J Clin Invest, 1988 Sep, 82(3), 872 - 9 Functional characterization of macrophage receptors for in vitro phagocytosis of unopsonized Pseudomonas aeruginosa; Speert DP et al.; The phagocytic receptor for unopsonized Pseudomonas aeruginosa was characterized functionally using human monocyte-derived macrophages . Freshly isolated human peripheral blood monocytes were unable to ingest unopsonized P . aeruginosa; ingestion did not occur until the cells had been in culture for 2 d and it became maximal after 4 d . Macrophages plated on coverslips derivatized with anti-BSA IgG or with human gamma-globulin lost the capacity to phagocytose unopsonized P . aeruginosa, unopsonized zymosan, and EIgG but bound C3bi-coated erythrocytes normally . Each of the four human IgG subclasses and Fc fragments of anti-BSA IgG inhibited phagocytosis of both unopsonized P . aeruginosa and EIgG . Phagocytosis of P . aeruginosa and zymosan was markedly impaired and EIgG minimally inhibited if macrophages were plated on coverslips derivatized with mannan or when mannan was added to the phagocytosis buffer . Phagocytosis of P . aeruginosa and zymosan, and binding of EC3bi was dependent on the presence of divalent cations, but phagocytosis of EIgG was not . The macrophage phagocytic receptor for unopsonized P . aeruginosa was inactivated by proteolytic enzymes . Phagocytosis of P . aeruginosa was inhibited by D-mannose, L-fucose, and alpha methyl mannoside, but not by L-mannose, D-fucose, or D-glucose . The same sugars inhibited phagocytosis of unopsonized zymosan . We conclude that phagocytosis of unopsonized P . aeruginosa by human monocyte-derived macrophages is facilitated by mannose receptors. J Infect Dis, 1988 Sep, 158(3), 542 - 8 Absence of a postantibiotic effect in experimental Pseudomonas endocarditis treated with imipenem, with or without gentamicin; Hessen MT et al.; We found an in vitro postantibiotic effect (PAE) of 3-4 h for imipenem and of approximately 5 h for imipenem plus gentamicin against Pseudomonas aeruginosa . We therefore evaluated these antibiotics in a rat model of pseudomonas endocarditis . A rapid bactericidal effect was initially observed in vegetations from rats treated with imipenem alone or in combination with gentamicin . Bacterial counts rose rapidly, however, as soon as levels of imipenem in vegetations fell below the minimal inhibitory concentration (i.e., no PAE was demonstrated) . Levels of gentamicin in vegetations were similar to those that had enhanced the bactericidal effect of imipenem and had resulted in a 5-h PAE in vitro . The presence in vitro of a PAE for imipenem, with or without gentamicin, does not necessarily predict its presence in vivo in pseudomonas endocarditis in the rat. J Infect Dis, 1988 Sep, 158(3), 537 - 41 Mechanism of ciprofloxacin resistance in Pseudomonas aeruginosa; Kaatz GW et al.; We studied the mechanism(s) of resistance to ciprofloxacin arising in a clinical strain of Pseudomonas aeruginosa and in a laboratory-derived isolate of that strain . Higher concentrations of ciprofloxacin were required to interfere with DNA synthesis in resistant isolates compared with the parent strain, a finding indicating a relative insensitivity of DNA gyrase to ciprofloxacin . Reduced uptake of ciprofloxacin was seen in one isolate and may have contributed to its ciprofloxacin resistance but was not associated with alterations in outer or cytoplasmic membrane proteins, a result suggesting that such changes are not required to decrease uptake of fluoroquinolones into cells . No evidence for plasmid-mediated resistance was found, and no ciprofloxacin-inactivating activity was detected in sonic extracts of resistant organisms . In these isolates, resistance to ciprofloxacin was likely the result of more than one mutation, because single-step mutations conferring such high-level resistance were not found in the parent strain. Infect Immun, 1988 Sep, 56(9), 2515 - 7 Measurement of Pseudomonas aeruginosa phenazine pigments in sputum and assessment of their contribution to sputum sol toxicity for respiratory epithelium; Wilson R et al.; The phenazine pigments pyocyanin and 1-hydroxyphenazine were resolved by high-pressure liquid chromatography from the sputum sol phase from 9 of 13 patients with cystic fibrosis or bronchiectasis colonized by Pseudomonas aeruginosa . The concentrations measured were each sufficient to inhibit ciliary beating in vitro and contributed a significant proportion of sol phase toxicity for respiratory epithelium. Pathol Biol (Paris), 1988 Sep, 36(7), 896 - 8 {Early bacterial infections after allogenic bone marrow grafts}; Guyotat D et al.; The bacterial infections occurring during the period of neutropenia have been reviewed in a series of 100 allogeneic bone marrow transplant patients . Forty episodes of septicaemia were observed, in 37 patients, with a large majority of Gram positive organisms (thirty cases) . Only one case of major local infection occurred (Gram negative meningitis) . A non-bacterial infection was seen in 10 patients, and 36 patients presented with fever of unknown origin . Three patients who failed to engraft died of bacterial infection . Surveillance cultures showed that upper respiratory tract was a frequent site of invasion, not only for Gram positive, but also for Gram negative organisms (Pseudomonas aeruginosa). Am J Med, 1988 Sep, 85(3), 391 - 8 Malignant external otitis: insights into pathogenesis, clinical manifestations, diagnosis, and therapy; Rubin J et al.; Malignant external otitis is an infection of the external ear canal, mastoid, and base of the skull caused by Pseudomonas aeruginosa . The condition occurs primarily in elderly patients with diabetes mellitus . Current theories on pathogenesis and anatomic correlations are reviewed . Severe, unrelenting otalgia and persistent otorrhea are the symptomatic hallmarks of the disease, whereas an elevated erythrocyte sedimentation rate is the only distinctive laboratory abnormality . Iatrogenic causes such as administration of broad-spectrum antibiotics and aural irrigation may play a predisposing role in high-risk populations . The disease can result in cranial polyneuropathies (with facial nerve {VII} paralysis being the most common) and death . The mainstay of treatment is administration of antipseudomonal antibiotics for four to eight weeks . Recurrence is common, and mortality remains at about 20 percent despite antibiotic therapy . Given the increasing longevity of diabetic patients, the frequency of this disease is increasing . Internists, family practitioners, and ambulatory care physicians must now be cognizant of the presenting symptoms, while infectious disease specialists and otolaryngologists need to be appraised of strides in diagnosis and therapy . The role of surgery should be minimized . Use of new diagnostic radiologic modalities and new antipseudomonal antibiotics discussed in this review should lead to improved outcome. J Infect, 1988 Sep, 17(2), 115 - 20 The anaerobic and aerobic bacterial flora of leg ulcers in patients with sickle-cell disease; Ademiluyi SA et al.; Leg ulcers in 26 patients with sickle-cell disease (SCD) were studied bacteriologically over a period of 6 months . The average age of the patients was 20.92 years and the mean duration of the ulcers was 3.43 years . In order of frequency, Staphylococcus aureus, Pseudomonas aeruginosa, and Bacteroides melaninogenicus were the predominant organisms . Anaerobes were isolated from 14 (54%) of 26 patients and represent 21% of the total 77 isolates . The presence of anaerobes correlated well with odorous ulcers . Isolation of anaerobes from leg ulcers of patients with SCD has added to knowledge of bacterial infection in SCD. FEMS Microbiol Rev, 1988 Sep, 4(3), 223 - 37 Molecular genetic analysis of bacterial plasmid promiscuity; Krishnapillai V; The molecular genetic basis of the promiscuity of the wide host range conjugative IncP-1 alpha plasmids has been investigated by transposon mutagenesis and by the construction of minireplicons . The former has identified the origin of plasmid vegetative replication, the replication genes needed for initiation of plasmid replication, the DNA primase gene and a gene encoding a polypeptide of 52 kDa and mapping near the origin of plasmid transfer as all contributing to promiscuity . Minireplicon constructions confirm this conclusion but in addition establish that the origins of replication, transfer and other genomic regions produce complex interactions with respect to host range . DNA sequence analysis within the origin of replication show that the first direct repeat of the cluster of five repeats and sequences immediately 5' to it appear to be required in some (Escherichia coli) but not in other (Pseudomonas aeruginosa) hosts for plasmid replication. Zh Mikrobiol Epidemiol Immunobiol, 1988 Sep, (9), 86 - 9 {The search for strains producing new restriction endonucleases}; Sokolov NN et al.; The search for restrictases in 154 strains belonging to 104 species of 32 genera of microorganisms has been carried out by the method of rapid toluene assay . In 10 strains the activity of endonucleases specifically fragmenting the DNA of phage lambda in the presence of Mg2+ ions has been detected . Restrictases Pae I and Pae II formed by two Pseudomonas aeruginosa strains have been identified as the true isoschizomers of restriction endonucleases Sph I and Sma I respectively . The results of the screening of restrictase-producing strains indicate that the production of restrictases is widely spread among microorganisms of the genus Bacillus. Genetika, 1988 Sep, 24(9), 1579 - 85 {Transcription of the genome of Pseudomonas aeruginosa phage-transposon D3112 in an homologous host}; Bidnenko EM et al.; In vivo transcription of Pseudomonas aeruginosa transposable phage D3112 was studied . The 3H-labelled RNA isolated from the lysogenic cells PAO1 (D3112cts) after heat induction was hybridized with D3112 DNA to estimate phage-specific RNA . Two main stages of D3112 transcription, including transcription of the early (first 6-8 min) and the late (after 8th min) genes, were revealed . The transcription rate of D3112 early genes achieves the maximum at 3-5 min, being reduced to the minimum at 6-8 min after heat induction . These data point to the existence of negative regulation of D3112 early genes' transcription . The influence of ts mutations in early A, B and C genes and in late ts25 gene on transcription of D3112 phage was studied . It is shown that the genes A and B have no effect on the late transcription, while gene C regulates transcription of the D3112 late genes. J Bacteriol, 1988 Sep, 170(9), 4309 - 14 Molecular characterization and nucleotide sequence of the Pseudomonas aeruginosa elastase structural gene; Bever RA et al.; The elastase structural gene (lasB) from Pseudomonas aeruginosa PAO1 has been previously cloned on an 8-kilobase (kb) DNA fragment . The lasB gene, cloned in both orientations in pUC18, produced elastase in Escherichia coli, indicating that its promoter and translation initiation sites are functional in E . coli . Deletion analysis further defined the location of the lasB gene to a 3.0-kb EcoRI-KpnI fragment (pRB1803) . Elastase prepared from E . coli TB1 (pRB1803) corresponded in molecular weight to mature P . aeruginosa extracellular elastase (33,000) . The lasB gene directed the synthesis of 54- and 50-kilodalton (kDa) proteins in a bacterial cell-free transcription-translation system . The 33-, 50-, and 54-kDa proteins reacted with elastase-specific antiserum . To further characterize the lasB gene, the nucleotide sequence of the 3.0-kb EcoRI-KpnI fragment was determined . This DNA fragment contained a 1,491-base-pair open reading frame encoding 498 amino acids, corresponding to a predicted molecular weight of 53,600 . The deduced amino acid sequence contained a putative signal sequence followed by a large polypeptide which preceded the mature 33-kDa elastase protein . Three zinc ligands and an active site were predicted for the mature elastase on the basis of its amino acid sequence homology with Bacillus thermoproteolyticus thermolysin. J Clin Oncol, 1988 Sep, 6(9), 1411 - 6 High-dose chemotherapy with autologous bone marrow transplantation in 50 advanced resistant Hodgkin's disease patients: an Italian study group report; Carella AM et al.; Fifty patients with recurrent Hodgkin's disease have been treated with high-dose therapy followed by autologous bone marrow transplantation . Forty-one patients had extranodal sites of relapse and 31 patients had constitutional symptoms . Two patients had been treated with mechlorethamine, vincristine, procarbazine, and prednisone (MOPP), lomustine, vinblastine, procarbazine, and prednisone (CcVPP), and radiation; 16 patients with MOPP, doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD), radiation, and lomustine, etoposide, and prednisone (CEP); 20 patients with alternating MOPP/ABVD, and 12 patients with alternating MOPP/ABVD followed by CEP and radiation . Eighteen patients had progressive disease during alternating MOPP/ABVD protocol alone or during conventional salvage therapy; 32 patients had had a complete remission with first-line therapy but later relapsed, 25 of them having received conventional salvage therapy; 12 achieved no response or progression ("resistant-relapse" patients); and 13 responded partially or completely ("sensitive-relapse" patients) . Complete remission occurred in 24 patients (48%) with a median duration of 24 months and 16 patients (32%) achieved partial response with a median duration of 9 months, for an overall response rate of 80% . Ten patients failed to respond and died in progressive disease 1 to 10 months (median, 6 months) after transplantation . Toxicity was significant including infections (20%), liver enzymes and alkaline phosphatase elevations (100%), and carmustine lung toxicity (7%) . There were two treatment-related deaths; one patient died of Pseudomonas aeruginosa septicemia and another patient died of cerebral hemorrhage . These results validate the procedure of high-dose therapy followed by autologous bone marrow transplantation in inducing remission in these advanced, highly-treated patients . Clearly, the question of whether high-dose therapy and transplantation will eventually supersede new conventional salvage therapies will be addressed after controlled clinical studies. J Biol Chem, 1988 Aug 15, 263(23), 11291 - 5 Structure of an extracellular cross-reactive polysaccharide from Pseudomonas aeruginosa immunotype 4; Kocharova NA et al.; A neutral small molecular mass (approximately 6.5 kDa) polysaccharide comprising a pentasaccharide repeat unit was isolated from culture supernatants of Pseudomonas aeruginosa immunotype 4 . The polysaccharide had a pentasaccharide repeating unit as follows (formula; see text) where Rha is rhamnose . The structure was determined using acid hydrolysis, solvolysis with anhydrous hydrogen fluoride, methylation analysis, and 1H and 13C nuclear magnetic resonance spectroscopy including nuclear Overhauser enhancement experiments . The polysaccharide bound antibody raised to the lipopolysaccharide of the seven P . aeruginosa Fisher-Devlin immunotype strains . Inhibition assays demonstrated the presence of a serologically similar polysaccharide in supernatants of these strains . Affinity-purified antibody to the polysaccharide bound to lipopolysaccharide and whole cells of the immunotype strains of P . aeruginosa in a Western immunoblot and colony blot assay, respectively . This polysaccharide seems to contain an antigenic determinant present in the core of the P . aeruginosa lipopolysaccharide or may represent another minor polysaccharide substituent on the lipopolysaccharide in addition to the O side chain. Epidemiol Infect, 1988 Aug, 101(1), 135 - 42 Washing with contaminated bar soap is unlikely to transfer bacteria; Heinze JE et al.; Recent reports of the isolation of microorganisms from used soap bars have raised the concern that bacteria may be transferred from contaminated soap bars during handwashing . Since only one study addressing this question has been published, we developed an additional procedure to test this concern . In our new method prewashed and softened commercial deodorant soap bars (0.8% triclocarban) not active against Gram-negative bacteria were inoculated with Escherichia coli and Pseudomonas aeruginosa to give mean total survival levels of 4.4 X 10(5) c.f.u . per bar which was 70-fold higher than those reported on used soap bars . Sixteen panelists were instructed to wash with the inoculated bars using their normal handwashing procedure . After washing, none of the 16 panelists had detectable levels of either test bacterium on their hands . Thus, the results obtained using our new method were in complete agreement with those obtained with the previously published method even though the two methods differ in a number of procedural aspects . These findings, along with other published reports, show that little hazard exists in routine handwashing with previously used soap bars and support the frequent use of soap and water for handwashing to prevent the spread of disease. Plast Reconstr Surg, 1988 Aug, 82(2), 267 - 76 Suppurative chondritis: its incidence, prevention, and treatment in burn patients; Mills DC 2nd et al.; A retrospective study of 317 cases of suppurative otochondritis occurring in a population of 4794 burned patients successively admitted to one institution between 1967 and 1984 is presented . During the study interval, the incidence of the disease decreased from 20 percent to less than 3 percent in patients with burns of one or both ears . The bacterial species associated with chondritis, principally Pseudomonas aeruginosa and Staphylococcus species, did not change . Patients admitted later than the second postburn day exhibited a significantly greater incidence of chondritis . The crucial factors in prevention of this complication appear to be avoidance of pressure on burned ears and topical chemotherapeutic control of local Pseudomonas infection . Systemic antibiotic prophylaxis did not appear to influence the incidence in the studied population . Several approaches to clinical management of suppurative chondritis are reviewed. J Med Chem, 1988 Aug, 31(8), 1586 - 90 Asymmetric synthesis and properties of the enantiomers of the antibacterial agent 7-(3-aminopyrrolidin-1-yl)-1-(2,4-difluorophenyl)-1,4-dihydro-6- fluoro-4-oxo-1,8-naphthyridine-3-carboxylic acid hydrochloride; Rosen T et al.; Compound 1 {7-(3-aminopyrrolidin-1-yl)-1-(2,4-difluorophenyl)-1,4-dihydro-6-f luoro-4-oxo-1,8-naphthyridine-3-carboxylic acid hydrochloride} is a potent member of the quinolonecarboxylic acid class of antibacterial agents and is currently undergoing clinical evaluation . We have developed efficient asymmetric syntheses of the enantiomers of this agent . The S-(+) enantiomer 1a is 1-2 log2 dilutions more active than the R-(-) enantiomer 1b against aerobic bacteria and 1-2 or more log2 dilutions more active against anaerobic bacteria in vitro . The enantiomer 1a shows significantly better in vivo activity in a Pseudomonas aeruginosa mouse protection model compared to racemic 1 . Coupled with the improved solubility profile of 1a relative to racemic material, these features may be of practical significance from a clinical standpoint. Chest, 1988 Aug, 94(2 Suppl), 140S - 145S Antimicrobial therapy against Staphylococcus aureus, Pseudomonas aeruginosa, and Pseudomonas cepacia; Geddes DM; The aims of antimicrobial therapy extend beyond short-term bacterial killing to long-term maintenance of weight and lung function . A review of antimicrobial drug trials shows that empiricism is still ahead of science and more studies are needed both to justify current practice and to make future changes logical. Chest, 1988 Aug, 94(2 Suppl), 120S - 125S Pharmacokinetics of antimicrobial drugs in cystic fibrosis . Aminoglycoside antibiotics; Horrevorts AM et al.; Patients with cystic fibrosis (CF) show abnormal aminoglycoside pharmacokinetics . After a conventional dose, the serum concentrations in CF patients are lower than those in nonCF patients . The lower serum concentrations in CF might be explained by increased total body clearance and/or a larger volume of distribution . The therapeutic range of aminoglycosides is narrow due to oto- and nephrotoxicity . The changed pharmacokinetics and the narrow therapeutic range make it difficult to ensure that patients with CF are adequately and safely treated with aminoglycosides . The mode of administration of aminoglycosides influences the antibacterial effect of these agents on Pseudomonas aeruginosa and the development of possible side effects . The therapeutic implications of these facts are discussed. Pediatrics, 1988 Aug, 82(2), 223 - 8 Respiratory failure in children with acquired immunodeficiency syndrome and acquired immunodeficiency syndrome-related complex; Vernon DD et al.; Acute respiratory failure has a high mortality in patients with acquired immunodeficiency syndrome (AIDS) . This study was undertaken to determine the etiology of acute respiratory failure and the outcome of children with AIDS and AIDS-related complex . Records of 31 children with AIDS or AIDS-related complex admitted to the pediatric intensive care unit for acute respiratory failure throughout a 46-month period were reviewed . Acute respiratory failure was due to Pneumocystis carinii pneumonia in 13, cytomegalovirus pneumonia in six, bacterial pneumonia in five, severe bacterial sepsis in four, Candida pneumonia in two, and a giant cell pneumonia in one patient . In addition, 11/19 patients with acute respiratory failure due to P carinii pneumonia or cytomegalovirus had superinfections with bacteria or Candida . Of the total of 19 primary and secondary bacterial infections, Pseudomonas aeruginosa was responsible in ten and Klebsiella pneumoniae in three children . Five children (16%) survived until pediatric intensive care unit discharge; three died within 6 months . The causes of acute respiratory failure were not significantly different in survivor and nonsurvivor groups . It is concluded that, in addition to P carinii pneumonia and cytomegalovirus pneumonia, bacterial infections (especially due to Pseudomonas and other Gram-negative organisms) are important causes of respiratory failure . The high mortality and grim ultimate prognosis seen may have implications for pediatricians attempting to identify the proper limits of medical intervention for this group of patients. J Pharmacokinet Biopharm, 1988 Aug, 16(4), 355 - 75 Microbial pharmacodynamics of piperacillin in neutropenic mice of systematic infection due to Pseudomonas aeruginosa; Zhi JG et al.; Mathematical solutions for two possible pharmacodynamic interactions (linear nonsaturable and nonlinear saturable) between antibiotics and microorganisms derived from the incorporation of clinically relevant antibiotic dosage regimens such as single bolus dosing, multiple doses, and constant infusion at steady state have been obtained . It is concluded that the saturable nonlinear interaction model between the tested antibiotic and microorganism appears appropriate . The model and its derived equations are capable of describing in vivo bacterial growth of P . aeruginosa after single bolus dosing and multiple doses of piperacillin as described by a linear one-compartment pharmacokinetic model . The activity of piperacillin against P . aeruginosa in the neutropenic mouse systemic infection model can be described by an equation with three dynamic parameters: the bacterial growth rate constant kapp, 0.02345 min-1, the bacterial killing rate constant k'kill, 0.02623 min-1, and the Michaelis-Menten type saturation constant Km, 0.05467 microgram/ml . The concept and derived equations for the optimal dosing interval and minimum critical concentration are of clinical importance for the proper selection of antibiotic dosage regimens. Antimicrob Agents Chemother, 1988 Aug, 32(8), 1278 - 81 Comparative in vitro activity of PD 127,391, a new fluorinated 4-quinolone derivative; Norrby SR et al.; PD 127,391, a newly developed 4-quinolone chemically similar to ciprofloxacin, was studied in vitro by using agar and broth dilution and two inoculum sizes . PD 127,391 was found to be highly active against gram-negative aerobes, including Pseudomonas aeruginosa and other Pseudomonas spp . Its activity against gram-positive aerobes was better than that of ciprofloxacin. Zentralbl Bakteriol Mikrobiol Hyg {B}, 1988 Aug, 186(5-6), 478 - 93 {The growth and lipopolysaccharide production of Escherichia coli and Pseudomonas aeruginosa}; Weber-Frick C et al.; The course of growth and LPS production of two strains of type cultures of Escherichia coli (ATCC 11229, ATCC 25922), one E . coli mutant strain P400 and one type strain of Pseudomonas aeruginosa (ATCC 15442), grown partly by repeated cultures in BHI and partly also in minimal medium or in 1:10 diluted PC broth in a gyratory shaker (60 rpm) at 30 degrees C, was monitored respectively by counting the cfu and by simultaneous determination of LPS by means of the three miniaturized LAL-tests, i.e . the capillary test, the "Mini" endotoxin test and the Coatest endotoxin method . All three tests yielded generally comparable and reproducible results . The LPS content for a defined number of cfu was virtually in the same order of magnitude in all cases, regardless of the nutrient content of the culture medium . The quantity of LPS was relatively high in the initial phases of growth but then decreased significantly to constant levels in the stationary phase . There was a remarkable increase in the yield of LPS in the mid- and late stages of the exponential phase in the three strains, in contrast to the mutant in which the LPS content declined continuously . A possible explanation for this variation could be due to the fact that specific cell membrane proteins, which are lacking in the mutant, react differently with the LPS and thus with the Limulus Amoebocyte lysate . When the LAL tests are used for the rapid determination of the gram negative bacterial load of in particular perishable fresh foods, in which generally bacterial cells are at different stages of the exponential growth phase, then it is necessary to standardize each method specifically both for the product and for the storage conditions. J Antimicrob Chemother, 1988 Aug, 22(2), 175 - 83 Interaction of aminoglycosides and cephalosporins against Pseudomonas aeruginosa . Correlation between interaction index and killing curve; Perea EJ et al.; Three multiresistant clinical isolates of Pseudomonas aeruginosa were evaluated, by the chequerboard and the killing curve methods, for in-vitro synergy between cephalosporins and aminoglycosides . The killing-curve method was standardized to give results comparable with those obtained with the chequerboard test (FIC index) . This was achieved with combinations showing synergy by chequerboard (FIC less than or equal to 0.75) by using in the killing curves subinhibitory concentrations: half the MIC in single antibiotic assay and one eighth the MIC in the combinations . For combinations that showed indifference by chequerboard (FIC = 1) half the MIC was used for antibiotics alone and in combination . For the antagonistic combinations by chequerboard (FIC greater than 2) concentrations equal to the MIC were used with single antibiotics and combinations in the killing curve experiments . Prediction of killing curve results could then be obtained with the FIC index . The killing curve results could not be explained by pH changes or inactivation of the antibiotics. Eur J Clin Microbiol Infect Dis, 1988 Aug, 7(4), 501 - 4 Survey of blood culture isolates in an area of Sweden from 1980 to 1986; Sjoberg L et al.; To demonstrate the local epidemiological situation with respect to septicemia in an area of Sweden, a survey was made of all blood cultures performed in the county of Orebro (270,000 inhabitants) from 1980 to 1986 . From these blood cultures 4,057 organisms presumed clinically significant were isolated which gives an annual incidence of 215 isolates per 100,000 inhabitants . The two most frequently isolated species were Escherichia coli (24.6%) and Staphylococcus aureus (16.9%) . Only minor annual variations in the frequency of different species was observed . An increasing tendency in the isolation rate of Pseudomonas aeruginosa and enterococci since 1980 was noted . Comparison of the findings of the present study with those of other studies demonstrated differences between geographical areas and time-periods . Repeated monitoring of the local spectrum of organisms isolated from blood cultures and, if necessary, adjustment of the routine antibiotic regimen is thus recommended. Appl Environ Microbiol, 1988 Aug, 54(8), 1923 - 9 Conjugal transfer of R68.45 and FP5 between Pseudomonas aeruginosa strains in a freshwater environment; O'Morchoe SB et al.; Recent concern over the release of genetically engineered organisms has resulted in a need for information about the potential for gene transfer in the environment . In this study, the conjugal transfer in Pseudomonas aeruginosa of the plasmids R68.45 and FP5 was demonstrated in the freshwater environment of Fort Loudoun Resevoir, Knoxville, Tenn . When genetically well defined plasmid donor and recipient strains were introduced into test chambers suspended in Fort Loudoun Lake, transfer of both plasmids was observed . Conjugation occurred in both the presence and absence of the natural microbial community . The number of transconjugants recovered was lower when the natural community was present . Transfer of the broad-host-range plasmid R68.45 to organisms other than the introduced recipient was not observed in these chambers but was observed in laboratory simulations when an organism isolated from lakewater was used as the recipient strain . Although the plasmids transferred in laboratory studies were genetically and physically stable, a significant number of transconjugants recovered from the field trials contained deletions and other genetic rearrangements, suggesting that factors which increase gene instability are operating in the environment . The potential for conjugal transfer of genetic material must be considered in evaluating the release of any genetically engineered microorganism into a freshwater environment. J Clin Microbiol, 1988 Aug, 26(8), 1565 - 70 Prediction and diagnosis of early Pseudomonas aeruginosa infection in cystic fibrosis: a follow-up study; Brett MM et al.; Immunoglobulin G (IgG) antibodies to Pseudomonas aeruginosa surface antigens in serum were estimated by enzyme-linked immunosorbent assay for all patients from whom P . aeruginosa was isolated for the first time during a study period of 3 years (33 patients) . The titer of IgG antibodies was greater than control values at or up to 24 months before the first isolation of P . aeruginosa in 24 patients . Another five patients had titers that were within the control range before isolation of P . aeruginosa but increased to above the control range within the following 2 months . In these 29 patients, continuing intermittent isolations of P . aeruginosa were accompanied by further increases in titer . The presence of a systemic immune response above the control range indicates tissue invasion and hence infection . Four patients were deemed to have no infection: one or two isolations of P . aeruginosa were accompanied by no increase in specific antibodies to above the control range throughout the entire study period . Fifteen patients received intravenous antipseudomonal chemotherapy . Eradication of the organism and a return of titer to control values, suggesting complete removal of the organisms, occurred in 5 patients, while continued isolations and only a partial decrease in titer occurred in 10 patients . The 15 patients who received treatment improved clinically, in contrast to untreated patients, whose clinical state worsened during the study period . Continuous steroid treatment, given to two patients, was accompanied by a dramatic decrease in both serum IgG concentration and titer, despite continuing intermittent isolations of P . aeruginosa . These results confirm and extend our earlier finding that this assay appears to detect P . aeruginosa infection at a very early stage and helps in differentiating between early infection and harmless colonization . It also appears to be a useful monitor of the progress of infection and the response to intravenous antibiotic treatment in these early stages of infection, before any clinical changes are sufficiently large to be detected, in patients who were not on continuous steroid therapy . The effect of steroid treatment on the immunological response and clinical outcome of patients with early P . aeruginosa infection requires further investigation. J Bacteriol, 1988 Aug, 170(8), 3668 - 74 Heat shock response of Pseudomonas aeruginosa; Allan B et al.; The general properties of the heat shock response in Pseudomonas aeruginosa were characterized . The transfer of cells from 30 to 45 degrees C repressed the synthesis of many cellular proteins and led to the enhanced production of 17 proteins . With antibodies raised against the Escherichia coli proteins, two polypeptides of P . aeruginosa with apparent molecular weights of 76,000 and 61,000 (76K and 61K proteins) were shown to be analogous to the DnaK and GroEL heat shock proteins of E . coli due to their immunologic cross-reactivity . The major sigma factor (sigma 87) of P . aeruginosa was shown to be a heat shock protein that was immunologically related to the sigma 70 of E . coli by using polyclonal antisera . A hybridoma was produced, and the monoclonal antibody MP-S-1 was specific for the sigma 87 and did not cross-react with sigma 70 of E . coli . A smaller 40K protein was immunoprecipitated with RNA polymerase antisera from cells that had been heat shocked . The 40K protein was also associated with RNA polymerase which had been purified from heat-shocked cells and may be the heat shock sigma factor of P . aeruginosa . Exposure to ethanol resulted in the production of seven new proteins, three of which appeared to be heat shock proteins. Infect Immun, 1988 Aug, 56(8), 1873 - 9 Human monoclonal antibodies that protect mice against challenge with Pseudomonas aeruginosa; Zweerink HJ et al.; Lymphocytes from healthy volunteers and from cystic fibrosis patients were transformed with Epstein-Barr virus and cultured at a limiting dilution to generate lymphoblastoid cell lines that secreted human monoclonal antibodies specific for lipopolysaccharide (LPS) from Pseudomonas aeruginosa . Three cell lines (RM5, FDD7, and 11F9) produced immunoglobulin M (IgM) antibody species that reacted specifically with P . aeruginosa Fisher immunotypes 2, 4, and 5, respectively, and with LPS extracted from these immunotypes . A fourth cell line (9H10) produced a single IgM antibody species that recognized P . aeruginosa immunotypes 3, 6, and 7 and LPS extracted from them . Monoclonal antibodies secreted by cell lines RM5, FDD7, and 11F9 protected neutropenic mice prophylactically against challenge with P . aeruginosa immunotypes 2, 4, and 5, and those secreted by 9H10 protected against P . aeruginosa immunotypes 3 and 6 but did not protect against immunotype 7 . In vivo experiments indicated that antibodies protected mice against infection by increasing the rate of bacterial clearance. Chest, 1988 Aug, 94(2 Suppl), 109S - 115S Immunologic aspects of cystic fibrosis; Doring G et al.; Bacterial infections determine life expectancy in the hereditary disease cystic fibrosis (CF) . The dominant pathogens are Staphylococcus aureus and Pseudomonas aeruginosa, which persist in the patient's respiratory tract . Current explanations of the chronicity of the infections in the apparently immunocompetent host are based on defective opsonophagocytosis . This may be caused by (1) bacterial exopolysaccharide production, leading to cryptic infection types; (2) cleavage of immunoglobulin, complement, and surface receptors on immunocompetent cells by host proteases; and (3) a change from opsonic to nonopsonic antibody isotypes . Continuous antigenic stimulation of the immune system leads to local immune complex formation and a high chronic hypersensitivity reaction as well as to temporary immune unresponsiveness . Progressive tissue damage caused by lysosomal enzymes and oxygen radicals from polymorphonuclear leukocytes is thought to be ultimately responsible for respiratory failure and death in CF . Besides antibiotic treatment, anti-inflammatory therapy is therefore currently considered beneficial. J Antimicrob Chemother, 1988 Aug, 22(2), 237 - 47 A randomized trial of empirical antibiotic therapy with one of four beta-lactam antibiotics in combination with netilmicin in febrile neutropenic patients; Sage R et al.; Over a two year period 174 evaluable episodes of fever in neutropenic patients were treated in a randomized study comparing four beta-lactam antibiotics, each given in combination with netilmicin . Exclusions included episodes due to viral or fungal infection, and trial violations . Most patients were receiving treatment for leukaemia, including 18% undergoing bone marrow transplantation . The overall response rate (EORTC criteria) was 66%, ranging from 56% for cefoperazone to 76% for mezlocillin . Microbial documentation was obtained in 31% of episodes; Gram-positive isolates were most frequent but Pseudomonas aeruginosa was found in 18 patients . In patients with microbiologically documented infection 70% improved, overall--from 40% with cefoperazone to 80% with piperacillin (P less than 0.05) . Nephrotoxicity was seen in 6.7% and was associated with severe documented sepsis . Hypokalaemia was seen in 29% and was most marked in patients receiving ticarcillin . Rashes occurred in 6.6% overall, with no difference between the groups . Ototoxicity, shown by serial audiograms, was seen in 4.7% of patients . No evidence of vestibular dysfunction was seen in 62 patients studied . Of thirteen deaths due to the primary infection, seven were caused by Ps . aeruginosa and five by fungi. J Infect Dis, 1988 Aug, 158(2), 355 - 9 Comparative pharmacokinetics and pharmacodynamics of amikacin and ceftazidime in tricuspid and aortic vegetations in experimental Pseudomonas endocarditis; Bayer AS et al.; A factor in the higher medical cure rates for endocarditis in the right as opposed to the left side of the heart in humans may be a difference in antimicrobial pharmacokinetics within vegetations . Rabbits with combined tricuspid and aortic endocarditis due to Pseudomonas aeruginosa received single intravenous doses of either ceftazidime (50 mg/kg) or amikacin (15 or 40 mg/kg) . For each antibiotic regimen, areas under the time-concentration curves and percent vegetation penetrances were significantly greater for tricuspid than aortic vegetations (P less than .001) . Time-concentration curves for aortic vegetations paralleled those for plasma; curves for the tricuspid vegetations resembled those for subcutaneous fibrin clots . The times above the minimum bactericidal concentration for tricuspid vegetations were significantly longer than those achieved within aortic vegetations for ceftazidime (P less than .01) and amikacin at 15 mg/kg (P less than .001) . Antimicrobial pharmacokinetics and pharmacodynamics may be more favorable within tricuspid than aortic vegetations; this difference may, in part, explain more salutary outcomes in bacterial endocarditis involving the right side of the heart. Clin Invest Med, 1988 Aug, 11(4), 247 - 52 Inhibition of polymorphonuclear leukocyte chemotaxis by the mucoid exopolysaccharide of Pseudomonas aeruginosa; Stiver HG et al.; We tested the effect on human polymorphonuclear leukocyte (PMN) chemotaxis of the mucoid exopolysaccharide (MEP) elaborated by Pseudomonas aeruginosa strain P1M recovered from the sputum of a patient with cystic fibrosis (CF) . A dose-related inhibition of chemotaxis under agarose was observed when mucoid exopolysaccharide at concentrations of 0.5 to 5.0 mg/ml was incorporated into the agarose . Similar inhibition was observed with the closely related polysaccharide alginic acid, but not with the neutral polymer Ficoll . The exopolysaccharide did not bind or inactivate the chemoattractant substance, nor was it toxic to PMN's . Digestion with alginase did significantly abrogate the inhibitory effect of alginic acid, but not of MEP . There was no difference in chemotaxis in the presence of the exopolysaccharide preincubated with heat-inactivated pooled antisera from CF patients chronically colonized with mucoid P . aeruginosa compared to MEP alone, or MEP preincubated with heat-inactivated pooled normal human sera . Inhibition of chemotaxis may represent yet another pathogenetic property of P . aeruginosa mucoid exopolysaccharide. J Hosp Infect, 1988 Aug, 12 Suppl D, 55 - 60 Recent experience with Pseudomonas aeruginosa immune globulin for the treatment of experimental pneumonia; Pennington JE; A guinea-pig model of experimental Pseudomonas aeruginosa pneumonia was used to determine what factors affect the efficacy of passive immune therapy with a hyperimmune P . aeruginosa globulin (PA-IGIV) . Animals treated 2 h after infection with a single iv infusion of PA-IGIV, 500 mg kg-1, demonstrated 33% survival . Lower dosages were less effective and there were no survivors among albumin-treated controls . Treatment with PA-IGIV was effective if given 2 or 8 h after infection but not if delayed until 24 h after infection . Animals rendered neutropenic with cyclophosphamide did not survive if treated with PA-IGIV alone . However, when PA-IGIV was added to tobramycin treatment, a significant increase in survival occurred (86%) as compared with that observed with tobramycin alone (43%) (P less than 0.05) . We conclude that PA-IGIV may offer a useful therapeutic option in management of P . aeruginosa pneumonia. Invest Ophthalmol Vis Sci, 1988 Aug, 29(8), 1277 - 84 Monoclonal antibodies provide protection against ocular Pseudomonas aeruginosa infection; Moon MM et al.; A panel of well characterized monoclonal antibodies (MAbs) directed against outer membrane proteins H2, or F (porin) of Pseudomonas aeruginosa were examined to determine whether they exhibited any protective effect against subsequent ocular challenge with the bacteria topically applied to the scarified corneal surface . Mice were observed macroscopically following bacterial challenge and the degree of ocular disease graded on a scale of 0 to 4 (0, normal, fully protected cornea; 4, corneal perforation or phthisis, not protected) . Mice treated intravenously with either MAb MA1-6 (anti-H2) or MA2-10 (anti-F), or a combination of these two MAbs and MAb MA4-4 (anti-F), two hours before corneal challenge with the viable bacteria, exhibited significantly less corneal disease than mice either not treated with the MAbs, treated with MA4-4 alone or treated with MAb MA1-3 (anti-I) . The latter MAb is directed against an outer membrane epitope that is not surface exposed . Light and transmission electron microscopic histopathology also was employed and provided confirmatory evidence to support the macroscopic analyses. J Bacteriol, 1988 Aug, 170(8), 3738 - 41 Two unusual pilin sequences from different isolates of Pseudomonas aeruginosa; Pasloske BL et al.; The pilin genes of two Pseudomonas aeruginosa strains isolated from two different patients with cystic fibrosis were cloned and sequenced . The predicted protein sequences of these two pilins had several unusual features compared with other published P . aeruginosa pilin sequences. Antimicrob Agents Chemother, 1988 Aug, 32(8), 1267 - 8 Selective imipenem resistance in Pseudomonas aeruginosa associated with diminished outer membrane permeability; Studemeister AE et al.; The permeability of the outer membranes of imipenem-susceptible and imipenem-resistant clinical isolates of Pseudomonas aeruginosa was investigated by the liposome swelling assay . Sugars and cephaloridine penetrated rapidly, whereas imipenem penetrated poorly into liposomes constructed from porin-rich outer membrane fractions of the resistant isolates. Am J Med, 1988 Jul 25, 85(1A), 56 - 8 Effect of dose and schedule on cefoperazone pharmacodynamics in an in vitro model of infection in a neutropenic host; Zinner SH et al.; Previous studies have shown that cefoperazone given in frequent, large doses is effective in the treatment of infection in patients with cancer . The pharmacodynamics of 2- and 4-g doses of cefoperazone administered either as a single dose or at 12-hour intervals were studied in an in vitro model that simulates infection in a neutropenic patient . One strain each of Pseudomonas aeruginosa (minimal inhibitory concentration {MIC} = 2 micrograms/ml), Staphylococcus aureus (MIC = 1 microgram/ml), Escherichia coli (MIC = 0.06 micrograms/ml), and Klebsiella pneumoniae (MIC = 0.25 micrograms/ml) was studied . The initial dose reduced the inoculum by approximately 3 logs for the Pseudomonas and the staphylococci and 3 to 5 logs for the other organisms . No significant differences in killing were found between the 2- and 4-g doses . Regrowth of Pseudomonas and staphylococci occurred with the single dose but not with the every-12-hour regimen . These data support the clinical use of cefoperazone in doses every 12 hours. Am J Med, 1988 Jul 25, 85(1A), 17 - 20 Comparison of cefoperazone and mezlocillin with imipenem as empiric therapy in febrile neutropenic cancer patients; Mortimer J et al.; Seventy-eight patients with cancer experienced 88 episodes of fever while neutropenic and were randomly assigned to receive empiric antibiotic therapy with cefoperazone 2 g intravenously every 12 hours and mezlocillin 4 g intravenously every six hours or imipenem/cilastatin 500 mg intravenously over 30 to 60 minutes every six hours . Within 96 hours of starting antibiotic treatment, 24 patients (57 percent) treated with cefoperazone and mezlocillin and 34 patients (74 percent) receiving imipenem/cilastatin became afebrile . One half of the patients in each arm required changes in the antibiotic regimen because of side effects, persistent fever with a site suspicious for infection, resistant organisms, or breakthrough bacteremias . Forty patients (95 percent) receiving cefoperazone and mezlocillin and 43 patients (93 percent) receiving imipenem/cilastatin recovered from the neutropenic episode . Two patients in each regimen group died of their underlying disease . One patient in the imipenem/cilastatin arm died of Pseudomonas aeruginosa sepsis . Although the two regimens are comparable in efficacy, the incidence of side effects favored the cefoperazone and mezlocillin group . No seizures or bleeding were seen in either arm; however, 19 patients (41 percent) receiving imipenem/cilastatin required pretreatment antiemetic drugs for nausea. Biochem J, 1988 Jul 15, 253(2), 323 - 8 Imipenem as substrate and inhibitor of beta-lactamases; Monks J et al.; The interaction between imipenem, a carbapenem antibiotic, and two representative beta-lactamases has been studied . The first enzyme was beta-lactamase I, a class-A beta-lactamase from Bacillus cereus; imipenem behaved as a slow substrate (kcat . 6.7 min-1, Km 0.4 mM at 30 degrees C and at pH 7) that reacted by a branched pathway . There was transient formation of an altered species formed in a reversible reaction; this species was probably an acyl-enzyme in a slightly altered, but considerably more labile, conformation . The kinetics of the reaction were investigated by measuring both the concentration of the substrate and the activity of the enzyme, which fell and then rose again more slowly . The second enzyme was the chromosomal class-C beta-lactamase from Pseudomonas aeruginosa; imipenem was a substrate with a low kcat . (0.8 min-1) and a low Km (0.7 microM) . Possible implications for the clinical use of imipenem are considered. Am J Ophthalmol, 1988 Jul 15, 106(1), 77 - 81 Topical imipenem therapy of aminoglycoside-resistant Pseudomonas keratitis in rabbits; Sawusch MR et al.; We used a rabbit model of bacterial keratitis to assess in vivo efficacy of topical imipenem, a highly potent beta-lactam antibiotic with an unusually broad spectrum of activity, including aminoglycoside-resistant Pseudomonas aeruginosa . Albino rabbits received intrastromal injections of 5 x 10(2) organisms of an aminoglycoside-resistant strain of P . aeruginosa . At five hours postinoculation, imipenem (5 mg/ml) therapy was initiated using one drop per 30 minutes for 12 hours . Corneal tissue was then excised for colony forming unit counts . Imipenem was highly effective in reducing colony forming unit counts to zero in comparison to 4.1 x 10(5) organisms for untreated controls . A second regimen beginning 24 hours postinoculation of one drop per hour for 24 hours was also successful in significantly reducing colony forming units vs controls (P less than .05) . These data suggest that topical imipenem may have clinical applicability in the treatment of P . aeruginosa keratitis. J Laryngol Otol, 1988 Jul, 102(7), 606 - 7 Necrotizing external otitis treated with ciprofloxacin . A case report; Sabater F et al.; Necrotizing (malignant) external otitis is a severe infection caused by Pseudomonas aeruginosa which occurs mainly in elderly diabetics or in immuno-depressed patients (Chandler, 1968) . The management of this condition requires the association between an aminoglycoside antibiotic and an antipseudomonal beta-lactamic, given parenterally during a 4 to 6 week period . Sometimes it is necessary to continue the therapy for months until there is no evidence of residual disease (Strauss et al., 1982) . Ciprofloxacin is a quinolone with antipseudomonal activity which can be taken orally, and it is a useful alternative to the current treatment . The authors report a case of necrotizing external otitis which was successfully treated with ciprofloxacin. J Laryngol Otol, 1988 Jul, 102(7), 579 - 81 Pseudomonas vaccination in chronic ear disease; Kent SE et al.; This paper presents the results of a pilot study of vaccination in thirteen patients with chronically infected mastoid cavities or central perforations of the tympanic membrane . In each patient Pseudomonas aeruginosa was repeatedly cultured from the ear and a long list of medical and surgical treatments had been tried, without success, to control the infection . The clinical, bacteriological and immunological responses were monitored over a six-month period . The results are extremely encouraging . Eight patients' ears became dry for the first time in many years . However, three months after vaccination three ears had become reinfected . All of the patients except one had a good immunological response to vaccination, with many-fold increases in the total serum immunoglobulin G, A and M levels. J Infect Dis, 1988 Jul, 158(1), 7 - 12 Single, large, daily dosing versus intermittent dosing of tobramycin for treating experimental pseudomonas pneumonia; Kapusnik JE et al.; Single, large, daily aminoglycoside doses in animals are less toxic than conventional dosing, and higher drug concentrations in vitro produce more-rapid bacterial killing . Thus, we compared various aminoglycoside dosing schedules in neutropenic (n = 153) and nonneutropenic (n = 192) guinea pigs with Pseudomonas aeruginosa pneumonia . Equivalent tobramycin dosages were given: 5 mg/kg every 4 h or 30 mg/kg every 24 h . Animals were serially killed during therapy, and quantitative lung cultures were performed . Bacterial titers in lungs dropped rapidly in all tobramycin-treated animals, both neutropenic and nonneutropenic, during the initial 16 h of therapy . In nonneutropenic guinea pigs, lung titers remained constant despite continued 4-h dosing . With subsequent 24-h dosing, titers continued to drop, and by 72 h there were a significant number of animals with sterile lungs (P less than .01) . In neutropenic guinea pigs given tobramycin every 24 h, bacterial regrowth occurred; thus, therapy was ineffective . Adding mezlocillin, however, suppressed regrowth; thus, combination therapy was superior (P less than .05). J Infect Dis, 1988 Jul, 158(1), 89 - 100 Fibronectin-cleaving activity in bronchial secretions of patients with cystic fibrosis; Suter S et al.; In cystic fibrosis, colonization of the airways with Pseudomonas aeruginosa follows colonization with Staphylococcus aureus and is related to accelerated deterioration of pulmonary function . Because P . aeruginosa adheres better to cell surfaces devoid of fibronectin, we searched for fibronectin-cleaving activity in bronchial secretions and saliva from 24 patients with cystic fibrosis who were followed up for 4.5 y and from two control groups . Proteolytic activity against 125I-labeled fibronectin was continuously present in cystic fibrosis bronchial secretions; significantly higher fibronectin-cleaving activity was found in older vs . younger patients, in patients with advanced disease stages determined by a five-stage scoring system, and in those colonized with P . aeruginosa . The fibronectin-cleaving activity was due to neutrophil elastase and cathepsin G . Cystic fibrosis bronchial secretions had proteolytic activity against surface fibronectin of airway mucosal cells . Thus fibronectin-cleaving activity of bronchial secretions rather than of saliva may favor P . aeruginosa colonization of the upper respiratory tract in individuals with cystic fibrosis. CLAO J, 1988 Jul-Sep, 14(3), 163 - 4 Delayed microbial keratitis following radial keratotomy; Insler MS et al.; A 21 year old man developed Pseudomonas aeruginosa keratitis 23 months after a reoperation for radial keratotomy . The ulcer developed in one of the inferior incision sites, presumably as a result of poor epithelial wound healing . Prompt medical therapy with fortified antibiotics led to normal visual recovery. CLAO J, 1988 Jul-Sep, 14(3), 148 - 50 True prophylactic gentamicin application in experimental Pseudomonas keratitis; Insler MS et al.; In an experimental model of Pseudomonas keratitis, 50 rabbit eyes were treated with gentamicin sulfate (3 mg/mL) prophylactically on four different treatment regimens . One group (13 eyes) received one drop of gentamicin sulfate every hour to a total of four drops, the last drop 25 minutes before inoculation . A second group (13 eyes) was given one drop of gentamicin sulfate one hour before inoculation . A third group (12 eyes) received one drop of gentamicin sulfate per minute to a total of 4 drops, the last drop 25 minutes prior to inoculation . A fourth group (12 eyes) received one drop of antibiotic 25 minutes before inoculation . Twelve control eyes received saline solution . Subsequently, a superficial corneal scratch was inflicted and each eye received one drop (0.05 mL) of a solution containing Pseudomonas aeruginosa . The infection rate in all four experimental groups was low, whereas all control eyes became infected . These results demonstrate the effectiveness of prophylactic antibiotic application in the prevention of Pseudomonas keratitis prior to superficial ocular trauma. FEMS Microbiol Immunol, 1988 Jul, 1(2), 97 - 102 Structural relationship between the polyagglutinable antigen and the core polysaccharide of Pseudomonas aeruginosa; Marx A et al.; It has been observed that each strain of the Pseudomonas aeruginosa species harbours the so-called polyagglutinable antigen (PA) . Some strains may produce it in a form which is linked to the core moiety of lipopolysaccharide (LPS) and this type of PA can thus be detected by passive haemagglutination using the isolated LPS as coating antigen . Other strains synthesize PA exclusively in a free form, which is also coextractable with LPS, its presence can, however, be demonstrated by the haemagglutination inhibition test . From a polyagglutinable strain of P . aeruginosa an R-type LPS was isolated having the core-linked PA . This LPS preparation was highly immunogenic with regard to its PA moiety . The core-bound PA seems to exert an immunosuppression on the core region, hence, the polyagglutinable strains isolated from cystic fibrosis patients only engender anti-PA antibodies, whereas antibodies against both, side chain and core region of LPS, are not engendered . The mucoid exopolysaccharide also contains the PA which could possibly play an important role in the patient by protecting P . aeruginosa cells against anti-PA antibodies. FEMS Microbiol Immunol, 1988 Jul, 1(2), 103 - 7 Antigenicity and immunogenicity of the polyagglutinable antigen of Pseudomonas aeruginosa; Marx A et al.; Pseudomonas aeruginosa strains isolated from cystic fibrosis patients agglutinate in antisera against anti-polyagglutinable antigen (PA) . Anti-PA antibodies were formed in rabbits when immunization was carried out with bacteria possessing core-bound PA, independently of whether the strains were of S or R phenotype . For bacterial agglutination with anti-PA antibodies two prerequisites are essential: the bacterial cell must be of R phenotype and must possess the core-linked PA . In contrast, the PA in the isolated LPS's can be demonstrated in passive haemagglutination for both (S or R) phenotypes, provided the PA is core-linked . Two PA forms have been recognized, one found only in P . aeruginosa species, both in free and bound form . The other one is shared by all members of Pseudomonas genus but is present only in a free, unbound form. ASAIO Trans, 1988 Jul-Sep, 34(3), 573 - 7 Enhanced bacterial adhesion on surfaces pretreated with fibrinogen and fibronectin; Mohammad SF et al.; The effect of certain plasma proteins on the adhesion of Pseudomonas aeruginosa and Staphylococcus epidermidis on polyurethane, polyvinylchloride, or glass was investigated . Test surfaces were treated with serum, plasma, albumin, immunoglobulin G, fibrinogen, or fibronectin . Using a specially designed test chamber, surfaces previously treated with test proteins were incubated with bacterial suspension . During the experiment, the test chamber was placed on a rotator to prevent settling of bacteria . At the end of the experiment, each test well was rinsed repeatedly to remove non-adherent bacteria . The number of bacteria adherent to the test surfaces was quantitated by a combination of methods including microscopic counting of cells, scintillation counting and autoradiography . It was noted that a greater number of bacteria adhered to surfaces coated with fibrinogen or fibronectin whereas surfaces treated with serum showed reduced bacterial adhesion . The inhibitory effect of serum appeared more pronounced with S . epidermidis when compared with P . aeruginosa under identical experimental conditions . Scanning electron microscopy revealed that adherent bacteria were randomly distributed on the test surfaces and appeared to replicate while still adherent . These observations suggested that bacterial adhesion to biomaterials can be significantly influenced by the composition of the adsorbed proteins at the interface. Antimicrob Agents Chemother, 1988 Jul, 32(7), 978 - 81 Iontophoretic application of tobramycin to uninfected and Pseudomonas aeruginosa-infected rabbit corneas; Hobden JA et al.; Pseudomonas aeruginosa keratitis was induced in rabbits to study the effects of corneal infection on the delivery of tobramycin by iontophoresis . Some rabbits were treated by use of an eye cup with no current as a control for iontophoresis, and others were treated with fortified drops (1.36%) delivered topically for comparison with results of earlier studies . One hour after treatment with tobramycin, the concentration of drug in the infected corneas was compared with that achieved in mock-infected and uninfected eyes . Iontophoresis of 25 mg of tobramycin per ml at 0.8 mA for 10 min delivered significantly more drug (P = 0.0001) to corneal tissue than did drops or use of an eye cup without current in P . aeruginosa-infected eyes mock-infected eyes, or uninfected eyes . Tobramycin concentrations in the infected corneas (605.9 micrograms/g) were not significantly different (P = 0.815) from the concentrations in mock-infected eyes (641.4 micrograms/g), but were lower (P = 0.007) than those obtained by iontophoresis in uninfected corneas (853.6 micrograms/g) . Use of an eye cup without current delivered tobramycin equally to infected, mock-infected, and normal eyes, i.e., 176.5, 171.0, and 163.1 micrograms/g, respectively (P greater than 0.709) . Tobramycin delivered by use of fortified drops delivered topically was detectable in mock-infected corneas (20 micrograms/g) and P . aeruginosa-infected corneas (6.0 micrograms/g) . These results suggest that iontophoresis has value as an ocular drug delivery system and that an eye cup could also be useful in a therapeutic regimen for ocular infections. Rev Infect Dis, 1988 Jul-Aug, 10 Suppl 2, S337 - 40 Polysaccharide antigens of Pseudomonas aeruginosa; Pier GB; Cell-surface polysaccharide antigens of Pseudomonas aeruginosa are important determinants of pathogenicity and represent target antigens for the development of vaccines . Two major and distinct cell-surface polysaccharides are expressed by different strains of P . aeruginosa: the serotype determinant located on the O polysaccharide side chain of the lipopolysaccharide and mucoid exopolysaccharide (MEP), an antigen usually made by isolates of P . aeruginosa from patients with cystic fibrosis . The serotype determinant has been isolated in a high-molecular-weight, non-toxic, immunogenic form known as high-molecular-weight polysaccharide . High-molecular-weight polysaccharide is safe and immunogenic in humans and elicits T cell immunity to P . aeruginosa infection in animals . Purified MEP antigen has been shown to be immunogenic in animals, and patients with cystic fibrosis colonized with mucoid P . aeruginosa respond to MEP with the production of high-titered serum antibody . In spite of brisk immune responses of patients with cystic fibrosis to MEP, they are unable to deal with the chronic respiratory colonization by mucoid P . aeruginosa . An association has been established between the presence of opsonic-killing antibody to MEP and the lack of colonization of older patients with cystic fibrosis with P . aeruginosa. Rev Infect Dis, 1988 Jul-Aug, 10(4), 892 - 8 Resistance to imipenem in Pseudomonas aeruginosa: clinical experience and biochemical mechanisms; Quinn JP et al.; Emergence of resistance to imipenem during therapy for Pseudomonas aeruginosa infections is common and may result in treatment failure . Resistance emerges most often during therapy for lower respiratory tract infections . There are several unique features of this resistance to imipenem . First, cross-resistance to other beta-lactam agents is not observed . Second, the mechanism of resistance in most of the isolates studied to date appears to be related to a selective permeability barrier across the bacterial outer membrane, usually associated with discrete alterations in the electrophoretic profiles of outer membrane proteins . These data suggest that imipenem may traverse the outer membrane of P . aeruginosa via a specific porin protein that is not critical for penetration of other beta-lactam antibiotics. Rev Infect Dis, 1988 Jul-Aug, 10(4), 691 - 8 Permeation of beta-lactam antibiotics into Escherichia coli, Pseudomonas aeruginosa, and other gram-negative bacteria; Livermore DM; Cell wall impermeability is a major determinant of the susceptibility of gram-negative bacilli to beta-lactam antibiotics . The outer membrane, which beta-lactam agents cross via pores composed of porin proteins, is the major individual barrier in the wall structure but does not of itself exclude these antibiotics . Rather, it slows their influx to a level that the periplasmic clearance mechanisms may manage to contain . The clearance mechanisms include hydrolysis and perhaps covalent binding by beta-lactamases and nonessential penicillin-binding proteins . The balance between uptake and clearance determines the fate of the cell, rather than one or the other factor alone . It is possible to represent this interplay mathematically for Escherichia coli, but Pseudomonas aeruginosa presents a more ambivalent picture . Moreover, the relations among porin quantity, permeability, and resistance are much better established for E . coli than for P . aeruginosa, and the possible existence of additional barrier layers--besides the outer membrane--in the latter species cannot be excluded. J Antimicrob Chemother, 1988 Jul, 22(1), 41 - 50 Synergy of ciprofloxacin with fosfomycin in vitro against Pseudomonas isolates from patients with cystic fibrosis; Figueredo VM et al.; Pseudomonas aeruginosa remains the most common respiratory pathogen causing morbidity and mortality in cystic fibrosis (CF) patients . The in-vitro activity of ciprofloxacin and fosfomycin (calcium and tromethamine salts) in combination against P . aeruginosa isolates from CF patients, all of whom had received previous courses of ciprofloxacin, was evaluated by agar plate dilution chequerboard technique . The concentrations of drugs used were those that would be achieved in patients by oral and intravenous (fosfomycin only) routes . Synergy, taking into account fosfomycin concentrations achievable intravenously, was found against 60% of P . aeruginosa isolates . With concentrations achieved after an oral dose, 17% of isolates were synergistically inhibited . Time-kill experiments confirmed these findings . Ciprofloxacin MICs against resistant P . aeruginosa were reduced to achievable sputum and serum levels in the presence of fosfomycin . However, in progressive resistance studies fosfomycin failed to delay the development of resistance to ciprofloxacin . The combinations of ofloxacin/fosfomycin and ciprofloxacin/fosmidomycin were also tested, but showed minimal synergy . No antagonism was observed with any combination . Fosfomycin, ciprofloxacin and azlocillin in triple combination did not show synergy . The antimicrobial combinations tested against P . cepacia isolates from CF patients were indifferent . The combination of ciprofloxacin and fosfomycin may be clinically useful in selected P . aeruginosa pulmonary exacerbations in cystic fibrosis patients, particularly as an oral out-of-hospital treatment alternative or in cases where MICs for ciprofloxacin are elevated. J Antimicrob Chemother, 1988 Jul, 22(1), 23 - 33 Postantibiotic effect of aminoglycosides on gram-negative bacteria evaluated by a new method; Isaksson B et al.; The in-vitro postantibiotic effect (PAE) of amikacin, gentamicin, netilmicin and tobramycin was investigated by a bioluminescent assay of bacterial ATP . Two strains each of Escherichia coli and Pseudomonas aeruginosa were exposed for 1 h to different concentrations of the aminoglycosides . The aminoglycoside was removed by a 10(-3) dilution, and regrowth of bacteria was followed at hourly intervals by monitoring bacterial ATP . This method simplified the PAE studies and made such studies possible at high aminoglycoside concentrations . The length of the PAE was dose-dependent for all the aminoglycosides studied . The PAEs ranged between three and seven hours for all four strains at the aminoglycoside concentrations normally reached in serum during standard dosing . The long PAE of aminoglycosides, especially after exposure to high drug concentrations, constitutes an argument in favour of administering aminoglycosides in higher-than-usual doses with longer intervals between doses . This proposal is also supported by recent pharmacokinetic, bacteriological and toxicity data. J Antimicrob Chemother, 1988 Jul, 22(1), 15 - 22 Comparative study on antagonistic effects of low pH and cation supplementation on in-vitro activity of quinolones and aminoglycosides against Pseudomonas aeruginosa; Blaser J et al.; The antagonistic effects of physiological levels of Ca++ and Mg++ on the in-vitro activity of aminoglycosides and quinolones against Pseudomonas aeruginosa were studied at both pH 7.4 and 5.5 . Adding Mg++ and Ca++ (100 mg/l) to commercial media deficient of these cations increased the MICs and MBCs of ciprofloxacin and enoxacin four-fold (2 P less than 0.01), which was significantly less than the 16-fold increase found for gentamicin and netilmicin (2 P less than 0.01) . However, the activity of both aminoglycosides and quinolones was similarly affected by reducing the pH to 5.5 (giving eight-fold increases in MICs) or by the combination of both low pH plus cation supplementation (giving 16-fold increases in MICs) . These data raise the question whether antagonizing factors should be considered not only for aminoglycosides, but also for quinolones during routine susceptibility tests on P . aeruginosa. Acta Paediatr Scand, 1988 Jul, 77(4), 576 - 82 Increased IgG2 and IgG3 concentration is associated with advanced Pseudomonas aeruginosa infection and poor pulmonary function in cystic fibrosis; Pressler T et al.; The concentrations of IgG subclass immunoglobulins were determined by radial immunodiffusion in serum from 126 patients with cystic fibrosis (CF) . The results were compared to values from age-matched healthy children and adults and correlated to patients age, duration of chronic Pseudomonas aeruginosa infection and lung function parameters . Fifty-two percent of the patients had an elevated concentration of at least one of the IgG subclasses; IgG1 28%, IgG2 16%, IgG3 18% and IgG4 48% . There was significant correlation between elevated serum levels of IgG2, and to a lesser extent IgG3, with decreased lung function (for FEV1; p = 0.0001, and p = 0.001 respectively) and high levels of antipseudomonas precipitins (p = 0.008, and p = 0.002) . A similar correlation was not found for IgG1 and IgG4 . IgG subclasses vary in their ability to promote phagocytosis and to activate complement and it is possible that individual differences in the IgG subclass pattern could explain the variable course of this disease. J Infect Dis, 1988 Jul, 158(1), 13 - 22 The influence of tobramycin dosage regimens on nephrotoxicity, ototoxicity, and antibacterial efficacy in a rat model of subcutaneous abscess; Wood CA et al.; The influence of dosage regimen on the nephrotoxicity, ototoxicity, and antibacterial efficacy of tobramycin was assessed in Fisher rats with Pseudomonas aeruginosa subcutaneous abscesses . A subcutaneous tobramycin dose of 10 mg/kg every 4 h resulted in peak and trough serum concentrations that approximated those currently recommended for patients . Subsequently, the influence of this subcutaneous dosage regimen was compared with three other regimens that administered the same total daily dose: 20 mg every 8 h, 30 mg every 12 h, and 60 mg every 24 h . Renal injury was assessed by measuring inulin clearance and in vivo synthesis of renal DNA and by histopathology . Cochlear histology was also assessed . The number of P . aeruginosa per abscess was quantitated . In animals with infected abscesses, there was a consistent trend of greater kidney injury with the more-frequent dosage regimens . There was no evidence of cochlear toxicity in any group . All regimens were equally effective in reducing the number of P . aeruginosa in subcutaneous abscesses. Infect Immun, 1988 Jul, 56(7), 1673 - 7 Inhibition of human lymphocyte proliferation and cleavage of interleukin-2 by Pseudomonas aeruginosa proteases; Theander TG et al.; This study was undertaken to determine the effect of Pseudomonas aeruginosa alkaline protease (AP) and elastase (ELA) on human lymphocyte function . AP at 50 micrograms/ml and ELA at 12 micrograms/ml caused a 50% inhibition of phytohemagglutinin-induced proliferation . There was no difference in the effect of proteases on CD4- and CD8-positive cells . To determine the effect of proteases on interleukin-2 (IL-2)-induced cell proliferation, the proteases and IL-2 were added to the IL-2-dependent CTLL-2 cell line . AP and ELA inhibited the proliferation of these cells . When IL-2 was added in excess, the inhibition was partly reversed . ELA at 10 micrograms/ml cleaved IL-2, as judged by size chromatography of a reaction mixture containing 125I-labeled IL-2 and the proteases . The ELA-digested IL-2 exhibited a reduced binding capacity to IL-2 receptors on the lymphocytes . Furthermore, treatment of phytohemagglutinin-stimulated lymphocytes with AP and ELA resulted in inhibition of binding of intact IL-2 to IL-2 receptors on the stimulated lymphocytes . These results indicated that P . aeruginosa-derived enzymes are able to interfere with human lymphocyte function in vitro and that this effect might be due to cleavage of IL-2. J Antimicrob Chemother, 1988 Jul, 22 Suppl A, 1 - 15 General mechanisms of resistance to antibiotics; Bryan LE; Resistance to antimicrobial agents may result from intrinsic properties of organisms, through mutation and through plasmid- and transposon-specified genes . beta-Lactam resistance is most frequently associated with one or more chromosomal- or plasmid-specified beta-lactamases . Recently, mutations modifying penicillin-binding proteins have been detected with increased frequency as a cause of beta-lactam resistance . Mixed mechanisms, reduced permeability and tolerance are other causes of resistance . Aminoglycoside resistance always involves some modification of drug uptake, most often due to a variety of enzymes modifying these compounds . Reduced uptake is a primary cause of resistance in anaerobic bacteria and bacteria growing anaerobically, some strains of Pseudomonas aeruginosa, and mutants that arise during antimicrobial therapy and are defective in energy-generation systems . Resistance to other antimicrobial agents is presented in tabular form. J Hosp Infect, 1988 Jul, 12(1), 43 - 9 The effects of antiperspirant on the perineal skin flora of patients with spinal cord injury; Montgomerie JZ et al.; The relationship of pH and moisture to Pseudomonas aeruginosa and Klebsiella pneumoniae colonization of the perineal skin was studied in male patients with spinal cord injury . The increased pH of the perineal skin was significantly associated with the presence of P . aeruginosa but not other bacterial species . No correlation between colonization and moisture or pH and moisture was found . An antiperspirant produced a significant reduction in the number of total aerobic bacteria, total gram-negative bacilli, P . aeruginosa and K . pneumoniae over a 24-h period . Long-term use of the antiperspirant for 10 days did not alter the persistence of P . aeruginosa of the same serotypes on the perineum. Mol Microbiol, 1988 Jul, 2(4), 489 - 95 The expression of mutant pilins in Pseudomonas aeruginosa: fifth position glutamate affects pilin methylation; Pasloske BL et al.; The expression within Pseudomonas aeruginosa PAO1 of three mutant pilin genes from P . aeruginosa PAK was studied to determine their effects on pilin stability, translocation into the membrane, leader peptide removal, and methylation of the mature N-terminal phenylalanine . The results revealed that a deletion of 4 or 8 amino acids within the immediate N-terminus of pilin had deleterious effects upon leader peptide cleavage . In addition, while the 4-amino-acid deletion did not affect pilin partitioning into the membrane, the 8-amino-acid deletion decreased the amount of pilin found within the membrane fraction . Of considerable interest was the finding that the mutation within the mature pilin of the glutamate at position 5 to a lysine did not prevent leader peptide removal but did inhibit the methylation of the N-terminal phenylalanine. J Bacteriol, 1988 Jul, 170(7), 3228 - 36 Use of a gene replacement cosmid vector for cloning alginate conversion genes from mucoid and nonmucoid Pseudomonas aeruginosa strains: algS controls expression of algT; Flynn JL et al.; Pseudomonas aeruginosa can convert to a mucoid colony morphology by a genetic mechanism called alginate conversion; this results in the production of copious amounts of the exopolysaccharide alginate . The mucoid phenotype of P . aeruginosa is commonly associated with its ability to cause chronic pulmonary tract infections in patients with cystic fibrosis . In this study we isolated the cis-acting locus involved in alginate conversion, called algS, from both mucoid and nonmucoid isogenic strains . We then examined the role of algS in the control of algT, a trans-active gene required for alginate production in P . aeruginosa . We used a new cosmid cloning vector, called pEMR2, that permitted both the cloning of large DNA fragments and their subsequent gene replacement in P . aeruginosa . To verify the predicted properties of this vector, we isolated and tested a pEMR2 hisI+ clone . Using cloned algS-containing DNA and a method for gene replacement, we constructed isogenic strains of P . aeruginosa that had Tn501 adjacent to algS on the chromosome . Two pEMR2 clone banks containing genomic fragments from isogenic algS(On) (exhibiting the alginate production phenotype) and algS(Off) (exhibiting the non-alginate production phenotype) strains were constructed, and Tn501 served as an adjacent marker to select for clones containing the respective algS allele . The pEMR2 algS(On) and pEMR2 algS(Off) clones were shown to contain the indicated algS allele by gene replacement with the chromosome of strains that carried the opposite allele . To test whether algS controls the expression of the adjacent algT gene, we constructed a pLAFR1 algS(Off)T clone and showed it to be unable to complement an algT::Tn501 mutation in trans . In contrast, a pLAFR1 algS(On)T clone did complement algT::Tn501 in trans . Thus, algS appears to control the activation of algT expression, bringing about alginate conversion. Rev Infect Dis, 1988 Jul-Aug, 10(4), 770 - 5 Roles of porin and beta-lactamase in beta-lactam resistance of Pseudomonas aeruginosa; Hancock RE et al.; Pseudomonas aeruginosa demonstrates high intrinsic resistance to most beta-lactam antibiotics . Two factors that are interrelated appear to be important in this intrinsic resistance: an inducible, chromosomally encoded type Id beta-lactamase and low outer-membrane permeability . beta-Lactamase-noninducible mutants are supersusceptible to many beta-lactam agents, whereas constitutively derepressed mutants are considerably more resistant even to so-called beta-lactamase-stable beta-lactams . For the latter mutants, by analysis of kinetics, it can be demonstrated that synergy between slow permeation across the outer membrane and slow hydrolysis of the beta-lactamase-stable beta-lactams can explain resistance . Wild-type P . aeruginosa allows outer membrane permeation of beta-lactam agents at rates 1%-8% of those measured for Escherichia coli . The majority of trans-outer-membrane channels formed by P . aeruginosa porin protein F are too small to allow passage of beta-lactam antibiotics . Nevertheless, this porin is apparently a conduit for beta-lactams, since protein F-deficient mutants have small changes in susceptibility to certain beta-lactam agents . This low outer-membrane permeability acting in synergy with beta-lactamase is probably responsible for intrinsic beta-lactam resistance in P . aeruginosa. Infect Immun, 1988 Jul, 56(7), 1829 - 30 Immunization with a Pseudomonas aeruginosa immunotype 5 O polysaccharide-toxin A conjugate vaccine: effect of a booster dose on antibody levels in humans; Cryz SJ Jr et al.; Healthy adult volunteers were vaccinated on day 0 and 28 and at 15 months with a Pseudomonas aeruginosa immunotype 5 O polysaccharide-toxin A conjugate vaccine . Immunization resulted in mild, transient local reactions in less than 20% of the subjects . Maximal immunoglobulin G (IgG) antibody titers to both toxin A and lipopolysaccharide (LPS) as determined by enzyme-linked immunosorbent assay were seen at day 42, at which time 50% of the vaccinees showed a fourfold or greater rise in toxin A-neutralizing titers . By 15 months postvaccination, both antitoxin A and anti-LPS IgG antibodies had markedly declined . A booster dose of vaccine administered at 15 months evoked a vigorous anti-toxin A IgG antibody response with 100% of the volunteers showing a fourfold or greater rise in neutralizing antibody titer compared with preimmunization levels . In contrast, there was no significant elevation of anti-LPS IgG antibody levels . At 24 months postimmunization, only anti-toxin A antibody levels were significantly higher than preimmunization levels. J Immunol, 1988 Jul 1, 141(1), 308 - 14 Cloning of a gamma 2b gene encoding anti-Pseudomonas aeruginosa H chains and its introduction into the germ line of mice; Tsang H et al.; A complete, functional gamma 2b gene (pVCM) was cloned from a mouse hybridoma (VD93) with antibody activity to Pseudomonas aeruginosa . DNA sequencing of the VDJ region of pVCM determined that the VH gene was a member of the J558 family rearranged to JH2 . Upon transfection into myeloma cells the gamma 2b gene gave rise to high levels of gamma 2b mRNA and gamma 2b protein . The gamma 2b protein had the same IEF pattern as the parent hybridoma protein VD93 and the antibodies formed from a combination of the pVCM gamma 2b chains and the myeloma lambda-chains bound weakly to P . aeruginosa . However, the hybrid antibodies did not discriminate between the serotypes 2 and 3, whereas the parent protein was specific for serotype 3 . Transgenic mice were produced with the pVCM gamma 2b gene which expressed the gamma 2b mRNA (both membrane and secreted forms) only in lymphoid organs . However, contrary to expectations, the gamma 2b mRNA levels were higher in T cells than in B cells in three different transgenic lines . The serum of the transgenic mice had no activity to P . aeruginosa indicating the importance of L chains for the conformation of the Ag binding site . These gamma 2b transgenic mice provide a convenient tool for the study of feedback inhibition of Ig gene rearrangement. Lancet, 1988 Jun 18, 1(8599), 1359 - 61 Novel modes of action of aminoglycoside antibiotics against Pseudomonas aeruginosa; Morris G et al.; Several mucoid strains and one non-mucoid strain of Pseudomonas aeruginosa were grown in subinhibitory concentrations of various aminoglycosides, beta-lactams, and ciprofloxacin . At antibiotic concentrations that did not affect bacterial growth, aminoglycosides inhibited the excretion of alginate capsule and the production of the iron-chelating siderophores . The same concentrations also disturbed other aspects of the bacterium's iron metabolism, manifested by reduced cellular levels of cytochromes and catalase . beta-lactams, except for cefotaxime, and ciprofloxacin did not reduce alginate excretion . These effects on virulence factors are likely to contribute to the efficacy of aminoglycosides. Biochem J, 1988 Jun 1, 252(2), 515 - 9 Inhibition of leucocyte elastase by heparin and its derivatives; Redini F et al.; Leucocyte proteinases, e.g . leucocyte elastase and cathepsin G, are inhibited by heparin . The activities of pig pancreatic and Pseudomonas aeruginosa elastases are unaffected by this polysaccharide . Heparin derivatives of known Mr and degree of sulphation were isolated . The inhibition of leucocyte elastase by these oligosaccharides can be classified as tight-binding hyperbolic non-competitive . Ki values ranged from 40 nM to 100 microM and were found to be inversely correlated with the chain length of the oligosaccharides . Desulphated compounds lacked inhibitory potential towards leucocyte elastase . Over-O-sulphated di- and tetra-saccharides are more potent inhibitors than their over-N-sulphated counterparts . It is proposed that the therapeutic use of heparin and its derivatives could be extended to disease states such as emphysema and rheumatoid arthritis, where the role of leucocyte elastase has been clearly established. J Antibiot (Tokyo), 1988 Jun, 41(6), 759 - 70 Synthesis and biological activity of 2-lactonyl penems; Capraro HG et al.; A series of potent antibacterial agents have been prepared . These agents are penems carrying a lactone ring in the C-2 position . Excellent activity against Gram-positive and Gram-negative organisms--except Pseudomonas aeruginosa--was found. Jpn J Antibiot, 1988 Jun, 41(6), 623 - 30 {Clinical research on transfer of cefpiramide to the blood and large intestinal tissue in patients with cancer of the large bowel}; Takahashi Y et al.; Cefpiramide (CPM) shows antibacterial activity against Staphylococci, Enterococci, Pseudomonas aeruginosa and anaerobic Bacteroides spp . CPM is a cephalosporin antibiotic which also exhibits antibacterial activity against Klebsiella and intestinal bacteria including Escherichia coli . Concentrations in blood of CPM which has antibacterial activity against main bacterial species detected during gastrointestinal surgeries and concentration transferred to the large intestinal tissue were measured in patients with cancer of the large intestine . Eighteen patients who were hospitalized and underwent large intestinal surgery from December, 1985 to March, 1987 were examined as subjects . CPM was administered at a dose 1 g each of 11 cases and 2 g to each of 7 cases . Concentrations in blood after administration of 1 g of CPM were in a range of 81.56-212.6 micrograms/ml between 25 minutes and 2 hours 20 minutes after administration, and concentrations in the large intestinal tissue were in a 14.17-66.95 micrograms/g range . Ratios of the tissue to the blood concentrations were 0.08-0.49, averaging 0.24 +/- 0.05 . Concentrations in blood after administration of 2 g of CPM were 128.4-253.5 micrograms/ml between 1 hour 10 minutes and 3 hours 50 minutes after administration . Tissue concentrations were 48.33-116.5 micrograms/g between 1 hour 10 minutes and 5 hours 15 minutes after administration . Ratios of the tissue to the blood concentrations were 0.21-0.55, averaging 0.42 +/- 0.05 between 1 hour 10 minutes and 3 hours 5 minutes.(ABSTRACT TRUNCATED AT 250 WORDS) HNO, 1988 Jun, 36(6), 230 - 3 {Ofloxacin in the treatment of Pseudomonas aeruginosa infections of the ear}; Oberascher G et al.; Twenty-three patients suffering from a Pseudomonas infection of the ear were treated with ofloxacin in a prospective study . The type of infection was as follows: external otitis 4, external otitis with furuncle 3, external otitis and otitis media 5, acute exacerbation of chronic otitis media 7, infection of radical mastoidectomy 3, malignant external otitis 1 . All patients had previously been unsuccessfully treated elsewhere . The average period of infection was 13.7 days . The patients received 200 mg of ofloxacin (Tarivid) twice daily for 3 weeks . At the same time the ear was cleaned by suction at 2-4 day intervals and a Normison ear strip applied . Twenty patients were completely cured, 2 patients stopped taking ofloxacin early as a result of improved symptoms but their condition subsequently deteriorated . After a further 3 weeks of consistent treatment both of the cases were also cured . Hence only one patient did not respond to our therapy; that is a success rate of 95% . These results are compared with those of our former therapeutic plan in which all patients with resistant Pseudomonas ear infections were treated as in-patients with selected intravenous antibiotics. J Gen Microbiol, 1988 Jun, 134 ( Pt 6), 1515 - 23 Integration of replication-defective R68.45-like plasmids into the Pseudomonas aeruginosa chromosome; Reimmann C et al.; R68.45 and other similar broad-host-range (IncP) plasmids carrying a tandem repeat of the 2.1 kb insertion element IS21 mobilize the chromosome of many different Gram-negative bacteria . To analyse the structure of R68.45-chromosome cointegrates, whose involvement in the mobilization process had been postulated previously, we selected for the stable integration of R68.45-like plasmids into the Pseudomonas aeruginosa chromosome . Two plasmids were chosen: pME28, a transfer-deficient, mobilizable RP1 derivative with an inactive replication control (trfA) gene, and pME487, an R68.45 derivative with a trfA(ts) mutation causing temperature-sensitive replication . Chromosomally integrated pME28 and pME487 were found to be flanked by single IS21 elements . This structure is in agreement with a 'cut-and-paste' mode of R68.45 transposition . pME28 and pME487 showed a low specificity of insertion but rarely (less than 0.1%) induced auxotrophic mutations . Hfr (high-frequency-of-recombination) donors of P . aeruginosa could be obtained by chromosomal integration of pME487 or pME28; in the latter case, the transfer functions lacking from pME28 had to be provided in trans on an autonomous plasmid . Hfr donors gave higher conjugational linkage and transferred longer stretches of the P . aeruginosa chromosome than did R68.45 donors . This suggests that the integration of R68.45 into the donor chromosome is short-lived in P . aeruginosa. Biol Chem Hoppe Seyler, 1988 Jun, 369(6), 431 - 9 Purification, crystallisation and characterization of quinoprotein ethanol dehydrogenase from Pseudomonas aeruginosa; Rupp M et al.; Pseudomonas aeruginosa ATCC 17933 when grown on ethanol produces high levels of a quinoprotein ethanol dehydrogenase, which amounts to 7% of the soluble protein . The enzyme has been purified to homogeneity and it crystallizes readily in the presence of polyethylene glycol 1550 or 6000 . The ethanol dehydrogenase (Km(ethanol) = 14 microM) resembles the dye-dependent quinoprotein methanol dehydrogenases of methylotrophic bacteria, but has a low affinity for methanol (Km (methanol) = 94mM) . In addition the enzyme oxidizes secondary alcohols . With its catalytic properties the ethanol dehydrogenase is similar to the enzyme isolated from P . aeruginosa LMD 80.53 (Groen, B., Frank, J . Jzn . & Duine, J.A . (1984) Biochem . J . 223, 921-924) . In contrast to this enzyme from P . aeruginosa LMD 80.53, which is a monomer, the ethanol dehydrogenase isolated from P . aeruginosa ATCC 17933 is a dimer of identical subunits of relative molecular mass 60,000 . The N-terminal amino acid is lysine . Inactivation with cyclopropanone ethylhemiketal reveals one molecule of pyrroloquinoline quinone per subunit . As shown by active enzyme sedimentation, the dimer is the enzymatically active form. Pathol Biol (Paris), 1988 Jun, 36(5 Pt 2), 671 - 4 {Preliminary study of the antagonistic effects between fosfomycin and beta-lactams on Pseudomonas aeruginosa observed on the antibiogram}; Buisson Y et al.; Antagonism patterns between beta-lactams (BL) and fosfomycin (FOS) were observed in antibiotic assay by agar diffusion method against the most of Pseudomonas aeruginosa strains . This antagonism was confirmed by antibiotic assays made on Mueller-Hinton agar containing subinhibiting concentrations of FOS . FOS + BL combinations, studied on the strain of Pseudomonas aeruginosa ATCC 27853 by means of checker board method, exhibit antagonism of FOS concentrations below 1 mg/l on ceftriaxone, piperacillin, ticarcillin and, less strongly, on imipenem . These results suggest that in vivo combination FOS + BL dosages must avoid low concentrations of FOS at the site of infection. Zh Mikrobiol Epidemiol Immunobiol, 1988 Jun, (6), 8 - 11 {Epidemiological study of the associations of Staphylococcus aureus and Pseudomonas aeruginosa in the pulmonology clinic}; Sologub VV et al.; The results of the five-year study of S . aureus and P . aeruginosa associations isolated from the sputa of pulmonological patients are presented . The incidence rate of these bacteria in monocultures and associations is estimated . The results of the phage typing and serotyping of S . aureus and P . aeruginosa strains suggest that the formation of the associations of these organisms occurs mainly due to the tendency of P . aeruginosa hospital strains to associate with S . aureus cultures present in the patients. Zh Mikrobiol Epidemiol Immunobiol, 1988 Jun, (6), 32 - 4 {Routes of the transmission of Pseudomonas aeruginosa infection in resuscitation and intensive therapy departments}; Svirskaia LM; To find out if the transfer of P . aeruginosa infection by droplet route is possible in resuscitation and intensive care units, the bacteriological study of air samples taken in different rooms of resuscitation units (altogether 234 air samples) was carried out with the subsequent identification and typing of isolated P . aeruginosa strains . In most cases (70.5%) the microbial contamination of the air in the main rooms of resuscitation units was found not to exceed 500 microbial cells per cu . m, and no P . aeruginosa strains were isolated . The identification and typing of six P . aeruginosa strains isolated from the air of an isolation ward for patients with infectious complications made it possible to find out intraspecific differences of these microorganisms, as all of them belonged to strains of different sero- and pyocinotypes . Thus, the results of these investigations indicate that the droplet route of the transfer of P . aeruginosa hospital infection is not characteristic of resuscitation and intensive care units, as no P . aeruginosa strains are isolated from the main rooms of such units; likewise, no circulation of this microorganism was observed in the air of an isolation ward for patients with infectious complications. Mol Gen Genet, 1988 Jun, 212(3), 510 - 3 Phosphate regulation in Pseudomonas aeruginosa: cloning of the alkaline phosphatase gene and identification of phoB- and phoR-like genes; Filloux A et al.; In Pseudomonas aeruginosa, phosphate limitation results in the synthesis of several protein species . We report the cloning of the P . aeruginosa alkaline phosphatase structural gene, phoA, and we show that this gene is regulated normally in Escherichia coli . We have also identified and cloned two P . aeruginosa genes which can complement phoB and phoR mutations in E . coli . This suggests that a pho regulon system similar to that in E . coli may exist in P . aeruginosa, using at least two similar regulatory factors. Appl Environ Microbiol, 1988 Jun, 54(6), 1587 - 90 Kelthane degradation by genetically engineered Pseudomonas aeruginosa BS827 in a soil ecosystem; Golovleva LA et al.; The aim of this study was to study the degradation of kelthane by Pseudomonas aeruginosa BS827, which carried the plasmid pBS3 . This plasmid encodes naphthalene oxidation . The strain was able to survive in the presence of kelthane and to retain its degradative ability . Kelthane also stabilized the biodegradative plasmid that was preserved by 70 to 100% of the cell population . Cells deficient in Nah or Sal characters were less effective in degrading kelthane, whereas plasmid-free cells lost this ability completely . Evidently, the degradative activity of P . aeruginosa BS827 was conditioned by plasmid determinants coupled with genes of the plasmid pBS3 Nah region. J Interferon Res, 1988 Jun, 8(3), 367 - 73 Effect of interferon-gamma treatment on the course of a burn wound infection; Hershman MJ et al.; Interferon-gamma (IFN-gamma) has been shown to have immunoregulatory properties and is able to modulate resistance to several microbial infections . This study was designed to determine the efficacy of IFN-gamma treatment in a murine burn wound infection model . Bacterial challenge consisted of Klebsiella pneumoniae (10(8) organisms/ml) or Pseudomonas aeruginosa (10(8) organisms/ml), applied topically immediately after burning . Groups of CBA/J mice received either IFN-gamma or RPMI-1640 medium (controls) subcutaneously . IFN-gamma was administered daily at a dose of 7,500 units for 5 days prior to bacterial challenge . Burn without bacterial challenge produced no mortality . Mice treated with IFN-gamma survived significantly longer than controls when the bacterial challenge was K . pneumoniae . There was no difference in survival when bacterial challenge was P . aeruginosa . The Ia antigen expression of peripheral blood mononuclear cells was severely reduced for 3 days post-burn . This drop was prevented on day 3 post-burn in mice treated with IFN-gamma . These data indicate that interferon was effective treatment in a murine model of Klebsiella burn wound infection and was associated with maintenance of Ia antigen expression that may have contributed to the action of the IFN-gamma. J Med Microbiol, 1988 Jun, 26(2), 133 - 41 Alteration of pulmonary structure by Pseudomonas aeruginosa exoenzyme S; Woods DE et al.; Intratracheal administration of purified Pseudomonas aeruginosa exoenzyme S elicited extensive, grossly observable damage in the rat lung within 2 h . Light and electronmicroscopy revealed injury and necrosis of bronchial epithelium, type I pneumocytes and capillary endothelial cells after 1 h; associated haemorrhage, fibrinous exudation and released type II cell lamellar bodies in alveolar lumina after 1-12 h; progressively increasing accumulations of polymorphonuclear leucocytes in the bronchi and alveoli and in alveolar septae (interstitial pneumonia) after 1-12 h; collapse of alveolar septal connective tissue and damage to pulmonary arterioles and venules . Treatment of monolayer cultures of bronchial fibroblasts with purified exoenzyme S elicited vacuolation of the cells with apparent membrane damage as revealed by light and electronmicroscopy . In-vivo production and activity of P . aeruginosa exoenzyme S may be an important pathogenicity determinant in the necrotising lung injury characteristic of P . aeruginosa pneumonia. J Clin Microbiol, 1988 Jun, 26(6), 1161 - 5 Evaluation of an immunofluorescent-antibody test for rapid identification of Pseudomonas aeruginosa in blood cultures; Counts GW et al.; An immunofluorescent-antibody test was developed for rapid detection of Pseudomonas aeruginosa in blood cultures . The test uses a murine monoclonal antibody specific for all strains of P . aeruginosa . In initial tests, bright uniform immunofluorescence signals were seen when each of the 17 international serotypes, as well as 14 additional isolates of P . aeruginosa, were examined . No immunofluorescent staining was observed when 37 other gram-negative and 15 gram-positive species were studied . In a clinical study, the assay was applied to broth smears of 86 gram-negative bacilli isolated from 74 bacteremic patients and 28 additional clinical isolates of Pseudomonas sp . and other oxidase-positive gram-negative bacilli recovered from various body sites . Smears were made directly from blood cultures which were positive for gram-negative bacilli by Gram staining . Eleven (15%) of 74 patients with gram-negative bacteremia had a positive test for P . aeruginosa . Including the results of these 11 isolates recovered in a prospective study and an additional 10 isolates from a retrospective study, we obtained a sensitivity and specificity of 100% (21 positive specimens and 103 negative specimens, respectively) . These preliminary results suggest that this is a useful reagent for rapid presumptive identification of P . aeruginosa in blood cultures . With the immunofluorescent-antibody test, P . aeruginosa could be identified within 1 h of Gram stain evidence of gram-negative bacteremia. J Lab Clin Med, 1988 Jun, 111(6), 701 - 7 Characterization of the human immune response to a Pseudomonas aeruginosa O-polysaccharide-toxin A conjugate vaccine; Cryz SJ Jr et al.; We characterized the immune response in humans after vaccination with a Pseudomonas aeruginosa immunotype 5 O-polysaccharide (O-PS)-toxin A conjugate vaccine . The majority of volunteers responded with an immunoglobulin G (IgG) antibody response on enzyme-linked immunosorbent assay (ELISA) to both the toxin A and the lipopolysaccharide (LPS) components of the vaccine . Maximal titers to both vaccine components were seen 42 days after immunization . Antibody levels remained elevated for at least 14 months after vaccination . The anti-LPS IgG response was predominantly within the IgG1 and IgG2 subclasses, whereas the antitoxin A response was within the IgG1 subclass . There was a gradual elevation of IgG4 antitoxin antibody, with maximal levels seen at 14 months after immunization . No concomitant IgG4 antibody rise to LPS was observed . Immunization evoked a vigorous IgA1 and IgA2 antibody response to LPS, which was long-lived . Only a modest, transient IgA antitoxin A response was noted . Both antitoxin A--neutralizing and opsonic antibodies were elicited by immunization and remained elevated over the 14-month postimmunization period studied. J Bacteriol, 1988 Jun, 170(6), 2725 - 34 Anabolic ornithine carbamoyltransferase of Pseudomonas aeruginosa: nucleotide sequence and transcriptional control of the argF structural gene; Itoh Y et al.; In Pseudomonas aeruginosa PAO the anabolic ornithine carbamoyltransferase (OTCase, EC 2.1.3.3) is the product of the argF gene and the only arginine biosynthetic enzyme whose synthesis is repressible by arginine . We have determined the complete nucleotide sequence of the argF gene including its promoter-control region . The deduced amino acid sequence of the anabolic OTCase consists of 305 residues (Mr 33,924), and this was confirmed by the N-terminal amino acid sequence, the total amino acid composition, and the subunit Mr of the purified enzyme . The native anabolic OTCase (Mr 110,000 to 125,000) was found to be a trimer by cross-linking experiments . P . aeruginosa also has a catabolic OTCase (the arcB gene product), which catalyzes the reverse reaction of the anabolic conversion . At the nucleotide sequence level, the P . aeruginosa argF gene had 52.4% identity with the arcB gene . The Escherichia coli argF and argI genes, which code for anabolic OTCase isoenzymes, had 47.3 and 44.9% identity, respectively, with the P . aeruginosa argF sequence . This suggests that these four genes have evolved from a common ancestral gene . The arcB gene appears to be more closely related to the E . coli argF gene than to the P . aeruginosa argF gene . Two transcripts (mRNA-1, mRNA-2) of the P . aeruginosa argF gene were identified by S1 mapping . The transcription initiation site for mRNA-1 was preceded by sequences having partial homology with the E . coli -35 and -10 consensus promoter sequences . No sequence similar to consensus promoters of enteric bacteria was found upstream of the 5' end of mRNA-2 . E . coli carrying a P . aeruginosa argF+ recombinant plasmid produced mRNA-1 with low efficiency but no (or very little) mRNA-2 . Arginine repressed argF transcription in P . aeruginosa . In the argF promoter region no sequence homologous to the "arg box" (arginine operator module) of E . coli was found . The mechanism of arginine repression in P . aeruginosa thus appears to be different from that in E . coli. Infect Immun, 1988 Jun, 56(6), 1641 - 6 Role of pili in adhesion of Pseudomonas aeruginosa to human respiratory epithelial cells; Doig P et al.; The ability of pili from Pseudomonas aeruginosa K (PAK) to act as an adhesin to human respiratory epithelial cells was examined using an in vitro adhesion assay . Equilibrium analysis of PAK binding to human buccal epithelial cells (BECs) and tracheal epithelial cells (TECs) by means of a Langmuir adsorption isotherm revealed that the maximum numbers of binding sites per epithelial cell (N) were 255 for BECs and 236 for TECs, with apparent association constants (Ka) of 2.8 x 10(-9) and 5.8 x 10(-9) ml/CFU, respectively . Trypsinization of the BECs before the binding assay increased N to 605 and decreased the Ka to 1.7 x 10(-9) ml/CFU . Addition of homologous pili to the binding assay with BECs or TECs or the addition of anti-pilus Fab fragments inhibited PAK adherence . Binding of purified pili to BECs was shown to reach saturation . Purified pili and PAK competed for the same receptor on the BEC surface . Further, by using peptide fragments of PAK pilin (derived from the native pili or produced synthetically) in the binding assay for PAK to BECs, we have presumptively identified the pilus binding domain in the C-terminal region of the pilin and shown that the C-terminal disulfide bridge is important in maintaining the functionality of the binding domain. J Bacteriol, 1988 Jun, 170(6), 2784 - 9 Nucleotide sequence and expression in Escherichia coli of the Pseudomonas aeruginosa lasA gene; Schad PA et al.; Pseudomonas aeruginosa PAO-E64 is a mutant which produces parental levels of elastase antigen but has no elastolytic activity at 37 degrees C . The lesion (lasA1) in PAO-E64 is not a mutation in the structural gene for P . aeruginosa elastase (P.A . Schad, R.A . Bever, T.I . Nicas, F . Leduce, L.F . Hanne, and B.H . Iglewski, J . Bacteriol . 169: 2691-2696, 1987) . A 1.7-kilobase segment of DNA that complements the lasA1 lesion was sequenced . Computer analysis of the DNA sequence showed that it contained an open reading frame which encoded a 41,111-dalton protein . The lasA gene was expressed under an inducible PT-7 promoter, and a 40,000-dalton protein was detected in Escherichia coli lysates . The lasA protein was localized in the outer membrane fraction of E . coli . This lasA protein produced in E . coli activated the extracellular elastase produced by the P . aeruginosa mutant, PAO-E64. J Bacteriol, 1988 Jun, 170(6), 2592 - 8 Construction and characterization of Pseudomonas aeruginosa protein F-deficient mutants after in vitro and in vivo insertion mutagenesis of the cloned gene; Woodruff WA et al.; Mutants with insertion mutations in the Pseudomonas aeruginosa protein F (oprF) gene were created in vivo by Tn1 mutagenesis of the cloned gene in Escherichia coli and in vitro by insertion of the streptomycin resistance-encoding omega fragment into the cloned gene, followed by transfer of the mutated protein F gene back to P . aeruginosa . Homologous recombination into the P . aeruginosa chromosome was driven by a bacteriophage F116L transduction method in the oprF::Tn1 mutants or Tn5-instability in the oprF::omega mutants . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western immunoblotting demonstrated that the resultant oprF insertion mutants had lost protein F, whereas restriction digestion and Southern blotting experiments proved that the mutants contained a single chromosomal oprF gene with either Tn1 or omega inserted into it . It has been proposed that protein F has a role in antibiotic uptake in P . aeruginosa . Measurement of antibiotic resistance levels showed small to marginal increases in resistance, compared with that of the parent P . aeruginosa strain, to a variety of beta-lactam antibiotics . Protein F-deficient mutants had altered barrier properties as revealed by a three- to fivefold increase in the uptake of the hydrophobic fluorescent probe 1-N-phenylnaphthylamine. Zh Mikrobiol Epidemiol Immunobiol, 1988 Jun, (6), 3 - 7 {Structure of the O-specific polysaccharides and the protective activity of the lipopolysaccharides in Pseudomonas aeruginosa of 7 immunotypes (after Fisher)}; Stanislavskii ES et al.; The structure of O-specific polysaccharides and the protective activity of lipopolysaccharides (LPS) obtained from seven P . aeruginosa immunotypes (according to Fisher's classification) have been studied . The structure of O-specific polysaccharides, immunotypes 2, 3, 4, 5 and 6, is identical to that of polysaccharides of serotypes 011; 0(2a), 2c; 01; 010a, 10b; 07a, 7d respectively . No structural analogs of O-polysaccharide characteristic of immunotypes 1 and 7 have been detected among serotypes classified according to the scheme of Lanyi-Bergan-Akatova-Smirnova . The specific character of O-polysaccharides is confirmed by the results of the passive hemagglutination inhibition test, but the data of the passive hemagglutination test indicate that LPS of different immunotypes are antigenically related . As revealed in experiments on the active immunization of mice, LPS of seven immunotypes possess more or less pronounced cross protective properties . The causes of the cross protective activity observed in experiments with these LPS are discussed. J Med Microbiol, 1988 Jun, 26(2), 107 - 14 Production and characterisation of mouse monoclonal antibodies reacting with the lipopolysaccharide core region of gram-negative bacilli; Appelmelk BJ et al.; Monoclonal antibodies to the lipopolysaccharide (LPS) core region were produced by immunising mice with Escherichia coli strain J5 (chemotype Rc) . One of these bound to the deepest part of the core, i.e., Lipid A, and reacted with other heat-killed but not live gram-negative bacilli, including E . coli, Klebsiella pneumoniae and Pseudomonas aeruginosa . Eight other monoclonal antibodies, binding to the terminal glucose residue of Rc LPS, reacted with live cells of E . coli strains only . Thus, the O antigen does not necessarily render the core inaccessible to antibody . However, despite binding to live bacteria, these monoclonal antibodies neither enhanced phagocytic killing, nor protected mice from dying from gram-negative infection or endotoxaemia . It is concluded that antibodies reacting with the most immunodominant parts of the J5 core are not protective. J Biol Chem, 1988 May 25, 263(15), 7364 - 9 An elastase inhibitor from equine leukocyte cytosol belongs to the serpin superfamily . Further characterization and amino acid sequence of the reactive center; Potempa J et al.; Horse leukocyte elastase inhibitor rapidly forms stable, equimolar complexes with both human leukocyte elastase and cathepsin G, porcine pancreatic elastase, and bovine alpha-chymotrypsin . Formation of the inhibitor-pancreatic elastase complex results in peptide bond cleavage at the reactive site of the inhibitor so that a small peptide fragment representing the carboxyl-terminal sequence of the inhibitor is released . Sequence analysis of both this peptide, as well as that of an overlapping peptide obtained by enzymatic inactivation of native inhibitor with either Staphylococcus aureus metalloproteinase, Pseudomonas aeruginosa elastase, or cathepsin B, yields data which indicate that the reactive site encompasses a P1-P1' Ala-Met sequence . However, unlike the human endothelial plasminogen activator inhibitor, which also has a Met residue in the P1' position, oxidation of the horse inhibitor only slightly reduces its association rate constant with either of the elastolytic enzymes tested or with chymotrypsin . Comparison of the amino acid sequence at or near the reactive site of the horse inhibitor (P2-P18') with members of the serpin superfamily of proteinase inhibitors indicates that it not only belongs in this class but also represents the first example of a functionally active intracellular serpin. Biochem Biophys Res Commun, 1988 May 16, 152(3), 1117 - 22 Cloning and characterization of elastase structural gene from Pseudomonas aeruginosa IFO 3455; Yamamoto S et al.; An 8.3 Kb DNA fragment was cloned from Pseudomonas aeruginosa IFO 3455 . This fragment-containing Escherichia clone, pEL2, produced a high level of elastase activity . A smaller EcoRI-KpnI fragment was subcloned into pUC118 and E . coli HB101 was transformed with the plasmid . A deletion mutant clone was also constructed in the same bacteria . These deletion mutants were tested for elastase activity and it became clear that the full length of the elastase gene was 1.0-1.3 Kb . DNA sequencing analysis revealed that this DNA fragment contains the DNA sequence coding N-terminal amino acid sequence of the elastase