J Clin Invest, 1991 Jun, 87(6), 2018 - 28 Differential adhesion of Pseudomonas aeruginosa to human respiratory epithelial cells in primary culture; Plotkowski MC et al.; Human nasal polyps in outgrowth culture were used to study the Pseudomonas aeruginosa adhesion to respiratory cells . By scanning electron microscopy, P . aeruginosa were seen associated with ciliated cells, but by transmission electron microscopy, bacteria were never seen at the interciliary spaces or attached along cilia, but were identified trapped at the extremities of cilia, usually as bacterial aggregates . A fibronectin-containing fibrillar material was seen associated with aggregated bacteria . By time-lapse video microscopy, bacteria were seen to aggregate in the culture medium following their addition to the culture wells . Progressively, these aggregates were trapped by cilia or attached to migrating cells of a lower cell layer that protruded beneath the upper layer cells, at the outgrowth periphery . P . aeruginosa adhesion to these lower cell layer migrating cells was significantly higher than to ciliated or nonciliated cells of the upper cell layer . Migrating cells were intensely labeled by the complexes Con A and arachis hypogea agglutinin (PNA)-FITC, in contrast to the other cells . The percentage of PNA-labeled cells with attached bacteria was significantly higher than that without bacteria . These results suggest that changes of cell surface glycoconjugates related with cell migration may favor P . aeruginosa adhesion to respiratory cells. Infect Immun, 1991 Jun, 59(6), 2152 - 7 Pseudomonas aeruginosa variants isolated from patients with cystic fibrosis are killed by a bactericidal protein from human polymorphonuclear leukocytes; Siefferman CM et al.; The susceptibility of paired mucoid and nonmucoid variants of Pseudomonas aeruginosa isolated from 13 patients with cystic fibrosis (CF) to killing by a 55,000-Da bactericidal protein (BP55) from human polymorphonuclear leukocytes was studied . Mucoid and nonmucoid variants were equally sensitive to killing by BP55 at both pH 5.6 and pH 7.2 . Eleven of the isolates were resistant to the bactericidal activity of 10% normal human serum but were as sensitive as the serum-sensitive isolates to BP55 . Similarly, the 15 isolates with lipopolysaccharides (LPS) containing O-polysaccharide side chains (smooth LPS) were as sensitive to BP55 as those isolates with rough LPS.P . aeruginosa isolates from patients in poor clinical condition were more likely to have LPS of the smooth type and to be resistant to killing by 10% human serum than the isolates from patients in good clinical condition . We have concluded that the susceptibility of the P . aeruginosa isolates from patients with CF to killing by BP55 does not correlate with mucoid or nonmucoid variations, with the presence or absence of smooth LPS, or with the sensitivity or resistance to killing by normal human serum. Infect Immun, 1991 Jun, 59(6), 1984 - 90 Serum antibody response to Pseudomonas aeruginosa antigens during corneal infection; Preston MJ et al.; Previous studies in our laboratory have indicated that naturally resistant, inbred DBA/2J mice mount a greater serum antibody response to Pseudomonas aeruginosa 19660 than susceptible C57BL/6J mice . However, the specificity of the antibody produced was not known . The present study examines the specificity and kinetics of the humoral response of these mouse strains to potential virulence factors produced by the organism during both a primary and a secondary corneal infection administered 4 weeks after the primary infection . Serum antibody levels specific for lipopolysaccharide (LPS), exotoxin A, phospholipase C (PLC), alkaline protease, elastase, and flagella were measured by enzyme-linked immunosorbent assay . Little or no antibody to either alkaline protease or elastase was detected during either primary or secondary infection . Immunoglobulin G (IgG) antibodies specific to exotoxin A, PLC, and flagella were detected 2 weeks after primary infection, and a rapid response to these antigens was measured 1 week after secondary infection . During primary infection, detectable LPS-specific antibody was only IgM, while IgG appeared only after secondary infection . The kinetics of the humoral response in susceptible C57BL/6J mice were similar to those in resistant DBA/2J mice, although the magnitude of the response varied according to the antigen tested . These results indicate that LPS, exotoxin A, PLC, and flagella are present or produced in amounts that are immunogenic during corneal infection by P . aeruginosa 19660 in the mouse strains tested. DICP, 1991 Jun, 25(6), 594 - 7 Aztreonam-induced myelosuppression during treatment of Pseudomonas aeruginosa pneumonia; Dallal MM et al.; Aztreonam is a synthetic, monobactam antibiotic structurally related to the beta-lactam class of drugs . It has inhibitory activity against many aerobic gram-negative bacteria, although it does not inhibit gram-positive or anaerobic bacteria . Administration of aztreonam occasionally is associated with minimal and transient adverse effects . This case report describes a patient we believe experienced bone marrow suppression approximately ten days after aztreonam was given for treatment of pneumonia caused by Pseudomonas aeruginosa . This untoward effect primarily was manifested as neutropenia, although normochromic, normocytic anemia and thrombocytopenia were noted as well . One week after aztreonam was discontinued, the patient's bone marrow suppression resolved spontaneously . Although the mechanism responsible for myelosuppression is unclear, aztreonam may be implicated as the offending agent based on the temporal relationship between the development of neutropenia and its administration, and the resolution of neutropenia upon its discontinuation. Mol Microbiol, 1991 Jun, 5(6), 1469 - 81 Identification and molecular characterization of a transcriptional regulator from Pseudomonas aeruginosa PAO1 exhibiting structural and functional similarity to the FNR protein of Escherichia coli; Sawers RG; A gene library of chromosomal DNA from Pseudomonas aeruginosa contained a DNA fragment which was able to restore anaerobic growth to an Escherichia coli fnr deletion mutant on glycerol/nitrate medium . The cloned gene (termed anr) was sequenced and shown to encode a protein of 244 amino acids with a calculated molecular weight of 27,129 . The deduced amino acid sequence of the anr gene product showed considerable similarity to the FNR protein from E . coli . Expression of the anr gene in a T7 promoter/polymerase system identified ANR as a 31 kDa protein . Transcriptional analysis of the anr gene showed that it is monocistronic but apparently lacks the equivalent sites for negative autoregulation which have been shown to be present in the promoter region of the E . coli fnr gene . The ANR protein was shown to activate transcription of the pfl gene in E . coli in response to anaerobiosis, as well as being able to restore the activity of three anaerobically inducible enzymes . A P . aeruginosa mutant incapable of growing anaerobically with nitrate or on arginine was fully complemented by the anr gene, indicating that it probably has a function in controlling anaerobic gene expression in Pseudomonas . Further corroboration for this assumption was provided by S1 nuclease analysis of transcription of the multiple promoters of the E . coli pfl operon in P . aeruginosa . Transcription was induced by oxygen limitation and was completely ANR-dependent in both aerobic and anaerobic cells . Removal of the upstream regulatory sequence of the pfl operon, which includes the sequences required for FNR-dependent regulation in E . coli, removed ANR-dependent transcriptional control of the remaining pfl promoters, irrespective of the cellular oxygen status . These results imply that the mechanisms by which ANR and FNR regulate transcription are fundamentally similar. Ann Ophthalmol, 1991 Jun, 23(6), 234 - 7 Infectious keratitis in Baltimore; Wahl JC et al.; One hundred thirty cases of ulcerative keratitis occurring from 1985 to 1989 at Sinai Hospital of Baltimore were reviewed . Positive corneal cultures were obtained from 40% of cases . The most common isolates were Staphylococcus epidermidis (28%), S . aureus (16%), and Pseudomonas aeruginosa . P . aeruginosa was the most common in contact lens wearers . Antibiotic pretreatment did not affect the rate of positive cultures significantly . Bilateral conjunctival and eyelid margin cultures also were examined . An association was found between corneal and ipsilateral conjunctival isolates (P = .05) . Gram stains were consistently negative . Most patients were treated successfully as outpatients . The value of simultaneous conjunctival and eyelid margin cultures and Gram stains was questioned. J Hosp Infect, 1991 Jun, 18 Suppl A, 535 - 42 Hydrotherapy pools of the future--the avoidance of health problems; Penny PT; The need to minimize the threat of infection to patients and the high incidence of health problems in hydrotherapy pool workers, have led to recommendations especially tailored to the design and operation of the water and air treatment plant of hydrotherapy pools . Hitherto unpublished surveys are detailed which confirm that pathogenic species of Pseudomonas aeruginosa in pools (in which ears may be wetted) cause a high incidence of otitis externa, but rarely cause body rashes (pseudomonas folliculitis) unless there has also been prolonged skin wetting . In brominated pools contact dermatitis is common and can be distinguished clinically from pseudomonas folliculitis by the onset of a pruritic rash less than 12 hours after exposure to the pool and reactivation of the rash on re-exposure to the brominated pool. J Hosp Infect, 1991 Jun, 18 Suppl A, 515 - 23 The role of barrier precautions in infection control; Goldmann DA; Barrier precautions are a fundamental component of any infection control strategy and a critical aspect of all isolation systems . Because many infections are transmitted from patient-to-patient via the hands of personnel, gloves and gowns are widely recommended to provide an extra measure of protection against cross-infection . It is not clear whether gloves are superior to handwashing (if performed obsessionally) in this respect, and there is little evidence that gowns confer additional benefit . These concerns notwithstanding, barrier precautions can substantially reduce the risk of some infections, such as respiratory syncytial virus disease . On the other hand, the modes of transmission of many infections are complex (e.g . with rotavirus) or controversial (e.g . with rhinovirus), and, even though hands are involved in transmission, barrier precautions alone may not suffice to prevent spread . Moreover, neither gloves nor gowns can prevent nosocomial infections caused by endogenous microbial flora; perhaps this explains the limited efficacy of barrier precautions in reducing the endemic rate of infection due to bacteria such as Pseudomonas aeruginosa in intensive care units . Barrier precautions may also fail if colonized patients are not identified promptly . One potential solution to this problem is 'body substance isolation' (BSI), in which all patients are considered to be potential carriers of nosocomial pathogens whether or not they have been cultured or have developed a clinical infection . In BSI barrier techniques are used when any potentially contaminated patient material is handled . BSI also provides barrier protection from bloodborne pathogens for personnel.(ABSTRACT TRUNCATED AT 250 WORDS) J Hosp Infect, 1991 Jun, 18 Suppl A, 341 - 54 Infectious consequences of continuous ambulatory peritoneal dialysis; Ludlam HA; The peritoneal cavity of patients undergoing CAPD is critically immunocompromised and infectious peritonitis is the most important complication of the technique . Nevertheless, recent research into the epidemiology and pathogenesis of infections caused by the most important microorganisms has enabled significant reductions in peritonitis rates to be made . Peritonitis caused by Staphylococcus aureus and Pseudomonas aeruginosa can be prevented by eliminating their principal source, an infected Tenckhoff catheter wound . The source of infection for coagulase-negative staphylococci and other pseudomonads cannot be eliminated, but peritonitis caused by these organisms may be prevented by interrupting their routes of entry into the peritoneal cavity . The identification of host factors predictive of enhanced susceptibility to infectious peritonitis offers the further possibility of prevention by immunological approaches . Although the main difficulties surrounding the diagnosis of infective peritonitis have been clarified, approximately 20% of episodes remain culture-negative, with multifactorial aetiology . Initial (empirical) combination antibiotic therapy can be both appropriate and effective in approximately 85% of cases . Intraperitoneal monotherapy with fluoroquinolones has been equally successful, and these agents may prove effective by the oral route, offering considerable advantages in cost and convenience . Approximately 5% of episodes of bacterial peritonitis are unresponsive to antibiotic therapy . These cases may be conveniently managed by the technique of Tenckhoff catheter removal and replacement at a single operation. Epidemiol Infect, 1991 Jun, 106(3), 531 - 9 Complexity of Pseudomonas aeruginosa infection in cystic fibrosis: combined results from esterase electrophoresis and rDNA restriction fragment length polymorphism analysis; Denamur E et al.; Esterase electrophoretic typing and restriction fragment length polymorphism of ribosomal DNA regions (ribotyping) were used to differentiate 102 Pseudomonas aeruginosa clinical isolates obtained from chronic lung infection in 23 patients with cystic fibrosis (CF) and two reference strains (including the type strain ATCC 10145) . Twenty-five zymotypes were obtained with the former method and 16 ribotypes with the latter . Combination of the two typing systems led to the finding of 30 different types . Our data highlights the physiopathological complexity of P . aeruginosa infection in CF as, in six individual cases, several types were found among isolates from a given patient . On the other hand, two unique types were found in two and three patients respectively, raising the possibility of cross-infections. Biofactors, 1991 Jun, 3(2), 103 - 7 Identification of a molybdopterin-containing molybdenum cofactor in xanthine dehydrogenase from Pseudomonas aeruginosa; Johnson JL et al.; Xanthine dehydrogenase has been purified from Pseudomonas aeruginosa cultured on a rich medium and induced with hypoxanthine . The enzyme was shown to contain FAD, iron sulfur centers and a molybdenum cofactor as prosthetic groups . Analysis of the molybdenum cofactor in this enzyme has revealed that the cofactor contains molybdopterin (MPT) rather than molybdopterin guanine dinucleotide or molybdopterin cytosine dinucleotide which have previously been identified in a number of molybdoenzymes of bacterial origin . The pterin cofactor in P.aeruginosa xanthine dehydrogenase was alkylated and the resulting product was identified as dicarboxamidomethyl molybdopterin . In addition, the pterin released from the enzyme by denaturation with guanidine-HCl was found to chromatograph on Sephadex G-15 with an apparent molecular weight of 350 . These results document the first example of a bacterial enzyme with a molybdenum cofactor comprising molybdopterin and the metal only. AIDS, 1991 Jun, 5(6), 761 - 3 Pseudomonas septicaemia associated with HIV; Nelson MR et al.; Between July 1985 and January 1990, pseudomonas scepticaemia occurred in 19 out of 584 patients with AIDS attending the Westminster and St Stephen's AIDS Unit, London, UK . Ten of these 19 were being treated for active cytomegalovirus infection . Fourteen of the 19 patients had a central venous catheter in situ, which was the source of infection in 11 . Seven patients died . Mortality was significantly greater in those patients infected with Pseudomonas aeruginosa, in those patients whose source of infection was not the central venous line, and in those patients whose central line was not removed. J Trauma, 1991 Jun, 31(6), 733 - 40; discussion 740-1 Delayed cyclo-oxygenase blockade reduces the neutrophil respiratory burst and plasma tumor necrosis factor levels in sepsis-induced acute lung injury; Carey PD et al.; Ibuprofen pretreatment attenuates the enhanced neutrophil (PMN) respiratory burst and reduces increased plasma tumor necrosis factor (TNF) activity in porcine sepsis-induced acute lung injury (ALI) . These septic responses have been linked to increased alveolar-capillary membrane (ACM) permeability . This study was designed to establish whether delayed ibuprofen treatment would have the same effect and to examine the relationship between PMN oxidant generation and TNF . Three groups of anesthetized, ventilated pigs (15-25 kg) were used . Group Ps received Pseudomonas aeruginosa (5 x 10(8) CFU/mL at 0.3 mL/20 kg/min) for one hour IV; The control group (Con) received 0.9% NaCl . Group D-Ibu received ibuprofen 12.5 mg/kg as a delayed bolus at 30 minutes and again at 120 minutes after Ps . Protein (BAL-P, microgram/mL) in harvested bronchoalveolar lavage fluid and extravascular lung water (EVLW, mL/kg) were used to estimate the integrity of the ACM . Superoxide anion (O2-) generation (ferricytochrome c reduction) from circulating PMNs and plasma TNF activity (L929 fibroblast bioassay) were measured . The EVLW increased significantly (p less than 0.05), as did BAL-P (p less than 0.01), in the P . aeruginosa-treated animals at 300 minutes . These increases were abolished in Group D-Ibu: EVLW, 6.6 +/- 1.0 baseline vs . 14.6 +/- 2.6 Ps 300 vs . 6.8 +/- 0.9 D-Ibu 300; BAL-P, 175 +/- 28 baseline vs . 984 +/- 186 Ps 300 vs . 284 +/- 42.8 D-Ibu 300 . Both enhanced PMN oxidant activity and increased plasma TNF activity were significantly attenuated by delayed ibuprofen treatment . These data support the efficacy of the nonsteroidal anti-inflammatory drug, ibuprofen, when used after the onset of a septic stimulus. Infect Immun, 1991 Jun, 59(6), 1893 - 8 Suppression by human recombinant gamma interferon of in vitro macrophage nonopsonic and opsonic phagocytosis and killing; Speert DP et al.; Although gamma interferon (IFN-gamma) exerts profound effects on the state of activation of macrophages, its influence on receptor-mediated phagocytosis and killing of extracellular bacteria is poorly understood . Human monocytes cultured in the presence of human recombinant IFN-gamma exhibited an enhanced capacity to produce superoxide anion . Although these cells bound greater numbers of particles via Fc receptors, their capacity to phagocytose by these receptors or to bind or ingest particles via receptors for C3bi, mannose, or unopsonized Pseudomonas aeruginosa was substantially depressed in a dose-dependent fashion (0.1 to 1,000 U of IFN-gamma per ml) . Macrophage phagocytosis of P . aeruginosa and Staphylococcus aureus opsonized with whole serum or with serum deficient in immunoglobulin or complement was also depressed . Macrophages cultured in the presence of IFN-gamma had a diminished capacity to kill both unopsonized and opsonized P . aeruginosa . We conclude that IFN-gamma inhibits macrophage nonopsonic and opsonic receptor-mediated phagocytosis and killing but enhances oxidative radical generation; its production may exacerbate host tissue damage during chronic infection with extracellular pathogens. J Biol Chem, 1991 May 25, 266(15), 9754 - 63 Characterization and regulation of the Pseudomonas aeruginosa algC gene encoding phosphomannomutase; Zielinski NA et al.; The nucleotide sequence of the Pseudomonas aeruginosa algC gene encoding phosphomannomutase (PMM; EC 5.4.2.8) was determined . The codon usage in algC in the wobble base position was 90.4% G+C, typical of Pseudomonas genes . The predicted amino acid sequence of phosphomannomutase (PMM) showed homology over a stretch of 112 amino acids in the carboxyl terminus with rabbit muscle phosphoglucomutase (PGM), an enzyme that catalyzes a reaction analogous to that catalyzed by PMM . In addition, a specific amino acid sequence within PMM showed homology with the catalytic site of PGM . DNA sequence analysis of a defective algC gene (algC') cloned from a mutant of P . aeruginosa that lacked PMM activity revealed one point mutation (a C to T transition) in the carboxyl terminus of PMM which resulted in an amino acid change from arginine 420 to cysteine 420 . The mutation identified in the algC' gene was not within the regions of homology with PGM . The algC promoter showed significant homology with the promoters of two other P . aeruginosa genes involved in alginate synthesis, algD and algR1 . Both the algD and algR1 promoters are activated by the product of the algR1 gene in P . aeruginosa . The upstream region of the algC gene contained a sequence identical to the algD upstream sequence that is known to be the binding site for the AlgR1 protein . Expression of algC was reduced 5.7-fold in an algR1 mutant of P . aeruginosa compared to its isogenic parent strain (lacking the algR1 mutation), suggesting that the algR1 gene product activates the transcription of the algC gene. Arch Biochem Biophys, 1991 May 15, 287(1), 160 - 6 Progesterone binding in a clinical isolate of Pseudomonas aeruginosa; Mosier S et al.; We have undertaken the characterization of progestin binding component(s) in the cytosol prepared from Pseudomonas aeruginosa isolated from an immunocompromised patient . Incubation of P . aeruginosa cytosol aliquots at 0 degrees C with 20 nM {3H}R5020 (a synthetic progestin) revealed the presence of saturable binding . The {3H}R5020 binding reached an equilibrium after 1 h at 0 degrees C and showed saturation at 30-50 nM with a Kd value of 7.7 nM . At 0 degrees C, beta-mercaptoethanol increased the {3H}R5020 binding by 20% but sodium molybdate had no effect . The {3H}R5020-macromolecular complex was stable for up to 4 h at 37 degrees C . Steroid binding specificity analysis revealed that {3H}R5020 binding could be eliminated in the presence of 2 microM progesterone, estradiol, or dihydrotestosterone but that the synthetic glucocorticoid, triamcinolone acetonide, did not compete . Postlabeling of the cytosol fractions obtained after 10-30% glycerol gradient analysis demonstrated association of the radioactivity with a molecule that sedimented as a 6-8 S protease-sensitive moiety which was unaltered in the presence of RNase or DNase . When cells were grown in the presence of 100 nM progesterone, a 50% inhibition in the number of resulting colonies was observed . In addition to its evolutionary significance, the presence of this steroid binding molecule suggests a potential in the endocrine manipulation in the treatment of infections caused by P . aeruginosa. FEMS Microbiol Lett, 1991 May 15, 64(2-3), 337 - 40 Factors that influence the permeability assay of the outer membrane of Pseudomonas aeruginosa; Lei Y et al.; Brief exposure of Pseudomonas aeruginosa to a temperature of 10 degrees C or lower caused a significant leakage of the periplasmic beta-lactamase into the medium . The extent of leakage increased as the incubation temperature was lowered to 4 degrees C and reached a maximum at 0 degrees C . Cells grown in the presence of beta-lactamase inducers were unsuitable for the permeability assay . It was found that the diffusion rates of beta-lactams through the outer membrane of P . aeruginosa were much lower than those previously reported, as assayed under refined conditions . The diffusion rates of beta-lactams in one of the mutants tested were an order of magnitude lower than those of the other strains, despite the fact that the outer membrane protein profile of the strain appeared to be indistinguishable from those of the others . These results suggest that beta-lactam antibiotics diffuse through the outer membrane of P . aeruginosa, at least partly, through a non-porin pathway. J Lab Clin Med, 1991 May, 117(5), 410 - 22 Airway adherence of Pseudomonas aeruginosa: mucoexopolysaccharide binding to human and bovine airway proteins; Hata JS et al.; The ability of Pseudomonas aeruginosa (PA) to adhere to cells of the upper and lower airways is considered the initial step in its colonization and subsequent infection . Many potential adhesins exist, but few eukaryote receptors have been reported . This study evaluates the adherence of PA and the well-recognized adhesin mucoexopolysaccharide, of the mucoid variant, to tracheal epithelial cells and airway secretions . A number of tools were used to systematically examine this process, each in increasing detail . Scanning electron microscopy of infected tracheal cell monolayers showed that although nonmucoid PA adhered most often as individual organisms, mucoid strains frequently were adherent in the form of clusters of microcolonies . Partially purified mucoexopolysaccharide was itself adherent to the tracheal cell monolayers . More quantitative studies of carbon 14-labeled PA adherence to tracheal cell monolayers demonstrated that (1) bacterial adherence was temperature dependent; (2) mucoid PA was more adherent than nonmucoid PA (p less than 0.005); (3) proteinase treatment of the intact monolayers increased adherence of mucoid PA; and (4) adherence of nonmucoid PA was partially inhibited by pretreatment with N-acetylglucosamine, a normal constituent of airway mucus (control, 9.5 +/- 2.0 bacteria/cell; N-acetylglucosamine, 5.8 +/- 1.5 bacteria/cell; p = 0.05) . These observations suggest that N-acetylglucosamine may play a role in the epithelial cell receptor for ligands on the surface of nonmucoid strains of PA . Pretreatment of mucoid PA with N-acetylglucosamine, D-arabinose, D-mannose, or N-acetylneuraminic acid did not affect adherence . Partially purified proteins isolated from the apical plasma membrane of bovine tracheal epithelial cells probed in a Western analysis with the iodine 125-labeled PA mucoexopolysaccharide revealed two binding proteins of 45,000 mol wt and 104,000 mol wt . Human airway secretions of patients with chronic PA infection were screened by the dot blot technique and demonstrated selective binding of mucoexopolysaccharide ligand . Western analysis of these biologically important secretions revealed that PA mucoexopolysaccharide bound to a 65,000 mol wt protein in airway secretions of infected individuals . This glycoprotein is similar to a previously described cell receptor for bacterial type 1 fimbriae. J Infect Dis, 1991 May, 163(5), 1040 - 5 Safety and immunogenicity of Escherichia coli O18 O-specific polysaccharide (O-PS)-toxin A and O-PS-cholera toxin conjugate vaccines in humans; Cryz SJ Jr et al.; O-specific polysaccharide (O-PS) isolated from serotype 18 Escherichia coli lipopolysaccharide (LPS) was covalently coupled to either Pseudomonas aeruginosa toxin A (TA) or or cholera toxin (CT) . The conjugates were nontoxic and nonpyrogenic . The conjugates were well tolerated on parenteral administration to human volunteers, with only mild, transient local reactions reported . Immunization engendered an IgG antibody response to both the O-PS and carrier protein . Anti-LPS antibody promoted the uptake and killing of an E . coli O18 strain bearing the K1 capsule by human polymorphonuclear leukocytes, which was complement dependent . Antibody to carrier protein neutralized the activity of native TA or CT in cell culture assays . Passively transferred IgG isolated from the serum of immunized donors provided a significant (P less than .01) degree of protection against fatal experimental E . coli O18 sepsis in mice . This study illustrates the potential use of such conjugates as vaccines against E . coli extraintestinal infections. Rev Infect Dis, 1991 May-Jun, 13 Suppl 7, S634 - 9 Efficacy of aztreonam in the treatment of skeletal infections due to Pseudomonas aeruginosa; Conrad DA et al.; Twenty-eight patients with skeletal infections due to Pseudomonas aeruginosa were treated with aztreonam during two open, noncomparative, multicenter clinical trials . Ten patients with septic arthritis received 2 g of aztreonam three times a day (modal) for 30 days (mean) . The mean follow-up period was 4 weeks . Microbiologic cure was achieved in all 10 patients; complete clinical cure, in eight; and partial clinical cure, in two . Eighteen cases of osteomyelitis were treated with 2 g of aztreonam three times a day (modal) for 40 days (mean), with a mean follow-up period of 6 months . Microbiologic cure was achieved in 17 patients . Relapse occurred 1 month after therapy in one patient . Complete clinical cure was achieved in 13 and partial clinical cure was achieved in five patients . The most common adverse reactions to aztreonam were a transient elevation in levels of hepatic enzymes and transient eosinophilia . Three superinfections and one subsequent infection occurred . These results support use of aztreonam for the treatment of skeletal infections due to P . aeruginosa. Rev Infect Dis, 1991 May-Jun, 13 Suppl 7, S608 - 11 Retrospective clinical study of hypersensitivity reactions to aztreonam and six other beta-lactam antibiotics in cystic fibrosis patients receiving multiple treatment courses; Koch C et al.; A total of 2,793 courses of treatment with seven beta-lactam antibiotics were administered to 121 cystic fibrosis patients chronically infected with Pseudomonas aeruginosa, and the patients were evaluated with respect to clinical hypersensitivity reactions . Seventy-five patients (62%) experienced 125 reactions, for an overall frequency (based on the number of courses) of 4.5% . Immediate reactions occurred in 34 patients (28.1%) during 53 courses (1.9%) . The highest rate of reactions involved piperacillin (50.9% of patients), and the lowest rate involved imipenem and aztreonam (4.0% and 6.5% of patients, respectively); intermediate reaction rates were noted for carbenicillin (23.6% of patients), azlocillin (20.8%), cefsulodin (17.1%), and ceftazidime (13.0%) . Cross-reactivity did not appear to be a major problem . Reactions to aztreonam seemed to be restricted to a small group of patients with a high propensity for beta-lactam hypersensitivity. Rev Infect Dis, 1991 May-Jun, 13 Suppl 7, S594 - 7 Safety of aztreonam in patients with cystic fibrosis and allergy to beta-lactam antibiotics; Jensen T et al.; In an open clinical trial, aztreonam was administered repeatedly to 15 cystic fibrosis patients who were chronically infected with Pseudomonas aeruginosa and had previously had severe hypersensitivity reactions to other beta-lactam antibiotics, including anaphylactic shock, generalized urticaria, and drug-associated fever . After negative results in a skin-prick test and a lack of reaction to an intravenous test dose of aztreonam, the patients were treated with aztreonam (150 mg/{kg.d}) in combination with tobramycin (10-20 mg/{kg.d}) for 14-day periods at 3- to 4-month intervals . To date, a total of 56 course of aztreonam have been administered to these patients (three to six courses per patient), and no type 1 hypersensitivity reactions have occurred . However, as a result of drug-associated fever, the administration of aztreonam had to be discontinued in two of 15 cases . The remaining 13 patients have tolerated treatment well. Am Rev Respir Dis, 1991 May, 143(5 Pt 1), 1076 - 82 Tumor necrosis factor . Alpha and beta subtypes appear in circulation during onset of sepsis-induced lung injury; Leeper-Woodford SK et al.; Tumor necrosis factor (TNF) may be involved in the pathogenesis of acute lung injury (ALI) associated with septicemia . Therefore, we measured plasma TNF activity during sepsis and development of lung injury in a porcine model of ALI . Plasma samples were obtained from anesthetized saline-infused control pigs (n = 10) and those infused for 1 h with live Pseudomonas aeruginosa (10(8) organisms/ml, 0.3 ml/20 kg/min) (n = 16) . TNF activity was measured in plasma using the L929 fibroblast cytolytic assay . L929 cytotoxicity caused by TNF-alpha or TNF-beta was determined in plasma by measuring the cytotoxicity neutralized by TNF antisera . No significant TNF activity was detected in control pig plasma . In septic pigs, TNF activity appeared in plasma 15 min after onset of septicemia and remained elevated throughout the experiment (6.1 +/- 10.2% to 80.0 +/- 5.0%, 15 and 300 min, respectively) . The appearance of pulmonary arterial hypertension, increased lung water, decreased lung compliance, and deteriorating gas exchange was correlated with the rise in plasma TNF activity, which reached a peak at 90 to 120 min in septic pigs . Our results provide evidence that both TNF subtypes are present in plasma during septicemia . Anti-TNF-alpha and anti-TNF-beta neutralized TNF activity in whole septic plasma at 15, 30, and 45 min after onset of septicemia, and the antibodies blocked TNF activity in serially diluted septic plasma at all time points up to 210 min of sepsis . TNF activity in septic plasma at 210 to 300 min was not neutralized entirely by TNF antisera.(ABSTRACT TRUNCATED AT 250 WORDS) Enferm Infecc Microbiol Clin, 1991 May, 9(5), 272 - 6 {Keratitis caused by Pseudomonas aeruginosa and secondary endophthalmitis}; Cuetara S et al.; We report three patients with Pseudomonas aeruginosa keratitis . Two patients in the Intense Care Unit had exogenous bacterial endophthalmitis assessed by histopathology; the infection had probably a nosocomial origin and the outcome was fatal . The third case was associated with the use of soft contact lenses and even though clinically endophthalmitis was suspected there was a favourable response to treatment . We review diagnostic aspects and discuss the management of bacterial keratitis and endophthalmitis. Am J Otolaryngol, 1991 May-Jun, 12(3), 161 - 4 Immunoglobulin-coated bacteria in effusions from secretory and chronic suppurative otitis media; Stenfors LE et al.; Samples of middle ear effusions from 10 children with secretory otitis media and from 10 children with chronic suppurative otitis media were subjected to qualitative and quantitative bacteriologic analysis . In addition, secretory immunoglobulin A (SlgA)- and IgG-coated bacteria were evaluated using the immunofluorescence technique . Secretory otitis media effusions harbored few, if any, immunoglobulin-coated bacteria, whereas chronic otitis media effusions as a rule had heavily IgG- and SlgA-coated bacteria . However, those chronic otitis media effusions that were culture positive for Pseudomonas aeruginosa had no immunoglobulin-coated bacteria . The effusions of very young children were completely devoid of SlgA-coated bacteria . This study demonstrates that, based on the immunoglobulin coating of bacteria obtained from the middle ear cleft, one can evaluate immunologic response during otitis media. Pharmazie, 1991 May, 46(5), 349 - 51 Enhancement of the cytotoxicity of mistletoe lectin-1 (ML-1) by high pH or perturbation in Golgi functions; Yoshida T et al.; We have studied the cytotoxicity of Mistletoe lectin-1 (ML-1), a cytotoxic protein produced by Viscum album, in CHO and V79 cells and in mutant cell lines altered in Golgi functions or in endosomal acidification . In wild-type CHO cells, cytotoxicity of ML-1 was greatly enhanced by ammonium chloride or nigericin . A CHO mutant defective in endosomal acidification (DMPR-2), which is resistant to diphtheria toxin, modeccin and Pseudomonas aeruginosa exotoxin A and hypersensitive to ricin, showed increased sensitivity to ML-1 . MonR-31 and MF-1 are monensin- and compactin-resistant mutants derived from CHO and V79 cell lines, respectively, and are presumably altered in Golgi functions . The cytotoxicity of ML-1 was found to be increased in both MonR-31 and MF-1 cells as compared with their parental cells . These results indicate that the effects of chemicals or mutations altering endosomal acidification and Golgi functions on the cytotoxicity of ML-1 are similar to those on ricin cytotoxicity . Our results suggest that the cytotoxicity of ML-1 is enhanced by an increase in endosomal pH, as well as by chemicals or mutations altering the structure/functions of the Golgi regions . Like ricin, the intoxication process of ML-1 may involve the Golgi regions. Zentralbl Hyg Umweltmed, 1991 May, 191(5-6), 494 - 505 Generation of Pseudomonas aeruginosa aerosols during handwashing from contaminated sink drains, transmission to hands of hospital personnel, and its prevention by use of a new heating device; Doring G et al.; Pseudomonas aeruginosa was isolated from sinks of washing basins, showers, toilets and bathtubs, from the personnel and patients of a mixed infectious disease ward in a German children's hospital during a prospective 4-week epidemiological study . 81% of all sinks were contaminated with P . aeruginosa strains . Upon entering the hospital, all personnel hand cultures were P . aeruginosa-negative . However, during duty, 42.5% of the personnel members carried different P . aeruginosa strains on their hands . Detection of P . aeruginosa strains in sinks preceding the isolation of identical genotypes from personnel hands suggested a transmission route from sinks to hands . Opening of water taps generated aerosols containing P . aeruginosa sink organisms which contaminated hands during hand washing . Survival times of various P . aeruginosa strains in aerosols was dependent on strain characteristics, light and humidity, and t 1/2 differed between 3-76 min . Heating of washing basin sinks to 70 degrees C with a new, safe and inexpensive device inhibited bacterial growth in sinks, generation of P . aeruginosa aerosols, and resulted in hand cultures negative for P . aeruginosa after washing. Pathol Biol (Paris), 1991 May, 39(5), 446 - 50 {Time-killing curves of Pseudomonas aeruginosa strains exposed to polymyxin B}; Peyret M et al.; The time-killing curves of three strains of Pseudomonas aeruginosa (PA01, ATCC10145 and ATCC27853) exposed to five concentrations of polymyxin B comprised: a latency phases, one or two decreasing phases and for the low polymyxin B concentrations a growth phasis . The five antibiotic concentrations were chosen to have a weak bactericidal effect such that decreasing exponential or biexponential models can be fitted to the data . In our experimental conditions, increasing Ca++ and Mg++ concentrations in the medium (Mueller-Hinton) reduced the bactericidal effect and increased the growth phases . Increasing inoculum (10(5) to 10(7) CFU/ml) decreased the bactericidal effect observed with polymyxin B. Naunyn Schmiedebergs Arch Pharmacol, 1991 May, 343(5), 538 - 41 Relationship of primary structure to functional properties of the cytotoxin protein from Pseudomonas aeruginosa; Xiong GM et al.; Deletion mutants of plasmid pSN3 carrying the Pseudomonas aeruginosa cytotoxin gene were prepared and expression in Escherichia coli of proteins with different molecular weights has been proved by Western blot . In addition, through PCR amplification of the cytotoxin gene and ligation into the vector, a single nucleotide change leading to an amino acid exchange from histidine to arginine has been constructed . The activities of the mutants were tested by binding of 125I-cytotoxin to rabbit erythrocyte ghosts and swelling of human granulocytes, showing that the cytotoxin activity is dependent on at least three different domains . Amino acids 12-20 from the C-terminus might be very important for proteolytic activation of protoxin to toxin. Vaccine, 1991 May, 9(5), 294 - 6 Oral immunization of mice with temperature-sensitive Pseudomonas aeruginosa enhances pulmonary clearance of the wild-type; Hooke AM et al.; DBA/2J mice were immunized daily for 3 days per os with 10(8)-10(9) colony forming units (c.f.u.) of two different temperature-sensitive (TS) mutants of Pseudomonas aeruginosa . At varying times after the final immunization the animals were exposed to aerosols of the parental immunotype 1, and the ability of the immunized and control mice to clear their lungs of the wild-type (WT) challenge was measured 4 h later . The number of c.f.u . remaining in the lungs of mice immunized with one mutant, D/1/8, was significantly less (p less than 0.01) than the number remaining in the lungs of control mice and mice immunized with a second TS mutant, E/9/9. J Appl Physiol, 1991 May, 70(5), 2259 - 67 Stimulation of secretion into human and feline airways by Pseudomonas aeruginosa proteases; Somerville M et al.; We have investigated the effect of elastase and alkaline protease from Pseudomonas aeruginosa on airway secretion into the trachea of anesthetized cats and from human bronchial mucosa in vitro . Secretory macromolecules were radiolabeled biosynthetically with two precursors in the cat, {3H}glucose and {35S}sulfate, and with {35S}-sulfate only in human tissue . Both enzymes (2.6 x 10(-9) to 1.3 x 10(-6)M elastase and 8 x 10(-9) to 2.4 x 10(-6)M alkaline protease) released radiolabeled macromolecules in a concentration-dependent manner from the two preparations . Purified elastase, 1.3 x 10(-6)M, released radiolabeled macromolecules (delta 3H = +397 +/- 72%, delta 35S 225 +/- 40% over control, P less than 0.001) and periodic acid-Schiff- (PAS) reactive glycoconjugates (delta PAS = +4.1 +/- 0.96 micrograms/min or +102 +/- 20%; P less than 0.01) from cat trachea, as did alkaline protease, 2.4 x 10(-6)M (delta 3H = +356 +/- 57%, delta 35S = +176 +/- 25%, delta PAS = +7.5 +/- 1.3 micrograms/min or 194 +/- 36%, P less than 0.001) . Increases in 3H exceeded those of 35S, suggesting surface epithelium as the main source of secretion . Inhibition of enzyme activity abolished secretory effects . Both enzymes also stimulated secretion from human bronchus (e.g., with elastase, 1.3 x 10(-6)M: delta 35S = +331 +/- 67%, delta PAS = +4.3 +/- 0.92 micrograms/min or +131 +/- 24%, P less than 0.001; with alkaline protease, 2.4 x 10(-6)M: delta 35S = +220 +/- 67%, delta PAS = +12.7 +/- 3.2 micrograms/min or +575 +/- 245%, P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS) J Antimicrob Chemother, 1991 May, 27 Suppl C, 15 - 9 Single daily dosing of amikacin in an in-vitro model; Dudley MN et al.; The pharmacodynamics of amikacin given as a single daily dose was compared with standard divided dosing in an in-vitro model of infection . This model allows the exposure of log phase bacteria to changing concentrations of antibiotics that simulate the kinetics of the drugs in human patients . Two strains of Pseudomonas aeruginosa, one sensitive and one resistant to azlocillin were studied (MICs for amikacin were 16 and 8 mg/l respectively) . Simulated drug regimens included: amikacin 400 mg q 8 h; amikacin 1.2 g q 24 h; and azlocillin 4 g q 12 h . Each regimen alone and both combinations of amikacin plus azlocillin were studied . With both amikacin regimens initial rapid killing was followed by regrowth of resistant subpopulations . Azlocillin alone produced minimal killing of the resistant strain and moderate killing with ultimate bacteriostasis of the susceptible strain . Bacterial regrowth was prevented with both combination regimens with the single daily dose of amikacin plus azlocillin producing the most rapid and complete killing, especially of the azlocillin resistant strain . These data support further clinical studies of single daily dosing of aminoglycosides. J Antimicrob Chemother, 1991 May, 27 Suppl C, 105 - 12 Clinical and pharmacokinetic study of a single daily dose of amikacin in paediatric patients with severe gram-negative infections; Kafetzis DA et al.; The efficacy and safety of a single 20 mg/kg daily dose of amikacin was studied in an open, uncontrolled trial in 56 infants and children . Most of the patients suffered from severe Gram-negative infection which had failed to respond to previous antibacterial therapy . Amikacin was given in combination with a beta-lactam antibiotic in 43 cases, with other antibiotics in five cases and monotherapy in eight cases . Pseudomonas aeruginosa was the most frequently isolated organism (27 cases) . Clinical results were satisfactory in 98% of the infections treated . Two patients died, one from the infection and the other 8 weeks after therapy from unrelated causes . Amikacin serum concentrations were high during the first hour after iv infusion, low after 8 h and undetectable at 24 h . Nephrotoxicity and ototoxicity was not detected in any patient . Amikacin when used as a single daily dose in combination with a beta-lactam antibiotic was effective and safe in treating infants and children with severe Gram-negative infection. FEMS Microbiol Lett, 1991 May 1, 64(1), 103 - 9 Functional analysis of exotoxin A-related protein of Pseudomonas aeruginosa lacking residues 225-412; Guidi-Rontani C; The crystal structure of the exotoxin A (ETA) of Pseudomonas aeruginosa showed that this protein is folded into three distinct domains . Domain I (Ia and Ib), the amino-terminal domain, is the receptor-binding domain of ETA and domain III, the carboxy-terminal domain, is responsible for the ADP-ribosyl transferase activity of the toxin . To elucidate the function(s) of domains 1b and II in the intoxication process and to define the region of the domain III necessary for ADP-ribosylating activity, a defined deletion in the structural gene of P . aeruginosa ETA encompassing residues 225-412 was constructed and an ETA-related product DeID, (from which all of domains II and Ib were deleted) was expressed . The ETA-related protein did not penetrate sensitive cells, but retained the same specific activity to ADP-ribosylate elongation factor-2 as wild-type toxin . This suggests that domain II is necessary to allow toxin internalization by sensitive cells and that the absence of domain Ib does not interfere with enzymic activity . The domain strictly involved in ADP-ribosylation activity encompasses residues 412-613. Antimicrob Agents Chemother, 1991 May, 35(5), 916 - 21 Invalidity for Pseudomonas aeruginosa of an accepted model of bacterial permeability to beta-lactam antibiotics; Livermore DM et al.; The accepted model for the penetration of beta-lactam antibiotics into gram-negative bacteria is that proposed by Zimmermann and Rosselet (Antimicrob . Agents Chemother . 12:368-372, 1977) . The model assumes (i) that diffusion of the antibiotic molecules across the outer membrane obeys Fick's law and can be characterized by a permeability constant for any given combination of organism and drug, (ii) that drug hydrolysis within the periplasm obeys Michaelis-Menten kinetics, and (iii) that a steady state is rapidly attained between drug uptake and hydrolysis . The model has allowed accurate prediction of antibiotic MICs for Escherichia coli strains from a knowledge of their beta-lactamase production and permeability characteristics . It has been suggested that the model is inappropriate for Pseudomonas aeruginosa, but attempts to confirm this have been bedevilled by experimental difficulties in estimating permeability coefficients for this species . In the present study, we tested a prediction of the model that the overall resistance of P . aeruginosa transconjugants containing a plasmid-encoded beta-lactamase should continue to depend partly on permeability . Transconjugants with PSE-4 beta-lactamase were constructed in host strains with widely different levels of intrinsic, presumably impermeability-determined resistance . Contrary to the prediction of the model, all the transconjugants developed identical overall levels of resistance to substrate beta-lactams, such as azlocillin and cefoperazone, irrespective of the initial levels of intrinsic resistance of the recipient strains . We conclude that the model is inappropriate for P . aeruginosa, and possible explanations for the organism's behavior are discussed. Pediatr Infect Dis J, 1991 May, 10(5), 381 - 6 Comparison of 6 and 8 hourly tobramycin dosing intervals in treatment of pulmonary exacerbations in cystic fibrosis patients; Winnie GB et al.; The efficacy and toxicity of a shortened tobramycin dosing interval in the treatment of exacerbations of Pseudomonas aeruginosa pulmonary infection in cystic fibrosis patients were evaluated prospectively . Patients ages 13 to 30 years received 34 treatment courses and were randomized by pairs to receive tobramycin administered either every 6 or 8 hours . Peak serum concentrations were adjusted to 8 to 10 micrograms/ml; thus a larger total daily dosage was administered to patients receiving tobramycin every 6 hours . The shorter dosing interval was associated with better pulmonary function at follow-up and significantly longer time before next hospital admission for a pulmonary exacerbation . During the study hospitalization there were no differences in pulmonary function tests, clinical score, sputum carriage of P . aeruginosa, toxicity or necessary length of hospitalization . A 6-hour tobramycin dosing interval was more efficacious than an 8-hour dosing interval in the treatment of cystic fibrosis patients. J Clin Microbiol, 1991 May, 29(5), 940 - 4 Effects of FP2 and a mercury resistance plasmid from Pseudomonas aeruginosa PA103 on exoenzyme production; Johnson J et al.; Plasmids encoding mercury resistance carried by Pseudomonas aeruginosa PAO1161 and PA103 were found to be involved in regulating the secretion of protease, phospholipase C, and alkaline phosphatase . Previously, mutations in Pseudomonas strains that caused pleiotropic effects on the production of extracellular enzymes were mapped to the bacterial chromosome . We show that pleiotropic changes in extracellular enzyme production can also be regulated by plasmids . In this study, the effects on secretion of exoenzymes by two mercury resistance plasmids, FP2 from PAO1161 and pRLW103 from PA103, were assayed in P . aeruginosa PAO1 and PAO18 . The introduction of either plasmid into PAO1 resulted in a significant decrease in exoprotease production . Additionally, pRLW103 significantly increased the production of alkaline phosphatase by both strains . Phospholipase C was produced only in strain PAO18 containing the pRLW103 plasmid . FP2 had no effect on alkaline phosphatase or phospholipase C production in either strain and was found to decrease exoprotease secretion only in strain PAO1 . The results indicate the P . aeruginosa mercury resistance plasmids vary in their ability to modify exoenzyme expression, and this ability is influenced by the host strain. J Laryngol Otol, 1991 May, 105(5), 341 - 2 Bacteriology of inadequately treated active chronic otitis media in paediatric age group; Amadasun JE; Haphazardly treated active otitis media was investigated bacteriologically in 214 children . Analysis showed that Pseudomonas aeruginosa was more prevalent than other micro-organisms . Candida was also found in appreciable quantity . The author contends that this was due to inadequate self treatment before these children reported in his hospital . He advocates that decision regarding the chemotherapeutic agent to be used should be based on culture and in vitro sensitivity. Infect Immun, 1991 May, 59(5), 1667 - 72 In vivo protective effect of lipopolysaccharide against Pseudomonas aeruginosa exotoxin A in mice; Zehavi-Willner T et al.; Lipopolysaccharide (LPS) treatment of mice 1 to 5 days prior to administration of Pseudomonas aeruginosa exotoxin A (PA) induced full or partial protection against PA intoxication . The optimal LPS dose that induced resistance was 50 to 100 micrograms per mouse . Simultaneous administration of LPS and PA to mice, however, increased their sensitivity to PA two- to fourfold . Mice pretreated with LPS demonstrated a markedly enhanced clearance rate of 125I-labeled PA from peripheral blood, livers, and kidneys . In mice exposed to LPS and PA simultaneously, the rate of elimination of labeled PA was lower than that in control mice . While protein synthesis was inhibited significantly in livers and other organs of PA-exposed mice, in LPS-pretreated mice, PA-induced inhibition of protein synthesis was either diminished or totally prevented and elongation factor 2 (EF2) levels were normal . In mice treated only with LPS, enhanced protein synthesis and increased levels of EF2 were observed, suggesting that LPS protection against PA intoxication was perhaps a consequence of excessive amounts of EF2 induced by LPS. Pathol Biol (Paris), 1991 May, 39(5), 361 - 6 {In vitro activity of tazobactam and piperacillin combination against 224 strains of Pseudomonas aeruginosa according to the production of beta-lactamase}; Thabaut A et al.; Piperacillin (PIP) alone and combined with 4 mg/l, 8 mg/l of tazobactam (TAZ) were tested by MIC determination on Mueller-Hinton agar against 224 clinical strains of P . aeruginosa: "wild type" (BLA-), 32 producing a constitutive cephalosporinase (CEP), 41 producing the PSE-1 type beta-lactamase, 7 PSE-2, 8 PSE-3, 9 PSE-4, 13 TEM-1, 24 TEM-2, 13 OXA-1, 22 OXA-2, 5 OXA-3 . The combination with 8 mg/l was more effective than that one with 4 mg/l . Combinations of PIP-TAZ 8 mg/l reduced the MICs of PIP for the resistant strains (MICs greater than 64 mg/l) to the susceptible ot the moderately susceptible range (MICs less than or equal to 64 mg/l) for 31% of the CEP producing strains, 63% of the PSE-1, 15% of the PSE-2, none of the PSE-3, 34% of the PSE-4, 39% of the TEM-1, 30% of the TEM-2, 23% of the OXA-1, 14% of the OXA-2, 27% of the OXA-3, TAZ is the first beta-lactamase inhibitor effective against the constitutive cephalosporinase of P . aeruginosa; it is also very effective against the most frequently found PSE-1 beta-lactamase in P . aeruginosa. J Gen Microbiol, 1991 May, 137 ( Pt 5), 1111 - 20 Chromosomal insertion of TOL transposons in Pseudomonas aeruginosa PAO; Sinclair MI et al.; Insertions of the TOL plasmid transposons Tn4651 and Tn4653 into the Pseudomonas aeruginosa PAO chromosome were isolated by a temperature selection technique . The locations and orientations of 16 insertions were determined by pulsed field gel electrophoresis and Southern hybridization with genomic and TOL DNA probes . All insertions occurred within a 334 kb region of the chromosome (representing less than 6% of the genome) with nine of the inserts clustered within a 10 kb area . Each transposon was able to insert in either orientation . An internal duplication of the 39 kb excisable region of pWW0 was seen in two independent insertions. Antimicrob Agents Chemother, 1991 May, 35(5), 892 - 7 Nucleotide sequence of the aacC3 gene, a gentamicin resistance determinant encoding aminoglycoside-(3)-N-acetyltransferase III expressed in Pseudomonas aeruginosa but not in Escherichia coli; Vliegenthart JS et al.; A chromosomal gentamicin resistance determinant from Pseudomonas aeruginosa was cloned on a 2.4-kb fragment in the broad-host-range vector pLAFR3 . Substrate profiles for eight aminoglycosides at three concentrations showed that resistance was due to aminoglycoside-(3)-N-acetyltransferase III . This enzyme was produced in Pseudomonas strains but not in an Escherichia coli strain bearing the aacC3 gene . Nucleotide sequencing revealed two contiguous open reading frames (ORFs) preceded by a potential promoter and a ribosome-binding site . ORF-1 was 642 bp in length and encoded a protein of unknown function with a molecular mass of 23.9 kDa . ORF-2 was 813 bp in length and encoded a protein of 29.6 kDa . From deletion mutagenesis, in vitro transcription-translation data, and protein analysis of bacterial lysates, it was inferred that this 29.6-kDa protein represents the aminoglycoside-(3)-N-acetyltransferase III enzyme . A polymerase chain reaction with two specific intragenic 20-mer primers was developed to detect the aacC3 gene . A BstEII restriction site in the amplified DNA region was used to demonstrate the specificity of the reaction . Tests of 23 reference strains, which produced 12 different aminoglycoside-modifying enzymes, confirmed the specificities of the primers . The gene proved to be absent from a collection of 50 gentamicin-resistant P . aeruginosa strains selected at random in The Netherlands. Enzyme Microb Technol, 1991 May, 13(5), 385 - 9 Alginate biosynthesis in mucoid recombinants of Pseudomonas aeruginosa overproducing GDP-mannose dehydrogenase; Martins LO et al.; The Pseudomonas aeruginosa algD gene, encoding GDP-mannose dehydrogenase (GMD) and cloned at Chakrabarty's Laboratory in the expression vector pMMB24 (plasmid pVD211), was mobilized into P.aeruginosa strains 8821 and 8821M . Strain 8821M was a high-alginate-producing variant, spontaneously obtained from mucoid strain 8821, with derepressed levels of GMD, a key enzyme in the regulation of alginate biosynthesis, leading to the irreversible oxidation of GDP-mannose to GDP-mannuronic acid . A slight increase in the level of GMD, in both strains harboring the plasmid pVD211 and batch-grown at 37 degrees C without IPTG induction, led to the increase of production rate and the final concentration of alginate produced by control strains harboring the cloning vector . However, the viscosity of the aqueous solutions prepared with the alginate (3 g l-1) produced by mucoid strains harboring pVD211 was lower than those with the alginate produced by the controls (shear rates in the range 0.6-12 s-1) . The specific activity of GMD assayed in crude extracts from cells harboring pVD211 and subjected to IPTG induction (0.5 and 3 mM) presented the highest values . However, either the rate of biosynthesis and final concentration of alginate or the viscosity of solutions prepared with the alginate produced by recombinants grown with IPTG were lower than that possible without overproduction . Therefore, the stimulation of the alginate pathway only by manipulating the rate of the step catalysed by GMD, although possible within certain levels, was at the expense of the final exopolysaccharide quality. Biochim Biophys Acta, 1991 Apr 29, 1077(3), 316 - 24 Pseudomonas aeruginosa alkaline proteinase might share a biological function with plasmin; Shibuya Y et al.; Pseudomonas aeruginosa alkaline proteinase, which is a zinc-dependent bacterial endopeptidase, preferentially hydrolyzed Boc-Val-Leu-Lys-methylcoumarylamide (MCA) which was originally designed as a specific substrate of plasmin, a plasma serine proteinase . The hydrolytic capacity was resistant to tosyl-lysine chloromethylketone at a concentration as high as 1 mM, but was blocked by a treatment with metal chelator such as o-phenanthroline at the concentration of 5 mM . Kinetic parameters of the amidolytic reaction were Km = 21 microM, kcat = 0.067 s-1 and kcat/Km = 3190 M-1 s-1 . A synthetic peptide inhibitor which bore a possible ligand for zinc atom at the carboxy terminal was designed . This inhibitor, Ac-Val-Leu-Lys-4-mercaptoanilide, blocked the amidolytic activity of the pseudomonal alkaline proteinase in a competitive manner with the dissociation constant (Ki) value of 24 microM . The results imply that P . aeruginosa alkaline proteinase must be an unusual zinc-dependent 'C (COOH)-type' endopeptidase, which hydrolyzes the peptide bond of certain amino acid residues at the carboxyl group side by specific recognition, like serine- and cysteine-proteinases . In comparison, P . aeruginosa elastase which is a typical 'N (NH2)-type' metalloproteinase did not hydrolyze all of the commercially available peptide-MCA substrates tested at the present study . P . aeruginosa alkaline proteinase also hydrolyzed natural substrates of plasmin, such as fibrin and fibrinogen, with similar specific activities to plasmin . The susceptible subunits of fibrinogen were the A-alpha and B-beta ones, in this order . P . aeruginosa alkaline proteinase also exhibited an anti-coagulant activity in human plasma attributed to the direct fibrinogenolytic function . Such potential anti-coagulant capacity of the P . aeruginosa alkaline proteinase might explain, at least partly, the most characteristic pathologic feature of the P . aeruginosa septicemia, hemorrhagic lesions with lacking thrombi (Fetzer, A.E . et al . (1967) Am . Rev . Respirat . Dis . 96, 1121-1130). J Immunol, 1991 Apr 15, 146(8), 2639 - 47 Recombinant human granulocyte colony-stimulating factor improves the compromised state of recipient mice without affecting the induction of specific tolerance in the cyclophosphamide-induced tolerance system; Nishimura Y et al.; The effects of recombinant human granulocyte CSF (rhG-CSF) on cyclophosphamide (CP)-induced tolerance was studied . In the recipient C57BL/10 Sn Slc (B10) mice given 1 x 10(8) B10.BR Sg Sn Slc (B10.BR) spleen cells (SC) on day -2 followed by 200 mg/kg CP on day 0, the number of leukocytes and neutrophils in the periphery declined to their minimum levels on day 4 . When rhG-CSF in a dose of 200 micrograms/kg was given daily for 5 days to the B10 mice, which had been treated with B10.BR SC and CP, starting one day after the administration of CP, the leukocyte and neutrophil counts declined to the same levels as those in the B10 mice treated with B10.BR SC and CP alone on day 2 . On day 4, however, the counts recovered to their normal levels . The nucleated cell count of the spleen in the B10 mice given B10.BR SC and CP followed by rhG-CSF decreased less and recovered faster than that in the B10 mice given B10.BR SC and CP . The case was found to be the same in bone marrow, and the difference did not reach statistical significance . When the recipient mice were inoculated i.p . with 4 x 10(4) Pseudomonas aeruginosa (GNB-139) on day 4, the survival of the B10 mice treated with B10.BR SC and CP was markedly improved by rhG-CSF administration . The administration of rhG-CSF did not affect either the prolongation or the specificity of skin allograft survival, as shown in an H-2 mis-matched combination of B10.BR----B10 and in an H-2 identical combination of AKR/J Sea(AKR)----C3H/HeN Crj (C3H) . The tolerant state, which was demonstrated by various immune responses, such as CTL, delayed footpad reaction, and antibody, was also not affected by rhG-CSF . Furthermore, the basic mechanisms for inducing a long-lasting skin allograft tolerance in this system--namely, the specific destruction of Ag-stimulated and then proliferating mature T cells in the periphery, the establishment of mixed chimerism, and the intrathymic clonal deletion of immature T cells--were preserved even when rhG-CSF was given to C3H mice previously made tolerant of AKR. J Biol Chem, 1991 Apr 5, 266(10), 6438 - 46 Pseudomonas aeruginosa exoenzyme S requires a eukaryotic protein for ADP-ribosyltransferase activity; Coburn J et al.; Pseudomonas aeruginosa exoenzyme S ADP-ribosylates several GTP-binding proteins of apparent Mr = 23,000-25,000 . Exoenzyme S absolutely requires a soluble eukaryotic protein, which we have named FAS (Factor Activating exoenzyme S), in order to ADP-ribosylate all substrates . The rate of ADP-ribosylation of all exoenzyme S substrates increases linearly with time and with the FAS concentration . FAS is wide-spread in eukaryotes but appears to be absent from prokaryotes . We have estimated the molecular mass of the protein to be approximately 29,000 daltons and its pI to be 4.3-4.5 . Several bacterial toxins share this sort of requirement for the presence of a eukaryotic protein for enzymic activity . In particular, FAS resembles ADP-ribosylation factor, a 21,000-dalton GTP-binding protein which performs an analogous function for cholera toxin . However, we can find no evidence that FAS binds GTP . In the presence of FAS, exoenzyme S ADP-ribosylates several proteins in lysates of P . aeruginosa . The requirement for a eukaryotic protein for enzymic activity, which is common to several bacterial toxins, may be a device to identify the eukaryotic environment and to ensure that the enzymes cannot function within and harm the toxin-producing bacteria. J Pediatr Surg, 1991 Apr, 26(4), 487 - 92; discussion 492-3 Management of anorectal/perineal infections caused by Pseudomonas aeruginosa in children with malignant diseases; Angel C et al.; The role of operation for anorectal infections associated with perineal gangrene and cellulitis in children with myelo-suppression from cancer chemotherapy is unclear . We evaluated anorectal/perineal infections caused by Pseudomonas aeruginosa in 16 children with malignant diseases seen over 27 years . In 12 of 16 patients, leukemia was the underlying malignancy (ALL 10, AML 2), and in 13 of 16, severe neutropenia (absolute neutrophil count less than 500/mm3) was present at diagnosis . Cultures of the lesions showed multiple organisms in 14 of 16 patients with Escherichia coli, Klebsiella species, and Enterococcus being the most frequent coexisting organisms . All positive blood cultures grew P aeruginosa exclusively . Of three patients with necrotizing infections, two had complete resolution with medical treatment alone; the other patient who developed this problem while on terminal care died . In none of the 16 patients was a major operation (debridement or diversion) performed . Five patients died, three of whom were considered terminally ill when the anorectal infections occurred . Four of the five deaths occurred before 1974 . Since then, only 1 of 7 patients died . Excluding the three terminally ill patients, the success rate of medical therapy alone is 85% (11/13) . The antibiotic regimen should include an aminoglycoside in synergistic combination with anti-Pseudomonas penicillin . These results suggest that operative management may have no role in the management of anorectal infections caused by P aeruginosa in children with cancer. Gene, 1991 Apr, 100, 65 - 73 Cloning and characterization of a chromosomal DNA region required for growth on 2,4,5-T by Pseudomonas cepacia AC1100; Haugland RA et al.; A series of spontaneous 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) nonmetabolizing mutants of Pseudomonas cepacia AC1100 were characterized to be defective in either 2,4,5-T uptake or conversion of this compound to 2,4,5-trichlorophenol (2,4,5-TCP) . Two of these mutants, RHC22 and RHC23, were complemented for growth on 2,4,5-T using an AC1100 genomic library constructed in the cosmid vector pCP13 . Recombinant cosmids isolated from the complemented mutants contained a 27.5-kb insert which frequently underwent various-sized deletions in Escherichia coli . Hybridization studies showed this DNA to be of chromosomal origin and totally deleted in RHC22, RHC23 and other similar mutants . Complementation analyses of RHC22 with a series of subcloned fragments and spontaneously deleted derivatives of the recombinant cosmid pRHC21 showed the 2,4,5-T (tft) genes to occur within an 8.9-kb region . Pseudomonas aeruginosa cells transformed with this DNA acquired the ability to convert 2,4,5-T to 2,4,5-TCP . The genetic determinant for this function was further localized within a 3.7-kb region . This DNA, in the absence of other sequences from the 8.9-kb tft gene region allowed RHC22 cells to metabolize 2,4,5-T, but at low rates which were insufficient to support growth . Copies of the insertions sequence element IS931 were identified either adjacent to or within this tft gene region in the genomes of two independent wild-type AC1100 isolates . Preliminary evidence suggests that these sequences either facilitate or are required for growth on 2,4,5-T and hence may be implicated in the genetic evolution of the 2,4,5-T metabolic pathway. Rinsho Byori, 1991 Apr, 39(4), 429 - 36 {Multi-center evaluation of Showa disk susceptibility to presumptively determine minimum inhibitory concentrations through linear regression analysis}; Sato Y et al.; To confirm the reliability of minimum inhibitory concentrations (MICs) determined by use of predefined linear regressions to bacterial growth inhibitory zone diameter on Showa disk susceptibility test, the multi-center evaluation along daily routines was performed in comparison with the standard agar dilution method . In total, 4,107 (89.0%) of 4,613 testings gave comparable MICs with 4-fold or less differences to those determined by the standard agar dilutions . The agreement of MICs (less than or equal to 4-fold differences) for gram-negative rods, excluding Pseudomonas aeruginosa, against 10 antimicrobial agents was estimated to 92.1%, and those for gram-positive cocci against 9 agents and for the strains of Pseudomonas aeruginosa against 8 agents were 84.5% and 81.7%, respectively . With these results, we can conclude that, under the well-controlled test procedures, the MIC correlates determined by Showa disk susceptibility test are enough comparable to those determined by the standard agar dilution method. Ann Thorac Surg, 1991 Apr, 51(4), 630 - 5 Phenotypic expression of bronchoalveolar lavage cells in lung rejection and infection; Shennib H et al.; The differentiation of episodes of lung allograft rejection from infection continues to be a problem . Bronchoalveolar lavage (BAL) has recently gained some success in the diagnosis and management of interstitial lung disease . To assess the usefulness of BAL in differentiating between lung allograft rejection and infection, we examined the differences in cellular subsets of BAL and peripheral blood (PBL) samples in a controlled canine model of rejection or pneumonia . Single-lung allotransplants were allowed to undergo rejection by withdrawing immunosuppressive agents (n = 6) . In another group of dogs (n = 5), pneumonia was induced by transbronchial injection of Pseudomonas aeruginosa and melted agar followed by bronchial fulguration . Cells obtained from bronchoscopic BAL and PBL samples were labeled with functionally characterized cross-reactive murine monoclonal antibodies . Transthoracic needle biopsies and transbronchial biopsies were done to assess their adequacy in examining the rejecting or infected lungs and were compared with open lung biopsies . We found the following: (1) the percentage of DT2-labeled cells was significantly higher (p less than 0.05) in BAL samples from rejecting lungs compared with infected lungs; (2) the PBL/BAL ratio of DT2-labeled cell percentages was significantly higher in pneumonia (1.7 +/- 0.3) than rejection (0.5 +/- 0.2) (p less than 0.004); (3) the percentage of E11-labeled cells in PBL samples was significantly higher (p less than 0.02) in rejection than in infection; and (4) the ratio of WIG4 to DT2 cellular subset percentages in BAL samples from rejection (26.8 +/- 9.9) was significantly lower than from infection (61.0 +/- 22.9) (p less than 0.03) . Transthoracic and transbronchial biopsies did not always yield representative specimens.(ABSTRACT TRUNCATED AT 250 WORDS) J Infect Dis, 1991 Apr, 163(4), 703 - 9 Anti-HIV activity of CD4-Pseudomonas exotoxin on infected primary human lymphocytes and monocyte/macrophages; Ashorn P et al.; CD4(178)-PE40 is a recombinant protein consisting of a portion of human CD4 linked to active domains of Pseudomonas aeruginosa exotoxin A . In previous experiments with human T cell lines, the hybrid toxin was found to selectively kill cells infected with human immunodeficiency virus type 1 (HIV-1), and to inhibit HIV-1 spread in mixtures of infected and uninfected cells . CD4(178)-PE40 inhibits HIV-1 spread in cultured primary human lymphocytes . Moreover, the hybrid toxin selectively kills HIV-1 chronically infected monocyte/macrophage cell lines and inhibits HIV-1 spread in primary macrophage cultures . Control experiments indicate that the protective effects of CD4(178)-PE40 against HIV-1 spread are due to selective killing of the infected cells rather than simply to neutralization by the CD4 moiety . Thus, for the major cell types susceptible to HIV infection in vivo, surface envelope glycoprotein is expressed at sufficient levels to enable binding and internalization of CD4(178)-PE40 and consequent selective cell killing. Infect Immun, 1991 Apr, 59(4), 1514 - 20 Morphological and immunohistochemical studies of the lungs and bronchus-associated lymphoid tissue in a rat model of chronic pulmonary infection with Pseudomonas aeruginosa; Iwata M et al.; Pseudomonas aeruginosa is one of the most frequently encountered bacterial pathogens in patients with chronic pulmonary infections, including cystic fibrosis and diffuse panbronchiolitis . Bronchus-associated lymphoid tissue (BALT), noted frequently in patients with cystic fibrosis and diffuse panbronchiolitis, is considered to play an important role in the local immunologic defense mechanisms in the respiratory tract . To investigate the role of BALT in chronic pulmonary infections, we developed an animal model for chronic pulmonary infection and studied the morphological and immunohistochemical characteristics of BALT . Experimental pneumonia was produced in rats by intratracheal inoculation of P . aeruginosa enmeshed in agar beads . The histological changes corresponded to those occurring in chronic bronchiolitis . Immunohistochemically, surface immunoglobulin M-positive (sIgM+) cells and sIgA+ cells were recognized in the inflamed bronchial walls from day 4, and sIgG+ cells were recognized from day 14, W3/25+ cells exceeded OX8+ cells in number until day 14 . In the BALT, there was a massive accumulation of lymphocytes in the lymphatics and high endothelial venules . The development of germinal centers was accompanied by increased numbers of sIgM+ and sIgA+ cells . W3/25+ cells exceeded OX8+ cells in number in the BALT until day 14 . On the other hand, OX8+ cells were predominant in comparison with W3/25+ cells at day 21, and then both sIgM+ and sIgA+ cells and inflammatory changes in the lung decreased at day 28 . These findings suggest that BALT regulates the local immune responses against chronic pulmonary infection due to P . aeruginosa. Zentralbl Bakteriol, 1991 Apr, 275(1), 100 - 11 Evaluation of protective mAbs against Pseudomonas aeruginosa outer membrane protein I by C1q binding assay; Eckhardt A et al.; Seven monoclonal antibodies (mAbs) against the outer membrane proteins (OPRs) F, H and I of Pseudomonas aeruginosa were prepared . Western blot analysis has shown the mAbs to cross-react with all 17 serotypes of P . aeruginosa according to the International Antigenic Typing Scheme . Two of the mAbs (2A1, 6A4) protected mice against fatal P . aeruginosa pneumonia . The protective potential of the mAbs did not correlate with the immunoglobulin isotype nor with the fine antigen specificity and the in vitro bactericidal activity of the mAbs . Only the binding of the first complement component C1q of the mAbs as estimated in vitro by an ELISA was significantly correlated with their protective potential. Planta Med, 1991 Apr, 57(2), 129 - 31 4-Hydroxy-2-cyclopentenone: an anti-Pseudomonas and cytotoxic component from Passiflora tetrandra; Perry NB et al.; 4-Hydroxy-2-cyclopentenone is responsible for the anti-bacterial activity of an extract of leaves from Passiflora tetrandra with minimum inhibitory doses (MID) of ca . 10 micrograms/disk against Escherichia coli, Bacillus subtilis, and Pseudomonas aeruginosa . 4-Hydroxy-2-cyclopentenone is also cytotoxic to P388 murine leukemia cells (IC50 of less than 1 microgram/ml). J Clin Microbiol, 1991 Apr, 29(4), 827 - 9 False resistance to imipenem with a microdilution susceptibility testing system; O'Rourke EJ et al.; Routine monitoring of antibiotic resistance at Children's Hospital, Boston, detected a dramatic increase in the prevalence of imipenem-resistant strains of Pseudomonas aeruginosa . Further studies documented that false resistance to imipenem was due, in part, to the loss of imipenem potency in customized MIC microdilution trays supplied by Sensititre Ltd . (West Sussex, United Kingdom) . Recognition of the problem was delayed by use of the quality control standard recommended by the manufacturer, which were higher and broader than those suggested by the National Committee for Clinical Laboratory Standards. Antimicrob Agents Chemother, 1991 Apr, 35(4), 783 - 4 Comparative activity of cefepime, alone and in combination, against clinical isolates of Pseudomonas aeruginosa and Pseudomonas cepacia from cystic fibrosis patients; Bosso JA et al.; The comparative in vitro activity and synergy of cefepime were evaluated with clinical isolates of Pseudomonas aeruginosa and Pseudomonas cepacia from cystic fibrosis patients . The activity of cefepime, both alone and in combination, was comparable to those of other antibiotics . The clinical efficacy of cefepime in cystic fibrosis patients merits investigation. Antimicrob Agents Chemother, 1991 Apr, 35(4), 753 - 5 Imipenem resistance in pseudomonas aeruginosa PAO: mapping of the OprD2 gene; Quinn JP et al.; Carbapenem antibiotics have been shown to penetrate the outer membrane of Pseudomonas aeruginosa through a unique porin protein, OprD2 . We mapped the OprD2 gene by conjugation using plasmid FP2 and by transduction using phage F116L . This gene maps between 71 and 75 min on the PAO1 chromosome. Antimicrob Agents Chemother, 1991 Apr, 35(4), 691 - 5 Hyperoxia prolongs the aminoglycoside-induced postantibiotic effect in Pseudomonas aeruginosa; Park MK et al.; The objective of this study was to determine whether hyperoxia enhances aminoglycoside activity against Pseudomonas aeruginosa . The existence of tobramycin-oxygen synergy was determined by using the in vitro postantibiotic effect (PAE) . P . aeruginosa strains were incubated for 1 h in medium containing tobramycin at four times the MIC in the following gas mixtures: normoxia (21% O2), hyperoxia (100% O2, 101.3 kPa), or hyperbaric oxygen (100% O2, 274.5 kPa) . Tobramycin was removed after 1 h and bacteria were incubated under normoxic conditions; growth rates were measured for 5 h . Exposure of three P . aeruginosa strains to hyperoxia prolonged the PAE of tobramycin approximately twofold compared with the PAE after exposure to normoxia (P less than 0.05) . Exposure of P . aeruginosa ATCC 27853 to tobramycin and hyperbaric oxygen prolonged the time required for bacteria to increase 1 log10 CFU/ml compared with the time after exposure for this increase to occur in tobramycin-treated, normoxic or hyperoxic groups (P less than 0.02) . Pulse-chase labeling of bacteria with L-{35S}methionine, immediately after removal of tobramycin, showed that protein synthesis rates were decreased compared with those in controls (P = 0.0001) . Moreover, in tobramycin-treated groups, hyperoxia and hyperbaric oxygen induced 2- and 16-fold decreases, respectively, in protein synthesis rates compared with normoxia; these results did not achieve statistical significance . In the absence of tobramycin, hyperoxia increased bacterial growth (134%; P less than 0.01) and protein synthesis (24%; not significant) compared with normoxia . Hyperbaric oxygen, however, delayed the growth recovery of bacteria (P less than 0.05) . We conclude that hyperoxia enhances the bacteriostatic effects of tobramycin in a synergistic manner.+ J Med Microbiol, 1991 Apr, 34(4), 213 - 7 Interaction between Pseudomonas aeruginosa and Staphylococcus aureus: description of an anti-staphylococcal substance; Machan ZA et al.; The presence of Pseudomonas aeruginosa in the sputum of 191 patients with cystic fibrosis was significantly related (p less than 0.0001) to the absence of Staphylococcus aureus . Cross-streaking tests showed that 40 of 50 clinical strains of P . aeruginosa produced substances that inhibited the growth of S . aureus . When incorporated into agar plates, this antibacterial substance(s) inhibited the growth of 177 of 189 strains of nine staphylococcal species, all of 16 methicillin-resistant S . aureus and 27 of 39 strains of six other gram-positive genera . The substance(s) did not inhibit 23 strains of seven gram-negative genera tested . The antibacterial activity was heat stable and could be extracted into chloroform; activity was retained on Sephadex G-15 (V/Vo approximately 2, Mr less than 500) and eluted as a single peak from high performance liquid chromatography, well separated from pseudomonic acid, pyocyanin and a number of other phenazines. J Med Microbiol, 1991 Apr, 34(4), 203 - 12 Serum antibodies to Pseudomonas aeruginosa outer-membrane proteins and iron-regulated membrane proteins at different stages of chronic cystic fibrosis lung infection; Shand GH et al.; Serum samples collected over periods up to 15 years from nine patients with cystic fibrosis (CF) were investigated by immunoblotting and crossed immuno-electrophoresis (CIE) for antibodies to Pseudomonas aeruginosa outer-membrane proteins (OMPs) . The earliest antibody response to OMPs was directed against proteins G, H1 and I . Detection by immunoblotting sometimes preceded the CIE response; the appearance of antibodies to the other major OMPs was co-incident with an increase in CIE precipitins . Isolation of the mucoid form of P . aeruginosa was associated with a rapid increase in both precipitin numbers and antibodies detected by immunoblotting . Antibodies to iron-regulated OMPs could be detected in all the serum samples that showed eight or more CIE precipitins but their presence became pronounced only in the advanced stages of disease . The clinical strain used in this study and other isolates from CF patients showed several atypical OMPs, perhaps as a consequence of antibiotic therapy or related to the serum sensitivity of mucoid P . aeruginosa . Their expression in vivo was confirmed by detecting antibodies to them in patients' serum. Bol Asoc Med P R, 1991 Apr, 83(4), 160 - 3 Unusual presentation of Pseudomonas aeruginosa infections: a review; Molina DN et al.; Pseudomonas aeruginosa is an opportunistic, gram negative bacillus that causes serious hospital acquired infections . However, it also causes infections with unusual presentations which are acquired in a non-hospital environment . This report will discuss the pathogenesis, clinical manifestations, and therapy of this uncommon infection, such as: 1) Pseudomonas folliculitis: a superficial or deep bacterial infection associated with the use of public hot tubs, whirlpools and swimming pools . 2) Invasive external otitis: an infection that can progress to skull base mostly associated to elderly diabetic patients . It is usually secondary to aural irrigation with contaminated water . 3) Pseudomonas osteomyelitis: an infection usually associated with nail puncture wounds especially if wearing tennis shoes . 4) Toe with infection: mostly associated with individuals using topical antibacterial agents . 5) Green nail syndrome: a non tender paronychia lesion that appears most often in persons whose hands are constantly exposed to water, soaps and detergents or are subject to mechanical trauma . 6) Corneal ulcer keratitis: mostly associated with the use of soft lenses, eye drops, mascara or contaminated whirlpools . This condition may terminate in panophthalmitis . 7) Endocarditis: most commonly associated with intravenous drug addicts. Rev Cubana Med Trop, 1991 Apr-Aug, 43(2), 132 - 5 {Epidemiologic markers in the study of Pseudomonas aeruginosa from a pediatric intensive care unit}; Azahares Ramirez L et al.; A study was made on 198 Pseudomonas aeruginosa strains isolated at a pediatric intensive care unit from January to August 1988, using pyocin typing and antibiotic typing as markers . The most frequent circulating pyocin types were 31.83 and atypical 23578 . The germ showed high resistance in vitro to the antimicrobial drugs used and the resistance patterns were distributed in 19 different antibiotic types. Pathology, 1991 Apr, 23(2), 145 - 8 Longitudinal studies of virulence factors of Pseudomonas aeruginosa in cystic fibrosis; Burke V et al.; Among 111 strains of Pseudomonas aeruginosa from 49 children with cystic fibrosis, duration of colonization correlated with bacterial phenotype . We confirmed that P . aeruginosa from chronically colonized patients tended to be less motile, produce lower levels of protease and elastase, to be more sensitive to normal serum and to be polyagglutinating or untypable with standard antisera . We also showed that phospholipase and heat-stable hemolysin, concerned in metabolism of inorganic phosphate, and exotoxin A, were lower in these isolates . In longitudinal studies there was a decrease in virulence properties when isolates from the same patient were compared . No reversion from altered phenotype to 'wild-type' characteristics was found. Jpn J Med Sci Biol, 1991 Apr, 44(2), 63 - 74 Growth and survival of Pseudomonas pseudomallei in acidic environments; Dejsirilert S et al.; A study was made on the growth and survival of Pseudomonas pseudomallei in culture environments differing in nutrients, initial pH, and aeration, in comparison with Pseudomonas cepacia and Pseudomonas aeruginosa . The observations led us to a view that P . pseudomallei has the highest adaptability to acidic environments among the three species . Unlike the other species, it grew in heart infusion broth of initial pH 4.5 under aeration and survived keeping a high level (10(9) per ml) of viable counts for as long as 30 days . This sort of adaptation was found to be more evident in the media of poor nutrition and under limited aeration. FEMS Microbiol Immunol, 1991 Apr, 3(2), 69 - 73 Detection of common polysaccharide antigen of Pseudomonas aeruginosa with O-antiserum to Pseudomonas cerasi; Kocharova NA et al.; The identity of the structures of common polysaccharide antigen (CPA) of Pseudomonas aeruginosa and O-antigen of Pseudomonas cerasi was used for immunochemical study of polysaccharide antigens of seven immunotypes (IT) of P . aeruginosa . ELISA performed with O-antiserum to P . cerasi showed that CPA is present in all seven ITs in different amounts . In SDS-PAGE this antigen behaves as a lipopolysaccharide (LPS) and is detected by immunoblotting technique in five of seven ITs. Infect Immun, 1991 Apr, 59(4), 1251 - 4 Protection of immunosuppressed mice against infection with Pseudomonas aeruginosa by recombinant P . aeruginosa lipoprotein I and lipoprotein I-specific monoclonal antibodies; Finke M et al.; Outer membrane protein I (OprI) is one of the major proteins of the outer membrane of Pseudomonas aeruginosa . The protective effect of OprI vaccination and that of three OprI-specific monoclonal antibodies (MAbs) against infection with P . aeruginosa were tested in immunosuppressed mice . The combination of Oprl and MAb 2A1 protected the mice against a challenge with a 96-fold 50% lethal dose . The binding site of MAb 2A1 was mapped, resulting in the identification of a protective epitope (amino acids 7 to 20). Curr Eye Res, 1991 Apr, 10(4), 351 - 62 Analysis of adhesion, piliation, protease production and ocular infectivity of several P . aeruginosa strains; Hazlett LD et al.; The role of bacterial piliation and protease production in Pseudomonas aeruginosa adhesion to the injured corneal epithelial surface and subsequent infectivity was examined using several bacterial strains, including three that were hyperpiliated . To initiate this study, bacteria were examined by transmission EM to confirm their piliation characteristics . The PAK strain, like pseudomonas ATCC 19660, possessed about 1-4 polar pili . The mutant PAK/PR11 lacked pili while PAK/PR1, DB2, a mutant of PAO1, and PA1244, a wild-type clinical isolate, were hyperpiliated . Ocular infectivity of these bacterial strains and mutants was examined macroscopically and histopathologically in mice and these data compared to the well-characterized ocular disease response of a murine model of infection with pseudomonas ATCC 19660 . The PAK strain was infective, but less virulent than strain 19660 by both macroscopic grading and histopathological analysis of infected eyes . Infectivity of the PR11 mutant was similar to the PAK parent strain, while PR1, DB2 and 1244, all hyperpiliated, were not infective . To explore the hypothesis that hyperpiliated bacteria bound less well to cornea and thus failed to induce corneal disease, in vitro quantitative studies of bacterial adhesion were done using an ocular organ culture model . The PR1 hyperpiliated mutant bound significantly less well to cornea than the PAK parent strain, PR11 mutant or pseudomonas 19660, while DB2 and 1244 binding did not differ significantly from 19660 or PAK . Examination of protease production, another factor which may influence adhesion, revealed that only 19660 and DB2 produced detectable protease . This study provides evidence that non-piliated, non-protease producing strains such as PAK/PR11 possess alternate virulence mechanisms to facilitate binding to and infectivity of corneal tissue. J Surg Res, 1991 Apr, 50(4), 323 - 9 CD18 adhesion receptors, tumor necrosis factor, and neutropenia during septic lung injury; Walsh CJ et al.; Sequestration of neutrophils (PMNs) in the pulmonary microvasculature and associated neutropenia are characteristic features of experimental models of septic lung injury . The etiology of altered PMN kinetics during septic lung injury is uncertain, but may be partially due to increased adhesiveness of activated PMNs to pulmonary endothelium . This study examines the relationship between the expression of PMN CD18 adhesion receptors, the evolving neutropenia, and plasma tumor necrosis factor (TNF) activity in a porcine model of septic lung injury . Acute lung injury was induced by infusion of live Pseudomonas aeruginosa (5 x 10(8) CFU/ml at 0.3 ml/20 kg/min) for 60 min (Group Ps, n = 6) . Control animals (Group C, n = 3) received a 60-min infusion of sterile 0.9% saline . CD18 expression of circulating PMNs was measured by quantitative immunofluorescent flow cytometry . Plasma TNF activity was measured by L929 fibroblast cytolytic assay . Group Ps developed a significant neutropenia by 30 min (14.9 +/- 2.5 vs 23.4 +/- 3.3 x 10(3) cells/microliter at baseline, P less than 0.05, ANOVA) with circulating neutrophils exhibiting significantly increased CD18 expression by 60 min (6.34 +/- 0.72 vs 5.01 +/- 0.52 equivalent soluble fluorescence molecules (ESFM) x 10(3) at baseline, P less than 0.05, ANOVA) . Group Ps demonstrated a significant increase in plasma TNF activity by 30 min (2.5 +/- 0.9 vs 0.7 +/- 0.3 U/ml at baseline) . There was no significant change in PMN count, PMN CD18 expression, or plasma TNF activity in Group C . In complimentary in vitro studies, porcine PMNs stimulated with recombinant human TNF-alpha (n = 5) demonstrated a time- and dose-dependent increase in CD18 expression.(ABSTRACT TRUNCATED AT 250 WORDS) Infect Immun, 1991 Apr, 59(4), 1307 - 11 Adhesion of Pseudomonas aeruginosa pilin-deficient mutants to mucin; Ramphal R et al.; Attachment of Pseudomonas aeruginosa to epithelial cells or tracheobronchial mucin is mediated by surface adhesins . Pili, composed of monomeric pilin subunits, make up one such class of adhesins . The formation of pili and flagella in P . aeruginosa is under the control of the alternative sigma factor rpoN . Isogenic mutant strains with insertionally inactivated rpoN genes were constructed with strains PAK, 1244, and CF613 and were tested for their ability to adhere to respiratory mucin . All rpoN mutants showed significant reduction of adherence to mucin relative to that of their wild-type parents . In contrast, the adherence of pilin structural gene mutants was similar to the adherence of wild types . These results provide suggestive evidence that P . aeruginosa also binds to mucin via adhesins that are distinct from pilin and are still under the genetic control of rpoN . Unlike the laboratory strain PAK, the clinical strains 1244 and CF613 are capable of agglutinating erythrocytes . The rpoN mutation had a minimal effect on the interaction of bacteria with erythrocytes, indicating that the transcription of a gene(s) specifying the agglutination phenomenon does not utilize rpoN . These findings collectively indicate the existence of several classes of adhesins on the surface of P . aeruginosa that may play an important role in colonization of the human respiratory tract. Hybridoma, 1991 Apr, 10(2), 297 - 307 Production and characterization of human monoclonal antibody recognizing the N-terminal residues of Pseudomonas aeruginosa exotoxin A; Ohtsuka H et al.; Human cell lines producing monoclonal antibodies (MAbs) against Pseudomonas aeruginosa exotoxin A were established by EBV transformation followed by cell fusion . Monoclonal antibody FK-001, IgM (mu, kappa), was demonstrated to be specifically reactive with exotoxin A in ELISA and immunoblotting, by recognizing N-terminal 16 amino acid residues of exotoxin A as an epitope . This epitope region belongs to domain I which is required for the binding of exotoxin A to the receptor on target cells . FK-001 showed a partial neutralizing activity for cell toxicity caused by exotoxin A and appeared to be effective against exotoxin A-producing P . aeruginosa infection in mice . A line of evidence suggests that monoclonal antibody FK-001 neutralizes exotoxin A-induced cell toxicity by the interference of accessibility and/or binding of exotoxin A to animal cell receptors. Appl Microbiol Biotechnol, 1991 Apr, 35(1), 23 - 31 Molecular cloning and characterization of the recA gene of Methylomonas clara and construction of recA deficient mutant; Ridder R et al.; The recA gene of the methylotrophic bacterium Methylomonas clara has been isolated from a genomic library by hybridization with the Escherichia coli recA gene . Its complete nucleotide sequence consists of 1029 bp encoding a polypeptide of 342 amino acids . Nucleotide sequence analysis of the M . clara recA gene revealed extensive homologies to recA genes from E . coli and Pseudomonas aeruginosa . Part of the physiological activity of the M . clara RecA protein has become evident in that E . coli recA mutant HB101 is complemented . The cloned recA gene has been modified in vitro by site-specific mutagenesis and by insertion of a kanamycin-resistance gene cassette into the recA coding sequence . M . clara recA mutants were obtained by replacement of the active recA gene by an in-vitro inactivated gene copy. Biochem Biophys Res Commun, 1991 Mar 29, 175(3), 1076 - 81 Glycolipid receptor binding specificity of exoenzyme S from Pseudomonas aeruginosa; Lingwood CA et al.; By use of the tlc overlay procedure we have shown that exoenzyme S extracted from cultures of Pseudomonas aeruginosa specifically binds to the glycolipids asialoGM1, asialoGM2 and to a lesser extent lactosyl ceramide . More significantly, strong binding was also observed to the glycerolipid receptor we have detected for Helicobacter pylori (Lancet ii, 238-241.1989) . Exoenzyme S can be extracted in a toxic and nontoxic form . Toxicity correlated with ability to bind the H . pylori receptor . This species was the only receptor detected in the most sensitive cell lines . The relative binding of exoenzyme S to the ganglio series glycolipids and the glycerolipid receptor was modified in a reciprocal manner in the presence of metal ions, suggesting that exoenzyme S has two interrelated receptor binding sites. J Mol Biol, 1991 Mar 20, 218(2), 349 - 64 DNA sequence of the filamentous bacteriophage Pf1; Hill DF et al.; The genome of the class II filamentous bacteriophage Pf1 has been sequenced by a combination of the chain termination and chemical degradation methods . It consists of 7349 nucleotides in a closed, circular loop of single-stranded DNA . The size and position of its open reading frames (ORFs) in general resemble those of other filamentous bacteriophage genomes . The size and position of the spaces between the ORFs have not been conserved, however, and six short reading frames (2 of which overlap) occupy a region corresponding to that filled by genes 2 and 10 in the Ff genome . Most of the ORFs are preceded by sequences resembling ribosome binding sites from the phage's host . Pseudomonas aeruginosa, that appear to differ somewhat from their counterparts in Escherichia coli . A search for sequences related to known pseudomonad promoters suggests that the promoters in this bacteriophage may well be ntr-dependent, with the two strongest preceding the gene for the major coat protein (gene 8) and another ORF (430) . Gene 8 is followed by a sequence with the properties of a rho-independent terminator of transcription, like that at the same position in the genome of Ff . The Pf1 genome contains no collection of potential stem-and-loop structures corresponding to those that initiate replication of Ff DNA and assembly of the Ff virion, although isolated structures of this kind are present . The available evidence suggests that at least 13 of the 14 major ORFs are expressed . Overall, the organization of the Pf1 genome differs from that of the other class II filamentous phage whose genome has been sequenced, Pf3, as much as it does from that of the class I phages Ff and IKe. Lancet, 1991 Mar 16, 337(8742), 631 - 4 Severity of cystic fibrosis in patients homozygous and heterozygous for delta F508 mutation; Johansen HK et al.; To assess the relation between genotype and severity of disease in cystic fibrosis (CF) the frequencies and extent of several features of its phenotypic expression were investigated in the 235 patients who attend the Danish CF Centre . 14 patients who attend irregularly and 3 who do not carry the delta F508 mutation at all were excluded . The case-reports of the remaining 218 patients (aged 4 months to 41 years) were carefully evaluated, and they were all analysed for the delta F508 mutation . 172 (79%) were homozygous for delta F508 and 46 (20%) were heterozygous . The mutation therefore occurs on 89% of the chromosomes analysed . There were no significant differences between the homozygous and heterozygous groups in the proportions with meconium ileus at birth, liver involvement, or chronic Pseudomonas aeruginosa infection . However, significantly more of the homozygous patients had onset of symptoms before the age of 6 months (p less than 0.025); they were significantly younger at diagnosis (p = 0.013) and centre referral (p = 0.006); they required greater pancreatic enzyme substitution (p = 0.0002) and had poorer lung function; and the calculated yearly incidence of chronic Ps aeruginosa infection and yearly mortality rates were greater than in heterozygous patients (p = 0.0001). FEMS Microbiol Lett, 1991 Mar 15, 63(1), 31 - 5 Cloning and characterization of a DNA fragment that complements the nfxB mutation in Pseudomonas aeruginosa PAO; Okazaki T et al.; The 2.2-kilobase DNA fragment that can complement the nfxB mutation in Pseudomonas aeruginosa PAO was cloned from chromosomal DNA of P . aeruginosa PAO1 . The nfxB mutants show an increase in resistance to quinolones and hypersusceptibility to beta-lactam and aminoglycoside antibiotics . In the mutants, a 54 kDa outer membrane protein newly appeared . The phenotype was restored to the wild-type by transformation with plasmid carrying the 2.2-kb DNA fragment. Minerva Chir, 1991 Mar 15, 46(5), 203 - 7 {Postoperative infections in otorhinolaryngologic surgery . Open study on efficacy and tolerability of antibiotic prophylactic treatment with sulbactam-ampicillin}; Borloni R et al.; The paper reports the results of an open study on the efficacy and tolerability of the association of sulbactam-ampicillin in antibiotic prophylaxis in ENT surgery . The study include 55 cases and confirmed both the total absence of major side-effects and a notable therapeutic efficacy of the drug in question . It is interesting to note the results obtained in ENT cancer surgery: out of 12 cases of partial or total laryngectomies there were 2 cases of infection in extra-operative sites (polmoniti ab ingestis from Pseudomonas aeruginosa) and none within the site of operation . Data regarding the efficacy of the drug were in line or better than previously published results and its low toxicity indicates the elective use of sulbactam/ampicillin in ENT surgical prophylaxis. J Biol Chem, 1991 Mar 15, 266(8), 4911 - 6 Immunochemical analysis of Pseudomonas aeruginosa exotoxin A . Analysis of the His426 determinant; McGowan JL et al.; This study describes a combined immunochemical and genetic approach defining a site on Pseudomonas aeruginosa exotoxin A (ETA) which is critical to the ADP-ribosyltransferase (ADPRT) activity of the toxin . The sequential epitope of a monoclonal antibody (TO-1) which binds to domain III (residues 405-613), containing the ADPRT activity of ETA, has been defined using a series of synthetic peptides . This epitope spans residues 422-432 which composes the major alpha-helical segment of domain III and includes His426 which has previously been shown to be essential for ADPRT activity (Wozniak, D.J., Hsu, L.-Y., and Galloway, D . R . (1988) Proc . Natl . Acad . Sci . U.S.A . 85, 8880-8884) . The critical His426 residue which projects into a major cleft becomes exposed when the ETA protein is in an ADPRT-active configuration . Since the TC-1 mAb does not block the binding of NAD+, it is possible that the alpha-helix site containing the TC-1 epitope and the His426 residue is associated with the interaction between ETA and its elongation factor 2 substrate. Biull Eksp Biol Med, 1991 Mar, 111(3), 285 - 7 {The role of a local infectious focus in the development of Pseudomonas aeruginosa sepsis (experimental research)}; Tepliakov VG et al.; The dynamics of sepsis development and the formation of metastatic foci have been studied using an experimental model . It has been established, that at the initial stages, the process manifests itself by bacteremia and endotoxemic shock, as well as by the complex of vascular disorders . The dependence of the formation of metastatic abscesses on the terms of the primary focus excision as well as the irreversibility of sepsis at the definite stage of infectious process have been studied . It is supposed, that the primary focus initiates the cascade activation mechanism of the factors, that determine the transition of bacteremia to the stage of septicopyemia. Clin Radiol, 1991 Mar, 43(3), 166 - 70 Computed tomography in malignant external otitis; Guy RL et al.; Malignant external otitis is a severe infection of the external auditory meatus occurring predominantly in diabetics and usually caused by Pseudomonas aeruginosa . The infection may spread along several routes: directly by bony erosion into the adjacent mastoid bone, anteriorly into the parotid gland and temporomandibular joint and inferiorly into the soft tissues of the infratemporal fossa . We present four cases of malignant external otitis that illustrate the typical patterns of spread of this disease and the role that radiology, and in particular computed tomography, plays in its diagnosis and management. Ann Thorac Surg, 1991 Mar, 51(3), 451 - 4 Hemodynamic effects of lobar pulmonary artery occlusion in a porcine sepsis model; Ward ME et al.; We induced severe pulmonary hypertension and acute lung injury in 6 pigs by Pseudomonas aeruginosa infusion . We studied the effect of pulmonary artery catheter inflation of a pulmonary artery catheter balloon in the left lower lobar pulmonary artery was accompanied by a significant (p less than 0.05, paired t test) increase in pulmonary artery pressure, a decrease in left atrial pressure, a decrease in cardiac output, and a decrease in mean arterial pressure . No significant changes occurred when the catheter was advanced into the wedged position without balloon inflation . Balloon inflation had no significant effect on these variables before bacterial infusion . We conclude that with sufficiently severe pulmonary hypertension in association with diffuse lung injury, lobar pulmonary artery occlusion may cause alterations in cardiac output and left atrial pressure . This may confuse interpretation of pulmonary artery catheter measurements. Microb Pathog, 1991 Mar, 10(3), 173 - 82 Extracellular lipase of Pseudomonas aeruginosa: biochemical characterization and effect on human neutrophil and monocyte function in vitro; Jaeger KE et al.; Lipase was isolated from P . aeruginosa by ultrafiltration of sterile-filtered culture supernatant . Gel filtration on Sepharose 4B yielded a broad peak corresponding to a molecular mass range of 100 to 1000 kDa . Electron microscopy of a negatively stained lipase preparation after Sepharose 4B chromatography revealed spherical particles with diameters ranging from 5 to 20 nm . Biochemical characterization and SDS polyacrylamide gel electrophoresis suggested that these particles consisted of protein and carbohydrate including lipopolysaccharide with the major enzyme activity being lipase . Various concentrations of this lipase preparation were preincubated with human peripheral blood neutrophils and monocytes . The chemotaxis and chemiluminescence of these cells were then determined . It was shown that lipase inhibited the monocyte chemotaxis and chemiluminescence, whereas it had no or very little effect on neutrophils . The inhibitory effect was concentration dependent and was abolished by heat treatment of the enzyme at 100 degrees C . Since monocytes are one of the important cells of the host defence system the inhibition of the function of these cells may contribute to the pathogenesis of infections caused by P . aeruginosa. Semin Respir Infect, 1991 Mar, 6(1), 11 - 8 Interaction between Pseudomonas aeruginosa and host defenses in cystic fibrosis; Marshall BC et al.; The major causes of morbidity and mortality in cystic fibrosis are chronic pulmonary obstruction and infection . Mucoid Pseudomonas aeruginosa is the primary pathogen in up to 90% of these patients . Once Pseudomonas organisms colonize the airways, they are virtually never eradicated . No defect in systemic host defense has been elucidated, however, several mechanisms contribute to the breakdown in host defenses that allow persistence of this organism in the endobronchial space . These mechanisms involve both bacterial adaptation to an unfavorable host environment and impaired host response . P aeruginosa adapts to the host by expressing excessive mucoid exopolysaccharide and a less virulent form of lipopolysaccharide . These features make it less likely to cause systemic infection, yet still enable it to resist local host defenses . Mucociliary clearance becomes impaired due to abnormal viscoelastic properties of sputum, squamous metaplasia of the respiratory epithelium, and bronchiectasis . Despite a brisk antibody response to a variety of Pseudomonas antigens, several defects in antibody-mediated opsonophagocytosis have been identified . These include (1) development of antibody isotypes that are suboptimal at promoting phagocytosis, (2) formation of immune complexes that inhibit phagocytosis, and (3) proteolytic fragmentation of immunoglobulins in the endobronchial space . Complement-mediated opsonophagocytosis is also compromised by proteolytic cleavage of complement receptors from the cell surface of neutrophils and complement opsonins from the surface of Pseudomonas . The resultant chronic inflammation and infection lead to eventual obliteration of the airways. Kansenshogaku Zasshi, 1991 Mar, 65(3), 286 - 92 {Influence of low level antibiotics on Pseudomonas aeruginosa}; Honma Y et al.; Erythromycine (EM) and chrolamphenicol (CP), the inhibitors of protein synthesis, were quantitatively examined for the growth of Pseudomonas aeruginosa EA83 and its production of extracellular proteins . The minimal inhibitory concentration (MIC) of EM for the strain was higher than 100 micrograms/ml and that of CP was 100 micrograms/ml . The growth curve of EA83 was not influenced by adding 5 micrograms/ml of EM to the broth culture, but proteolytic activity and the total protein in the culture supernate went down to 60% of the control . Number of organisms in 20 hour culture was almost constant regardless of EM concentration ranging from 0 to 50 micrograms/ml, however, the suppression of proteolytic activity and total protein in the culture supernate was seen even at 1 micrograms/ml of EM concentration . The degree of suppression was inversely proportional to EM concentration . This phenomenon was also seen in the substitution of CP for EM . All extracellular proteins separated on SDS-PAGE decreased the amount as increasing EM in the culture media . These results suggested that EM inhibited the production of not only protease including elastase, but also any other extracellular proteins including well known pathogenic factors such as exotoxin A and phospholipase C. Pathol Biol (Paris), 1991 Mar, 39(3), 171 - 6 {Determination of factors promoting, in vitro, the expression of adhesion of Pseudomonas aeruginosa to buccal cells}; Agueda L et al.; The composition of the bacterial flora in the upper respiratory tract is closely correlated with the type of pathogens recovered from the respiratory tract in patients . In intensive care patients, colonization of the oral cavity with Gram-negative organisms increases the risk of Gram-negative respiratory tract infection; the ability of bacterial cells to attach to buccal cells seems to play a central role in this correlation . Similar findings have been reported in chronic respiratory tract infections, including bronchiectasis and cystic fibrosis, with Pseudomonas aeruginosa colonization . This study was undertaken to determine the conditions best suited to in vitro detection of adhesion of P . aeruginosa to buccal cells . Use of brain-heart-infusion medium, incubation at 35 degrees C for 2 hours, and a bacterial concentration of 2 x 10(9) cells/ml were the factors correlated with improved detection of adhesion to buccal cells . Furthermore, attachment of bacteria to buccal cells was not found to vary across donors or over time in a given donor . Adhesion was independent of cell viability. J Burn Care Rehabil, 1991 Mar-Apr, 12(2), 127 - 31 Bacterial virulence and host selection: bacteria "select" patients to infect; Ward CG et al.; A clinically lethal strain of Pseudomonas aeruginosa was tested and its growth patterns in normal plasma and in normal whole blood clotted with thrombin were compared . Two stock cultures were used; one was maintained in liquid nitrogen and one was passed from plate to plate 24 times on blood agar plates at room temperature . The results showed that plasma alone and whole blood controlled the growth of a clinically pathogenic strain of P . aeruginosa consistently and uniquely for each donor, dependent on size of inoculum, length of incubation, and means by which the culture was maintained . The changing virulence of an organism and its unique growth patterns in different individuals' plasma and whole blood may explain why patients exposed to the same organisms within the same environment vary in susceptibility to clinical infection. J Burn Care Rehabil, 1991 Mar-Apr, 12(2), 106 - 15 Evaluation of a synthetic wound dressing capable of releasing silver sulfadiazine; Kuroyanagi Y et al.; A silver sulfadiazine-impregnated poly-L-leucine wound dressing, AgSD-medicated wound dressing, was evaluated for antibacterial capacity against Pseudomonas aeruginosa and cytotoxicity to human fibroblasts and human epidermal keratinocytes . This wound dressing contained 0.4 mg AgSD/cm2 . Antibacterial capacity was examined on experimentally infected wound surfaces (3.4 x 10(4) P . aeruginosa organisms/gm) on the dorsum of mice . The AgSD-medicated wound dressing showed effective bacterial control . Cytotoxicity was examined on a monolayer of cells formed in culture dishe