tyg Lek, 1989 Oct 23-Nov 6, 44(43-45), 924 - 7 {Results of using polyvalent Pseudomonas aeruginosa vaccine in children with burns by various medical centers}; Bbukowska D et al.; Polyvalent Pseudomonas aeruginosa vaccine, prepared at the Institute of Hematology from 10 hospital strains isolated from burn wounds, was administered to 32 children with extensive and deep burns . The vaccine was well tolerated . The vaccine produced a high degree of the immunity against Pseudomonas aeruginosa infection . Agglutinin serum titre increased significantly . Vaccination either prevented or inhibited the infection of burn wounds with Pseudomonas aeruginosa in all immunized children . The symptoms of Pseudomonas aeruginosa infection usually disappeared following one or two vaccinations . Bacteriemia caused by P . aeruginosa was not observed in 31 out of 32 children . In the remaining child transient bacteriemia was noted . No septicemia caused by P . aeruginosa was seen . Due to the high efficiency of the polyvalent P . aeruginosa vaccine all burned children with burns exceeding 10% of the total body surface should by vaccinated to prevent the life-threatening infections with Pseudomonas aeruginosa. Infect Immun, 1989 Oct, 57(10), 3066 - 71 Differences in adhesion of Pseudomonas aeruginosa to mucin glycopeptides from sputa of patients with cystic fibrosis and chronic bronchitis; Ramphal R et al.; Pseudomonas aeruginosa is the most prominent colonizer of the respiratory tract of patients with cystic fibrosis, but it is not known why this occurs . P . aeruginosa adheres to mucins from normal individuals, but mucins from cystic fibrosis patients have not been studied . To compare adhesion to mucins from cystic fibrosis with other mucins, we prepared highly glycosylated mucin glycopeptides from cystic fibrosis and chronic bronchitis patients by ion-exchange and gel-filtration chromatography and measured the adhesion of P . aeruginosa 1244 to these glycopeptides . We found (i) that the most mucinlike glycopeptides from P . aeruginosa-infected cystic fibrosis sputa showed less bacterial adhesion than did the corresponding bronchitis samples, (ii) that the most adhesive activity in cystic fibrosis samples came from a fraction that contains O and N glycopeptides and may be in part a degradation product of P . aeruginosa infection, and (iii) that highly glycosylated glycopeptides of the most acidic species (sialylated and sulfated) showed no adhesion at all . A single cystic fibrosis sample not infected by P . aeruginosa showed better binding in the adhesion-positive fractions than did the infected sputa . These studies suggest that cystic fibrosis mucins may be altered after infection is established, resulting in less binding to some fragments . However, since the clinical picture shows heavy mucus colonization, other receptors, such as cellular glycolipids which have been shed into mucus, may be contributing to this colonization. J Antimicrob Chemother, 1989 Oct, 24(4), 591 - 604 Prospective comparative trial of short course (four day) and continuous tobramycin in combination with cefoperazone or mezlocillin in febrile, granulocytopenic patients; Pegram PS et al.; In a prospective, randomized trial of 195 febrile episodes in granulocytopenic patients short course aminoglycoside treatment (initial tobramycin and cefoperazone followed by tobramycin discontinuation at day four of therapy) was compared with two regimens (tobramycin plus cefoperazone and tobramycin plus mezlocillin) in which both drugs were continued for up to 26 days . All regimens were successful as empirical therapy with comparable response rates of just over seventy per cent . Fifty-three per cent of the initial episodes of fever were related to documented infections which responded less well (P = 0.007) than unexplained fever . Patients with bacteraemia, pneumonia or Gram-positive aerobic or Pseudomonas aeruginosa infections responded poorly to all regimens . The recovery from granulocytopenia was the most important determinant of successful response . Aminoglycoside discontinuation followed by cefoperazone monotherapy after day four was statistically as effective as the combination regimens . Short course tobramycin therapy eliminated the nephrotoxicity seen in the combination limbs . The use of cefoperazone was not associated with an increased incidence of hypoprothrombinemia; however, the only three bleeding episodes occurred in patients given cefoperazone but not vitamin K . Short course aminoglycoside therapy will reduce cost and nephrotoxicity when compared with prolonged combination therapy and should be further explored in this setting, with use of different agents and comparison with monotherapy. Rev Ig Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Bacteriol Virusol Parazitol Epidemiol, 1989 Oct-Dec, 34(4), 325 - 36 {A nonspecific immunostimulant effect with Cantastim on the cellular and humoral immune responses in mice evaluated by in vivo and in vitro tests}; Olinescu A et al.; The stimulatory effect was evaluated, of an ethanol extraction obtained from a pathogenic strain of Pseudomonas aeruginosa type XV . It was noted that this extract, commercially known as "Cantastim" has mitogenic effects in vitro for mouse lymphocytes, but not for those of humans or of guinea pigs . It activates the cytostatic functions of macrophages . It is thermoresistant, nonimmunogenic and it is not allergic . A low amount of proteins and carbohydrates are contained in this extract, but it contains over 80% phospholipids . It has a transient stimulating effect of the cytotoxic effects of NK cells, and it also activates the synthesis of antibodies from the IgM and the IgG classes . It probably stimulates the expression of receptors for PHA in the membrane of T lymphocytes, and retards the development of the Ehrlich ascites tumour, protecting at the same time the animals from severe infections with conditionally pathogenic germs . These data suggests that "Cantastim" is a potent immunomodulating agent that could be used successfully in the fight against certain chronic diseases with a bacterial or neoplastic etiology of humans. J Infect Dis, 1989 Oct, 160(4), 657 - 61 Pseudomonas osteochondritis complicating puncture wounds of the foot in children: a 10-year evaluation; Jacobs RF et al.; From 1978 to 1988, microbiologically proven Pseudomonas osteochondritis and septic arthritis following nail puncture wound to the foot was diagnosed in 77 children aged 18 mo-19 y (77 and 17 cases, respectively) . The syndromes were found in children with a history of wearing tennis shoes (70 cases), other shoes (5), and no shoes (2) . All cases had surgical debridement of the infected cartilage or bone and drainage of infected joints . Pseudomonas aeruginosa was isolated in 38 cases and in conjunction with Staphylococcus aureus in 18 . Anti-Pseudomonas antibiotics were initiated in all cases before surgical exploration; the average duration of treatment was 7.5 +/- 1.2 d postoperatively . Patient follow-up was 5.2 +/- 3.4 y (median, 4.8 y; range, 3 mo-10 y) . Two relapses occurred; both patients had a previously undetected septic arthritis . These data suggest that with aggressive surgical management, Pseudomonas osteochondritis and septic arthritis can be treated effectively with postoperative antibiotics for 7 d. Jpn J Exp Med, 1989 Oct, 59(5), 189 - 96 Thermal injury-induced non-specific resistance to fatal Pseudomonas aeruginosa burn-infection in mice; Pinto M et al.; Nonlethal thermal injury in mice results in rapid death by immediate injection of 10(3) viable P . aeruginosa in the skin of the burn sites . Resistance to the lethal burn combined with P . aeruginosa infection developed 24 h after initial thermal injury and reached maximal effect 7 days later; it then continued for at least 21 days . The optimal survival was achieved when the first thermal injury was made for 7 seconds at 350 degrees C . Increased resistance, but for a short period could also be obtained by injection of lipopolysaccharide (LPS) 1-4 days prior to the burn-P . aeruginosa infection . However, when the LPS was injected immediately after the burn-infection, the lethal effect was increased . The induction of late protection after thermal injury and bacterial infection was demonstrated with P . aeruginosa organisms only . Under similar schedule of thermal injury resistance was not induced by infection with Semliki forest virus . On the contrary viral infection increased the susceptibility of burned mice to a fatal outcome . Immune or natural antibodies were not elevated in the sera of post burn mice . Furthermore, delayed type hypersensitivity response, as evaluated by a footpad weight assay was inhibited and this inhibition persisted at least for 7 days post burn . The thymus weight and its lymphoid cell content in thermally injured mice decreased significantly 7 days post burn, whereas the weight of the spleen increased and it contained fewer lymphocytes per gram tissue . We suggest that endotoxin entering the systemic circulation post-burn might be one of the factors contributing to the early sensitivity and the late protection against the fatal P . aeruginosa burn-infection. Antimicrob Agents Chemother, 1989 Oct, 33(10), 1724 - 8 Fatty acid alterations and polymyxin B binding by lipopolysaccharides from Pseudomonas aeruginosa adapted to polymyxin B resistance; Conrad RS et al.; Lipopolysaccharides were extracted from freeze-dried cells of Pseudomonas aeruginosa PAO1 (polymyxin B susceptible), isolate A (polymyxin B resistant), and isolate A-reverted (polymyxin B intermediate resistance) by either the phenol-chloroform-petroleum ether or the modified phenol-water method . Isolate A and isolate A-reverted had drastic losses of 2-hydroxydodecanoic acid and significant decreases in 3-hydroxydecanoic acid . Concentrations of amide-linked 3-hydroxydecanoic acid were similar in all three strains . Minor alterations were noted in the composition of 3-deoxy-D-manno-2-octulosonic acid, heptose, phosphate, neutral sugars, and amino compounds . The concentrations of rhamnose in isolate A and of rhamnose and glucose in isolate A-reverted were significantly different from those in PAO1 . Trace amounts of mannose and other minor unidentified carbohydrates were detected in all strains . Polymyxin B included in isolate . A growth medium complexed with lipopolysaccharides and remained bound throughout purification . PAO1 lipopolysaccharides bound more polymyxin B than did isolate A lipopolysaccharides . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated minor differences in smooth- and rough-form lipopolysaccharides of the different strains . We propose that loss of hydroxy fatty acids from lipopolysaccharides perturbs outer membrane hydrophobicity and is a contributing factor to polymyxin B adaptive resistance. Wei Sheng Wu Xue Bao, 1989 Oct, 29(5), 378 - 82 {Studies on the adherent factors of Pseudomonas aeruginosa with bioluminescence}; Sun JY et al.; A bioluminescence method was established for quantifying the adhesion of P . aeruginosa to polystyrene and the adherent components were investigated . The results indicated that the slime polysaccharide (SPS) is an important adherent factor of some slime strains of P . aeruginosa . The adhered amount of washed slime strains could be increased by pre-coating of polystyrene with SPS obtained from PA3 . The activity of PA3SPS could be inhibited by anti-PA3SPS antiserum and blocked by N-acetylglucosamine. Wiad Lek, 1989 Oct 1-Nov 1, 42(19-21), 1028 - 32 {Sensitivity to antibiotics of Pseudomonas aeruginosa isolated from patients in the years 1986-1988}; Gosciniak G et al.; Pseudomonas aeruginosa strains (240 in all) isolated from various clinical materials in the years 1986-1988 showed a high sensitivity to amikacin (92.3-100%), colistin (84.6-100%) and ceftazidim (84.6-100%) . Netilmycin acted on 75.4% of the strains, and penicillins (carbenicillin, ticarcillin and azlocillin) similarly as gentamicin, tobramycin, cefotaxim, cefoperazon and ceftriaxon were active only against 51.3-66.7% of the tested strains. FEMS Microbiol Lett, 1989 Oct 1, 52(1-2), 33 - 6 Outer membrane proteins of gentamicin induced small colony variants of Pseudomonas aeruginosa; Langford PR et al.; Small colony variants (SCVs) of Pseudomonas aeruginosa NCTC 6750 (WT) were repeatedly isolated in an in vitro kinetic model after exposure to gentamicin (GM) . There were minor differences biochemically and in phage and serotyping between the wild type (WT) strain and SCVs . Changes in outer membrane protein profiles were found . SCVs were more resistant to polymixin and to a range of aminoglycosides (except kanamycin), but were more susceptible to a range of other antibiotics (hydrophilic and hydrophobic) with differing modes of action. Eur J Clin Microbiol Infect Dis, 1989 Oct, 8(10), 917 - 9 Comparative in vitro activity of cefepime (BMY 28142) against multiresistant nosocomial isolates of Pseudomonas aeruginosa; Voutsinas D et al.; The in vitro activity of a new parenteral cephalosporin cefepime (BMY 28142) was compared with that of ceftazidime, cefotaxime, piperacillin, imipenem, gentamicin, amikacin and ciprofloxacin against 173 recent multiresistant Pseudomonas aeruginosa isolates of nosocomial origin using an agar dilution technique with an inoculum of 10(4) CFU per spot . The activity of cefepime was comparable to that of ceftazidime, superior to that of cefotaxime, piperacillin, gentamicin and amikacin, but inferior to that of imipenem and ciprofloxacin . Cross-resistance of Pseudomonas aeruginosa to ceftazidime and cefepime occurred in nearly 50% of the cefepime resistant strains and 61.5% of the ceftazidime resistant strains respectively. Eur J Clin Microbiol Infect Dis, 1989 Oct, 8(10), 858 - 65 Antipseudomonal therapy in cystic fibrosis: aztreonam and amikacin versus ceftazidime and amikacin administered intravenously followed by oral ciprofloxacin; Schaad UB et al.; In order to determine the optimal antipseudomonal therapy in patients with cystic fibrosis aztreonam plus amikacin was compared to ceftazidime plus amikacin, and these two-week hospital regimens were followed by oral ciprofloxacin given for four weeks . Fifty-six cases of acute pulmonary exacerbation of the disease in 42 patients associated with isolation of Pseudomonas aeruginosa from the sputum were randomly treated with either aztreonam or ceftazidime (300mg/kg/day i.v.; maximum daily dose 12g) in combination with amikacin (36mg/kg/day i.v.; maximum daily dose 1,500mg) . Other aspects of the two-week treatment were constant . The two therapy groups were comparable in all respects . Both regimens were well tolerated and resulted in similar improvements in clinical, bacteriologic, radiologic and laboratory findings, and pulmonary function . Fifty patients could be reevaluated after subsequent outpatient therapy consisting of oral ciprofloxacin (30mg/kg/day; maximum daily dose 1,500mg) given for four weeks . During this period, the clinical and laboratory improvements persisted, and the rate of eradication of Pseudomonas aeruginosa from sputum decreased from 62% to 34% . Ciprofloxacin was well tolerated and there was no drug toxicity or serious adverse effect . In the 25 prepubertal patients there was neither subjective nor objective evidence of skeletal drug toxicity . In patients with cystic fibrosis, aztreonam or ceftazidime in combination with amikacin represents an effective and safe systemic anti-pseudomonal therapy . Subsequent oral ciprofloxacin therapy for four weeks prolongs the beneficial effects and is well tolerated. Antimicrob Agents Chemother, 1989 Oct, 33(10), 1741 - 7 Sodium hexametaphosphate sensitizes Pseudomonas aeruginosa, several other species of Pseudomonas, and Escherichia coli to hydrophobic drugs; Vaara M et al.; Many gram-negative bacteria are known to be remarkably resistant to hydrophobic noxious agents by virtue of their outer membranes (OM) . We investigated, by using four different assay methods, the ability of sodium hexametaphosphate (HMP) to disrupt this OM barrier . (i) In the growth inhibition assay, HMP was found to sensitize strains of Pseudomonas aeruginosa to all the hydrophobic probes tested (rifampin, fusidic acid, dactinomycin, sodium dodecyl sulfate, and Triton X-100) . A concentration of 0.3% HMP decreased the MICs of the probes by a factor of approximately 10, and maximally even a 30-fold sensitization was found with 1% HMP . (ii) In the bactericidal assay, 0.3% HMP decreased the MBC of the hydrophobic probe rifampin by a factor of approximately 30 . (iii) In the bacteriolytic assay, 0.1% HMP sensitized the target bacteria to lysis by sodium dodecyl sulfate and Triton X-100 . (iv) In the fluorescent-probe binding assay, HMP drastically enhanced the binding of fluorescent N-phenyl naphthylamine to the membranes of the target cells . In addition to P . aeruginosa, P . fluorescens, P . putida, P . fragi, and Escherichia coli were susceptible to the OM permeability-increasing action of HMP, while P . cepacia was resistant. J Natl Med Assoc, 1989 Oct, 81(10), 1061 - 4 Saudi Arabian-American differences in antimicrobial resistance of Escherichia, Klebsiella, and Pseudomonas; Qadri SM et al.; Increasing microbial resistance to newly developed antibiotics has been a limiting factor in the therapeutic use of these agents . To determine the extent of the problem, in vitro antimicrobial susceptibility of 7,140 clinical isolates of Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa to seven commonly used antibiotics was established at the 1,700-bed Riyadh Central Hospital in Saudi Arabia and compared with 5,513 isolates at the Oklahoma Memorial Hospital and Veterans Administration Medical Center, Oklahoma City, Oklahoma . Escherichia coli and Pseudomonas aeruginosa at Riyadh Central Hospital were generally more resistant to ampicillin, carbenicillin, gentamicin, and trimethoprim-sulfamethoxazole than those at Oklahoma Memorial Hospital and the Veterans Administration Medical Center . All 1,022 isolates of Klebsiella pneumoniae at Oklahoma Memorial Hospital were more sensitive to the antibiotics than those at Riyadh Central Hospital or the Veterans Administration Medical Center . The sensitivity pattern of Klebsiella pneumoniae at Riyadh Central Hospital and the Veterans Administration Medical Centers was similar. Infect Control Hosp Epidemiol, 1989 Oct, 10(10), 447 - 50 Pseudomonas aeruginosa infections associated with use of povidone-iodine in patients receiving continuous ambulatory peritoneal dialysis; Goetz A et al.; Fifteen episodes of infection due to Pseudomonas aeruginosa, including peritonitis and catheter site infections, occurred in nine patients receiving continuous ambulatory peritoneal dialysis over a 27-month period . Eight episodes were associated with catheter loss . Occurrence of P aeruginosa infection was significantly associated with use of povidone-iodine solution to cleanse the catheter site . There was no association with use of povidone-iodine solution to disinfect tubing connections, use of other skin care products or exposure to other environmental sources of P aeruginosa . Cultures of available povidone-iodine products were negative . Local irritation and alteration in skin flora caused by antiseptic solution or low-level contamination of povidone-iodine solution are potential mechanisms of infection. J Clin Invest, 1989 Oct, 84(4), 1302 - 13 Complement receptor expression on neutrophils at an inflammatory site, the Pseudomonas-infected lung in cystic fibrosis; Berger M et al.; Activation of human neutrophils (PMN) is accompanied by rapid upregulation of CR1, the C3b receptor, and CR3, the iC3b receptor, which also serves as the PMN's major adherence protein . This is necessary for migration and phagocytosis, but the extent of expression of these proteins on PMN at inflammatory sites has not been determined . We used monoclonal antibodies and flow cytometry to assess CR1 and CR3 expression on PMN in bronchoalveolar lavage (BAL) fluid of cystic fibrosis (CF) patients chronically infected with pseudomonas and in sterile joint fluid of arthritis patients . Resting peripheral blood PMN from these patients and normals expressed similar low levels of CR1 and CR3, and the patients' PMN increased CR1 and CR3 expression normally when stimulated in vitro . CR3 expression on CF BAL PMN was 90 +/- 12% of that on the same patient's blood cells stimulated in vitro with FMLP . In contrast, CR1 expression on BAL PMN was only 27 +/- 8% of that on stimulated blood cells . Similar results were obtained for joint PMN . This pattern could be reproduced in vitro by treating FMLP-stimulated blood cells with BAL supernatants or with pseudomonas or PMN elastase . The serine protease inhibitors, PMSF and alpha 1-antitrypsin prevented the lavage supernatant from reducing CR1 expression, while metalloprotease inhibitors had no effect . Treatment of PMN with elastase in vitro decrease their ability to kill opsonized Pseudomonas aeruginosa . These results suggest that PMN at inflammatory sites have maximally upregulated expression of their complement receptors, but that CR1 is then cleaved by proteolysis in situ . Although not related to the basic defect in CF, this may interfere with efficient phagocytosis and contribute to the CF patient's inability to eradicate chronic lung infection. J Bacteriol, 1989 Oct, 171(10), 5304 - 13 Differential regulation by iron of regA and toxA transcript accumulation in Pseudomonas aeruginosa; Frank DW et al.; Iron regulation of toxA and regA transcript accumulation was examined in Pseudomonas aeruginosa PA103 containing the regA gene on a multicopy plasmid . The patterns of transcript accumulation for toxA and regA were found to be positively correlated . Dot blot and Northern (RNA) blot analysis of total RNA isolated throughout the bacterial growth cycle indicated that multiple copies of the regA gene uncoupled iron repression of the first phase of transcript accumulation for both regA and toxA genes . However, regulation by iron of the second phase of transcript accumulation for each gene was unaffected by several regA gene copies . Total toxin production was increased in cells with multiple copies of regA grown in either low- or high-iron medium . Primer extension analysis of regA mRNA extracted from cells grown in high- and low-iron medium and examined at different points in the cell growth cycle supported the hypothesis that iron regulation of regA transcription occurs at the level of transcriptional initiation . Two start sites were shown for regA transcription at -164 and -75 base pairs from the ATG start codon . The differential regulation of regA transcript accumulation when regA is present in single or multiple copy and the mapping of independent start sites for regA mRNA support the evidence that regA transcription is directed by independently regulated promoter regions. Biochem Biophys Res Commun, 1989 Sep 29, 163(3), 1312 - 8 Identification of RegA protein from Pseudomonas aeruginosa using anti-RegA antibody; Zimniak L et al.; The product of Pseudomonas aeruginosa regA gene acts as a positive regulator of exotoxin A expression . The protein, RegA, was overproduced in E . coli transformed with an expression vector containing the regA gene . The overproduced RegA accumulated in E . coli in the form of inclusion bodies . The latter were isolated and served as a source of antigen for raising polyclonal antibodies . The antibodies reacted specifically with a P . aeruginosa protein whose molecular weight corresponded to that predicted for RegA from its known DNA sequence, and whose response to modulating factors matched that expected for RegA . The immunodetectable RegA was localized in the membrane fraction of P . aeruginosa strain PA103. J Biol Chem, 1989 Sep 15, 264(26), 15569 - 73 The structure and serologic distribution of an extracellular neutral polysaccharide from Pseudomonas aeruginosa immunotype 3; Kocharova NA et al.; Previous work has described small molecular weight neutral polysaccharides from isolates of Pseudomonas aeruginosa that appear to be associated with the lipopolysaccharide (LPS) and distributed across serologic barriers defined by antibody to the O side chain . We have isolated and characterized another of these structures obtained from culture supernatants of an immunotype 3 strain of P . aeruginosa . The isolated neutral polysaccharide has a tetrasaccharide repeat unit: (formula; see text) where Rha is rhamnose . The structure was determined by 1H and 13C nuclear magnetic resonance spectroscopy including nuclear Overhauser enhancement experiments, acid hydrolysis, methylation analysis, Smith degradation, and optical rotation determinations . Polyclonal antibodies raised to intact and alkali-treated (0.1 N NaOH, 56 degrees C, 2 h) LPS from the seven Fisher immunotype strains of P . aeruginosa bound well to the neutral polysaccharide . Antibodies affinity purified from these sera using immobilized neutral polysaccharide as well as a neutral polysaccharide-specific monoclonal antibody, E87, reacted with an antigenically similar structure found among many isolates of different LPS serotypes in a colony blot and with LPS from the seven Fisher immunotypes in an immunoblot . In an immunoblot assay, the neutral polysaccharide inhibited binding of the monoclonal antibody, E87, to material present in LPS preparations from a variety of serotypes . This structure may represent another P . aeruginosa neutral polysaccharide variant found associated with the LPS. Hosp Pharm, 1989 Oct, 24(10), 814 - 20, 823-6, 842 Focus on oral ciprofloxacin; clinical and economic considerations; Pastel D; Ciprofloxacin, a recently released oral fluorinated quinolone structurally related to nalidixic acid, joins norfloxacin as the second drug of this class to be released . Ciprofloxacin has a wide spectrum of antimicrobial activity and importantly demonstrates little cross resistance to non-quinolone drug classes (e.g . ureidopenicillins, cephalosporins, monobactams, carbapenems, aminoglycosides) . Unlike other antibacterial classes such as the beta-lactams or aminoglycosides, ciprofloxacin does not suffer from transferable plasmid-mediated (i.e . R-factor) antibiotic resistance . Against gram-positive (including penicillin-resistant and methicillin-resistant staphylococci aureus) and gram-negative aerobic bacteria including Pseudomonas aeruginosa, ciprofloxacin demonstrates excellent activity . Ciprofloxacin is inactive against Trichomonas sp., treponemes, and fungi and anaerobes are considered resistant . Ciprofloxacin is rapidly absorbed from the gastrointestinal tract (i.e . 70-80% bioavailable), demonstrates extensive extravascular distribution, and its 3.5-5 hour half-life allows twice daily dosing . The bacteriologic and clinical efficacy of oral ciprofloxacin was shown to be comparable to third generation cephalosporins or aminoglycosides for osteomyelitis, cefotaxime for skin structure infections, and to a combination of tobramycin with azlocillin for pulmonary exacerbation of cystic fibrosis . Adverse events associated with ciprofloxacin are related mostly to gastrointestinal disturbance and consist of nausea/vomiting or diarrhea . Concomitant administration of ciprofloxacin and theophylline may lead to decreased theophylline clearance and necessitates periodic measurements of theophylline levels to avoid toxic levels . Treatment with oral ciprofloxacin should offer substantial cost savings over a variety of parenteral antimicrobial regimens (e.g . aminoglycoside + beta-lactams) for difficult to treat infections such as chronic pyelonephritis, osteomyelitis, and skin structure infections . Consideration of important precautions (e.g . contraindications, drug interactions) and potential disadvantages (e.g . emergence of resistance) must also guide the rational use of oral ciprofloxacin. J Biol Chem, 1989 Sep 5, 264(25), 15151 - 6 A dipeptide insertion in domain I of exotoxin A that impairs receptor binding; Chaudry GJ et al.; Deletions within the structural exotoxin A gene of 27 or 119 amino acids in domain I of the mature polypeptide, or of 88 or 105 amino acids in domains I and II, resulted in the synthesis of exotoxin A (ETA) polypeptides that were not secreted from Pseudomonas aeruginosa hosts but were localized in the cell membrane . Insertions of a hexanucleotide sequence, either pCGAGCT or pCGAATT, at TaqI sites within the gene resulted in variant exotoxin A polypeptides which were secreted normally . pCGAGCT causes insertion of either Glu-Leu or Ser-Ser in the amino acid sequence of the toxin, while pCGAATT causes insertion of either Glu-Phe or Asn-Ser dipeptides . Although the cytotoxicity of eight variants was unimpaired, that of four others was reduced, and one variant which had a Glu-Phe insert between residues 60 and 61 (ETA-60EF61) was 500-fold less cytotoxic than wild-type exotoxin A . Purified ETA-60EF61 dissociated much faster from mouse LMTK- cells than wild-type ETA, suggesting that the insertion impaired the ability of ETA-60EF61 to interact with exotoxin A receptors . The location of the insert is within a major concavity on the surface of domain I of the exotoxin A molecule, suggesting that this concavity is important for toxin-receptor interaction. J Biol Chem, 1989 Sep 5, 264(25), 14869 - 73 Biochemical analysis of CRM 66 . A nonfunctional Pseudomonas aeruginosa exotoxin A; Galloway DR et al.; Pseudomonas aeruginosa exotoxin A (ETA) is an ADP-ribosyltransferase which inactivates protein synthesis by covalently attaching the ADP-ribose portion of NAD+ onto eucaryotic elongation factor 2 (EF-2) . A direct biochemical comparison has been made between ETA and a nonenzymatically active mutant toxin (CRM 66) using highly purified preparations of each protein . The loss of ADP-ribosyltransferase activity and subsequent cytotoxicity have been correlated with the presence of a tyrosine residue in place of a histidine at position 426 in CRM 66 . In the native conformation, CRM 66 demonstrated a limited ability (by a factor or at least 100,000) to modify EF-2 covalently and lacked in vitro and in vivo cytotoxicity, yet CRM 66 appeared to be normal with respect to NAD+ binding . Upon activation with urea and dithiothreitol, CRM 66 lost ADP-ribosyltransferase activity entirely yet CRM 66 retained the ability to bind NAD+ . Replacement of Tyr-426 with histidine in CRM 66 completely restored cytotoxicity and ADP-ribosyltransferase activity . These results support previous findings from this laboratory (Wozniak, D . J., Hsu, L.-Y., and Galloway, D . R . (1988) Proc . Natl . Acad . Sci . U . S . A . 85, 8880-8884) which suggest that the His-426 residue of ETA is not involved in NAD+ binding but appears to be associated with the interaction between ETA and EF-2. Jpn J Antibiot, 1989 Sep, 42(9), 1926 - 37 {Clinical evaluation of a combination therapy with aztreonam and clindamycin for severe infections in leukemia and related disorders}; Tsuda S et al.; A combination of aztreonam (AZT) and clindamycin (CLDM) was evaluated for severe infections associated with leukemia and related disorders . AZT is a monobactam antibiotic which has strong bactericidal effect against Gram-negative bacteria including Pseudomonas aeruginosa . CLDM which has strong antibacterial spectrum against Gram-positive and anaerobic bacteria, was chosen as a partner of AZT in order to complement the weak points of AZT . Fifty six patients were treated with the combination therapy . Among them, 51 patients were evaluable for the effectiveness . Five patients were excluded from the evaluation because 3 patients were subjected to additional therapy with other agents such as amikacin, miconazole and pulse therapy of a large dose of methylprednisolone, one had a fever episode apparently due to primary disease, and the remaining one was discontinued of the combination therapy of administration due to mild nausea after 3 days . Excellent responses were obtained in 17 (33.3%) patients and good responses in 20 (39.2%) patients, with a total rate of effectiveness of 72.5% . One patient with whom the combination therapy was stopped due to nausea, was included in the final evaluation of side effects . Side effects were observed in 2 patients of 50 and 40 years of ages (2/52, 3.8%), both of whom suffered with nausea . In the 50 years patient of acute myeloblastic leukemia, nausea occurred in a slight degree in 3 hours after the combined regimen was started . But, it disappeared during the continuation of the combination therapy . In the 40 years patient of acute myelomonocytic leukemia, mild nausea occurred after 3 day administration of the combined regimen . It disappeared soon after the cessation of the treatment . These results indicated that a combination of AZT and CLDM was an effective and safe regimen for the treatment of severe infections in patients with hematological disorders. J Antimicrob Chemother, 1989 Sep, 24(3), 447 - 53 A prospective study of ofloxacin in acute exacerbations of chronic respiratory disease associated with Pseudomonas aeruginosa; Meek JC et al.; Forty patients (32 inpatients and 8 outpatients) with acute purulent exacerbations of obstructive respiratory disease associated with Pseudomonas aeruginosa were treated orally with 800 mg ofloxacin twice daily for seven days . Sputum was cultured before, during and after treatment . The drug was extremely well tolerated, only one patient discontinuing because of alleged nausea . The infection was completely eradicated, with excellent clinical results and negative sputum cultures, in 23 patients . In six others the Ps . aeruginosa infection was cleared although reinfection with different organisms followed . Five patients showed recurrence of the infection after apparent initial eradication, but Ps . aeruginosa persisted in three others . Three patients were not evaluable because of death, transfer elsewhere, or discontinuation . There was a slight tendency for the mean MICs to rise during treatment, owing to eradication of all highly sensitive strains . Resistance did not develop . These results suggest that purulent exacerbations of chronic respiratory disease caused by Ps . aeruginosa can safely be treated orally with 800 mg ofloxacin twice daily, but that further studies in outpatients should be performed. J Burn Care Rehabil, 1989 Sep-Oct, 10(5), 421 - 4 Therapeutic efficacy of timentin and augmentin versus silvadene in burn wound infections; Heggers JP et al.; Successful closure of thermal injuries, by either skin graft or delayed wound closure, largely depends on the ability to control the number of bacteria in the wound . The purpose of this study was to investigate the efficacy of two new antimicrobial agents, ticarcillin and clavulanate (Timentin) and amoxicillin and clavulanate (Augmentin), in the infected thermal injury . The therapeutic results were compared with the model treated with the standard topical silver sulfadiazine (Silvadene) . Seventy-six Sprague-Dawley rats received a 20% full-thickness thermal injury and were then divided into six treatment groups . Three of the groups were inoculated topically with 10(8) Pseudomonas aeruginosa/ml, and three of the groups received topical inoculation of 10(8) Staphylococcus aureus/ml . The groups inoculated with P . aeruginosa received either intraperitoneal Timentin, topical Silvadene, or placebo treatment . The groups inoculated with S . aureus were treated with either enteral Augmentin, topical Silvadene, or placebo . The animals received 10 days of therapy and underwent tissue biopsies on alternate days . Statistical analysis showed that the level of bacteria in the wounds compared with the control group was significantly (p less than 0.05) decreased for both antibiotics tested as measured by quantitative wound biopsies . These studies demonstrate the efficacy of systemic Timentin and Augmentin in the infected thermal injury. Cornea, 1989 Sep, 8(3), 195 - 9 Gentamicin-resistant pseudomonal infection . Rationale for a redefinition of ophthalmic antimicrobial sensitivities; Ormerod LD et al.; Eight pseudomonal species were involved in 106 invasive infections of the eye; all were community acquired . Eighteen percent of the total and 9% of the Pseudomonas aeruginosa strains were gentamicin resistant, as defined using conventional criteria . All 10 cases of "resistant" pseudomonal (nine P . aeruginosa) keratitis responded satisfactorily to treatment with gentamicin . The resistance breakpoint (defined by safe serum levels in parenteral therapy) for most P . aeruginosa is much lower than ocular gentamicin levels achievable by optimal local application . We argue for a specific ophthalmologic definition of antibiotic resistance in infections of the cornea and external eye . MIC quantitative determinations of ocular isolates would provide more useful information to ophthalmologists than conventional qualitative disc sensitivity testing. J Infect Dis, 1989 Sep, 160(3), 483 - 9 Treatment of experimental Pseudomonas aeruginosa pneumonia with a human IgM monoclonal antibody; Hector RF et al.; A human IgM monoclonal antibody (MA-1C1) to Fisher immunotype 3 Pseudomonas aeruginosa lipopolysaccharide antigen was evaluated for in vivo activity in a guinea pig model of experimental pneumonia . Pharmacokinetics of MA-1C1 were compared in infected and noninfected animals . Intravenous bolus infusion of MA-1C1, 1 mg/kg, resulted in peak serum antibody concentrations of 3.8 +/- 0.08 and 3.7 +/- 0.05 micrograms/ml in infected and noninfected animals, respectively . Serum half-lives were 25 and 22 h in infected and noninfected groups . Treatment with a single intravenous infusion of MA-1C1 improved survival from pneumonia and was effective over a broad dose range (0.1-2.5 mg/kg) . Cumulative survivals were 18 of 47 in the MA-1C1 group and 0 of 31 in controls (P less than .001) . Treatment with MA-1C1 also resulted in fewer positive blood cultures 12 h after infection (P = .04) . Although MA-1C1 penetrated into inflamed bronchial fluids, local concentrations were only 5% of the concentrations achieved in serum . Thus, MA-1C1 seems to provide significant therapeutic activity against experimental P . aeruginosa pneumonia by preventing dissemination of infection from the lung. Pediatr Infect Dis J, 1989 Sep, 8(9 Suppl), S117 - 9; discussion S128-32 The use of aztreonam in the cystic fibrosis patient; Matsen JM et al.; Eradication of pulmonary infection by Pseudomonas aeruginosa in cystic fibrosis (CF) patients has long presented a significant challenge to the medical community . Many antimicrobial agents have proved incompletely effective against this persistent pathogen, and even the aminoglycosides, which represent the traditional therapy for such infections, have been associated with considerable toxicity and resistance . The monobactam antibacterial agent aztreonam is used both as single-agent therapy and in combination with other drugs . Several controlled, clinical trials have demonstrated the efficacy of aztreonam in the treatment of CF patients with pulmonary exacerbations caused by P . aeruginosa . The only side effect of aztreonam therapy commonly encountered in these studies was elevation of hepatic transaminase concentrations; this effect was of uncertain significance . It was concluded that aztreonam may offer clinical efficacy comparable to that provided by the combination of tobramycin plus azlocillin . Further, there does not seem to be any appreciable difference in the development of resistance to aztreonam compared with traditional therapies. Eur J Cancer Clin Oncol, 1989 Sep, 25(9), 1359 - 64 Immunotherapy of gram-negative infections in oncological patients; Glauser MP; While it is widely recognized that gram-negative bacteria (GNB) are a leading cause of morbidity and mortality among patients whose immune defenses are compromised both by their underlying malignancy as well as by its treatment, efforts to prevent or to treat such infections by immunological means have been hampered for several reasons . First, the natural anatomical barriers are often disrupted either by the tumor itself, by the chemotherapeutic agents, or, as recognized more recently, by viral (herpetic) or fungal infections, so that entrance of the gram-negative flora into the host tissue and blood stream is greatly facilitated . Efforts to diminish the bacterial load by means of oral decontamination, be it selective or not, to prevent such ingress have brought limited success . Second, active immunization is not likely to elicit a useful response in these patients, so that mainly passive immunotherapy has been considered and studied . However, since the most immunocompromised cancer patients are very often leucopenic, the opsono-phagocytic function of passively administered antibodies is not likely to help the patients to get rid of the invading gram-negative bacteria . This latter observation has been particularly well established in the case of Pseudomonas aeruginosa infections in leukemic patients (Young LS, Stevens P, Ingram J . J Clin Invest 1975, 56, 850-861). Infection, 1989 Sep-Oct, 17(5), 311 - 5 Comparative efficacy of ciprofloxacin, azlocillin, imipenem/cilastatin and tobramycin in a model of experimental septicemia due to Pseudomonas aeruginosa in neutropenic mice; Ulrich E et al.; The in vivo activity of ciprofloxacin against Pseudomonas aeruginosa was studied in a septicemia model in neutropenic mice and compared to that of other antibiotics with established activity against P . aeruginosa . When given as a single agent, ciprofloxacin proved to be as effective as imipenem/cilastatin, whereas azlocillin and tobramycin were rather ineffective . After infection with higher challenge inocula, combinations of two (synergistic) antibiotics were more effective than single agent therapy in most instances . The combination of ciprofloxacin with azlocillin was at least as effective as that of imipenem/cilastatin with tobramycin . Selection of mutants with decreased sensitivity to ciprofloxacin occurred during therapy, however, post-therapy MICs of ciprofloxacin did not exceed a level of 1 mg/l and rises of MICs did not detrimentally influence treatment outcome . Taken together with the results of earlier studies, our data encourage the use of ciprofloxacin in gram-negative septicemia in neutropenic patients. Thorax, 1989 Sep, 44(9), 739 - 42 Infective respiratory exacerbations in young adults with cystic fibrosis: role of viruses and atypical microorganisms; Ong EL et al.; Thirty six adults with cystic fibrosis were studied over one year to determine the incidence of infection with respiratory viruses and atypical organisms . Nineteen patients entered the study during an acute exacerbation of respiratory symptoms with an increase in purulent sputum production, cough, or breathlessness accompanied by a fall in FEV1 (group 1); 17 patients entered when they were stable both clinically and in terms of lung function values (group 2) . Group 1 patients had a mean of 2.6 (range 1-4) infective exacerbations during the year and group 2 patients a mean of 1.1 (0-2) exacerbations . Eleven patients developed serological evidence of viral (influenza virus A and B, cytomegalovirus, human rhinovirus 2, adenovirus) or Mycoplasma pneumoniae infection . There was no difference in seroconversion rates between group 1 (five patients) and group 2 (six patients) . There was a weak association between viral seroconversion and the isolation of Pseudomonas aeruginosa from sputum, though this was not significant. Gene, 1989 Sep 1, 81(1), 25 - 34 The activity of the Pseudomonas aeruginosa pilin promoter is enhanced by an upstream regulatory site; Pasloske BL et al.; Recently, a comparison of the nucleotide sequences upstream of the structural pilin genes (pil) from five different isolates of Pseudomonas aeruginosa identified a sequence homologous to a NifA-binding site, a positive regulatory sequence in Klebsiella pneumoniae which controls the expression of the genes which fix nitrogen {Pasloske et al., J . Bacteriol . 170 (1988) 3738-3741} . This sequence was located between -98 and -114 bp relative to the transcription start point . To establish whether this putative recognition sequence exerted any regulatory effect on pil gene expression, we assayed for the expression of pilin and pili, and assessed the strength of the pilin promoter with and without this upstream sequence . A 48-bp deletion, which removed the putative recognition sequence, decreased the levels of pilin and pilus expression . Also, the equivalent deletion reduced pilin promoter activity by 90% when it was measured using a promoter probe vector . These results provide evidence that a nucleotide sequence upstream of the pilin promoter positively regulates the transcription of the P . aeruginosa pil gene. Infect Immun, 1989 Sep, 57(9), 2764 - 70 Cross-reactivity of Pseudomonas aeruginosa antipilin monoclonal antibodies with heterogeneous strains of P . aeruginosa and Pseudomonas cepacia; Saiman L et al.; Much of the morbidity and mortality in patients with cystic fibrosis (CF) is secondary to pulmonary infections with Pseudomonas aeruginosa and, more recently, with Pseudomonas cepacia . Prevention of colonization and subsequent infection would be a useful therapeutic strategy . The pili (fimbriae) of P . aeruginosa are a potential vaccine antigen, as they have been implicated in binding to respiratory epithelium and appear to have limited antigenic diversity . Monoclonal antibodies (MAbs) raised to P . aeruginosa pilin demonstrated significant cross-reactivity, as four of five P . aeruginosa strains with known pilin sequences and 10 of 15 P . aeruginosa clinical isolates hybridized by immunoblot with at least one of the three MAbs tested . The P . cepacia strains demonstrated minimal cross-reactivity with these MAbs, as only 2 of 16 strains hybridized immunologically . The three MAbs decreased the adherence of 35S-labeled P . aeruginosa PA1244 to bovine tracheal cells by 56, 45, and 31% . One of these MAbs decreased the adherence of strains P . aeruginosa PAO1 and P . cepacia 249 to CF epithelial cells by 46 and 25%, respectively . While antibodies to Pseudomonas pili must be shown to be protective in patients with CF, these studies give support for a multivalent vaccine strategy using P . aeruginosa pilin as the immunogen. Antimicrob Agents Chemother, 1989 Sep, 33(9), 1435 - 42 Cell surface changes in Pseudomonas aeruginosa PAO4069 in response to treatment with 6-aminopenicillanic acid; Godfrey AJ et al.; Pseudomonas aeruginosa PAO4096 was induced for beta-lactamases with 6-aminopenicillanic acid . Surface changes concomitant with beta-lactamase induction were monitored . The surface hydrophobicity of the culture increased during exposure to 6-aminopenicillanic acid . The increase was associated with a change in the distribution of the O antigen in the lipopolysaccharide of treated cells . The hydrophobicity change was reversible and partially inhibited by depressed protein synthesis . The susceptibility of induced cells to rifampin was increased transiently, suggesting increased permeability of the induced cells. J Antimicrob Chemother, 1989 Sep, 24 Suppl A, 149 - 59 Comparative activity of meropenem against Pseudomonas aeruginosa strains with well-characterized resistance mechanisms; Livermore DM et al.; Four major mechanisms cause resistance to beta-lactams in Pseudomonas aeruginosa: (i) cell wall impermeability gives broad-spectrum intrinsic resistance to all beta-lactams except imipenem, (i) loss of D-group outer membrane proteins correlates with narrow spectrum imipenem resistance, (iii) plasmid mediated beta-lactamases compromise antipseudomonal penicillins, cefoperazone and cefsulodin, and (iv) chromosomal beta-lactamase hyper-production compromises most beta-lactams except carbenicillin and imipenem . Meropenem was tested in vitro against P . aeruginosa isolates, mutants and transconjugants with these mechanisms . Meropenem had impaired activity (MIC 1-2 mg/l compared to 0.25 mg/l for sensitive isolates) for organisms with broad-spectrum intrinsic resistance . MICs of meropenem also were elevated (to 1-2 mg/l) for mutants with D2-protein-deficiency-associated imipenem resistance . Most plasmids encoding TEM, OXA or PSE beta-lactamases did not increase the MIC (0.12 mg/l) of meropenem for P . aeruginosa PU21 . Decreased susceptibility (MIC 4 mg/l), however, was observed when plasmids coding the uncommon NPS-1, PSE-2 and OXA-3 enzymes were present in this strain . MICs of meropenem remained identical for chromosomal beta-lactamase-inducible P . aeruginosa strains and their enzyme-derepressed and basal mutants, indicating that the chromosomal beta-lactamase could not protect against the new carbapenem, regardless of its mode of expression. Infect Immun, 1989 Sep, 57(9), 2591 - 6 Effects of pyocyanine, a phenazine dye from Pseudomonas aeruginosa, on oxidative burst and bacterial killing in human neutrophils; Muller PK et al.; The effects of pyocyanine (phenazinium, 1-hydroxy-5-methyl-hydroxide, inner salt) on oxidative burst in human polymorphonuclear leukocytes were studied by several different approaches . In a cell- and enzyme-free system, pyocyanine oxidized NADPH . The reduced pyocyanine could be measured by its reaction with ferricytochrome c . It was shown by this assay that resting as well as phorbol myristate acetate- or zymosan-stimulated granulocytes reduced pyocyanine . The effect was independent of mitochondria, as cytoplasts were similarly active . Measurement of the hexose monophosphate shunt in intact granulocytes in the presence of pyocyanine indicated a concentration-dependent activation of the shunt without the generation of O2-, suggesting that pyocyanine oxidizes NADPH to NADP+ when it enters granulocytes . Intracellular NADPH in granulocytes was indeed lowered by almost 40% after incubation with pyocyanine . It is by this shuttling of reduction equivalents, leading to the partial depletion of NADPH, that pyocyanine affects the observed concentration-dependent partial inhibition of the phorbol myristate acetate- and zymosan-stimulated generation of O2- . A further consequence was that the intracellular killing of Staphylococcus aureus was also partially suppressed, particularly at higher loads of granulocytes with bacteria . Phagocytosis was not inhibited by pyocyanine concentrations as high as 500 microM . Pyocyanine did not affect the intracellular killing of Pseudomonas aeruginosa . The possible relevance of these findings to the course of mixed hospital infections in immunocompromised patients is discussed. G Ital Dermatol Venereol, 1989 Sep, 124(9), 415 - 7 {Splash rash dermatitis . A case}; Schiazza L et al.; The Authors report a case of a folliculitis developed in a woman who used a spa with hydrojet circulation . This dermatitis is due to Pseudomonas aeruginosa, which can colonizes the closed-cycle water systems. Antimicrob Agents Chemother, 1989 Sep, 33(9), 1553 - 6 In vitro bactericidal activities of gentamicin, cefazolin, and imipenem in peritoneal dialysis fluids; Halstead DC et al.; Continuous ambulatory peritoneal dialysis is an important modality of therapy for patients with renal disease . However, peritonitis continues to be a major risk factor and is usually treated by intraperitoneal administration of antimicrobial agents . Few data are available concerning the stability of antimicrobial agents in peritoneal dialysis solution beyond 48 h . Our investigation was designed to establish the chemical and biological stability of gentamicin alone and in combination with cefazolin in peritoneal dialysis solution at 6 and 72 h by an immunoassay and by an in vitro bactericidal test against American Type Culture Collection (Rockville, Md.) strains of Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis . In addition, uninfected peritoneal dialysis effluent was inoculated with three American Type Culture Collection strains and gentamicin or imipenem . Gentamicin alone or in combination with cefazolin was not altered chemically and was bactericidal for Staphylococcus spp . but not P . aeruginosa . In contrast, imipenem was active against both Staphylococcus spp . and P . aeruginosa . Undefined factors other than inactivation of gentamicin may be responsible for the lack of bactericidal activity and treatment failure of Pseudomonas infections. Antimicrob Agents Chemother, 1989 Sep, 33(9), 1500 - 5 Antipseudomonal activity of simulated infusions of gentamicin alone or with piperacillin assessed by serum bactericidal rate and area under the killing curve; Tisdale JE et al.; The objectives of this study were to (i) determine which of three simulated dosing regimens (gentamicin alone, simultaneous infusions of gentamicin and piperacillin, or staggered infusions of gentamicin and piperacillin) produced the fastest killing rate of Pseudomonas aeruginosa in serum, using the serum bactericidal rate (SBR) assay; and (ii) describe an alternative method of analysis of killing curves, the area under the killing curve (AUKC) . Gentamicin alone or combined with piperacillin was added to heat-inactivated human serum to approximate drug concentrations achieved after the above-mentioned types of infusion . By a microdilution technique, seven strains of P . aeruginosa were exposed to no drug (control) and gentamicin alone or with piperacillin; colony counts were determined at hourly intervals for 5 h, and log10 CFU per milliliter was plotted versus time . Linear regression was used to calculate the slope (SBR) of each timed killing curve for each drug concentration tested alone or in combination . In addition, the AUKC for each curve was calculated . To compare simulated infusion regimens further, the cumulative AUKC (the sum of AUKCs for specific time points along the serum concentration-time curve for each simulated regimen) was calculated . With the SBR assay or AUKC determination, there was a significant increase in the rate of killing of all test strains by the combination compared with gentamicin alone only at gentamicin concentrations which exceeded the MIC (8, 5, and 2.5 micrograms/ml) . Mean cumulative AUKC of the simultaneous-infusion regimen was significantly less (indicating faster killing) than either the staggered-infusion regimen or the gentamicin infusion alone . Both the SBR and AUKC have the potential for integration of in vitro microbiologic effects and in vivo pharmacokinetics of antimicrobial agents. J Antimicrob Chemother, 1989 Sep, 24 Suppl A, 233 - 8 The antibacterial activity of meropenem in combination with gentamicin or vancomycin; Wise R et al.; The kinetics of bacterial killing by meropenem alone and in combination with gentamicin (for Pseudomonas aeruginosa) and vancomycin (for Staphylococcus aureus) were studied for two strains of each species . Against the two strains of P . aeruginosa, meropenem at concentrations up to 4 x MIC was rapidly bactericidal--but regrowth occurred by 24 h . The addition of half the MIC of gentamicin to the MIC of meropenem led to a more rapid decline of the colony count and to the prevention of regrowth of the strain which was gentamicin-susceptible . Similar results were obtained for a methicillin-susceptible strain of S . aureus when vancomycin was added at a concentration of half the MIC . A methicillin-resistant strain also was killed by a combination of vancomycin at half the MIC plus meropenem at the MIC . The study showed that the killing action of meropenem against P . aeruginosa and staphylococci is enhanced by the addition of gentamicin or vancomycin respectively. J Antimicrob Chemother, 1989 Sep, 24 Suppl A, 169 - 74 Bactericidal activity of meropenem against Pseudomonas aeruginosa; Yourassowsky E et al.; Ten strains of Pseudomonas aeruginosa that were susceptible to imipenem (MICs 2 mg/l) were exposed to a new parenteral carbapenem, meropenem (MIC 0.25 mg/l) . Kinetic turbidometry showed that, as with other beta-lactam antibiotics, there was a prelytic increase in the culture OD following exposure to meropenem . The maximal value of the prelytic increase in the OD was higher for meropenem than for imipenem at concentrations 0.5, 1, 2, 4 and 8 x MIC . This corresponded to the formation of short filaments during exposure to low concentrations of meropenem . These filaments remained viable for 1-2 h, according to the drug concentration . For this reason, the killing began later with meropenem than with imipenem . After this delay, the killing rate for meropenem was the same as with imipenem, but occurred with lower concentrations of meropenem. J Antimicrob Chemother, 1989 Sep, 24 Suppl A, 161 - 7 Emergence of resistance to carbapenem antibiotics in Pseudomonas aeruginosa; Margaret BS et al.; Meropenem is a new carbapenem antibiotic with activity similar to that of imipenem . Development of resistance to imipenem by Pseudomonas aeruginosa has occurred in therapy and is associated with loss of a 45,000-48,000 molecular weight outer membrane protein (OMP) and decreased imipenem penetration . Seven pairs of isolates of P . aeruginosa obtained before treatment and after emergence of resistance were examined for changes in MIC of both imipenem and meropenem, and for changes in OMP profile . Resistant mutants were obtained also from the pre-therapy strains on Mueller-Hinton agar containing increasing amounts of either drug, and maintained under antibiotic pressure . OMP preparations of all pre- and post-therapy strains and laboratory mutants were separated by polyacrylamide gel electrophoresis . Loss of a 45,000-48,000 molecular weight protein occurred in variants selected under carbapenem pressure and in all post-therapy isolates . Meropenem remained four-fold more active than imipenem against the resistant strains and mutants. Can J Microbiol, 1989 Sep, 35(9), 890 - 4 Flagellar antibody stimulated opsonophagocytosis of Pseudomonas aeruginosa associated with response to either a- or b-type flagellar antigen; Anderson TR et al.; Pseudomonas aeruginosa exhibit one of two flagella types: a homogeneous b type, with molecular weight of 53,000, or a heterogeneous a type (subtypes a0, a1, a2, a3, and a4), with molecular weights ranging from 45,000 to 52,000 . Pseudomonas aeruginosa flagellar antiserum was shown to promote uptake of radiolabeled bacteria by mouse polymorphonuclear leukocytes . Bacteria were detected directly associated with washed leukocytes and visualized, by electron microscopy, internalized in polymorphonuclear leukocytes . Phagocytosis was specific for the flagella type (a or b) in that homologous flagella serum enhanced uptake three to four times greater than heterologous serum or normal rabbit serum . An a-type antiserum was shown to enhance phagocytosis of four different a-type strains with varying subantigen types, indicating the presence of a common cross-reactive a0 antigen in this flagella type . Phagocytic killing of internalized bacteria was not seen with the addition of only flagellar antiserum. Zentralbl Bakteriol, 1989 Sep, 271(3), 364 - 71 Lectin typing of Pseudomonas aeruginosa strains of different serogroups, Habs and Fisher types; Chatterjee BP et al.; Sixteen Habs and three Fisher types of Pseudomonas aeruginosa were typed with lectins of know specificity resulting from their interaction with bacterial cell surface carbohydrates as evidenced by agglutination-inhibition assay with simple carbohydrates . Lipopolysaccharides of few strains of Pseudomonas are precipitated with different lectins and the results are corroborated by those of agglutination suggesting that Pseudomonas aeruginosa can be characterized intraspecifically by lectins. Photochem Photobiol, 1989 Sep, 50(3), 413 - 8 Photo-induced electron ejection from the reduced copper of Pseudomonas aeruginosa azurin; Corin AF et al.; The reduced form of Pseudomonas aeruginosa azurin exhibits an enhanced absorbance in the UV compared to that of the oxidized protein . This enhancement has also been observed for azurins from other bacterial species and for another type I copper protein, plastocyanin . Pulsed laser excitation of the reduced azurin in the region of enhanced absorbance at 308 nm results in single ph