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Pseudomonas pachastrellae sp. nov., isolated from a marine sponge.

Two Gram-negative, non-fermentative, non-denitrifying, non-pigmented, rod-shaped bacteria that were motile by means of polar flagella, designated strains KMM 330(T) and KMM 331, were isolated from a deep-sea sponge specimen and subjected to a polyphasic taxonomic study. The new isolates exhibited 16S rRNA gene sequence similarity of 99.9 %, and their mean level of DNA-DNA relatedness was 82 %. Phylogenetic analysis based on their 16S rRNA gene sequences placed the strains within the genus Pseudomonas as an independent deep clade. Strain KMM 330(T) shared highest sequence similarity (96.3 %) with each of Pseudomonas fulva NRIC 0180(T), Pseudomonas parafulva AJ 2129(T) and Pseudomonas luteola IAM 13000(T); sequence similarity to other recognized species of the genus Pseudomonas was below 95.7 %. The marine sponge isolates KMM 330(T) and KMM 331 could be distinguished from the other recognized Pseudomonas species based on a unique combination of their phenotypic characteristics, including growth in 8 or 10 % NaCl, the absence of pigments, the inability to denitrify and lack of carbohydrate utilization. On the basis of phylogenetic analysis, physiological and biochemical characterization, strains KMM 330(T) and KMM 331 should be classified as a novel species of the genus Pseudomonas, for which the name Pseudomonas pachastrellae sp. nov. is proposed. The type strain is KMM 330(T) (=JCM 12285(T)=NRIC 0583(T)=CCUG 46540(T)).

Animals↗

Biochemical and genetic analyses of ferulic acid catabolism in Pseudomonas sp. Strain HR199.

The gene loci fcs, encoding feruloyl coenzyme A (feruloyl-CoA) synthetase, ech, encoding enoyl-CoA hydratase/aldolase, and aat, encoding beta-ketothiolase, which are involved in the catabolism of ferulic acid and eugenol in Pseudomonas sp. strain HR199 (DSM7063), were localized on a DNA region covered by two EcoRI fragments (E230 and E94), which were recently cloned from a Pseudomonas sp. strain HR199 genomic library in the cosmid pVK100. The nucleotide sequences of parts of fragments E230 and E94 were determined, revealing the arrangement of the aforementioned genes. To confirm the function of the structural genes fcs and ech, they were cloned and expressed in Escherichia coli. Recombinant strains harboring both genes were able to transform ferulic acid to vanillin. The feruloyl-CoA synthetase and enoyl-CoA hydratase/aldolase activities of the fcs and ech gene products, respectively, were confirmed by photometric assays and by high-pressure liquid chromatography analysis. To prove the essential involvement of the fcs, ech, and aat genes in the catabolism of ferulic acid and eugenol in Pseudomonas sp. strain HR199, these genes were inactivated separately by the insertion of omega elements. The corresponding mutants Pseudomonas sp. strain HRfcsOmegaGm and Pseudomonas sp. strain HRechOmegaKm were not able to grow on ferulic acid or on eugenol, whereas the mutant Pseudomonas sp. strain HRaatOmegaKm exhibited a ferulic acid- and eugenol-positive phenotype like the wild type. In conclusion, the degradation pathway of eugenol via ferulic acid and the necessity of the activation of ferulic acid to the corresponding CoA ester was confirmed. The aat gene product was shown not to be involved in this catabolism, thus excluding a beta-oxidation analogous degradation pathway for ferulic acid. Moreover, the function of the ech gene product as an enoyl-CoA hydratase/aldolase suggests that ferulic acid degradation in Pseudomonas sp. strain HR199 proceeds via a similar pathway to that recently described for Pseudomonas fluorescens AN103.

Acetyl-CoA C-Acyltransferase↗

Input of protein to lake water microcosms affects expression of proteolytic enzymes and the dynamics of Pseudomonas spp.

The objective of this study was to determine how an input of protein to lake water affects expression of a proteolytic potential and influences the abundance and composition of a specific group of bacteria. Pseudomonas spp. were chosen as a target group that can be recovered on selective growth media and contain both proteolytic and nonproteolytic strains. Amendment with 2 mg of casein per liter increased total proteinase activity (hydrolysis of [(3)H]casein) by 74%, leucine-aminopeptidase activity (hydrolysis of leucine-methyl-coumarinylamide) by 133%, bacterial abundance by 44%, and phytoplankton biomass (chlorophyll a) by 39%. The casein amendment also increased the abundance of culturable Pseudomonas spp. by fivefold relative to control microcosms but did not select for proteolytic isolates. Soluble proteins immunochemically related to the Pseudomonas fluorescens alkaline proteinase, AprX, were detected in amended microcosms but not in the controls. The expression of this class of proteinase was confirmed exclusively for proteolytic Pseudomonas isolates from the microcosms. The population structure of Pseudomonas isolates was determined from genomic fingerprints generated by universally primed PCR, and the analysis indicated that casein amendment led to only minor shifts in population structure. The appearance of AprX-like proteinases in the lake water might thus reflect a general induction of enzyme expression rather than pronounced shifts in the Pseudomonas population structure. The limited effect of casein amendment on Pseudomonas population structure might be due to the availability of casein hydrolysates to bacteria independent of their proteinase expression. In the lake water, 44% of the total proteinase activity was recovered in 0.22-microm-pore-size filtrates and thus without a direct association with the bacteria providing the extracellular enzyme activity. Since all Pseudomonas isolates expressed leucine-aminopeptidase in pure culture, proteolytic as well as nonproteolytic pseudomonads were likely members of the bacterial consortium that metabolized protein in the lake water.

Bacterial Proteins↗

Longitudinal study of antibody response to lipopolysaccharides during chronic Pseudomonas aeruginosa lung infection in cystic fibrosis.

Antibodies to Pseudomonas aeruginosa from 10 cystic fibrosis patients with chronic P. aeruginosa lung infections were quantitatively and qualitatively analyzed. The development of specific antibodies in patient serum was evaluated in a longitudinal study (1972 to 1987). The concentrations and specificities of immunoglobulin G (IgG) and IgM antibodies to purified lipopolysaccharides (LPS) from clinical isolates of P. aeruginosa and to a variety of other gram-negative bacteria were studied by immunoblotting and enzyme-linked immunosorbent assay techniques. Results were compared with the number of immunoprecipitates to P. aeruginosa whole-cell extracts detected by crossed immunoelectrophoresis. IgG, but not IgM, anti-Pseudomonas LPS concentrations increased significantly at the onset of chronic infection and continued to increase during the course of the infection. There was a good positive correlation between the concentration of IgG anti-Pseudomonas LPS antibodies and the number of crossed-immunoelectrophoresis precipitins. The increases in IgG anti-LPS antibody concentrations were much higher to Pseudomonas LPS than to other LPSs. Binding studies demonstrated an increase in binding of IgG anti-Pseudomonas LPS during infection, whereas the binding of other anti-LPS antibodies decreased. Immunoblotting studies confirmed that antibodies reacted strongly with Pseudomonas LPS and weakly with Escherichia coli core-lipid A. The specificity of the reaction with Pseudomonas LPS increased with the duration of infection. It is concluded that anti-LPS response in cystic fibrosis patients during chronic P. aeruginosa infection demonstrates a marked increase in IgG anti-Pseudomonas LPS antibody concentration, specificity, and affinity. The anti-LPS enzyme-linked immunosorbent assay is proposed as a routine test to diagnose and to follow the course of chronic P. aeruginosa lung infection in patients with cystic fibrosis.

Adolescent↗

Cloning and nucleotide sequence analysis of xylE gene responsible for meta-cleavage of 4-chlorocatechol from Pseudomonas sp. S-47.

Pseudomonas sp. S-47 expresses catechol 2,3-dioxygenase (C230) catalyzing the conversion of 4-chlorocatechol (4CC) as well as catechol to 5-chloro-2-hydroxymuconic semialdehyde and 2-hydroxymuconic semialdehyde, respectively, through meta-ring cleavage. The xylE gene encoding C230 for meta-cleavage was cloned from strain S-47 and its nucleotide sequence was analyzed. The pRES101 containing the xylE gene exhibited high C230 activity toward catechol and 4CC without altering the substrate specificity from natural strain. The xylE gene was composed of 924 bp and encoded polypeptide of molecular mass 35 kDa containing 307 amino acids. A deduced amino acid sequence of the C230 from strain S-47 exhibited over 80% identity with those of Pseudomonas putida mt-2, Pseudomonas putida G7, and Pseudomonas sp. CF600. However, it shows below 45% identity with those of Pseudomonas cepacia LB400 and Pseudomonas sp. KKS102. The C230 of strain S-47 was conserved in the amino acids (His150, His214, Glu261) for metal binding ligands and those (His199, His242, and Tyr251) for catalytic sites. Therefore, Pseudomonas sp. S-47 can be explained as acting by degrading catechol as well as 4CC by xylE-encoding C230 which was fused by N domain of nahH and C domain of dmpB from other Pseudomonas strains.

Amino Acid Sequence↗

[In vitro bacteriostatic activity of piperacillin sodium against Pseudomonas].

One hundred and twenty-two strains of Pseudomonas isolated from diverse pathological processes were typified. In vitro activity of Piperacillin (antibiogram and MIC) was studied and compared with another two semisynthetic penicillins, Carbenicillin and Mezlocillin, and two aminoglycosides, Amikacin and Gentamicin. The strains isolated corresponded to: Pseudomonas aeruginosa (116), Pseudomonas cepacia (3), Pseudomonas fluorescens (1) and Pseudomonas putida (1). It was found that 110 Pseudomonas aeruginosa, one Pseudomonas cepacia and one Pseudomonas fluorescens were susceptible to Piperacillin. The susceptibility to this drug was quite higher than to the other penicillins tested. The MIC50 and MIC90 of Piperacillin were lower than those of semisynthetic penicillins and the minimum susceptibility value. In comparison with aminoglycosides it was found that, although Amikacin and Gentamicin did not reach the minimum value, Piperacillin exhibited higher MICs.

Humans↗

Repetitions in the NH2-terminal amino acid sequence of beta-ketoadipate enol-lactone hydrolase from Pseudomonas putida.

Muconolactone delta-isomerase (EC 5.3.3.4) and beta-ketoadipate enol-lactone hydrolase (EC 3.1.1.24) mediate consecutive reactions in the beta-ketoadipate pathway of bacteria. An earlier investigation (Yeh, W.K., Davis, G., Fletcher, P., and Ornston, L.N. (1978) J. Biol. Chem. 253, 4920-4923) revealed that the respective NH2-terminal amino acid sequences of Pseudomonas putida muconolactone isomerase and Acinetobacter calcoaceticus beta-ketoadipate enol-lactone hydrolase II are evolutionarily homologous. In this report, we describe the purification of Pseudomonas beta-ketoadipate enol-lactone hydrolase and present evidence indicating that the protein is a trimer composed of identical 11,000-dalton subunits. The NH2-terminal amino acid sequences of Pseudomonas muconolactone isomerase and Pseudomonas enol-lactone hydrolase have diverged widely from each other, yet the two sequences contain different fragments of an ancestral sequence which is represented in Acinetobacter enol-lactone hydrolase II. The widely divergent Pseudomonas muconolactone isomerase and Pseudomonas enol-lactone hydrolase sequences each contain unique sets of repeated peptides. In principle, the repetitive sequences might have been introduced by elongation mutations which occurred early in the evolution of the proteins. However, the divergence of Pseudomonas muconolactone isomerase and Pseudomonas enol-lactone hydrolase is so extreme that the observed sequence repetitions cannot have been conserved from ancestral duplication mutations. Rather, the data favor the interpretation that copies of DNA were substituted into structural genes for the enzymes as they diverged.

Adipates↗

Activity of 4-quinolones against Pseudomonas aeruginosa.

The minimum inhibitory concentrations (MICs), bactericidal activities and mechanisms of action of ofloxacin (CAS 82419-36-1), levofloxacin (CAS 100986-85-4) and ciprofloxacin (CAS 86393-32-0) were investigated against Pseudomonas aeruginosa. All three 4-quinolones were found to possess higher MICs against Pseudomonas aeruginosa than against other Gram-negative bacteria. Despite this, however, all three drugs were more rapidly bactericidal and produced a greater level of kill against Pseudomonas aeruginosa than against any other bacterial species previously tested. Thus MIC tests cannot be used to predict the bactericidal activity of 4-quinolones. Furthermore, MIC tests showed ciprofloxacin to be more potent than ofloxacin or levofloxacin against Pseudomonas aeruginosa. However, bactericidal tests showed levofloxacin to be about 10 times more bactericidal than either ciprofloxacin or ofloxacin. Thus MIC tests cannot predict the relative bactericidal potency of 4-quinolones against Pseudomonas aeruginosa. Therefore, MIC tests should not be used as the sole measure for the efficacy of 4-quinolones, as is often the case. Surprisingly, the characteristic biphasic dose response curve, normally shown by 4-quinolones against other bacteria, was absent when Pseudomonas aeruginosa was tested. This unusual effect was explained by the presence of bactericidal mechanism B associated with the unique loss of bactericidal mechanism A at high 4-quinolone concentrations. This loss of bactericidal mechanism A may explain the recent high incidences of chromosomally-mediated 4-quinolone resistance with Pseudomonas aeruginosa because it may be easier for Pseudomonas aeruginosa to mutate to resist one mechanism of action than to mutate to resist two or more mechanisms of action.

Anti-Infective Agents↗

Vaccines for preventing infection with Pseudomonas aeruginosa in people with cystic fibrosis.

OBJECTIVES: To assess the effectiveness of vaccination against Pseudomonas aeruginosa in patients with cystic fibrosis. SEARCH STRATEGY: The Cochrane Cystic Fibrosis and Genetic Disorders Group specialist trials register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. Date of the most recent search of the Group's specialised register: November 1999. SELECTION CRITERIA: All randomised or pseudorandomised trials (published or unpublished) comparing Pseudomonas aeruginosa vaccines (oral, parenteral or intranasal) with control vaccines or no intervention in patients with cystic fibrosis. DATA COLLECTION AND ANALYSIS: We planned to assess the following outcomes: time to infection with Pseudomonas aeruginosa, pulmonary function, body mass index, Schwachman score, frequency of pulmonary infective exacerbations, days of antibiotic usage, days unable to carry out normal daily activities, adverse events, mortality, antibody levels to Pseudomonas aeruginosa and T cell proliferation and cytokine production in response to Pseudomonas aeruginosa. MAIN RESULTS: One trial which included 17 vaccinated patients, with follow up reported to 10 years met the inclusion criteria. Finding only a single trial, and the lack of information on our predefined outcomes limited analysis. REVIEWER'S CONCLUSIONS: There is a paucity of randomised controlled trials assessing the effectiveness of vaccination against Pseudomonas aeruginosa in cystic fibrosis. Increased understanding of modulation of the immune response by vaccination has led to the development of alternative vaccines. We suggest that there is an urgent need for newer vaccines to be evaluated in adequately-powered, multicentre randomised controlled trials examining clinically relevant end-points in addition to immunological variables. Such a trial should assess effectiveness over several years, and include follow-up of vaccinees who become colonised with Pseudomonas aeruginosa.

Bacterial Vaccines↗

[Automated micromethod for the determination of the utilization of carbon sources by clinically significant Pseudomonas species].

The assimilation of 43 different carbon substrates by 93 clinical strains of Pseudomonas aeruginosa was studied by a new miniaturized rapid method. Reading of assimilation results was done photometrically after 18-20 h incubation and the resulting data were captured and stored by a microcomputer. The differentiating capacity of the assimilation tests were verified by comparing the results of 41 strains of Pseudomonas fluorescens, 48 strains of Pseudomonas putida, 52 strains of Pseudomonas maltophilia and respectively 10 strains of Pseudomonas pseudomallei and Pseudomonas cepacia. The assimilation pattern obtained from the Pseudomonas aeruginosa strains agreed to those described in literature and because of miniaturization, standardisation, facility of use and automatic reading the method seems to be suitable for routine laboratory work.

Amino Acids↗

Pseudomonas aeruginosa and Burkholderia cepacia cannot be detected by PCR in the breath condensate of patients with cystic fibrosis.

The collection of sputum for microbiological examination in young cystic fibrosis patients can be very difficult. However, a knowledge of bacterial flora colonizing the patient's airways is of paramount importance for proper antimicrobial therapy. It is also known that cystic fibrosis patients colonized by Pseudomonas species have a poorer prognosis than Pseudomonas-negative patients. Noninvasive ways of diagnosing airway inflammation that require only minimal cooperation of the patient might yield new possibilities for early detection of airway colonisation. The breath condensate method as a noninvasive diagnostic technique seems especially appropriate for use in children. Therefore, the aim of this study was to evaluate whether the breath condensate method could be used for detection of Pseudomonas species in children with cystic fibrosis. In total, 32 breath condensate and seven sputum samples were obtained from 13 cystic fibrosis patients with Pseudomonas aeruginosa- or Burkholderia cepacia-positive sputum culture (20 samples were obtained during forced expiration). PCR for combined detection of Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Burkholderia cepacia was performed. PCR results of all breath condensate samples were negative for Pseudomonas aeruginosa, Stenotrophomonas maltophilia, or Burkholderia cepacia, while all sputum sample results were positive. A minimum DNA quantity of 10 fg could be detected in dilution series of the positive control group. We conclude that the breath condensate method cannot be used as a tool for detection of Pseudomonas species.

Adolescent↗

Detection of Pseudomonas aeruginosa from ovine fleece washings by PCR amplification of 16S ribosomal RNA.

Two oligonucleotides were selected from the variable regions of the 16S rRNA gene of P. aeruginosa and used as PCR primers for the detection of P. aeruginosa. The specificity of the primers was tested against the following bacterial species; Pseudomonas putida, Pseudomonas cepacia, Xanthamonas maltophilia, Pseudomonas mendocina, Pseudomonas stutzeri, Pseudomonas fluorescens, Pseudomonas alcaligenes and Pseudomonas diminuta. These primers had a sensitivity of detection of 1 pg of chromosomal DNA or 1 x 10(5) cfu/microliters and were species-specific. The sensitivity of detection was increased to 1 fg or less than 10 cfu/microliters using a non-radioactively labelled probe. Using these PCR primers it was possible to detect the presence of P. aeruginosa from fleece washings collected from a flock of 100 sheep. Correlation between single PCR and bacteriological isolation showed agreement in 89% of fleece samples tested, 2% of the samples contained organic PCR inhibitors in the fleece washings, 3% were below the level of sensitivity of the test and the remaining 6% were culture negative for P. aeruginosa but PCR positive. Use of nested PCR did not increase the sensitivity of detection over single round PCR combined with the use of non-radioactively labelled probe.

Animals↗

Adhesins and receptors of Pseudomonas aeruginosa associated with infection of the respiratory tract.

Substantial progress has been made in defining a number of Pseudomonas adhesins which may be involved in the pathogenesis of respiratory infection. As yet, it is unclear which of these adhesins are primarily responsible for initiating infection in CF. The findings that CF epithelial cells have increased numbers of receptors for P. aeruginosa attachment and that CF epithelia are less highly sialylated than normal epithelial cells is consistent with a role for Pseudomonas pili in the initial recognition of asialoganglioside receptors on epithelial cells. In addition, there is ample evidence supporting the presence of several classes of non-pilus adhesins. Adherence properties of P. aeruginosa clearly vary from strain to strain and it appears likely that all potential adhesions are not equally expressed. More importantly, the regulation of the expression of these adhesins is unlikely to be constitutive. Some may be expressed only when triggered by the appropriate environmental conditions as found in vivo. In reviewing the pathogenesis of Pseudomonas infection in the CF lung, several classes of receptors must be considered. Pseudomonas infection is limited to the bronchi in CF. The organisms do not invade the bronchial tissue, but remain in the airways forming a biofilm with associated microcolonies. Thus, it would seem reasonable to expect Pseudomonas receptors within respiratory mucin. However, to date, there is little confirmatory data to support the presence of specific receptors in mucin. Alternatively, it is possible that the failure of bacterial binding to mucin components may contribute to colonization as organisms which are not efficiently cleared by muco-ciliary function may persist in the airways long enough to find or expose cryptic epithelial binding sites. This hypothesis is supported by binding studies which demonstrate decreased Pseudomonas attachment to CF as compared with normal respiratory mucins. Based on the available data, there appears to be a hierarchy of adhesin expression. Multiple ligand-receptor interactions may occur in the respiratory tract and it may be difficult to analyze the effect of secondary adhesins in the presence of what appears to be the dominant ligand, i.e. pilin. Thus, the failure to find the expected sialylated receptor for Pseudomonas attachment may be due to methodologic problems such as studying strains under conditions in which pili are well expressed and affinities for asialylated receptors predominate. This may not be the situation in vivo after the initial contact of the infecting organisms with the epithelial surface. Not only must the organism attach initially, but it must then be able to persist within the lung. Further studies, based on genetically defined mutants should help define which P. aeruginosa gene products and which components of the CF but not the normal epithelium are responsible for this unique but ultimately fatal host/bacterium interaction.

Adhesins, Bacterial↗

Pseudomonas aeruginosa in cystic fibrosis: cross-infection and the need for segregation.

Evidence-based reasons for segregation of patients colonized with Pseudomonas aerugionsa in the outpatient setting are unclear. To clarify local decisions, Pseudomonas genotyping of the local environment, patients and patient contacts was undertaken in 1993. The hospital environment was re-swabbed in 1997. Pseudomonas genotyping of old and new patients attending the North Staffordshire cystic fibrosis clinic has subsequently been undertaken and more recently been repeated on an annual basis to assess whether the same Pseudomonas genotypes can be found in both the environment and in patients, and whether the same Pseudomonas genotype can be transferred from one patient to another. No Pseudomonas genotype found in the local environment in 1993 or in 1997 has been found in any of our patients. Nine children attending the same special school for many years and sharing the same physiotherapy facilities showed no evidence of cross-infectivity. Except for siblings living in the same household our cross-infectivity rate is very low and where cross-infection has potentially occurred the level of contact between these patients has been minimal. This study does not support the suggestion that patients with cystic fibrosis attending the North Staffordshire clinic and colonized with Pseudomonas aeruginosa should be segregated from non-colonized patients.

Adolescent↗

Complicated urinary tract infection caused by Pseudomonas aeruginosa in a single institution (1999-2003).

BACKGROUND: Pseudomonas aeruginosa has been an important uropathogen that causes complicated urinary tract infection. We investigated the clinical characteristics of complicated urinary tract infection caused by Pseudomonas aeruginosa in a single institution. METHODS: We studied those patients who had basal disease in their urinary tract that was diagnosed as urinary tract infection caused by more than 10(4) colony forming units (CFU)/mL of Pseudomonas aeruginosa isolated from their urine. In those patients, we analysed infectious risk factors, treatment methods including the use of antimicrobial agents, the presence of a urinary tract catheter, and the relationship between febrile infection and urinary tract catheter. In addition, we examined the various antimicrobial susceptibilities against Pseudomonas aeruginosa. RESULTS: We studied 76 patients (59 men and 17 women). Of their basal diseases of the urinary tract, bladder tumor was the most prevalent (42.1%). Of the 39 patients who had an indwelling urinary tract catheter, 26 (66.7%) experienced a high-grade fever, a higher rate than that of the 37 patients who were not catheterized (40.5%). Seven patients were treated with anticancer chemotherapy drugs and 31 cases of urinary tract infection caused by Pseudomonas aeruginosa were diagnosed in the perioperative period. Piperacillin showed lower susceptibility against Pseudomonas aeruginosa in these 2 years (P<0.05). CONCLUSIONS: Our results indicated that those patients with urinary tract catheterization had a higher incidence of fever than patients without catheterization. Therefore, we must improve not only the antimicrobial treatment of Pseudomonas aeruginosa but also our management of catheters.

Adult↗

Evaluation of a new polyvalent Pseudomonas vaccine in respiratory infections.

A new polyvalent, cell wall extract, Pseudomonas aeruginosa vaccine (PEV-01), was evaluated by using a guinea pig model of experimental Pseudomonas pneumonia. Guinea pigs routinely developed fourfold rises in serum hemagglutinating Pseudomonas antibodies after four vaccine injections given over 2 weeks. Vaccinated animals survived an intratracheal Pseudomonas challenge (1 X 10(8) colony-forming units) significantly better (13 of 14 survived) than did a control group (5 of 14 survived) (P less than 0.01). Clearance of viable Pseudomonas from lung tissue was significantly better in vaccinees than controls at both 3 h (P less than 0.02) and 6 h (P less than 0.05) after infection. Both gross and histological examinations of lung tissue revealed less pulmonary tissue damage in vaccinated animals following Pseudomonas infection. Thus, PEV-01 Pseudomonas vaccine appears capable of eliciting a specific protective response in the guinea pig respiratory tract.

Animals↗

Headspace analysis of volatile metabolites of Pseudomonas aeruginosa and related species by gas chromatography-mass spectrometry.

Gas chromatographic-mass spectrometric analysis of headspace volatiles was performed on cultures of 11 strains of Pseudomonas aeruginosa and 1 strain each of Pseudomonas cepacia, Pseudomonas putida, Pseudomonas putrefaciens, Pseudomonas fluorescens, and Pseudomonas maltophilia. All strains of Pseudomonas aeruginosa produced a distinctive series of odd-carbon methyl ketones, particularly 2-nonanone and 2-undecanone, and 2-aminoacetophenone. The other strains failed to produce 2-aminoacetophenone. Two sulfur compounds, dimethyldisulfide and dimethyltrisulfide, were present in strains of P. aeruginosa and in variable amounts in other species. Butanol, 2-butanone, 1-undecene, and isopentanol were also detected in P. aeruginosa cultures.

Alcohols↗

Functional importance of cystic fibrosis immunoglobulin G fragments generated by Pseudomonas aeruginosa elastase.

We examined the functional importance of immunoglobulin polypeptide fragments generated by Pseudomonas aeruginosa elastase (Pseudomonas elastase). The purpose of this study was to determine whether the elastase produced by Pseudomonas aeruginosa cleaves human IgG into immune fragments that functionally inhibit opsonophagocytosis. Our results confirm that IgG isolated from patients with cystic fibrosis (CF) incubated with purified pseudomonas elastase results in the generation of two major polypeptide fragments and that, furthermore, these fragments significantly inhibit bacterial uptake by human neutrophils. After 75 minutes bacterial uptake was six times greater when intact IgG was used as an opsonin (uptake 90.2% +/- 18.6% SEM) compared with a IgG was used as an opsonin (uptake 90.2% +/- 18.6% SEM) compared with a mixture of pseudomonas-lipopolysaccharide-reactive Fab and F(ab')2 fragments generated by pseudomonas elastase (uptake 15.4% +/- 0.8% SEM, p less than 0.001). Hydrolyzed CF IgG antibodies consistently resulted in a level of bacterial uptake less than that of normal saline negative controls (NS): (at 10 minutes, NS 26.6% vs CF 16.8%, p less than 0.05; at 75 minutes, NS 28.2% vs CF 15.4%, p less than 0.01. This suggests that the immune polypeptides are active inhibitors of the essential neutrophil phagocyte-bacterial cell interaction. Intact immune IgG reversed the defect in opsonophagocytosis. When intact IgG was mixed with IgG fragments the phagocytic rates increased directly with increasing amounts of intact IgG. We conclude that the elastase exoproduct secreted by Pseudomonas aeruginosa is capable of cleaving IgG into functionally important fragments that inhibit bacterial uptake. Furthermore, this inhibition can be overcome by increasing amounts of a commercially available preparation of intact immune IgG.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Bacterial↗