Pseudomonas aeruginosa

Science, 1995 Jun 30, 268(5219), 1899 - 902
Common virulence factors for bacterial pathogenicity in plants and animals; Rahme LG et al.; A Pseudomonas aeruginosa strain (UCBPP-PA14) is infectious both in an Arabidopsis thaliana leaf infiltration model and in a mouse full-thickness skin burn model . UCBPP-PA14 exhibits ecotype specificity for Arabidopsis, causing a range of symptoms from none to severe in four different ecotypes . In the mouse model, UCBPP-PA14 is as lethal as other well-studied P . aeruginosa strains . Mutations in the UCBPP-PA14 toxA, plcS, and gacA genes resulted in a significant reduction in pathogenicity in both hosts, indicating that these genes encode virulence factors required for the full expression of pathogenicity in both plants and animals.

Science, 1995 Jun 23, 268(5218), 1733 - 5
Electron tunneling in proteins: coupling through a beta strand; Langen R et al.; Electron coupling through a beta strand has been investigated by measurement of the intramolecular electron-transfer (ET) rates in ruthenium-modified derivatives of the beta barrel blue copper protein Pseudomonas aeruginosa azurin . Surface histidines, introduced on the methionine-121 beta strand by mutagenesis, were modified with a Ru(2,2'-bipyridine)2(imidazole)2+ complex . The Cu+ to Ru3+ rate constants yielded a distance-decay constant of 1.1 per angstrom, a value close to the distance-decay constant of 1.0 per angstrom predicted for electron tunneling through an idealized beta strand . Activationless ET rate constants in combination with a tunneling-pathway analysis of the structures of azurin and cytochrome c confirm that there is a generally efficient network for coupling the internal (native) redox center to the surface of both proteins.

Commun Dis Rep CDR Rev, 1995 Jun 23, 5(7), R102 - 4
Whirlpool baths in nursing homes: use, maintenance, and contamination with Pseudomonas aeruginosa; Hollyoak V et al.; Transmission of Pseudomonas aeruginosa wound infection was associated with the use of a whirlpool bath in a nursing home . The nursing home inspection unit asked for guidance on whirlpool baths in nursing homes and advice for proprietors about their use, cleaning, disinfection, and maintenance . Seventeen whirlpool baths in 16 nursing homes in two health districts were examined for the presence of P . aeruginosa . A survey was made of the use made of whirlpool baths, methods used to clean and disinfect them, and the occurrence of P . aeruginosa wound infection in users . P . aeruginosa were found in large numbers in water samples from all whirlpool baths after agitation . Only one of the 253 residents who used whirlpool baths was known to have a P . aeruginosa wound infection . The local nursing home inspection unit was advised that whirlpool baths could continue to be used in nursing homes but only by continent residents with intact skin . The bath should be cleaned and disinfected, preferably with hypochlorite, after each use; the bath should be more thoroughly cleaned and disinfected daily and the bath should be fully serviced at least once a year . Suspected or confirmed cases of P . aeruginosa infection in residents of nursing homes should be reported to the consultant in communicable disease control . The prevalence of known infection with P . aeruginosa was low in the residents of the nursing homes, but the unguided and unregulated use of whirlpool baths in nursing homes may present an infection hazard to residents who use the bath and to hospitals that admit residents from such nursing homes.(ABSTRACT TRUNCATED AT 250 WORDS)

Commun Dis Rep CDR Rev, 1995 Jun 23, 5(7), R100 - 2
Pseudomonas aeruginosa wound infection associated with a nursing home's whirlpool bath; Hollyoak V et al.; Whirlpool baths are fitted with hydrojet circulation and/or air induction bubble systems . Water in a whirlpool bath, unlike a spa pool, is not filtered or chemically treated but the bath is drained and cleaned between each bather . This is, we believe, the first report of Pseudomonas aeruginosa wound infection associated with the use of a whirlpool bath in a nursing home . Microbiologically confirmed infections with P . aeruginosa of identical antibiotic sensitivity patterns arose in one week in wounds of four of 24 residents who used a whirlpool bath from which P . aeruginosa was also isolated . P . aeruginosa was not isolated from the wounds of a further seven residents who did not use the whirlpool bath . The incident control team advised that use of the whirlpool bath should be restricted to continent residents with intact skin, and that the bath should be cleaned with a degreasing agent and disinfected with hypochlorite between use by individual residents . The hazard of infection posed by whirlpool baths, particularly in nursing homes, needs to be assessed . National guidance for their cleaning, maintenance, and disinfection is required.

FEMS Microbiol Lett, 1995 Jun 15, 129(2-3), 251 - 4
Outer membrane permeability of beta-lactamase inhibitors in Pseudomonas aeruginosa; Satake S et al.; Evaluation of four beta-lactamase inhibitors in terms of their outer membrane permeability in Pseudomonas aeruginosa revealed that sulbactam and tazobactam diffused most efficiently and equally well . That of BRL42715 appeared to be a factor of ten lower than that of the above two, but it showed the strongest beta-lactamase inhibitory activity . This is most likely due to its better beta-lactamase inactivating activity . BRL42715 at 1.56 micrograms ml-1 lowered the minimum inhibitory concentrations of ceftazidime and imipenem in a strain producing fully derepressed beta-lactamase and an undetectable level of the outer membrane protein OprD2.

J Chemother, 1995 Jun, 7 Suppl 2, 165 - 73
Evaluation of the efficacy and safety of isepamicin compared with amikacin in the treatment of nosocomial pneumonia and septicaemia; Beaucaire G; Isepamicin is a new aminoglycoside antibiotic which possesses greater stability to aminoglycoside-inactivating enzymes compared with other available aminoglycosides . In this prospective, randomised, open trial, the safety and efficacy of intravenous administration of isepamicin was compared with that of intravenous amikacin in seriously ill adults with nosocomial pneumonia or septicaemia . Each study aminoglycoside was administered concurrently with ceftazidime or imipenem . Patients were randomised to receive isepamicin 15 mg/kg once daily, isepamicin 7.5 mg/kg twice daily or amikacin 7.5 mg/kg twice daily . For patients with nosocomial pneumonia, the proportions of patients in the intent-to-treat population (n = 130) who were clinically cured at the end of treatment were similar in each treatment group: 18/44 (41%) isepamicin once daily; 19/45 (42%) isepamicin twice daily; and 17/41 (42%) amikacin . Corresponding results for the efficacy population (n = 58) were: 12/20 (60%) isepamicin once daily; 14/21 (67%) isepamicin twice daily; 9/17 (53%) amikacin . In patients with septicaemia, clinical cure was achieved in 8/10 (80%) patients treated with isepamicin once daily, compared with 8/13 (62%) patients who received isepamicin twice daily, and 7/12 (58%) patients treated with amikacin . For both diagnoses, there were no statistically significant differences between the treatment groups in clinical cure rate . The most commonly isolated target pathogen was Pseudomonas aeruginosa . For both nosocomial pneumonia and septicaemia, the proportion of patients in the intent-to-treat population whose pretreatment valid target pathogens were eliminated was similar in each treatment group . In total, 51 patients (30%) died during study, mostly due to disease progression or complications, or concurrent illness . All three treatment regimens were well tolerated . The proportion of patients experiencing at least one adverse event was 11%, 25% and 9% for isepamicin once daily, isepamicin twice daily and amikacin, respectively . The incidence of ototoxicity and nephrotoxicity was relatively low in both treatment groups.

J Chemother, 1995 Jun, 7 Suppl 2, 137 - 42
Comparison of the efficacy and safety of isepamicin and amikacin in the treatment of acute lower respiratory tract infections caused by gram-negative organisms; Covi M et al.; In a prospective multicentre open trial, hospitalised adult patients with acute lower respiratory tract infections, mainly pneumonia or bronchitis, were randomised to receive either isepamicin 8 or 15 mg/kg once daily depending on the severity of the infection or amikacin 7.5 mg/kg twice daily . Patients with infections known to be caused by Pseudomonas aeruginosa were to be given concomitant treatment with ceftazidime . In the intent-to-treat population, i.e . patients who received at least one dose randomised treatment, a clinical cure or improvement at the end of treatment was seen in 112/125 (90%) isepamicin patients and 55/60 (92%) amikacin patients . The corresponding rates for patients with a primary diagnosis of pneumonia were 45/52 (87%) and 25/28 (89%) . Cure/improvement rates for patients with P . aeruginosa as the causative pathogen (34 of whom also received ceftazidime) were 28/30 (93%) and 16/18 (89%), respectively . In the efficacy population (patients who had a valid pretreatment culture and who met other evaluability criteria), total elimination (documented or presumed if infection had resolved) of target pathogens occurred in 54/63 (86%) of isepamicin patients and 25/30 (83%) of amikacin patients . P . aeruginosa, Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus were commonly isolated pathogens . Treatment-related adverse were mainly mild or moderate in severity and occurred in 10% of isepamicin patients and 13% of amikacin patients . Four patients (3 isepamicin and 1 amikacin) discontinued treatment because of severe adverse events and a further isepamicin patient withdrew because of a mild adverse event . Nephrotoxicity and ototoxicity occurred infrequently.

J Chemother, 1995 Jun, 7 Suppl 2, 129 - 35
The efficacy and safety of isepamicin and ceftazidime compared with amikacin and ceftazidime in acute lower respiratory tract infection; Colardyn F; Isepamicin is a new aminoglycoside antibiotic which has a superior stability to aminoglycoside-inactivating enzymes compared with other available aminoglycosides . In this multicentre, randomised, open study, the safety and efficacy of isepamicin plus ceftazidime was compared with that of amikacin plus ceftazidime in adults with acute lower respiratory tract infection . Patients with severe infections received intravenous administration of isepamicin 15 mg/kg once daily + ceftazidime 2g twice daily (n = 121) or amikacin 7.5 mg/kg twice daily + ceftazidime 2g twice daily (n = 61) . Those with less severe infection received intramuscular or intravenous administration of isepamicin 8 mg/kg once daily + ceftazidime 1g twice daily (n = 56) or amikacin 7.5 mg/kg twice daily + ceftazidime 1g twice daily (n = 28) . In the efficacy populations, the proportion of patients clinically cured in the isepamicin group (87/100; 87%) was similar to that in the amikacin group (36/47; 77%) . Significantly more patients in the isepamicin group were cured or improved compared with the amikacin group (97% vs 89%; p = 0.042) . The difference between treatment groups was also significant in patients with pneumonia (p = 0.05) . The most commonly isolated target organisms were Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Klebsiella pneumoniae . The proportion of patients in the efficacy population whose pretreatment valid target organisms were eliminated was similar in each treatment group (90% isepamicin vs 89% amikacin) . A retrospective analysis showed there were slightly fewer clinical successes and a higher death rate in patients with nosocomial rather than community acquired pneumonia . Both treatments were well tolerated . Fourteen per cent of isepamicin and 11% of amikacin patients experienced adverse events . The incidence of ototoxicity and nephrotoxicity was low.

Pathol Biol (Paris), 1995 Jun, 43(6), 551 - 3
{Effects of azithromycin on the virulence of Pseudomonas aeruginosa}; Reinert P; Pseudomonas aeruginosa is a saprophyte opportunistic bacteria which frequently colonizes the respiratory tract of patients presenting a severe chronic bronchitis pathology . Secreting a number of exotoxins and enzymes inducing an inflammation and necrosing of the surrounding tissues, it provokes irreversible pulmonary lesions . Different experimental in vitro works evidenced macrolides activity on the production and/or secretion of these factors, with a diminution of elastase, protease, lecithinase and D-nase synthesis . Among the macrolides, azithromycin seems to have the most pronounced activity . In vivo, some patients suffering from bronchiolitis or cystic fibrosis have been clinically improved with a treatment using erythromycin, or clarithromycin or azithromycin . These very preliminary results demand to be confirmed but the macrolides could allow a decrease of Pseudomonas aeruginosa pathogenicity and thus stop the deterioration of pulmonary functions.

Appl Environ Microbiol, 1995 Jun, 61(6), 2172 - 9
Copper as a signal for alginate synthesis in Pseudomonas syringae pv . syringae; Kidambi SP et al.; Plant-associated pseudomonads are commonly exposed to copper bactericides, which are applied to reduce the disease incidence caused by these bacteria . Consequently, many of these bacteria have acquired resistance or tolerance to copper salts . We recently conducted a survey of 37 copper-resistant (Cur) Pseudomonas spp., including P . cepacia, P . fluorescens, P . syringae, and P . viridiflava, and found that a subset of the P . syringae strains showed a dramatic increase in exopolysaccharide (EPS) production on mannitol-glutamate medium containing CuSO4 at 250 micrograms/ml . A modified carbazole assay indicated that the EPS produced on copper-amended media contained high levels of uronic acids, suggesting that the EPS was primarily alginic acid . Uronic acids extracted from selected strains were further confirmed to be alginate by demonstrating their sensitivity to alginate lyase and by descending paper chromatography following acid hydrolysis . Subinhibitory levels of arsenate, cobalt, lithium, rubidium, molybdenum, and mercury did not induce EPS production, indicating that alginate biosynthesis is not induced in P . syringae cells exposed to these heavy metals . A 200-kb plasmid designated pPSR12 conferred a stably mucoid phenotype to several P . syringae recipients and also increased their resistance to cobalt and arsenate . A cosmid clone constructed from pPSR12 which conferred a stably mucoid phenotype to several P . syringae strains but not to Pseudomonas aeruginosa was obtained . Results obtained in this study indicate that some of the signals and regulatory genes for alginate production in P . syringae differ from those described for alginate production in P . aeruginosa.

J Med Microbiol, 1995 Jun, 42(6), 415 - 20
Electronmicroscopic investigation of the effects of biocides on Pseudomonas aeruginosa PAO bacteriophage F116; Maillard JY et al.; Electronmicroscopy was used to observe morphological changes of the Pseudomonas aeruginosa PA0 bacteriophage F116 when treated with various biocides commonly used as antibacterial and antifungal agents . Because of its large size (145 nm) and its organised structure (an isometric head and a tail), it was possible to classify structural damage into eight categories . The morphological changes induced depended on the type of biocide used and its concentration . Glutaraldehyde increased the number of phages with empty heads . Peracetic acid and phenol altered the appearance of the viral genome packaged inside the head, produced fractured heads, and damaged the tail . Peracetic acid also induced folding of the phage heads . The alcohols tested also altered the head structure . Cetylpyridinium chloride induced mainly fractured head damage . Chlorhexidine had little effect on the structure of F116.

Lett Appl Microbiol, 1995 Jun, 20(6), 357 - 60
Energy dispersive analysis of X-rays study of the distribution of chlorhexidine diacetate and cetylpyridinium chloride on the Pseudomonas aeruginosa bacteriophage F116; Maillard JY et al.; Using an energy dispersive analyser of X-rays fitted to a scanning electron microscope, chlorhexidine was shown not to bind onto F116 bacteriophage, unlike cetylpyridinium chloride, which possibly penetrated the phage . This could explain the difference in viricidal activity between the two compounds.

Am J Ophthalmol, 1995 Jun, 119(6), 738 - 43
The effect of cleaning and disinfection of soft contact lenses on corneal infectivity in an animal model; Aswad MI et al.; PURPOSE: Bacterial contamination of previously worn soft contact lenses, especially at sites of lens deposits, might play a role in the pathogenesis of lens-associated bacterial keratitis . We studied the effects of three commercial contact lens cleaners and disinfectants in a rabbit model to determine whether cleaning and disinfection reduced infectivity . METHODS: Duragel 75 soft contact lenses, designed to fit the eyes of rabbits, were worn by rabbits under tarsorrhaphies, then were removed and cleaned in one of three cleaner and disinfectant solutions according to the manufacturers' instructions . The lenses were contaminated by overnight incubation in a suspension of 10(8) Pseudomonas aeruginosa/ml and were placed under tarsorrhaphies on the eyes of fresh rabbits . The rabbits were observed for two weeks for signs of infection . Control rabbits wore new, uncleaned but contaminated lenses or worn, uncleaned but contaminated lenses . RESULTS: The rates of infection with the three commercial cleaner and disinfectant solutions ranged from 18% (two of 11) to 31% (four of 13); these incidences were not significantly different from one another or from the 19% (three of 16) incidence with new, contaminated but uncleaned lenses . By contrast, when worn, uncleaned but contaminated lenses were placed in rabbits' eyes, seven of eight were infected, a rate that is significantly higher than that of the other four groups (P = .0003) . CONCLUSIONS: These data indicate that the three commercial lens cleaner and disinfectant solutions were of similar efficacy in reducing the infectivity of contaminated contact lenses to a level similar to that of new, unworn lenses.(ABSTRACT TRUNCATED AT 250 WORDS)

Epidemiol Infect, 1995 Jun, 114(3), 395 - 402
Epidemiology of Pseudomonas aeruginosa keratitis in contact lens wearers; Stapleton F et al.; This study evaluated the epidemiology of Pseudomonas aeruginosa keratitis in contact lens (CL) wearers; the relationships between CL storage case contamination and CL hygiene practice and between CL hygiene and the development of keratitis . Sixteen CL wearers with keratitis were compared with 44 asymptomatic controls . Lens hygiene practice was assessed and CL care materials, domestic water sites and endogenous sites were evaluated microbiologically . Poor CL hygiene was not associated with Ps . aeruginosa keratitis . There was an association between keratitis and bacterial contamination of the CL and storage case (P < 0.0005) . Lens and storage case contamination were not significantly associated with poor hygiene . No domestic or endogenous source for Ps . aeruginosa was found . Causative organisms may be derived from other sources, but CLs and CL storage cases provide a favourable environment for Ps . aeruginosa colonization . Changing the CL care environment to one less favourable for Ps . aeruginosa may help to eliminate this problem.

Invest Ophthalmol Vis Sci, 1995 Jun, 36(7), 1371 - 8
In vivo bacterial protease production during Pseudomonas aeruginosa corneal infection; Kernacki KA et al.; PURPOSE . To establish if active pseudomonal proteases are present in vivo during corneal infection with Pseudomonas aeruginosa and to determine if the mouse strains used in these and previous studies have the ability to mount a nonocular antibody response to the purified proteases because antibodies to the bacterial proteases were not detected previously during in vivo ocular infection . METHODS . At certain times after corneal infection with P . aeruginosa, corneas were harvested and supernatants from the corneal homogenates were analyzed for proteolytic activity by zymography and immunoreactivity by immunoblotting . The efficiency of the extraction procedures used in these studies was determined by incubating uninfected corneal homogenates with the purified proteases . The resultant supernatants were analyzed for alkaline protease and elastase activity . Additionally, mice were immunized intraperitoneally with the purified proteases with and without adjuvant to determine if the animals could mount a nonocular antibody response . RESULTS . Corneas infected with P . aeruginosa demonstrated the presence of alkaline protease, but not elastase, by the two methods examined . The kinetics of the in vivo alkaline protease response closely parallels previously reported bacterial clearance studies in that peak alkaline protease activity was detected in corneal tissue when peak bacterial numbers also were observed in the eye, and it was absent when the eyes were sterile or nearly sterile . In addition, C57BL/6J mice were capable of mounting a nonocular antibody response to microgram quantities of both proteases only in the presence of adjuvant . CONCLUSIONS . In the model described, enzymatically active alkaline protease, but not elastase, was demonstrated in corneal tissues during in vivo infection . Concentrations of these proteases were much lower than those required to stimulate an antibody response.

J Bacteriol, 1995 Jun, 177(11), 3021 - 6
The OprB porin plays a central role in carbohydrate uptake in Pseudomonas aeruginosa; Wylie JL et al.; Using interposon mutagenesis, we have generated strains of Pseudomonas aeruginosa which lack or overexpress the substrate-selective OprB porin of this species . A marked decrease or increase in the initial uptake of glucose by these strains verified the role of OprB in facilitating the diffusion of glucose across the outer membrane of P . aeruginosa . However, we also demonstrated that the loss or overexpression of OprB had a similar effect on the uptake of three other sugars able to support the growth of this bacterium (mannitol, glycerol, and fructose) . This effect was restricted to carbohydrate transport; arginine uptake was identical in mutant and wild-type strains . These results indicated that OprB cannot be considered strictly a glucose-selective porin; rather, it acts as a central component of carbohydrate transport and is more accurately described as a carbohydrate-selective porin.

J Hosp Infect, 1995 Jun, 30(2), 125 - 31
Investigation of a nosocomial outbreak of Pseudomonas aeruginosa pneumonia in an intensive care unit by random amplification of polymorphic DNA assay; Kerr JR et al.; From July to September 1993 in the intensive care unit of the Royal Victoria Hospital there were 10 cases of pneumonia associated with sputum culture of Pseudomonas aeruginosa . The isolates had an identical biotype and pyocine typing profile . The same strain of P . aeruginosa was recovered from the sink plug-hole in two rooms, and the tap handles and ventilator tubing in a third room . All strains were retrospectively typed by the random amplification of polymorphic DNA (RAPD) method using a 26-mer oligonucleotide primer, and were identical in profile . Recommendations to medical and nursing staff included secretion isolation precautions, terminal disinfection after patient discharge, use of disposable vinyl gloves by hospital staff for all body substance contacts, thorough handwashing with 4% chlorhexidine gluconate before and after dealing with all patient contacts, and prompt, appropriate antibiotic treatment for P . aeruginosa pneumonia . RAPD is a simple and effective method to determine the relatedness of P . aeruginosa isolates, and typing results are available within a single working day; thus dramatically increasing its clinical relevance over existing molecular methods.

Jpn J Antibiot, 1995 Jun, 48(6), 741 - 8
{Annual number of Pseudomonas aeruginosa isolated inpatients, and antibacterial activity of various antibiotics against P . aeruginosa isolated from clinical specimens of inpatients}; Kouda M et al.; In recent years, isolation rate of methicillin-resistant Staphylococcus aureus (MRSA) has decreased, while the rate of Pseudomonas aeruginosa has increased . This phenomenon may be a result of regulation of use of antibiotics which belongs to the third cephems and extensive preventive measures against hospital acquired infections due to MRSA . Based on our investigation in which we determined MICs of various antibiotics against P . aeruginosa strains isolated from inpatients, the antibacterial activities of cefclidin (CFCL) was superior to those of other antibiotics . The antibacterial activity of CFCL against P . aeruginosa tested was similar to or stronger than that of tobramycin.

Pacing Clin Electrophysiol, 1995 Jun, 18(6), 1272 - 5
Removal of infected dual chambered transvenous pacemaker and implantation of a new epicardial dual chambered device with cardiopulmonary bypass: experience with seven cases; Abad C et al.; Seven patients with infected transvenous dual chambered pacemakers have undergone removal of the device using cardiopulmonary bypass . There were four women and three men with a mean age of 58 years . Six patients had localized infection in the generator pocket (mean of 4.6 previous unsuccessful operations for surgical sterilization) . Four infections were due to Staphylococcus epidermidis, two to Staphylococcus aureus, and one patient presented septicemia caused by Staphylococcus epidermidis and Pseudomonas aeruginosa . The atrial and ventricular transvenous electrodes were removed under direct vision using cardiopulmonary bypass . A new dual chambered epicardial pacemaker was implanted . The procedure was well-tolerated, and all patients are infection free with working pacemakers after a mean follow-up of 25.4 months.

Can J Microbiol, 1995 Jun, 41(6), 461 - 9
Survival of and lacZ expression in recombinant Pseudomonas strains introduced into river water microcosms; Leung K et al.; The lacZY gene cassette inserted into the chromosome of Pseudomonas aeruginosa UG2Lr and Pseudomonas aureofaciens Ps3732RNL11 was used as a genetic marker to study the fate of the Pseudomonas strains in river water microcosms . Expression of the lacZ marker in UG2Lr and Ps3732RNL11 was detected in microcosms containing as few as 12 and 14 colony-forming units (cfu)/mL of river water, respectively, by fluorimetric measurement of the beta-galactosidase activity against 4-methylumbelliferyl-beta-D-galactoside as the substrate . The persistence of and lacZ expression in the pseudomonas strains were monitored in sterile and nonsterile river water in the presence and absence of added nutrients by dilution plating and fluorimetry, respectively . After incubation for 10 d at 10 degrees C, culturable populations of strain UG2Lr in sterile water samples, with and without nutrient added, decreased from an initial density of 1.5 x 10(4) to 1.7 x 10(3) and 4.6 x 10(3) cfu/mL, respectively . Despite similar numbers of UG2Lr cells in the two treatments, expression of the lacZ marker in the surviving cells of the nutrient-supplemented sample was 24 times higher than in the cells of the unamended sample . In nonsterile water samples, the density of UG2Lr declined to less than 6 cfu/mL in 30 d regardless of the nutrient conditions . A nutrient supplement increased the growth of the native bacterial population but did not enhance growth of and lacZ expression in the bacteria . Polymerase chain reaction analysis showed a decrease in amplification signal indicating a genuine decline in viable UG2Lr cell density in the water samples.(ABSTRACT TRUNCATED AT 250 WORDS)

Planta Med, 1995 Jun, 61(3), 275 - 6
An antibacterial vitamin E derivative from Tovomitopsis psychotriifolia; Setzer WN et al.; The crude ethanol extract from the leaves of Tovomitopsis psychotriifolia (Clusiaceae) exhibits antibacterial activity against Bacillus cereus, Staphylococcus aureus, and Pseudomonas aeruginosa . The biologically active agent in the extract has been isolated by chromatographic techniques and identified by NMR spectroscopy as trans-delta-tocotrienoloic acid.

Kansenshogaku Zasshi, 1995 Jun, 69(6), 711 - 22
{Study on azithromycin in treatment of diffuse panbronchiolitis}; Kobayashi H et al.; In the treatment of diffuse panbronchiolitis, azithromycin (AZM), a new macrolide antibiotic with 15-membered lactone ring, was studied for its efficacy and safety . AZM, 250 mg, was intermittently administered to a total of 60 patients twice a weeks, for 3 months as a rule, and its efficacy was clinically evaluated in 52 patients and the safety in 55 . The rate of efficacy was 84.6% (44/52) . Clinical findings 12 weeks after the start of administration showed a decrease in sputum volume in 30 of 46 patients and amelioration of dyspnea on exertion in 23 of 46 patients, and no worsening of symptoms was observed in the patients . Vital capacity (4/22), FEV1.0 (6/21), cold agglutination reaction (22/28), and CRP (16/36) were also improved . The rate of eradication of organisms isolated from the sputum except for indigenous organisms was 39.5% (15/38); 4 of the 22 strains of Pseudomonas aeruginosa were eradicated . Adverse reactions were observed in 4 of the 55 patients (7.3%), 1 patient each with rash, itching, diarrhea, and a gastric symptom (heavy feeling in the stomach) . 4 of the 54 patients (7.4%) exhibited abnormal changes in clinical laboratory test values values . These were an increase in eosinophil count in 2, elevation of GOT in 1, and elevation of Al-P in 1 . These adverse reactions and abnormal changes in laboratory tests were mild or moderate . Therefore, long-term intermittent administration of AZM, twice a week, is expected to have the same effect in the treatment of diffuse panbronchiolitis as long-term small-dose administration of 14-membered macrolides such as erythromycin and clarithromycin, whose effects have already been established.

Kansenshogaku Zasshi, 1995 Jun, 69(6), 696 - 700
{Effectiveness of antibacterial clothes and wall material against methicillin resistant Staphylococcus aureus and Pseudomonas aeruginosa}; Okuda K et al.; To evaluate the antibacterial activities of antibacterial clothes and wall material, we measured the reduction rates of MRSA and P . aeruginosa using four products of antibacterial clothes and one wall material . Antibacterial activities are judged by the reduction rates of bacteria after 18 hours incubation at 35.0 degrees C . In the 50 strains of MRSA tested, two of four brands of clothes showed over 99% reduction rates, while only one brand showed a reduction rates over 99% in the 13 strains of P . aeruginosa . The wall material tested showed a reduction rates over 99% for the 50 strains of MRSA and over 90% for the 50 strains of P . aeruginosa . These data indicate that usage of some products of antibacterial clothes and wall material in the hospital may be useful the prevention of nosocomial infection.

FEMS Immunol Med Microbiol, 1995 Jun, 11(3), 191 - 5
Purified Pseudomonas aeruginosa PA-I lectin induces gut growth when orally ingested by rats; Grant G et al.; The effects of PA-I lectin isolated from the human pathogen Pseudomonas aeruginosa upon cellular metabolism in vivo have been studied using the rat gut as a model system . Orally ingested PA-I lectin stimulated metabolic activity and induced polyamine accumulation and growth in the small intestine, caecum and colon . The nature and extent of the changes induced by PA-I lectin were similar to those caused by dietary kidney bean lectin and were likely to lead to impaired epithelial cell function and integrity . This finding contributes to our understanding of the possible roles of these lectins in Pseudomonas aeruginosa infection.

Antimicrob Agents Chemother, 1995 Jun, 39(6), 1324 - 8
Noncytotoxic combinations of topical antimicrobial agents for use with cultured skin substitutes; Boyce ST et al.; Cultured skin grafts are destroyed more easily than split-thickness skin grafts by common burn wound organisms, including gram-negative and gram-positive bacteria and fungi . To increase the survival and engraftment of cultured skin grafts, formulations of antimicrobial agents were tested for cytotoxicity to cultured human keratinocytes and fibroblasts and for activity against common organisms from burn wounds . On the basis of previous studies, a base formulation containing neomycin (40 micrograms/ml), polymyxin B (700 U/ml), and mupirocin (40 micrograms/ml) was prepared, to which ciprofloxacin (20 micrograms/ml) or norfloxacin (20 micrograms/ml) and amphotericin B (0.25 microgram/ml) or nystatin (100 U/ml) were added . Toxicity to cultured human cells was determined by the growth response of cell cultures (n = 6) to each drug combination over 4 days . Activity against clinical isolates (n = 40) of Staphylococcus aureus, Pseudomonas aeruginosa, other gram-negative bacteria, and Candida spp . was determined by the wet disc assay . Analysis of variance testing showed no significant differences in the growth of keratinocytes or fibroblasts under control or experimental conditions . Medium without antimicrobial agents was not effective against any of the 40 microbial strains tested . The base formulation was effective against all bacterial strains tested but against none of the fungi, while all experimental formulations were effective against all microbial strains tested . These findings suggest that neomycin, mupirocin, and polymyxin B may be combined with a quinolone and an antimycotic agent to provide broad antimicrobial activity for a formulation for topical use with cultured skin on burns . However, the formulations described here are strictly experimental and are not recommended for clinical use without further evaluation.

Antimicrob Agents Chemother, 1995 Jun, 39(6), 1314 - 9
Different patterns of bacterial DNA synthesis during postantibiotic effect; Gottfredsson M et al.; Studies on bacterial metabolism during the postantibiotic effect (PAE) period are limited but might provide insight into the nature of the PAE . We evaluated the rate of DNA synthesis in bacteria during the PAE period after a 1-h exposure of organisms in the logarithmic growth phase to various antibiotics . Staphylococcus aureus ATCC 25923 was exposed to vancomycin, dicloxacillin, rifampin, and ciprofloxacin; Escherichia coli ATCC 25922 was exposed to gentamicin, tobramycin, rifampin, imipenem, and ciprofloxacin; and Pseudomonas aeruginosa ATCC 25783 was exposed to imipenem, tobramycin, and ciprofloxacin . DNA synthesis was determined by measuring the rate of {3H}thymidine incorporation in S . aureus and E . coli and {3H}adenine incorporation in P . aeruginosa . DNA synthesis in S . aureus was suppressed during the PAE phase with vancomycin, dicloxacillin, and rifampin, it was suppressed in E . coli with rifampin, and it was suppressed in P . aeruginosa after exposure to tobramycin . Conversely, DNA synthesis was relatively enhanced in the gram-negative bacilli after exposure to imipenem and in all three species after exposure to ciprofloxacin . However, DNA synthesis in E . coli was only minimally affected after exposure to tobramycin and gentamicin . The differences in DNA synthesis observed after exposure to various antimicrobial agents suggest multiple mechanisms for the PAE.

J Chemother, 1995 Jun, 7(3), 179 - 83
Comparative in vitro killing activity of meropenem versus imipenem against multiresistant nosocomial Pseudomonas aeruginosa; Giamarellos-Bourboulis EJ et al.; In order to compare the in vitro killing activity of meropenem and imipenem against multiresistant P.aeruginosa 14 strains were used . All nosocomial isolates were susceptible to meropenem and imipenem minimum inhibitory concentration (MIC < or = 4 micrograms/ml) and resistant to at least two other antimicrobial agents of diverse chemical class with antipseudomonal activity . Forty-two killing curves were performed by exposing a 5 x 10(5) CFU/ml log-phase inoculum to 1x minimum bactericidal concentration (MBC) of each carbapenem . Meropenem was found to possess a slower killing rate than imipenem over the first 5 hours of P.aeruginosa exposure, but to be equally effective as imipenem after 24 hours of incubation . Forty percent and 11.1% of P.aeruginosa strains developed resistance to imipenem and meropenem respectively after a 24-hour exposure to carbapenem . The authors speculate about the underlying mechanisms explaining the higher rate of resistance development to imipenem than to meropenem.

J Hosp Infect, 1995 Jun, 30 Suppl, 275 - 81
Contact lens wear by hospital health care staff: is there cause for concern?
Hay J, Seal DV.
Microbial keratitis can occur in association with contact lens wear . The absolute risk of infection is low but may be enhanced as a consequence of increased exposure to potentially pathogenic microbes in a hospital setting . There is variation in risk depending on type of lens worn and its modality of use . Extended-wear lenses carry the greatest risk . Pseudomonas aeruginosa and Acanthamoeba are causes of potentially devastating ocular infections in contact lens wearers . The risk of these infections could be reduced by fastidious hygiene practice . Hydrogen peroxide disinfection is recommended when a storage case is included in the care regimen . This should be cleaned thoroughly and dried prior to disinfection and never exposed to tap water . Daily wear of one-day 'disposable' soft contact lenses or use of rigid gas permeable lenses is recommended for hospital staff . Contact lenses should be removed immediately and discarded or disinfected if the eye becomes contaminated and/or use of an eyewash is required.

Intern Med, 1995 Jun, 34(6), 564 - 8
Ruptured subcapsular giant hematoma of the spleen as a complication of chronic pancreatitis; Kuramitsu T et al.; We present a 63-year-old man with chronic pancreatitis and the rare complication of giant subcapsular splenic hematoma . The hematoma showed no size reduction for 6 weeks . Then, the hematoma was infected with pseudomonas aeruginosa after the recurrence of the pancreatitis, and it finally ruptured . This case suggested that in cases of giant subcapsular splenic hematoma with chronic pancreatitis reductive pressure treatment should be administered as early as possible.

APMIS, 1995 Jun, 103(6), 447 - 59
Tumor necrosis factor alpha plus interleukin 1 beta treatment protects granulocytopenic mice from Pseudomonas aeruginosa lung infection: role of an unusual inflammatory response; Amura CR et al.; We have recently demonstrated that treatment with interleukin 1 beta (IL-1 beta) plus tumor necrosis factor alpha (TNF alpha) protects granulocytopenic hosts from Pseudomonas aeruginosa aerosol challenge . In this study we characterized the inflammatory response induced by P . aerugionsa in granulocytopenic mice treated with 2,000 U IL-1 beta plus 2,000 U TNF alpha . Treatment with the nonsteroidal anti-inflammatory agent piroxicam abolished both the protective effect of cytokine treatment and the increase in myeloperoxidase (MPO) pulmonary activity . Histopathological studies revealed that, after aerosol challenge with P . aeruginosa, treatment with these cytokines induced migration and extravasation of mononuclear cells of immature appearance into the lung parenchyma . These cells contained MPO in their cytoplasm and displayed phagocytic capacity . Resident alveolar macrophages exhibited signs of activation and appeared in reduced numbers in bronchoalveolar lavage fluid . We suggest that the inflammatory response promoted by low TNF alpha plus IL-1 beta doses may be one mechanism responsible for protection of granulocytopenic hosts from P . aeruginosa aerosol challenge.

J Clin Microbiol, 1995 Jun, 33(6), 1461 - 6
Genetic diversity of Pseudomonas aeruginosa strains isolated from patients with cystic fibrosis revealed by restriction fragment length polymorphism of the rRNA gene region; Martin C et al.; The restriction fragment length polymorphism patterns of rDNAs from Pseudomonas aeruginosa strains isolated from the respiratory tracts of patients suffering from cystic fibrosis were obtained to evaluate the genetic polymorphism of this population of strains . Eighty-seven P . aeruginosa strains isolated from 87 patients from diverse areas of France and the ATCC 10145 strain were examined . Four restriction enzymes were used: BamHI, ClaI, EcoRI, and PstI . Forty-nine strains (56%) were in the three most frequent ribotypes (ribotypes R1 to R3) . In addition, hierarchical clustering analysis of the data showed that 71 of the 88 strains (81%) clustered at a distance of less than one-third of the greatest distance observed in the total population . This indicates that clinical isolates implicated in the pathology of cystic fibrosis present a low degree of heterogeneity of rRNA operons, in contrast to the heterogeneity of strains of P . aeruginosa isolated from patients with various other pathologies . This relative homogeneity of rRNA genes was observed independently of the clinical status of the patient and the colony morphology.

J Bacteriol, 1995 Jun, 177(11), 3052 - 7
Transcriptional analysis of the amidase operon from Pseudomonas aeruginosa; Wilson SA et al.; The transcriptional start point for the amidase structural gene (amiE) of Pseudomonas aeruginosa has been identified, and the promoter (pE) has been shown to function constitutively, as predicted for a system regulated by transcription antitermination . Northern (RNA) analysis results show that in cells grown under inducing conditions, a major 1.3-kb amiE transcript arises from pE, and in addition, a larger transcript of approximately 5.0 kb in length has been shown to derive from the same promoter, encoding all of the genes of the operon . DNA sequencing and S1 nuclease mapping have located a transcription terminator downstream of amiE, which terminates approximately half of the pE transcripts . Previously, two RpoN-dependent promoter-like sequences (pN1 and pN2) were identified upstream of the negative regulator gene, amiC, and we have now constructed a promoter probe vector which shows weak constitutive promoter activity within this region . This promoter would be expected to provide basal levels of expression of the amiC and amiR regulatory genes to allow induction of the system.

Clin Immunol Immunopathol, 1995 Jun, 75(3), 214 - 24
Role of the low zinc bioavailability on cellular immune effectiveness in cystic fibrosis; Mocchegiani E et al.; An altered cellular immune response as a secondary phenomenon has been suggested to be probably involved in the bronchopulmonary infections by Pseudomonas aeruginosa in cystic fibrosis (CF) . The difficulty to eradicate with modern anti-pseudomonal antibiotics the bronchopulmonary infections has led us to further investigate the possible existence of other cellular immune defects and their cause . Alterations in zinc turnover are present in CF . Zinc is relevant for good immune functioning . In particular, zinc is required to confer biological activity to thymulin (ZnFTS), a biochemically defined thymic hormone with a modulating action on cell-mediated immunity . The zinc-unbound form (FTS) is inactive and it can be unmasked by in vitro zinc addition to the plasma samples revealing the total amount of circulating thymulin (active + inactive) . Marginal zinc deficiencies may prevent peripheral biological activation of active thymulin . Total zinc-saturable thymulin fractions in CF are similar to those observed in normal subjects, whereas the active quota is strongly reduced associated with concomitant high plasma levels of inactive thymulin compared to the values of healthy children (P < 0.01) . A strict correlation exists between zinc and thymic hormone-saturable fraction (r = 0.87, P < 0.01) in CF . These findings suggest that the defect is not due to a thymic failure but to a reduced peripheral saturation of thymulin by zinc ions . This defect might depend on augmented plasma concentration of alpha 2-macroglobulin, which has a higher binding affinity for zinc than thymulin . T cell subsets are normal in CF . Reduced NK cell number and activity are present . Also, plasma IL-2 levels are reduced . The existence of positive correlations between zinc and IL-2 (r = 0.79, P < 0.01) and between zinc or active thymulin and NK activity (r = 0.70, P < 0.01 and r = 0.88, P < 0.01, respectively) suggest a close link among zinc failure, impaired IL-2 activity, low thymulin level, and reduced NK activity in CF patients with both normal and growth retardation . Although the role of NK cells is unknown in CF, a zinc supplementation, in order to induce a complete saturation of thymulin molecules, to correct some cellular immune defects and to improve the growth, may be suggested.

Mol Microbiol, 1995 Jun, 16(5), 931 - 41
Membrane topology and site-specific mutagenesis of Pseudomonas aeruginosa porin OprD; Huang H et al.; Pseudomonas aeruginosa OprD is a 420-amino-acid protein that facilitates the uptake of basic amino acids, imipenem and gluconate across the outer membrane . OprD was the first specific porin that could be aligned with members of the non-specific porin super-family . Utilizing multiple alignments in conjugation with structure predictions and amphipathicity calculations, an OprD-topology model was proposed . Sixteen beta-strands were predicted, connected by short loops at the periplasmic side . The eight external loops were of variable length but tended to be much longer than the periplasmic ones . Polymerase chain reaction (PCR)-based site-specific mutagenesis was performed to delete separately short stretches (4-8 amino acid residues) from each of the predicted external loops . The mutants with deletions in the predicted external loops L1, L2, L5, L6, L7 and L8 were tolerated in both Escherichia coli and P . aeruginosa . The expressed mutant proteins maintained substantial resistance to trypsin treatment in the context of isolated outer membranes . Proteins with deletions in loops L1, L5, L6, L7 and L8 reconstituted similar imipenem supersusceptibility in a P . aeruginosa OprD:: omega background . The L2-deletion mutant only partially reconstituted super-susceptibility, suggesting that loop L2 is involved in imipenem binding . These data were generally consistent with the topology model.

Mol Microbiol, 1995 Jun, 16(5), 1001 - 9
Importance of lipooligosaccharide structure in determining gonococcal resistance to hydrophobic antimicrobial agents resulting from the mtr efflux system; Lucas CE et al.; Levels of gonococcal resistance to antimicrobial hydrophobic agents (HAs) are controlled by the mtr (multiple transferrable resistance) system, composed of the mtrRCDE genes . The mtrR gene encodes a transcriptional repressor that appears to regulate expression of the upstream and divergent mtrCDE operon . The mtrCDE genes encode membrane proteins analogous to the MexABOprK proteins of Pseudomonas aeruginosa that mediate export of structurally diverse antimicrobial agents . In this study we found that a single base pair deletion in a 13 bp inverted repeat sequence within the mtrR promoter resulted in increased resistance of gonococci to both crystal violet (CV) and erythromycin (ERY) as well as to the more lipophilic non-ionic detergent Triton X-100 (TX-100) . However, this cross-resistance was contingent on the production of a full-length lipooligosaccharide (LOS) by the recipient strain used in transformation experiments . Introduction of this mutation (mtrR-171) into three chemically distinct deep-rough LOS mutants by transformation resulted in a fourfold increase in resistance to TX-100 compared with a 160-fold increase in an isogenic strain producing a full-length LOS . However, both wild-type and deep-rough LOS strains exhibited an eightfold increase in resistance to CV and ERY as a result of the mtrR-171 mutation . This suggests that gonococci have different LOS structural requirements for mtr-mediated resistance to HAs that differ in their lipophilic properties . Evidence is presented that gonococci exclude HAs by an energy-dependent efflux process mediated by the mtr system.

FEBS Lett, 1995 May 29, 365(2-3), 152 - 4
Nucleotide sequence encoding the di-haem cytochrome c551 peroxidase from Pseudomonas aeruginosa; Ridout CJ et al.; The nucleotide sequence of the gene encoding cytochrome c551 peroxidase from Pseudomonas aeruginosa is reported . The translated amino acid sequence differs from the sequence reported earlier by peptide mapping most significantly by the presence of a section containing an additional 20 residues . A number of minor differences are also evident . The new sequence translates to a protein containing 346 amino acids, the first 23 being typical of a hydrophobic leader peptide with a characteristic protease cleavage site.

Gene, 1995 May 26, 158(1), 15 - 22
An improved system for gene replacement and xylE fusion analysis in Pseudomonas aeruginosa; Schweizer HP et al.; A novel pUC19-based gene replacement vector has been developed . This vector incorporates (i) the counterselectable sacB marker, (ii) a lacZ alpha allele for blue-white screening, (iii) an oriT for conjugation-mediated plasmid transfer and (iv) unique cloning sites for SmaI and the rare-cutting meganuclease I-SceI . These rare restriction sites are also present on the helper plasmid pUC19Sce . The replacement vector is engineered to contain few restriction sites to gain greater access to restriction sites within cloned DNA fragments, thus facilitating their genetic manipulation . The usefulness of the system was demonstrated by chromosomal integration of a newly constructed xylE::GmR fusion cassette into the glpD gene of Pseudomonas aeruginosa.

Gene, 1995 May 26, 158(1), 143 - 4
Nucleotide sequence of a small plasmid isolated from Acetobacter pasteurianus; Fomenkov A et al.; A 1440-bp plasmid named pAP12875 was isolated from Acetobacter pasteurianus and its nucleotide sequence determined . An open reading frame was found capable of coding for a protein that has similarity with the replication protein of pVT736-1 from Actinobacillus actinomycetemcomitans and the 32-kDa protein of phage Pf3 from Pseudomonas aeruginosa.

Gene, 1995 May 26, 158(1), 55 - 60
Pseudomonas aeruginosa outer membrane protein OprF as an expression vector for foreign epitopes: the effects of positioning and length on the antigenicity of the epitope; Wong RS et al.; OprF, the major outer membrane (OM) protein of Pseudomonas aeruginosa, has been proposed to be comprised of a series of beta-strands separated by periplasmic or surface-exposed loop regions . In this study, a simple malarial epitope was used to demonstrate that OprF can be used as an expression vector to present foreign peptide sequences, namely, the 4-amino-acid (aa) repeating epitope (Asn-Ala-Asn-Pro = NANP) of the circumsporozoite protein of the human malarial parasite Plasmodium falciparum . Eight permissive sites, that allowed the expression and surface exposure of the malarial epitope, were identified throughout OprF . Using a monoclonal antibody (mAb) specific for the malarial epitope, we investigated the effects of positioning and length of the epitope on its antigenicity in the OprF expression vector system . It was demonstrated that the malarial epitope inserted at aa26 was significantly more reactive with the epitope-specific mAb (i.e., more antigenic) when assayed in the context of whole cells whereas those at aa213 and aa290 were more antigenic when assayed in the OM . The malarial epitope inserted at aa188 and aa196 was moderately antigenic, while this epitope inserted at aa215 and aa310 showed low antigenicity with the same mAb in both whole cell and OM assays . For two insertion sites, aa26 and aa213, we demonstrated that the insertion of multiple copies of the epitope enhanced reactivity with the malarial epitope-specific mAb . These data are discussed with respect to the local OprF sequences into which the epitope was inserted.

Biochem Biophys Res Commun, 1995 May 16, 210(2), 356 - 62
Expression of genes associated with antibiotic extrusion in Pseudomonas aeruginosa; Morshed SR et al.; We cloned the gene(s) associated with multiantibiotic resistance in Pseudomonas aeruginosa from a mutant with elevated drug extrusion . The strain harboring cloned gene accumulated less amount of ofloxacin than the strain without the clone . By using Southern and northern blot analyses, we investigated whether multiantibiotic resistance is caused by gene amplification or overexpression . We found that all our multiantibiotic resistance is caused by gene amplification or overexpression . We found that all our multiantibiotic resistant mutants isolated earlier overexpressed mRNA homologous to the cloned gene(s) . Overexpression of the 41 KDa and 110 KDa inner and the 50 KDa outer membrane proteins were detected . Other multiantibiotic resistant mutants including the nalB mutant overexpressed the mRNA and the membrane proteins, but the nfxB and nfxC mutants did little . We concluded that low antibiotic accumulation in the multiantibiotic resistant mutants is attributable to overexpression of the antibiotic extrusion machinery.

Biochim Biophys Acta, 1995 May 11, 1244(1), 185 - 90
B/PI-derived synthetic peptides: synergistic effects in tethered bactericidal and endotoxin neutralizing peptides; Gray BH et al.; Human neutrophil bactericidal protein (B/PI) is known for its ability to kill bacteria and to neutralize the action of endotoxin . Short linear peptides derived from residues 80-109 have been synthesized and their bactericidal and endotoxin neutralizing activities have been assayed . A series of 'walk-through' decapeptides, overlapping 3 to 4 residues, indicates that endotoxin neutralizing and partial bactericidal activities can be localized within the N- and C-terminal portions, respectively, of the 80-109 sequence . Bactericidal activity toward Pseudomonas aeruginosa was localized in central peptides of the walk-through series and greatest in peptide 90-99 . By using longer peptides, residues 86-104 and 82-108, both bactericidal and endotoxin neutralizing activities are significantly enhanced . Bactericidal activity of peptide 82-108 is now only 6-fold less than that of parent B/PI and 9-fold more potent than peptide 86-104 . The 82-108 peptide was 7-fold more active at endotoxin neutralization than 86-104 but showed less enhanced activity, being approx . 470-times less active than B/PI . Cyclized 82-108 peptide retained bactericidal activity but did not improve in capacity to neutralize endotoxin.

Biochim Biophys Acta, 1995 May 10, 1229(3), 393 - 7
The Na(+)-translocating ATPase of Acetobacterium woodii is a F1F0-type enzyme as deduced from the primary structure of its beta, gamma and epsilon subunits; Forster A et al.; A 4.5 kbp EcoRI fragment hybridizing to a fragment of uncD (coding for subunit beta of F1F0-ATPases) was cloned from chromosomal DNA of Acetobacterium woodii . The nucleotide sequence was determined and revealed five open reading frames (ORF), four of which were identified to code for subunits of the Na(+)-ATPase . The deduced amino acid sequences of these ORF's are homologous to subunit alpha (partial coding sequence, C-terminal end), gamma, beta and epsilon of F1F0-ATPases from various organisms; furthermore, the organization of the genes in the order uncA (alpha), uncG (gamma), uncD (beta), uncC (epsilon) is identical to the structure of unc operon as present in most bacteria . Downstream of uncC is an ORF whose deduced amino acid sequence has 53% sequence homology to AlgD from Pseudomonas aeruginosa . The structure and organization of the unc genes are the final proof that the Na(+)-ATPase from A . woodii is a member of the family of F1F0-ATPases.

Dtsch Med Wochenschr, 1995 May 5, 120(18), 646 - 8
{Chronic granulomatosis: a rare differential diagnosis in liver granulomas in adulthood}; Glaser J et al.; A now 43-year-old man was known since childhood to have mesenteric and subcutaneous lymphadenopathy . Histological examination of liver biopsies and excision of some lymph nodes when an adult showed epithelioid granulomas, in places with Langhans giant cells . Diagnostic splenectomy revealed no pathological findings . His present admission to hospital was for an infection with high fever . On auscultation moist rales were audible over the apex of the left lung . The chest radiography showed pneumonic infiltration . Blood culture grew Pseudomonas aeruginosa . Ultrasound demonstrated hypoechogenic homogeneous and smoothly circumscribed round foci in the liver hilus and around the coeliac trunk . The upper lobe pneumonia healed under antibiotic treatment . As chronic granulomatosis was suspected, the nitroblue tetrazolium and superoxide production tests were performed . They demonstrated that the capacity of the granulocytes to form oxygen radicals was markedly diminished . Chronic granulomatosis is an inherited disorder of granulocyte function linked to the X-chromosome . It must be included in the differential diagnosis of any unclear granulomatous disease even in adults.

Ear Nose Throat J, 1995 May, 74(5), 360 - 3
The management of sinusitis in patients infected with the human immunodeficiency virus (HIV); Tami TA; As the Human Immunodeficiency Virus (HIV) has extended its influence across the United States, otolaryngologists have been increasingly called upon to manage its various head and neck manifestations . Sinusitis is a very prevalent, yet difficult, management problem in this patient population . The pathophysiology of sinusitis in this setting relates to altered helper T-lymphocyte function, an abnormal inflammatory response as well as increased IgE-mediated inflammation . Chronic HIV-related sinusitis is often due to Pseudomonas aeruginosa, Staphylococcus aureus, or anaerobic bacteria, and empiric antibiotic therapy must include these potential pathogens . Early cultures can facilitate organism-specific antibiotic therapy . Aggressive treatment with decongestants, topical nasal steroids, mucoevacuants and occasionally antihistamines should be included at maximal tolerated doses . When medical therapy fails, surgical drainage can be a safe and effective management option . Appropriately directed medical, and occasionally surgical, therapy can lead to a dramatic clinical response and provide an improved quality of life in this patient population.

Microbiology, 1995 May, 141 ( Pt 5), 1247 - 54
pilO, a gene required for glycosylation of Pseudomonas aeruginosa 1244 pilin; Castric P; Nucleotide sequencing of a region downstream from the Pseudomonas aeruginosa 1244 pilin structural gene, pilA, revealed an ORF potentially able to code for a protein of M(r) 50,862 . This ORF, called pilO, was flanked by a tRNAthr gene, which was followed by a transcriptional termination sequence . The tRNAthr gene and the termination sequence were nearly identical to sequences found immediately adjacent to the pilA gene of several P . aeruginosa strains . A 2200 base mRNA strand, which contained both the pilO and pilA transcripts, was produced from this region, while a 650 base transcript containing only pilA was present in a 100-fold excess over the longer transcript . Hyperexpression of the pilA gene in a PilO- strain resulted in normal pilus-specific phage sensitivity and twitching motility . The pilin produced by this strain had a lower apparent M(r) and a more neutral pl compared to that produced by a strain containing a functional pilO gene . This pilin failed to react with a sugar-specific reagent which recognized pilin produced by the strain containing a functional pilO gene.

J Bacteriol, 1995 May, 177(10), 2744 - 50
Cloning and characterization of pvdS, a gene required for pyoverdine synthesis in Pseudomonas aeruginosa: PvdS is probably an alternative sigma factor; Cunliffe HE et al.; Cells of Pseudomonas aeruginosa secrete a fluorescent yellow-green siderophore, pyoverdine, when grown under iron-deficient conditions . We describe here the cloning and characterization of a gene, pvdS, which is required for this process . The pvdS gene is required for expression from promoters of at least two pyoverdine synthesis genes and can cause expression from these promoters in Escherichia coli, where they are otherwise inactive . Sequencing of pvdS revealed that it is a member of a subfamily of RNA polymerase sigma factors which direct the synthesis of extracellular products by bacteria . The pvdS gene is expressed only in iron-starved bacteria, and in E . coli cells at least, expression is regulated by the Fur repressor protein . We propose that in iron-rich cells of P . aeruginosa, Fur binds to the pvdS promoter and prevents expression of the gene; under conditions of iron starvation, repression is relieved and PvdS is made, reprogramming the cells for pyoverdine synthesis.

Chest, 1995 May, 107(5), 1317 - 22
The pulmonary nodule after lung transplantation . Cause and outcome; End A et al.; In the immunocompromised patient, the pulmonary nodule remains a diagnostic and therapeutic challenge . We studied the incidence, cause, diagnosis, and therapy of pulmonary nodules after lung transplantation (LTx) . Eight out of 64 patients (12.5%) developed pulmonary nodules after a median follow-up of 5.8 months (range, 1 to 10 months) . The median age was 30.5 years (range, 21 to 62 years) . Solitary pulmonary nodules (n = 2) disappeared spontaneously within 3 weeks and were suspected to be of infectious origin . The cause of multiple nodules (n = 6) was posttransplant lymphoproliferative disorder (PTLD {n = 3}), aspergillosis (n = 2), and abscesses caused by Pseudomonas aeruginosa and Staphylococcus aureus (n = 1) . After an initial chest radiograph, CT with fine-needle biopsy was the most valuable diagnostic tool . In six patients, nodules resolved within 10 weeks (median, 8 weeks) . Two patients, however, died of sepsis (P aeruginosa and S aureus and Aspergillus, respectively) . The differential diagnosis of pulmonary nodules after LTx primarily comprises PTLD and infection (bacterial or fungal) . To improve the outcome, early, aggressive treatment is mandatory; therefore, serial CT scans are strongly recommended to be part of the diagnostic armamentarium in LTx recipients.

Am J Respir Cell Mol Biol, 1995 May, 12(5), 513 - 9
Lymphocyte accumulation during Pseudomonas aeruginosa-induced pneumonia in rodents does not require CD11a and intercellular adhesion molecule-1; Wiebke JL et al.; During Pseudomonas aeruginosa-induced pneumonia in rodents, the acute infiltrate of neutrophils is followed by accumulation of lymphocytes in the perivascular connective tissue . The roles of the adhesion molecules CD11a/CD18 and intercellular adhesion molecule-1 (ICAM-1) in this accumulation of lymphocytes were investigated . The numbers of lymphocytes in P . aeruginosa-induced pneumonia were compared in animals treated with blocking antibodies to either CD11a, ICAM-1, IgG, or no antibody . In other experiments, the lymphocyte accumulation during P . aeruginosa-induced pneumonia in ICAM-1 mutant mice was compared with that in wild-type mice . In rats, both a murine anti-rat CD11a antibody and nonspecific murine IgG partially inhibited the lymphocyte accumulation by 30 to 40% compared with animals that received no antibodies . In mice, blocking antibodies to either CD11a or ICAM-1 did not decrease the lymphocyte accumulation compared with mice given IgG or no antibody . Further, there was no attenuation of the lymphocyte accumulation induced by P . aeruginosa in the ICAM-1 mutant mice compared with wild-type mice, either in the total number of lymphocytes or the number of CD4+, CD8+, or B cells . We conclude that neither CD11a/CD18 nor ICAM-1 are required for lymphocyte accumulation during P . aeruginosa-induced pneumonia in rodents . The partial inhibition of the lymphocyte accumulation in both the anti-CD11a- and IgG-treated rats may be due to nonspecific effects of foreign proteins on cellular functions.

J Bacteriol, 1995 May, 177(9), 2469 - 74
Regulation of nucleoside diphosphate kinase and secretable virulence factors in Pseudomonas aeruginosa: roles of algR2 and algH; Schlictman D et al.; Alginate is an important virulence factor for Pseudomonas aeruginosa during infection of the lungs of cystic fibrosis patients . The genes encoding enzymes for alginate production by P . aeruginosa are normally silent . They are activated in response to several environmental conditions, including high osmolarity, exposure to ethanol, or long-term growth under conditions of nutrient deprivation . Several genes which participate in the activation of alginate gene promoters have been identified; among these is the algR2 (algQ) gene . AlgR2 is an 18-kDa protein which has been shown to regulate the critical algD gene encoding GDP-mannose dehydrogenase as well as to regulate the levels of a tricarboxylic acid cycle enzyme, i.e., succinyl coenzyme A synthetase, and nucleoside diphosphate kinase (Ndk), an enzyme involved in nucleoside triphosphate synthesis . Succinyl coenzyme A synthetase and Ndk form a complex in P . aeruginosa . While algR2 is required for alginate synthesis at 37 degrees C, an algR2 insertion mutant was still able to make alginate slowly at 37 or at 30 degrees C . We used this observation to identify and clone a gene, termed algH . A strain with mutations in both algR2 and algH is unable to produce alginate at either 37 or 30 degrees C, and it is fully defective in Ndk production.

Invest Ophthalmol Vis Sci, 1995 May, 36(6), 1107 - 14
Aged mice fail to upregulate ICAM-1 after Pseudomonas aeruginosa corneal infection; Hobden JA et al.; PURPOSE . In young Swiss (HSD:ICR) outbred mice, corneal clarity is restored after Pseudomonas aeruginosa ocular infection, whereas disease in aged outbred mice progresses to corneal perforation . This study was conducted to elucidate further the mechanism responsible for this age-related disparity in disease response . METHODS . Corneas of young (6 to 8 weeks of age) and aged (1.5 to 2 years) female mice were scarified and inoculated with 1.0 x 10(8) colony-forming units of P . aeruginosa ATCC 19660 . Eyes were scored for corneal pathology (0 to +4) at 6, 12, 24, 48, 72, 96, and 120 hours after infection . At each time point, six mice were killed from each age group, and both eyes were enucleated . Eyes (three infected, three uninfected) were embedded in OCT compound, frozen in liquid nitrogen, sectioned on a cryostat, and stained for ICAM-1 and LFA-1 immunoreactivity . The remaining six eyes (three infected, three uninfected) were embedded in eponaraldite resin, thick sectioned, and stained for light microscopic histopathologic examination . RESULTS . Immunostaining of slight to moderate intensity for ICAM-1 was seen on conjunctival fibroblasts, stromal keratocytes, corneal epithelium, and endothelium and conjunctival blood vessel endothelium of uninfected contralateral eyes in both age groups . In response to P . aeruginosa infection, only young animals were capable of upregulating ICAM-1 (as evidenced by an increase in the intensity of immunostaining) on these cells when compared to aged mice . Conversely, the intensity of immunostaining for LFA-1, a ligand for ICAM-1 on infiltrating leukocytes, was similar despite animal age . On gross observation, corneal pathology was more severe in young mice 24 to 96 hours after infection . Histopathologically, in contrast to young mice, eyes of aged animals 24 to 48 hours after infection had significantly fewer inflammatory cells, such as polymorphonuclear leukocytes (PMNs), infiltrating the corneal stroma and adhering to the endothelium near wound sites . CONCLUSION . These data suggest that the disparate response to ocular P . aeruginosa infection in young versus aged mice is due, at least in part, to the inability of aged animals to upregulate ICAM-1 above constitutively expressed levels . Consequently, the migration of inflammatory cells (PMNs) into infected corneas of aged mice is delayed, perhaps facilitating bacterial growth and contributing to a poor prognosis.

Infect Immun, 1995 May, 63(5), 1855 - 62
Protection of immunocompromised mice against lethal infection with Pseudomonas aeruginosa by active or passive immunization with recombinant P . aeruginosa outer membrane protein F and outer membrane protein I fusion proteins; von Specht BU et al.; Recombinant outer membrane proteins (Oprs) of Pseudomonas aeruginosa were expressed in Escherichia coli as glutathione S-transferase (GST)-linked fusion proteins . GST-linked Oprs F and I (GST-OprF190-350 {GST linked to OprF spanning amino acids 190 to 350} and GST-OprI21-83, respectively) and recombinant hybrid Oprs (GST-OprF190-342-OprI21-83 and GST-OprI21-83-OprF190-350) were isolated and tested for their efficacy as vaccines in immunodeficient mice . GST-OprF-OprI protected the mice against a 975-fold 50% lethal dose of P . aeruginosa . Expression of GST-unfused OprF-OprI failed in E . coli, although this hybrid protein has been expressed without a fusion part in Saccharomyces cerevisiae and used for immunizing rabbits . The immune rabbit sera protected severe combined deficient (SCID) mice against a 1,000-fold 50% lethal dose of P . aeruginosa . Evidence is provided to show that the most C-terminal part of OprF (i.e., amino acids 332 to 350) carries an important protective epitope . Opr-based hybrid proteins may have implications for a clinical vaccine against P . aeruginosa.

Infect Immun, 1995 May, 63(5), 1718 - 24
Endobronchial inflammation following Pseudomonas aeruginosa infection in resistant and susceptible strains of mice; Morissette C et al.; The early endobronchial inflammation induced by Pseudomonas aeruginosa infection varies in resistant and susceptible strains of mice . Mice of the DBA/2 strain are severely afflicted by the infection, with a high bacterial burden accumulating rapidly following inoculation and a high mortality rate occurring . Mice of the BALB/c strain are resistant to infection and clear the bacteria within 3 to 7 days . Infection of (BALB/c x DBA/2)F1 hybrid mice showed that the resistance to lung P . aeruginosa infection is inherited as a dominant trait . Mice of the A/J and C57BL/6 strains were found to have an intermediate phenotype to Pseudomonas aeruginosa infection when compared with BALB/c and DBA/2 strains . The decrease in the bacterial load seen early after infection coincided with a steady and strong recruitment of inflammatory cells to the bronchoalveolar spaces of mice of the resistant BALB/c strain . On the other hand, the recruitment of inflammatory cells to the lungs of mice of the susceptible DBA/2 strain was deficient, resulting in the failure to control bacterial multiplication . Chemotactic factors, proinflammatory cytokines, and the number and function of recruited inflammatory cells may play major roles in the determination of the genetic resistance to lung infection with P . aeruginosa in a normal immunocompetent host.

Drug Saf, 1995 May, 12(5), 305 - 13
Adverse effects of monobactams and carbapenems; Alvan G et al.; Monobactams and carbapenems are 2 classes of beta-lactam antibiotics that were introduced in the 1980s . This review considers the monobactam aztreonam and the carbapenems imipenem and meropenem . Imipenem is administered together with cilastatin, which inhibits the enzymatic breakdown of imipenem in the kidney . The antibacterial activities of these drugs are quite different from older beta-lactams . Aztreonam is directed towards aerobic Gram-negative bacteria, especially Pseudomonas aeruginosa, while imipenem and meropenem are active against both aerobic and anaerobic Gram-positive and Gram-negative bacteria . Thus, these drugs should be reserved for patients who have a special need for them . They are also structurally different from older beta-lactams and possess different adverse drug reaction profiles . It was initially suggested that aztreonam would be less immunogenic than previous beta-lactams because reactive breakdown products acting as haptens are less likely to be formed . Clinical reports now support this assumption, and, in particular, cross hypersensitivity between aztreonam and other beta-lactams seems to be rare which makes the drug a useful therapeutic alternative . However, hypersensitivity to aztreonam does occur . The predominant concern in terms of adverse reactions to imipenem/cilastatin is the increased tendency to cause seizures compared with other beta-lactams . The risk of producing a seizure is highly associated with inadequate dose adjustment in relation to kidney function . If appropriate care is taken, seizures occur in less than 1% of patients treated . However, it is possible that concomitant administration of other drugs with neurotoxic profiles (e.g . theophylline and cyclosporin) given in overdose, may increase the risk of seizures.(ABSTRACT TRUNCATED AT 250 WORDS)

Am J Vet Res, 1995 May, 56(5), 613 - 8
Pharmacokinetics, nephrotoxicosis, and in vitro antibacterial activity associated with single versus multiple (three times) daily gentamicin treatments in horses; Godber LM et al.; Once-daily administration of aminoglycosides may be a safe and effective therapeutic regimen, on the basis of the microbiologic and pharmacokinetic characteristics of these antibiotics . This study was designed to determine serum and tissue concentrations following i.v . administration of gentamicin, at dosages of 6.6 mg/kg of body weight, every 24 hours, and 2.2 mg/kg, every 8 hours, for 10 days in adult horses . Nephrotoxicosis from these dosage regimens also was compared, and microbiologic effects, including postantibiotic effects, were determined with various concentrations of gentamicin against an equine clinical isolate of Pseudomonas aeruginosa . Treatment at the 6.6-mg/kg dosage resulted in maximal serum concentrations (77.93 +/- 19.90 micrograms/ml, mean +/- SEM) and area under the concentration-vs-time curves (83.79 +/- 14.97 micrograms.h/ml) that were significantly (P < 0.05) greater than those following treatment at the 2.2-mg/kg dosage (5.05 +/- 0.50 micrograms/ml and 6.03 +/- 0.66 micrograms.h/ml, respectively) . Nephrotoxicosis was not induced with either dosage regimen, and postantibiotic effects were prolonged with a higher gentamicin concentration . This study provided evidence to support the use of once-daily gentamicin treatment in adult horses.

Zh Mikrobiol Epidemiol Immunobiol, 1995 May-Jun, (3), 89 - 92
{The safety, reactogenicity and antigenic activity of a Pseudomonas aeruginosa anatoxin studied in volunteers}; Mikhailova NA et al.; The safety, reactogenicity and immunogenic activity of P . aeruginosa toxoid were studied in 46 volunteer donors . Systemic reactions to the injection of the preparation were absent in all vaccinees, in 2 subjects (4.3%) insignificant reactions at the site of injection in the form of hyperemia, sized 13-14 mm and disappearing within 48 hours, were registered . The preparation was found stimulate humoral immunity, which was manifested by an increase in the number of B-rosette-forming lymphocytes and the level of antitoxic IgG in the blood of the vaccinees . Besides, immune sera obtained from the blood of the volunteers were found to possess protective properties.

APMIS, 1995 May, 103(5), 367 - 74
Experimental chronic Pseudomonas aeruginosa lung infection in rats . Non-specific stimulation with LPS reduces lethality as efficiently as specific immunization; Lange KH et al.; In a rat model of chronic Pseudomonas aeruginosa lung infection mimicking cystic fibrosis, we investigated the possibility of preventing chronic lung inflammation or decreasing the progression of the infection . We compared the lethality, pathology, bacterial clearance, and immunogenicity after stimulation of the non-specific defence mechanisms by Escherichia coli lipopolysaccharide (LPS) or P . aeruginosa sonicate, or the acquired specific immune response by vaccination with the same bacterial antigens . One day prior to challenge with P . aeruginosa embedded in alginate beads, rats were stimulated with either E . coli LPS or P . aeruginosa sonicate . Four and two weeks prior to challenge other rats were vaccinated with either E . coli LPS or P . aeruginosa sonicate . Controls did not receive any stimulation or vaccination . The lethality after challenge was lower in rats stimulated with E . coli LPS (p = 0.02) or vaccinated with P . aeruginosa sonicate (p = 0.03) as compared to controls . The histopathology of the surviving rats showed an acute inflammation dominated by polymorphonuclear leukocytes (PMNs), but the offending bacteria were not completely eliminated in any group . The increased survival was probably due to earlier recruitment of PMNs most likely mediated by either cytokines and other chemotactic factors (stimulated group) or the immune response in concert with the complement cascade (vaccinated group) . The results of the present and previous vaccination studies show that it is possible to improve survival but not to prevent the chronic P . aeruginosa lung infection and inflammation caused by alginate-embedded bacteria.

Appl Environ Microbiol, 1995 May, 61(5), 2020 - 2
Emergence of nylon oligomer degradation enzymes in Pseudomonas aeruginosa PAO through experimental evolution; Prijambada ID et al.; Through selective cultivation with 6-aminohexanoate linear dimer, a by-product of nylon-6 manufacture, as the sole source of carbon and nitrogen, Pseudomonas aeruginosa PAO, which initially has no enzyme activity to degrade this xenobiotic compound, was successfully expanded in its metabolic ability . Two new enzyme activities, 6-aminohexanoate cyclic dimer hydrolase and 6-aminohexanoate dimer hydrolase, were detected in the adapted strains.

Appl Environ Microbiol, 1995 May, 61(5), 1739 - 44
Construction and use of a nontoxigenic strain of Pseudomonas aeruginosa for the production of recombinant exotoxin A; Wozniak DJ et al.; To express recombinant forms of Pseudomonas aeruginosa exotoxin A in high yield, we have developed a nontoxigenic strain of P . aeruginosa derived from the hypertoxigenic strain PA103 . The nontoxigenic strain, designated PA103A, was produced by the excision marker rescue technique to replace the toxA structural gene in PA103 with an insertionally inactivated toxA gene . The PA103A strain (ToxA-) was used subsequently as the host strain for the expression and production of several recombinant versions of exotoxin A, and the results were compared with exotoxin A production in other P . aeruginosa and Escherichia coli strains . Use of the PA103A strain transformed with the high-copy-number pRO1614 plasmid bearing various toxA alleles resulted in final purification yields of exotoxin A averaging 23 mg/liter of culture . By comparison, exotoxin A production in other expression systems and host strains yields approximately 1/4 to 1/10 as much toxin.

Minerva Pediatr, 1995 May, 47(5), 175 - 85
{Pseudomonas aeruginosa colonization in Turin CF center . Microbiological and therapeutic observations}; Castello M et al.; We reviewed 4,277 sputum cultures performed in our FC patients during antipseudomonas courses of antibiotic therapy . The median age of colonization is 8.6 years, and the chronically infected subjects are 33.65% of all our patients . The most efficient antibiotics were imipenem, aztreonam, ceftazidime and amikacin . Ceftazidime gave the best results in terms of antibiotic resistance.

Pediatr Infect Dis J, 1995 May, 14(5), 367 - 71
Pseudomonas aeruginosa infection in very low birth weight infants: a case-control study; Leigh L et al.; The perinatal histories and hospital courses of all neonates born at Grady Memorial Hospital who developed Pseudomonas aeruginosa sepsis or meningitis in the 5-year period 1989-1993 were reviewed . In addition a case-control study was performed to evaluate selected risk factors for this infection . Twenty-one patients had one or more blood cultures positive for P . aeruginosa . An additional patient had P . aeruginosa meningitis without bacteremia . All infections occurred after 5 days of age . The overall incidence of P . aeruginosa infection was 0.7/1000 live births . All cases occurred in infants < 1500 g at birth, for a birth weight-specific rate of 19.5/1000 livebirths in this weight class . Clinical manifestations of disease did not distinguish P . aeruginosa from other causes of fulminant neonatal sepsis . Fifty percent of cases died . Mortality was inversely related to postnatal age at diagnosis . The 22 cases were compared with 44 controls matched for birth weight, gestational age, sex, duration of hospital stay and admission date . Cases were more likely than controls to have a history of feeding intolerance, interrupted enteral intake and prolonged parenteral hyperalimentation . Case infants received intravenous antibiotics for a significantly longer period of time than did controls . There was an association between P . aeruginosa sepsis and necrotizing enterocolitis (36% cases vs . 7% of controls had prior or concurrent necrotizing enterocolitis, P < 0.01) . In summary P . aeruginosa sepsis is primarily a late onset nosocomial infection in very low birth weight infants . The case fatality rate of 50% in this series emphasizes its continued importance.

ORL J Otorhinolaryngol Relat Spec, 1995 May-Jun, 57(3), 148 - 52
Are ABH antigenic determinants on human outer ear canal epithelium responsible for Pseudomonas aeruginosa infections?
Steuer MK, Hofstadter F, Probster L, Beuth J, Strutz J.
The outer ear canal expression of ABH human blood group antigens has been analyzed with a standardized routine histological procedure by monoclonal antibodies in the case of blood groups A and B, and a corresponding lectin in the case of blood group 0, respectively . In all 20 cases the blood groups were histochemically confirmed . Furthermore, Pseudomonas aeruginosa-specific inhibition experiments were performed with different sugar solutions as well as A-like substance incubating outer ear canal tissue sections with P . aeruginosa strain (No . 60) presenting lectin specificity for N-acetyl-galactosamine (GalNAc) . P . aeruginosa lectins with GalNAc specificity apparently adhere to GalNAc as terminal blood group A determinant and indicate that patients presenting with blood group A may have a genetic predisposition to this form of otitis externa.

Kansenshogaku Zasshi, 1995 May, 69(5), 553 - 67
{Study on the pathogenetic role of alginate produced by mucoid Pseudomonas aerugiosa in diffuse panbronchiolitis}; Ohtami H; It has scarcely been known that the pathogenetic role of mucoid Pseudomonas aeruginosa being colonised on the airway surface in diffuse panbronchiolotis DPB . The role of alginate, a main component of mucoid substance of Pseudomonas aeruginosa was demonstrated in reference to DPB . 1 . Clinical Observation Serum titer of anti-alginate antibody IgG in the group of Pseudomonas-positive DPB was higher than that of the groups of Pseudomonas-negative DPB (p < 0.01) and healthy volunteers (p < 0.01) . The concentration of immune complex in Pseudomonas-positive DPB group at serum level was also higher than that of the above two groups (p < 0.01), and it was well correlated between activity in the patient's symptom and the patient's prognosis . 2 . Experimental Observation In the immunised mice made by free alginate injections, a lymphocyte infiltration around small vessels and small airways in lung was characteristically found at the early stage after the inhalation with mucoid Pseudomonas aeruginosa PT1252 . It disappeared 14-15 days after the inhalation . By repeating the inhalation for 6 days, such a lymphocyte infiltration had been persisted and lymphocyte granulomatous change was formed around small airways . The transformation and narrowing of the small airway occurred by lymphocyte granulomatous change . At the same time, some degree of neutrophil infiltration into the airway was also observed . These findings were closely similar to that of human diffuse panbronchiolitis . 3 . Conclusive Words From the above, the pathogenetic role of alginate in mucoid Psuedomonas aerginosa colonised on airway surface in DPB patient will be explained as follows . Local immunoreaction of antigen and antibody through alginate evolved lymphocyte infiltration around small airway area . A persistent localization of antigen (alginate) makes such an immunoreaction repeat . Consequently, lymphocyte-granulomatous change is formed around the small airway . On the other hand, a state of excess antigen introduced by long term colonization of mucoid Pseudomonas aeruginosa forms an immune complex in the host side . Neutrophil combined with the immune complex deposited on the airway surface may act in a destructive manner for the lung tissue of DPB patient.

Thorax, 1995 May, 50(5), 548 - 50
Recovery of Pseudomonas aeruginosa in respiratory specimens from HIV positive patients being evaluated for Pneumocystis carinii pneumonia; Doyle RL et al.; BACKGROUND--Despite the immune suppression, frequent hospital admissions, and many intercurrent illnesses associated with HIV infection, Pseudomonas aeruginosa has been cited relatively infrequently as a respiratory pathogen in HIV positive patients . METHODS--The microbiological isolates, medical records, radiographic reports, and laboratory data from 224 patients undergoing sputum induction and/or bronchoalveolar lavage for evaluation of respiratory symptoms suspicious for Pneumocystis carinii pneumonia (PCP) from 1989 to 1992 were reviewed retrospectively . RESULTS--An increasing number of respiratory isolates with Pseudomonas aeruginosa was found over this time period . Eighteen of the 224 patients were identified in whom P aeruginosa was recovered on at least one occasion . These patients were more likely to have a history of smoking and prior PCP than those in whom Pseudomonas was not recovered . Mean CD4 counts were also significantly lower in these patients . CONCLUSIONS--Pseudomonas aeruginosa may be recovered from a substantial number of respiratory isolates from HIV positive patients suspected of having PCP . The prevalence of this phenomenon may be increasing.

Mol Microbiol, 1995 May, 16(3), 565 - 74
Molecular cloning and characterization of the rfc gene of Pseudomonas aeruginosa (serotype O5); de Kievit TR et al.; Previous work from our laboratory has shown that cosmid clone pFV100, containing a 26 kb insert, is able to restore O-antigen synthesis in serotype O5 rough mutants of Pseudomonas aeruginosa . Mobilization of pFV100 into two P . aeruginosa semi-rough (SR) mutants, AK14O1 and rd7513, resulted in O-antigen expression, indicating that pFV100 may contain an O-polymerase (rfc) gene . pFV.TK6, a subclone of pFV100 that contains a 5.6 kb chromosomal insert, was able to complement O-antigen expression in these SR mutants . Mutagenesis of pFV.TK6 using Tn1000 exposed a 1.5 kb region that was essential for complementing O-antigen expression in AK14O1 . A 2.0 kb XhoI-HindIII fragment, containing this region, was cloned into vector pUCP26 and the resulting plasmid called pFV.TK8 . In Southern analysis of the 20 P . aeruginosa serotypes using a probe generated from the 1.5 kb XhoI fragment of pFV.TK8, the rfc probe hybridized to a common fragment of the cross-reactive O2-O5-O16-O18-O20 serogroup, suggesting that these serotypes may share a common O-polymerase gene . In functional studies of the rfc gene, the PAO1 (serotype O5) chromosomal rfc was mutated using a gene-replacement strategy . These knockout mutants expressed the SR lipopolysaccharide (LPS) phenotype, which indicated that they were no longer producing a functional O-polymerase enzyme . Nucleotide sequence analysis of the insert DNA of pFV.TK8 revealed one open reading frame (ORF), designated ORF48.9, which could code for a 48.9 kDa protein . In comparisons of the P . aeruginosa rfc nucleotide and amino acid sequences with DNA and protein databases, no significant homology was found . However, the deduced structure of the P . aeruginosa Rfc protein indicated that it is very hydrophobic and contains 11 putative membrane-spanning domains . Therefore, the predicted structure is similar to that of other reported Rfc proteins . Furthermore, comparison of the amino acid composition and codon usage of the P . aeruginosa Rfc with other Rfc proteins revealed significant similarity between them.

Mol Microbiol, 1995 May, 16(3), 497 - 508
Characterization of a five-gene cluster required for the biogenesis of type 4 fimbriae in Pseudomonas aeruginosa; Martin PR et al.; The opportunistic pathogen Pseudomonas aeruginosa produces type 4 fimbriae which promote adhesion to epithelial cells and are associated with a form of surface translocation called twitching motility . Transposon mutagenesis was used to identify loci required for fimbrial assembly or function by screening for mutants that lack the spreading colony morphology characteristic of twitching motility . Six mutants were isolated that contain transposon insertions upstream of the previously characterized gene pilQ . This region contains four genes: pilM-P, which encode proteins with predicted sizes of 37.9, 22.2, 22.8 and 19.0 kDa, respectively . pilM-P appear to form an operon and to be expressed from a promoter in the intergenic region between pilM and the divergently transcribed upstream gene ponA . PilM-P were found to be required for fimbrial biogenesis by complementation studies using twitching motility and sensitivity to fimbrial-specific phage as indicators of the presence of functional fimbriae . This was confirmed by electron microscopy . PilO and PilP did not have homologues in the sequence databases, but the predicted PilN amino acid sequence displayed similarity to XpsL from Xanthamonas campestris, a protein required for protein secretion . PilP contained a hydrophobic leader sequence characteristic of lipoproteins, while PilN and PilO have long internal hydrophobic domains which may serve to localize them to the cytoplasmic membrane . PilM has shared sequence motifs with the cell division protein FtsA from Bacillus subtilis and Escherichia coli, as well as the rod-shape-determining protein MreB from E . coli . These motifs are also conserved in eukaryotic actin, in which they are involved in forming an ATPase domain . Deletion mutants of pilM and pilQ displayed a dominant negative phenotype when transformed into wild-type cells, suggesting that these genes encode proteins involved in multimeric structures.

Mol Microbiol, 1995 May, 16(3), 485 - 96
Identification of a gene, pilV, required for type 4 fimbrial biogenesis in Pseudomonas aeruginosa, whose product possesses a pre-pilin-like leader sequence; Alm RA et al.; Type 4 fimbriae are important colonization factors in Pseudomonas aeruginosa and other pathogens that mediate attachment to epithelial cells of the host . They are also responsible for a form of translocation termed 'twitching motility' and are implicated in the susceptibility to fimbrial-specific bacteriophage . Analysis of a transposon mutant which lacks functional fimbriae has identified a new gene which is required for fimbrial biogenesis . This gene, termed pilV, is located on chromosomal SpeI fragment E, 2 kb downstream of the previously characterized pilSR genes involved in transcriptional activation of the fimbrial subunit gene . The pilV gene encodes a 20 kDa membrane-located protein with considerable amino-terminal homology to the type 4 consensus pre-pilin leader sequence, suggesting that it is processed by a leader peptidase . Site-directed mutagenesis has shown that PilV requires such cleavage to be functional . PilV also exhibits close similarity to a group of proteins involved in extracellular protein secretion from a number of Gram-negative bacteria, suggesting that the biogenesis of type 4 fimbriae may have a similar basis.

Infect Immun, 1995 May, 63(5), 1800 - 5
Pseudomonas aeruginosa exoenzyme S induces proliferation of human T lymphocytes; Mody CH et al.; Pseudomonas aeruginosa is a gram-negative bacterium that is responsible for devastating acute and chronic infections, which include bronchiectasis in cystic fibrosis, nosocomial pneumonia, and infection of burn wounds . Previous studies have demonstrated that these patients have impaired host responses, including cell-mediated immune responses, which are important in anti-Pseudomonas host defense . The P . aeruginosa exoproduct, exoenzyme S, has a number of characteristics which suggest that it might be important in cell-mediated immunity . To determine whether exoenzyme S activates lymphocytes to proliferate, peripheral blood mononuclear cells (PBMC) from normal volunteers were stimulated with purified exoenzyme S, and the lymphocyte response was assessed by measuring {3H}thymidine uptake and by counting the number of cells after various times in culture . Ninety-five percent of healthy adult donors had a lymphocyte response to exoenzyme S . The optimal lymphocyte response occurred on day 7, with 4 x 10(5) PBMC per microtiter well when cells were stimulated with 10 micrograms exoenzyme S per ml . {3H}thymidine uptake correlated with an increase in the number of mononuclear cells, indicating that proliferation occurred . In unseparated PBMC, T cells, and to a lesser extent B cells, proliferated . Purified T cells proliferated, while purified B cells proliferated only after the addition of irradiated T cells . Thus, T lymphocytes are necessary and sufficient for the proliferative response to exoenzyme S . We speculate that exoenzyme S from P . aeruginosa is important in T-lymphocyte-mediated host defense to P . aeruginosa . In strategies to enhance impaired cell-mediated immunity, exoenzyme S should be considered as a potential stimulant.

Infect Immun, 1995 May, 63(5), 1674 - 80
Identification of rfbA, involved in B-band lipopolysaccharide biosynthesis in Pseudomonas aeruginosa serotype O5; Dasgupta T et al.; Previous work from this laboratory has shown that a 26-kb insert in cosmid clone pFV100, isolated from a Pseudomonas aeruginosa gene library, contained genes that could restore serotype-specific B-band lipopolysaccharide (LPS) expression in rough mutant ge6 . In this study, subclones from pFV100 were made to identify genes responsible for B-band LPS synthesis . Transformation of Escherichia coli HB101 with cosmid clone pFV100 resulted in expression of P . aeruginosa serotype O5 B-band LPS, indicating the presence of an rfb cluster in pFV100 . Expression of P . aeruginosa LPS could not be achieved in E . coli HB101 transformed with any of the subclones . Complementation studies of well-characterized rough mutants of P . aeruginosa PAO1 deficient in B-band LPS biosynthesis were performed with the various subclones . Subclone pFV110, containing a 1.4-kb XbaI-HindIII insert, restored B-band LPS biosynthesis in mutant AK44 (A+B-; complete core) . Probing chromosomal DNA from the 20 International Antigenic Typing Scheme serotypes with the 1.4-kb insert from pFV110 in Southern hybridizations revealed a positive reaction to restriction fragments in serotypes O2, O5, O16, O20, and O18 . LPS of serotypes O2, O5, O16, and O20 were shown earlier to have a similar backbone structure in their O antigen . The insert in pFV110 was sequenced, and the deduced amino acid sequence was compared with sequences of protein databases . No significant homology could be detected with any sequences in the database . Open reading frame analysis identified one region, ORF303, which could encode a 33-kDa protein . Using E . coli maxicells for protein expression, orf303 mediated the expression of a unique polypeptide with an apparent molecular mass of 32.5 kDa . The deficiency in the synthesis of B-band LPS biosynthesis in mutant AK44 is apparently complemented by the 33-kDa protein encoded by orf303 . We have designated this ORF rfbA . This investigation is the first report on cloning and sequencing of an rfb gene involved specifically in O-antigen biosynthesis in P . aeruginosa PAO1.

Tohoku J Exp Med, 1995 May, 176(1), 25 - 33
Daily single-dose regimen and alternate-two-week triple-dose/day regimen of oral ofloxacin for the prophylaxis and control of exacerbations of chronic respiratory tract infections; Watanabe A et al.; Two different oral ofloxacin regimens, a daily single-dose regimen with 200 mg (Regimen I) and an every-two-week multidose regimen with 3 x 200 mg/day (Regimen II) was compared as to the efficacy in controlling repeated acute exacerbations of chronic respiratory tract infections . Fifty-eight patients consisting of 19 patients each of bronchiectasis and pulmonary emphysema, 10 patients of chronic bronchitis, 5 patients of old pulmonary tuberculosis, 4 patients of diffuse panbronchiolitis and 1 patient of multiple pulmonary bullae were evaluated: 32 patients in Regimen I and 26 patients in Regimen II . The corrected mean incidence of exacerbations per case decreased from pre-study 2.47 to intra-study 0.59 in Regimen I, and from pre-study 2.66 to intra-study 0.95 in Regimen II, respectively, with a statistically significant difference (p < 0.05, respectively) . Only one of 12 persistent isolates of Pseudomonas aeruginosa acquired a certain degree of resistance to ofloxacin . Adverse reactions were found in six of 66 patients . We conclude that long-term administration of an new-quinolone, especially a daily single-dose regimen with ofloxacin, is useful to control acute exacerbations of chronic respiratory tract infections.

Biochemistry, 1995 Apr 18, 34(15), 5066 - 74
Escherichia coli UMP-kinase, a member of the aspartokinase family, is a hexamer regulated by guanine nucleotides and UTP; Serina L et al.; The pyrH gene, encoding UMP-kinase from Escherichia coli, was cloned using as a genetic probe the property of the carAB operon to be controlled for its expression by the concentration of cytoplasmic UTP . The open reading frame of the pyrH gene of 723 bp was found to be identical to that of the smbA gene {Yamanaka, K., et al . (1992) J . Bacteriol . 174, 7517-7526}, previously described as being involved in chromosome partitioning in E . coli . The bacterial UMP-kinase did not display significant sequence similarity to known nucleoside monophosphate kinases . On the contrary, it exhibited similarity with three families of enzymes including aspartokinases, glutamate kinases, and Pseudomonas aeruginosa carbamate kinase . UMP-kinase overproduced in E . coli was purified to homogeneity and analyzed for its structural and catalytic properties . The protein consists of six identical subunits, each of 240 amino acid residues (the N-terminal methionine residue is missing in the expressed protein) . Upon excitation at 295 nm, the bacterial enzyme exhibits a fluorescence emission spectrum with maximum at 332 nm which indicates that the single tryptophan residue of the protein (Trp119) is located in a hydrophobic environment . Like other enzymes involved in the de novo synthesis of pyrimidine nucleotides, UMP-kinase of E . coli is subject to regulation by nucleotides: GTP is an allosteric activator, whereas UTP serves as an allosteric inhibitor . UTP and UDP, but none of the other nucleotides tested such as GTP, ATP, and UMP, enhanced the fluorescence of the protein . The sigmoidal shape of the dose-response curve indicated cooperativity in binding of UTP and UDP.(ABSTRACT TRUNCATED AT 250 WORDS)

Eur J Biochem, 1995 Apr 15, 229(2), 385 - 94
Purification and characterization of the arylsulfatase synthesized by Pseudomonas aeruginosa PAO during growth in sulfate-free medium and cloning of the arylsulfatase gene (atsA); Beil S et al.; An arylsulfatase (EC 3.1.6.1) was extracted from Pseudomonas aeruginosa PAO1 and purified 2700-fold to homogeneity . Synthesis of this enzyme was repressed when sulfate, cysteine or thiocyanate was supplied as the sole sulfur source for growth, but derepressed with all other sulfur sources tested . The apparent molecular mass was determined by SDS/PAGE to be 57 kDa, and the enzyme was presumed to be a monomer after gel filtration chromatography . The arylsulfatase showed maximal activity at 57 degrees C and pH 8.9, and a Km of 105 microM for 4-nitrocatecholsulfate . Despite previous reports that both inducible and derepressible forms of arylsulfatase exist in P . aeruginosa, we found only one enzyme under a variety of growth conditions: a sulfate-repressed enzyme with a native isoelectric point of 4.76 . The gene encoding this enzyme (atsA) was isolated by complementation of a Tn5-751 mutant of P . aeruginosa PAO1 . Sequencing revealed a 1602-bp reading frame encoding a 534-amino-acid protein with sequence similarity to known bacterial and eukaryotic arylsulfatases (30-40% and 25-30% identity, respectively), but lacking the signal peptide which is present in all known sequences . The lack of this signal peptide suggests that the P . aeruginosa arylsulfatase is neither periplasmic nor membrane-associated, unlike other known arylsulfatases . The atsA gene was located at 15-17' on the P . aeruginosa genome by Southern hybridization . Only a single copy was observed under moderate stringency conditions.

Gene, 1995 Apr 14, 156(1), 63 - 7
Genetic linkage in Pseudomonas aeruginosa of algT and nadB: mutation in nadB does not affect NAD biosynthesis or alginate production; DeVries CA et al.; The 68-min region of the chromosome of Pseudomonas aeruginosa (Pa) contains the gene algT, encoding a putative alternate sigma factor similar to sigma E in Escherichia coli, that is required for the expression of several genes in the alginate biosynthetic regulon . Sequences immediately upstream from algT were found to contain a divergently expressed open reading frame encoding a 60-kDa protein with 64 and 36% identity to the nadB gene products of E . coli and Bacillus subtilis, respectively . The nadB gene encodes L-aspartate oxidase and has been shown in several bacteria to be essential for de novo nicotinamide-adenine dinucleotide (NAD) biosynthesis . Pa nadB complemented the growth requirement for nicotinic acid in a nadB mutant strain of E . coli, suggesting that this gene encodes a functional homologue of L-aspartate oxidase . A nadB::Tn501 mutant was constructed by gene replacement in the alginate-producing strain, Pa FRD . This NadB- mutant still produced alginate and appeared normal with respect to the regulation of alginate synthesis . Interestingly, the NadB- mutant did not have an auxotrophic phenotype for nicotinic acid, indicating that this nadB was not essential for NAD biosynthesis in Pa . These results suggest the possibility that Pa has an alternate mechanism for de novo NAD biosynthesis.

J Biol Chem, 1995 Apr 14, 270(15), 8920 - 7
Molecular characterization of pyocin S3, a novel S-type pyocin from Pseudomonas aeruginosa; Duport C et al.; The genetic determinant for the soluble pyocin S3 was isolated from a genomic library constructed in the plasmid pGV1122, of Pseudomonas aeruginosa strain P12 isolated from a cystic fibrosis patient . The nucleotide sequence of a 3270-base pair DNA fragment was determined, and the two structural genes, pyoS3A and pyoS3I, and the 3'- and 5'-flanking regions were localized . Transcription (Northern blot) analysis showed that the two genes were co-transcribed . The genes pyoS3A and pyoS3I code for polypeptides of 767 and 154 amino acids, respectively, with calculated molecular weights of 81,385 and 17,047 . Pyocin S3 was produced in Escherichia coli from a plasmid and purified as a complex of two components (S3A and S3I) corresponding to the pyoS3A and pyoS3I gene products, respectively . The S3A component, like pyocin S3, had a killing effect involving DNase activity and was inhibited by the S3I protein . Comparisons of the predicted amino acid sequence of the two components of pyocin S3 to those of pyocins S1, S2, and AP41 indicate that pyocin S3 is a new type of S-type pyocin.

Cas Lek Cesk, 1995 Apr 5, 134(7), 212 - 3
{Detection of the G551D mutation in a patient with nasal polyps}; Kuchynkova Z et al.; Atypical forms of cystic fibrosis can be manifested by a mild affection of the airways in adult age . They are the phenotypic manifestation of less frequent mutations for CF . The authors present the case-history of a man who developed the first symptoms of respiratory disease at the age of 16 years . He had relapsing nasal polyps, sinusitis, bronchial asthma, intolerance of acetylsalicylic acid . Bacteriological examination revealed repeatedly Pseudomonas aeruginosa on the respiratory mucosal membranes . The levels of the chloride ion in sweat were normal, genetic examination revealed a mutation of the gene for cystic fibrosis, G 551 D.

Biochim Biophys Acta, 1995 Apr 4, 1261(2), 279 - 84
The structural genes for nitric oxide reductase from Pseudomonas aeruginosa; Arai H et al.; The genes for nitric oxide reductase (norCB) from Pseudomonas aeruginosa were identified and sequenced . They are located about 2 kb upstream of nirS, the structural gene for nitrite reductase . norC and norB encode cytochrome c (16 kDa) and cytochrome b (52 kDa) subunits of the enzyme, respectively . norCB is immediately followed by an open reading frame encoding a protein of 612 residues.

J Immunol, 1995 Apr 1, 154(7), 3420 - 8
Nonopsonic phagocytosis of Pseudomonas aeruginosa requires facilitated transport of D-glucose by macrophages; Barghouthi S et al.; Pseudomonas aeruginosa is the predominant respiratory tract pathogen in patients with cystic fibrosis (CF), and its resistance to phagocytosis may contribute to its virulence . P . aeruginosa ingestion by macrophages occurs only in the presence of D-glucose or D-mannose, sugars present in low concentrations in the endobronchial space . Here we show that only isolates of P . aeruginosa and not other bacterial species were ingested by murine macrophages in a glucose-dependent manner . Glucose transport inhibitors blocked both {3H}-2-deoxy-glucose (2dG) uptake and phagocytosis of P . aeruginosa . P . aeruginosa pretreated with 2dG or 5-thio-D-glucose (5TG) was efficiently ingested . Macrophages pretreated with 2dG or 5TG were able to bind but unable to ingest P . aeruginosa in the presence of glucose; however, they efficiently ingested zymosan or IgG-coated sheep erythrocytes . Macrophages produced lactate only from glucose or mannose . The facilitative glucose transporter GLUT1 mRNA transcript was detected by PCR in preparations from purified macrophages . The nucleotide sequence of the PCR product was identical to that published for murine GLUT1 . GLUT1 protein was detected with anti-GLUT1-peptide polyclonal Abs . We conclude that glucose exerts its effect on the macrophage, not on the bacterium, in the glucose-dependent nonopsonic phagocytosis of P . aeruginosa and that glucose transport via GLUT1 by the macrophages is required to trigger ingestion . The unique glucose dependency for phagocytosis of P . aeruginosa by macrophages may contribute to the pathogenicity of this bacterial species in CF patients.

J Cell Physiol, 1995 Apr, 163(1), 1 - 8
Impaired alveolar macrophage function in smoke inhalation injury; Herlihy JP et al.; The high incidence of both bacterial pneumonia and the adult respiratory distress syndrome (ARDS) associated with smoke inhalation injury (SII) may result, at least in part, from smoke-induced injury to the alveolar macrophage (AM) . Specifically, we hypothesized that AM antimicrobial function, ability to phagocytose apoptotic PMNs, and capacity to prevent apoptosis in PMNs are impaired by smoke . To test these hypotheses, AMs were harvested by bronchoalveolar lavage from sheep before and after the animal was exposed to cotton smoke . The two populations of AMs were incubated with Pseudomonas aeruginosa (PSA) in vitro . Normal AMs (NAMs) phagocytosed a mean of 99 +/- 11% of the PSA placed in their wells, whereas smoke-exposed AMs (SAMs) ingested only 60 +/- 8% . NAMs killed 80 +/- 8% of PSA ingested, whereas SAMs killed only 56 +/- 16% (P < 0.05) . When sheep PMNs, allowed to undergo apoptosis, were incubated with the two AM populations, 66 +/- 3% of the NAMs and 40 +/- 6% of the SAMs demonstrated phagocytosis of these apoptotic PMNs (P < 0.05) . Fresh sheep PMNs were incubated in unconditioned media, NAM and SAM-conditioned media, and followed over 48 hr for the development of apoptosis and maintenance of viability . The NAM-conditioned media markedly prevented apoptosis and augmented PMN survival relative to the unconditioned and SAM-conditioned media (P < 0.05) . The poor antimicrobial function known to be characteristic of apoptotic PMNs, together with the directly impaired antimicrobial function of AMs, may contribute to the infectious complications of SII . If the PMNs recruited to the lung in SII are not properly supported by the AMs following smoke injury, large numbers may undergo apoptosis . If not properly disposed of by these SAMs, the apoptotic PMNs could eventually lyse, releasing tissue toxins, resulting in escalation of lung injury and leading to ARDS.

Radiology, 1995 Apr, 195(1), 247 - 52
Ventilator-associated Pseudomonas aeruginosa pneumonia: radiographic findings; Winer-Muram HT et al.; PURPOSE: To characterize the radiographic features of ventilator-associated Pseudomonas aeruginosa pneumonia (PAP) . MATERIALS AND METHODS: In 56 patients (40 men and 16 women), PAP was documented with fiberoptic bronchoscopy . All patients underwent mechanical ventilation for at least 48 hours before diagnosis . The findings on chest radiographs were recorded . In eight patients in whom computed tomography (CT) was performed, results were compared with radiographic findings . RESULTS: Twenty-six patients with adult respiratory distress syndrome (ARDS) had diffuse bilateral confluent opacities; 30 patients without ARDS had multifocal opacities . In 13 patients, cavities were detected at chest radiography, CT, or both . Seven of 29 patients with pleural abnormalities had empyema . CT provided important additional information (presence of cavities or effusions) in four cases . CONCLUSION: Findings on chest radiographs are nonspecific for PAP . The frequencies of cavities and empyema are surprisingly low, perhaps owing to prompt diagnosis and therapy.

Infect Immun, 1995 Apr, 63(4), 1541 - 51
Characterization of Pseudomonas aeruginosa-induced MDCK cell injury: glycosylation-defective host cells are resistant to bacterial killing; Apodaca G et al.; As a model for bacterium-induced epithelial cell injury, we have studied the interaction of Pseudomonas aeruginosa with polarized Madin-Darby canine kidney (MDCK) cells grown on filters . Following an initial period of bacterial adhesion, foci of injured host cells, which consisted of a central region of cell debris, surrounded by cells that were permeable and apparently necrotic, were formed . Host cell death was quantified by measuring the increased permeability of the monolayer to the macromolecular tracer {14C}inulin . Using this MDCK model system, we have identified bacterial and host cell factors necessary for the host cell damage . The ability of P . aeruginosa to cause MDCK cell damage was independent of elastase or exotoxin A production . In contrast, bacteria with a mutation in the regulatory locus exsA (which are deficient in exoenzyme S production) neither bound to nor caused host cell injury . MDCK cells with defects in cell surface glycosylation were resistant to cell injury, indicating that bacteria may require host cell glycolipids and/or glycoproteins as points of adhesion to cause subsequent host cell injury.

Infect Immun, 1995 Apr, 63(4), 1278 - 85
Role of Pseudomonas aeruginosa pili in acute pulmonary infection; Tang H et al.; The role of piliation in the development and course of acute pulmonary infection was examined using infant BALB/cByJ mice inoculated by intranasal instillation of isogenic Pil+ and Pil- mutants of Pseudomonas aeruginosa PA1244, PAK, and PAO1 . The piliated strains caused more cases of pneumonia, bacteremia, and mortality than the nonpiliated strains (chi-square analysis, alpha = 0.001) . The piliated strains were more often associated with severe diffuse pneumonias, while the nonpiliated organisms resulted in less severe, focal pneumonias, although these differences did not achieve statistical significance . Purified pilin protein used to inoculate the mice resulted in local inflammatory changes . The nonpiliated strain PA1244-NP was as virulent as the piliated strain PAO1, suggesting that expression of other virulence factors are also important in the development of acute pneumonia . This infant mouse model of pulmonary infection appears to be a useful system for the analysis of P . aeruginosa virulence factors involved in the pathogenesis of pneumonia.

J Clin Microbiol, 1995 Apr, 33(4), 924 - 9
Production of elastase, exotoxin A, and alkaline protease in sputa during pulmonary exacerbation of cystic fibrosis in patients chronically infected by Pseudomonas aeruginosa; Jaffar-Bandjee MC et al.; Secretion of Pseudomonas aeruginosa elastase, exotoxin A, and alkaline protease in sputum during bronchopulmonary exacerbations was examined in 18 cystic fibrosis patients chronically infected with this microorganism . The patients were studied during one or several exacerbation periods necessitating hospitalizations of 12 to 20 days . In all cases, P . aeruginosa was present in bronchial secretions at admission and was not eradicated after treatment . The P . aeruginosa density decreased significantly after antibiotic therapy but remained greater than 10(6) CFU/g of sputum in most cases . Significant amounts of P . aeruginosa exoproteins were measured in total homogenized bronchial secretions by immunoenzymatic assays . The detection of higher levels of exoproteins at admission, the significant decrease after treatment, and the absence of exoproteins during intercrisis phases constituted arguments for a renewal of virulence of P . aeruginosa during exacerbations . Nevertheless, the concomitant changes in bacteria load and the triggering of the inflammatory process and immune complex formation could also contribute to pulmonary exacerbations.

Antimicrob Agents Chemother, 1995 Apr, 39(4), 887 - 93
Primary structure of OXA-3 and phylogeny of oxacillin-hydrolyzing class D beta-lactamases; Sanschagrin F et al.; We determined the nucleotide sequence of the blaOXA-3(pMG25) gene from Pseudomonas aeruginosa . The bla structural gene encoded a protein of 275 amino acids representing one monomer of 31,879 Da for the OXA-3 enzyme . Comparisons between the OXA-3 nucleotide and amino acid sequences and those of class A, B, C, and D beta-lactamases were performed . An alignment of the eight known class D beta-lactamases including OXA-3 demonstrated the presence of conserved amino acids . In addition, conserved motifs composed of identical amino acids typical of penicillin-recognizing proteins and specific class D motifs were identified . These conserved motifs were considered for possible roles in the structure and function of oxacillinases . On the basis of the alignment and identity scores, a dendrogram was constructed . The phylogenetic data obtained revealed five groups of class D beta-lactamases with large evolutionary distances between each group.

Antimicrob Agents Chemother, 1995 Apr, 39(4), 839 - 45
Quantitation of slow drug release from an implantable and degradable gentamicin conjugate by in vivo magnetic resonance imaging; Weissleder R et al.; A biodegradable model hydrogel containing a covalently bound aminoglycoside in which drug release can be monitored by magnetic resonance imaging (MRI) in vivo was developed . The hydrogel consists of the bishydroxysuccinimide ester of polyethylene glycol disuccinate cross-linked albumin, to which gentamicin and Gd-diethylenetriaminepentaacetic acid are covalently attached in stochiometric quantities . MRI allowed us to depict the three-dimensional structure of implanted gels, to accurately calculate their volumes, and thus to calculate the concentration of hydrogel-bound gentamicin . The correlation coefficient for the concentration of released gentamicin and the hydrogel volume was 0.965 . Free and hydrogel-released gentamicin conjugates had similar antibiotic efficacies when tested in microbiological agar diffusion assays . In vivo, hydrogel-released gentamicin had a longer half-life in plasma than unaltered gentamicin (5.6 versus 0.7 h), presumably because of residual bound polyethylene glycol residues . Hydrogel implants into rats resulted in a prolonged (7 to 10 days) release of gentamicin and a decreased 24-h mortality in mice infected with a lethal dose of Pseudomonas aeruginosa . The results indicate the feasibility of imaging and quantitating therapeutic drug concentrations in vivo by MRI.

J Appl Bacteriol, 1995 Apr, 78(4), 373 - 7
Antibiotic activity of isoxazolylnaphthoquinone imines on mice infected with Staphylococcus aureus; Albesa I et al.; The antibiotic activity of new synthetic isoxazolylnaphthoquinone imines was studied . Pseudomonas aeruginosa ATCC 27853 and Escherichia coli ATCC 25922 were resistant to the four compounds studied (MIC > 128 micrograms ml-1), but Staphylococcus aureus ATCC 25923, ATCC 29213 and 30 clinical isolates of Staph . aureus were inhibited by 2-hydroxy-N-(3,4-dimethyl-5-isoxazolyl)-1,4-naphthoquinone-4-imine (I) . This compound diminished bloodstream infection of mice injected i.m . with Staph . aureus; septicaemia decayed significantly when I was applied at the beginning of the infection while when I was given 3 d after bacterial challenge, a significant protection was afforded . Bactericidal activity in serum increased during the 5 h after I was administered i.p . The acetyl derivative of I had a high MIC but when inoculated orally in mice decreased the Staph . aureus counts in circulation . This protection occurred only when the schedule of administration started close to the bacterial challenge . Antibiotic activity in vivo may be associated with in vitro effects.

J Appl Bacteriol, 1995 Apr, 78(4), 359 - 65
Separation and concentration of bacteria with immobilized antibody fragments; Molloy P et al.; New methods to quantitatively remove bacteria from food and water samples are required to meet modern safety standards . The recent development of techniques to make Fab/Fv/scFv fragments in bacteria has provided the opportunity to exploit antibodies as specialized chemicals for affinity removal technologies . Single-chain fragments against Pseudomonas aeruginosa have been expressed in Escherichia coli, purified via a fused poly-histidine tail and immobilized upon polystyrene beads . The resulting immunoaffinity columns have been shown to effectively remove greater than 90% of an applied 10 million bacteria after a single passage through the column . Column material in the absence of single-chain retained less than 10% of the bacteria . Pseudomonas were also removed from milk, mixed bacterial cultures and when present at low cell densities.

Genitourin Med, 1995 Apr, 71(2), 73 - 7
Bronchopulmonary infection with Pseudomonas aeruginosa in patients infected with human immunodeficiency virus; Ali NJ et al.; BACKGROUND--Pseudomonas aeruginosa infection is uncommon in HIV infected patients and is usually nosocomially acquired and associated with risk factors such as neutropenia or central lines . We have recently noted an increase in the number of respiratory isolates of Ps aeruginosa in hospitalised HIV positive patients and sought to describe the clinical correlates of this observation . METHODS--A retrospective case notes review of HIV positive patients admitted to a specialist unit for respiratory investigations from January 1989 to December 1993 was undertaken in order to identify those with Ps aeruginosa respiratory infection and to describe associated risk factors, patterns of presentation and radiographic abnormalities . RESULTS--Of 617 patients admitted 38 (6%) had Ps aeruginosa respiratory infection (notes were incomplete in 1 patient) . All patients had advanced HIV disease; median CD4 = 0.02 x 10(9)/l . Two distinct presentations were seen; 9 patients had a fulminant course as part of a sepsis syndrome, 28 patients had an indolent presentation (18 had a single episode and 10 relapsed on one or more occasions, despite successful treatment of the initial episode) . Infection was community acquired in 24 patients . Many patients had risk factors traditionally associated with Ps aeruginosa including neutropenia or indwelling central venous catheters, but 13 had no obvious risk factor . Most patients were receiving systemic pneumocystis prophylaxis and/or broad spectrum antibiotics; 20 had co-existent symptomatic sinus disease . A wide variety of chest radiographic abnormalities were seen including interstitial shadowing, mimicking pneumocystis pneumonia in 12 patients, lobar pneumonia in 2 and bronchial wall thickening in 13 patients . CONCLUSIONS--Ps aeruginosa respiratory infection occurs with increased frequency in patients with advanced HIV disease; in a significant proportion infection is community acquired . Although recognised risk factors were present in two thirds of patients it appears that advanced HIV immunosuppression, use of systemic pneumocystis prophylaxis and/or broad spectrum antibiotics and sinus disease are important risk factors . The diagnosis should be considered in patients with advanced HIV disease who present with new respiratory symptoms.

Int Arch Allergy Immunol, 1995 Apr, 106(4), 357 - 65
Effect of Pseudomonas aeruginosa on interleukin-8 release from human phagocytes; Konig B et al.; Pseudomonas aeruginosa infections are commonly observed in sepsis, burns, as well as cystic fibrosis (CF) . Among the professional phagocytes neutrophils and monocytes are recruited by various chemotactic factors from the cellular environment . Although they provide the first line of host defense excessive neutrophil accumulation seems to be a major cause of pathogenesis during P . aeruginosa infection . Interleukin-8 (IL-8) represents one important chemoattractant for professional phagocytes . To evaluate IL-8 releasability by phagocytes in the context of P . aeruginosa infection and especially of CF, we stimulated human polymorphonuclear neutrophilic granulocytes (PMN) and peripheral blood mononuclear cells (PBMC) as a source for monocytes with clinical P . aeruginosa isolates, with mucoid P . aeruginosa strain (CF3M) and its nonmucoid revertant (CF3), and with purified P . aeruginosa mucoid exopolysaccharide (alginate) . A significant increase in IL-8 release as compared to unstimulated cells was observed after an incubation time of 90 min for PMN and after 60 min for PBMC which increased (PMN: up to 60-fold; PBMC: up to 40-fold) over time (up to 4 h) . In contrast of PBMC, when PMN were studied, intracellular IL-8 exceeded the IL-8 release in unstimulated as well as in stimulated cells by up to 10-fold . All clinical P . aeruginosa isolates, independent of the clinical source, induced IL-8 release from human PBMC and PMN in a dose- and time-dependent manner.(ABSTRACT TRUNCATED AT 250 WORDS)

J Pediatr, 1995 Apr, 126(4), 515 - 23
A multicenter study of alternate-day prednisone therapy in patients with cystic fibrosis . Cystic Fibrosis Foundation Prednisone Trial Group; Eigen H et al.; The purpose of this study was to evaluate the efficacy and safety of alternate-day prednisone therapy in treating patients with mild-to-moderate cystic fibrosis during a 4-year period . In 15 North American cystic fibrosis centers, we screened 320 patients and enrolled 285 patients from April 1986 to December 1987 . Patients were randomly assigned to take prednisone, 1 mg/kg per dose or 2 mg/kg per dose, or a matching placebo given on alternate days . Lung function, clinical status, hospitalizations, growth, and steroid side effects were monitored . During the first 24 months the percentage of the predicted forced vital capacity was greater in the 1 mg/kg group (p < 0.0001) and the 2 mg/kg group (p < 0.01) when each was compared with placebo . Patients in the 1 mg/kg group had a higher percentage of predicted forced vital capacity than placebo patients during the entire 48 months (p < 0.0025), but only in the group of patients who were colonized with Pseudomonas aeruginosa at baseline . For 48 months, the 1 mg/kg group had a higher percentage of predicted forced expiratory volume in 1 second than patients given placebo (p < 0.02) . The prednisone-treated groups had a reduction in serum IgG concentrations (1 mg/kg vs placebo, p < 0.007; 2 mg/kg vs placebo, p < 0.003) . From 6 months onward, height z scores fell in the 2 mg/kg group compared with those given placebo (p < 0.001) . For the 1 mg/kg group, height z scores were lower from 24 months . An excess of abnormalities in glucose metabolism was seen in the 2 mg/kg group compared with the placebo group (p < 0.005) . Our findings suggest a role for alternate-day prednisone therapy at a dose of 1 mg/kg in patients with mild to moderate cystic fibrosis . The benefit of improved lung function appears to outweigh the potential for adverse effects when the treatment period is less than 24 months.

Am J Respir Crit Care Med, 1995 Apr, 151(4), 983 - 5
Parenteral application of a Pseudomonas aeruginosa flagella vaccine elicits specific anti-flagella antibodies in the airways of healthy individuals; Doring G et al.; To test the hypothesis that a Pseudomonas aeruginosa flagella (FLA) vaccine, intramuscularly administered, elicits specific antibodies in the respiratory tract, antibody titers against FLA were determined in sera and bronchoalveolar lavage (BAL) fluids of 10 healthy human adults before and after immunization . Immunization produced significantly increased anti-FLA antibody titers in all sera (mean reciprocal titers: IgG: 26,711; IgA: 2,767; IgM: 421) and on respiratory epithelial lining fluids (mean reciprocal titers: IgG: 112; IgA: 76; sIgA: 26) . Significant differences in class-specific serum and BAL anti-FLA titers suggested airway antibody production . The study shows that parenteral immunization provides not only high systemic antibody titers but also local antibody production in the human respiratory tract.

Am J Respir Crit Care Med, 1995 Apr, 151(4), 1255 - 8
The ciliary transportability of sputum is slow on the mucus-depleted bovine trachea; Wills PJ et al.; Mucus retention in the lungs is a feature of several chest diseases . It is unclear to what extent suboptimal mucus transportability is responsible for the poor clearance of lung secretions . We described a new model, the mucus-depleted bovine trachea, for measurement of the ciliary transportability of respiratory mucus . Mucus depletion was demonstrated microscopically and functionally, and it was accomplished by simple physical means without impairing ciliary action . Control mucus from the tracheas of humans and animals was transported rapidly on this system . However, sputum from 54 patients with bronchiectasis was transported slowly, at a mean of 15% of the rate of control mucus . There was no correlation between sputum transportability and either purulence or the presence of Pseudomonas aeruginosa . This work suggests that there is a serious defect in the ciliary transportability of sputum that is unrelated to the presence of infection . The model should allow in vitro assessment of agents designed to aid mucociliary clearance.

Am J Respir Crit Care Med, 1995 Apr, 151(4), 1093 - 100
Resistance of the alveolar epithelium to injury from septic shock in sheep; Pittet JF et al.; Experimentally, the intravenous administration of a bolus dose of Escherichia coli endotoxin in sheep or a bolus dose of live Pseudomonas aeruginosa in rats is insufficient to cause injury to the alveolar epithelial barrier . Therefore, the first objective of these studies was to maximize the injury caused by live bacteria to the lung by administering a large dose of live P . aeruginosa into the lung perfusate of goat lungs in situ . P . aeruginosa (2.4 x 10(10) colony-forming units {cfu}) and {131I}albumin (vascular protein tracer) were added to the lung perfusate . Even though the bacterial inoculum remained very high in this isolated perfused lung system, there was no change in the permeability to protein or clearance of fluid across the alveolar epithelium, although there was an increase in lung endothelial protein permeability . Therefore, since systemic factors have been implicated in the severity and pathogenesis of septic lung injury, the second objective was to administer a continuous intravenous infusion of live P . aeruginosa over 8 h in intact anesthetized sheep . The eight sheep so treated exhibited an intact, functional alveolar barrier, even though there was an increase in lung endothelial permeability to protein and an increase in extravascular lung water . In fact, in these eight sheep, alveolar epithelial fluid transport was significantly greater than in control sheep . In the three other septic sheep there was injury to the alveolar epithelial barrier with an increase in permeability of the barrier to protein, an inability to transport fluid out of the airspaces, and an even greater increase in extravascular lung water.(ABSTRACT TRUNCATED AT 250 WORDS)

Singapore Med J, 1995 Apr, 36(2), 232 - 4
Scleral necrosis and infection 15 years following pterygium excision; Au Eong KG et al.; Scleral necrosis and infection are serious late complications of pterygium treatment and are difficult to manage . We describe a 70-year-old Chinese male who presented with scleral necrosis and Pseudomonas aeruginosa infection 15 years after the excision of a pterygium . The infection was treated early and aggressively with intensive topical and intravenous antibiotics and the thin necrotic sclera was reinforced with a donor scleral patch graft when the scleral infection was clinically controlled . The integrity of the globe was maintained by a thin layer of sclera anterior to the graft after the graft gradually shrunk in size and retracted posteriorly . The eye was saved from possible scleral perforation and endophthalmitis . This case is reported to highlight the importance of early aggressive treatment of infection and the value of prophylactic repair of scleral necrosis in the management of these late complications of pterygium treatment.

Eur J Epidemiol, 1995 Apr, 11(2), 193 - 7
Evaluation of in vitro efficacy of the disinfectant Virkon; Gasparini R et al.; A study was conducted on a new acid peroxygen system based disinfectant (Virkon), in order to assess its in vitro efficacy . The chemical was tested on different bacteria (Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli), spores (Bacillus subtilis) and on the Hepatitis B surface antigen (HBsAg), and compared in its activity with phenol and glutaraldehyde (calculation of the 'phenol coefficient' and the 'glutaraldehyde coefficient') . The constancy of speed of disinfection, the coefficient of concentration, the minimal inhibitory concentration (MIC) were also determined, and the destruction of the HBsAg antigenic activity was studied using an ELISA kit . The sporicidal efficacy of Virkon was assessed by cultivating spores in agar nutrient after contact with different dilutions of the disinfectant . The results of the tests showed that Virkon has a high concentration coefficient (mean value of k: 0.374/min) and a wide range of action . The low MIC demonstrates how little concentrations of Virkon can inactivate all studied bacteria . The disinfectant was also able to destroy the hepatitis B surface antigen, and it demonstrated good activity against spores, especially if used in physiologic solution . These characteristics, coupled with the absence of initiation or toxic effects on animals showed by other studies, make wide fields of application for the new disinfectant foreseeable.

Eur J Clin Microbiol Infect Dis, 1995 Apr, 14(4), 291 - 6
Correlation between activity of beta-lactam agents in vitro and bacteriological outcome in acute pulmonary exacerbations of cystic fibrosis; Gaillard JL et al.; A study was conducted to determine whether a direct relationship exists between beta-lactam and/or aminoglycoside activity measured in vitro and bacteriological outcome in acute pulmonary exacerbations of cystic fibrosis . Twenty-seven patients, aged between 6 months and 24 years (mean age 10 1/2 years), were included in the study and received 41 i.v . courses of a beta-lactam agent combined with an aminoglycoside . A total of 63 Pseudomonas aeruginosa strains were found in sputum taken on admission at densities exceeding 10(6) cfu/g of sputum . For each episode, the serum inhibitory quotient (SIQ) and the serum bactericidal quotient (SBQ) of the beta-lactam agent and of the aminoglycoside administered were determined for the Pseudomonas aeruginosa isolate(s) . The SIQs and SBQs were calculated by dividing the average peak serum levels achievable in the patients by the minimal inhibitory concentrations and minimal bactericidal concentrations, respectively . The SIQs and SBQs were compared to bacteriological outcome . Bacteriological success was defined as a decrease of 2 log10 counts or more in the Pseudomonas aeruginosa density in sputum between days 0 and 7 of therapy . The SIQ and SBQ of beta-lactam agents were good predictors of bacteriological outcome: SIQs of < 1:16 were 100% predictive of failure (chi 2 28; p < 0.001) and of > or = 1:64 were 92.9% predictive of success (chi 2 35.68; p < 0.001); SBQs of < 1:8 were 100% predictive of failure (chi 2 42.78; p < 0.001) and of > or = 1:32 were 95.8% predictive of success (chi 2 31.5; p < 0.001) . Aminoglycoside SIQs and SBQs were not predictive of outcome.

Vaccine, 1995 Apr, 13(5), 449 - 53
Synthesis and characterization of a polyvalent Escherichia coli O-polysaccharide-toxin A conjugate vaccine; Cryz SJ Jr et al.; A 12-valent Escherichia coli O-polysaccharide (O-PS)-toxin A conjugate vaccine was formulated . Nonpyrogenic, low-molecular-weight O-PS was derived from lipopolysaccharides (LPS) of the following serotypes: O1,O2,O4,O6,O7,O8,O12, O15,O16,O18,O25, and O75 . Individual O-PS were covalently coupled to Pseudomonas aeruginosa toxin A using adipic acid dihydrazide as a spacer molecule and carbodiimide as a coupling agent . On a weight basis, the final multivalent vaccine was composed of 43% O-PS and 57% toxin A . The vaccine was nontoxic nad nonpyrogenic in anti-LPS immunoglobulin G (IgG) antibody titers . When passively transferred to mice, immune rabbit IgG conferred statistically significant (p < 0.05) protection against a challenge with 9 of the 12 vaccine serotypes . For two serotypes, although the mortality rate declined by > or 50% in the passively immunized versus the control group, the difference did not reach statistical significance . The degree of protection provided by passively transferred IgG was influenced by both the anti-LPS antibody levels in the IgG preparation and the virulence of the challenge strain . Active immunization of mice with either conjugate vaccine or killed E . coli whole cells did not confer protection . This was most probably due to the fact that these antigens induced a meagre anti-LPS IgG antibody response.

J Antimicrob Chemother, 1995 Apr, 35(4), 521 - 34
National survey of susceptibility to antimicrobials amongst clinical isolates of Pseudomonas aeruginosa; Chen HY et al.; Between September and December 1993, each of 24 hospitals in the UK collected up to 100 consecutive clinical isolates of Pseudomonas aeruginosa and sent these to the London Hospital Medical College (LHMC) . Of 2184 cultures received, 1991 contained viable P . aeruginosa . Minimum inhibitory concentrations (MICs) of antimicrobials were determined by agar dilution . The frequencies of resistance to low breakpoints were as follows: gentamicin, MIC > 2 mg/L, 11.7%; amikacin, MIC > 4 mg/L, 10.5%, carbenicillin, MIC > 128 mg/L, 11.7%; azlocillin, MIC> 16 mg/L, 10.9%; ceftazidime, MIC > 4 mg/L, 9.6%; ciprofloxacin, MIC > 1 mg/L, 8.1%; imipenem, MIC > 4 mg/L 2.5% and meropenem, MIC > 4 mg/L, 1.1% . Resistance to each antimicrobial except amikacin was commoner among the 134 isolates from patients in intensive care units (ICUs) than amongst the 1042 isolates from other in-patients (P < 0.01) . Resistance to penicillins and ceftazidime, though not to other agents, was rarer among the 797 isolates from out-patients than amongst those from non-ICU in-patient (P < 0.01) . Compared to a similar study in 1982, during which 1866 isolates had been examined, the frequency of resistance to the aminoglycosides increased (P < 0.05) as had those to the penicillins and ceftazidime (P < 0.01) . Ciprofloxacin and the carbapenems were not tested in 1982 . Cross-resistance patterns suggested that the increases in resistance to aminoglycosides and beta-lactams were largely a reflection of greater numbers of isolates with barrier or efflux mechanisms and were not due to an increase in isolates with antibiotic-degrading enzymes . The participating hospitals mostly employed Stokes' disc diffusion method and, when the results were compared to the MICs determined at the LHMC, fewer than 9% of the isolates reported as susceptible were found to be resistant . However, up to 72% of those reported by the hospitals as resistant were found to be susceptible.

Proc Natl Sci Counc Repub China B, 1995 Apr, 19(2), 105 - 12
Production of chitinase by Pseudomonas aeruginosa K-187 using shrimp and crab shell powder as a carbon source; Wang SL et al.; Shrimp and crab shell powder prepared by treating shrimp and crab processing waste with boiling and crashing was used as a substrate for isolating alkali-tolerant chitinolytic microorganisms . Strain K-187 appeared to be the chitinase-producing strain with the most potential . The organism was identified as a strain of Pseudomonas aeruginosa . Maximum chitinase activity was obtained when the strain was grown aerobically in a medium consisting of 3.0% shrimp and crab shell powder, 0.1% CMC, 0.1% (NH4)2SO4, 0.1% K2HPO4, 0.1% MgSO4.D7H2O and 0.1% ZnSO4 (pH 9), at 45 degrees C after 3 days . The optimum pH and temperature of the enzyme reaction were 7 and 40 degrees C, respectively . The chitinase was stable at pH from 5 to 10 and was stable under 60 degrees C.

APMIS, 1995 Apr, 103(4), 286 - 92
An easy microtiter assay for quantitation of cytokine induction by lipopolysaccharide (LPS) and activity of LPS-binding serum components; Garde AH et al.; Lipopolysaccharide (LPS, endotoxin) is a major inducer of cytokines, such as interleukin 1 (IL1), IL6, IL8 and tumor necrosis factor (TNF) . A convenient microtiter assay was developed to measure such activity . LPS coated onto a plastic surface was used to stimulate purified human mononuclear cells (MNC) in microtiter plates . Following stimulation the supernatants were assayed for presence of TNF by ELISA . Purified rough and smooth LPS from Pseudomonas aeruginosa gave a dose-dependent TNF release over a range of 0.1-1.0 microgram LPS/well . The assay was subsequently used to investigate the biological activity of anti-LPS antibodies and other LPS-specific serum components in sera from patients with cystic fibrosis (CF) . As a group, sera from 10 CF patients chronically infected with P . aeruginosa did not affect the LPS-induced TNF release, while sera from normal controls inhibited this biological activity . When individual CF patients with or without chronic lung infection are considered, the antibodies appear to either enhance or inhibit the LPS-stimulated TNF release (range: 73-120%), while all antibodies from healthy controls inhibit the activity of LPS (range: 76-97%) . Only a weak correlation (rho = 0.491, p = 0.037, n = 19) was found between the antibody titer in ELISA and the biological activity of sera . This new assay is suggested for convenient measurement of interference with cytokine induction from human MNC by patient or therapeutic anti-LPS antibodies and other LPS-specific serum components.

Protein Expr Purif, 1995 Apr, 6(2), 164 - 8
Plasmid mutagenesis by PCR for high-level expression of para-hydroxybenzoate hydroxylase; Moran GR et al.; We report a PCR deletion mutagenesis method for the exact positioning of a foreign gene (pobA) in the lac operon of an expression plasmid in place of the lacZ protein code . This method requires the synthesis of four oligonucleotides and three PCR reactions to delete unwanted bases and retain the nucleotide sequence naturally found between the lac promoter and the protein code . The engineered plasmid results in the production of at least 40% of the cellular protein as the foreign polypeptide . In the example presented the expression of the protein is high even with a substantial difference in codon usage between the host (Escherichia coli) and a foreign gene from Pseudomonas aeruginosa . Some of the polypeptide produced has the ame properties as native protein and is easily purified . The remainder is present as insoluble inclusion bodies . This method of plasmid refinement may be applicable to the expression of many proteins.

Pathol Biol (Paris), 1995 Apr, 43(4), 343 - 51
{Bacteriological monitoring of the treatment of Pseudomonas aeruginosa infections with amikacin administrated at once-daily dosis in patients with cystic fibrosis}; Canis F et al.; The interest of the treatment with a single daily dose of amikacin (AMK) in cystic fibrosis (CF) patients with P . aeruginosa infections has been much debated . The aim of work was to study the efficiency of this treatment on (CF) patients with chronic bronchopulmonary P . aeruginosa infections previously treated for two weeks with the combination ceftazidime (CAZ 200 mg/day in 3 inj . IVD) and AMK (35 mg/day in one IV perf . of 30 minutes) . The bacteriological supervision of this treatment was performed 1: by the determination of MICs before and after treatment, 2: by the decrease of P . aeruginosa colonization immediately after this treatment and during 11 months, 3: by the identification of P . aeruginosa strains with phenotypic methods (serotyping and antibiotyping) and with genotypic method (pulsed field gel electrophoresis) . The use of AMK in a single daily dose in order to treat chronic lung infections colonized with P . aeruginosa susceptible to this antibiotic shows encouraging results as far as bacteriology is concerned: this treatment has given means to reduce colonization for a month in 15 of 18 patients . For 9 of the 18 patients, no P . aeruginosa strains were isolated for nine months . The serotyping and antibiotyping systems do not enable us to study the P . aeruginosa epidemiology . Genome macrorestriction fingerprinting of P . aeruginosa in pulsed field gel electrophoresis confirms that patient with CF were colonized with one or several clones . In our study no variation of these clones was noticed for the first eleven months . Genome macrorestriction fingerprinting appears to be one of the most effective methods for delineate strains of P . aeruginosa colonizing CF patients.

Mol Microbiol, 1995 Apr, 16(2), 309 - 20
The Escherichia coli genes sspA and rnk can functionally replace the Pseudomonas aeruginosa alginate regulatory gene algR2; Schlictman D et al.; The algR2 (also known as algQ) gene of Pseudomonas aeruginosa has previously been identified as being necessary for alginate production at 37 degrees C . We have cloned two genes, from a cosmid library of Escherichia coli, which can restore mucoidy to an algR2 mutant of P . aeruginosa . The complementing regions of both cosmids were localized by subcloning restriction fragments . One of the E . coli genes identified here has not previously been described; we have named this gene rnk (regulator of nucleoside diphosphate kinase) . It encodes a 14.9 kDa protein with no homology to any other protein . The other gene, sspA, is a regulator involved in stationary-phase regulation in E . coli . Either gene will restore mucoidy to an algR2-deficient strain of P . aeruginosa . AlgR2 has been shown to regulate at least two enzymes, succinyl-CoA synthetase (Scs) and nucleoside diphosphate kinase (Ndk), which form a complex in P . aeruginosa . When we examined the ability of the E . coli analogues to regulate Ndk, we found that rnk but not sspA was able to restore Ndk activity to the P . aeruginosa algR2 mutant . Furthermore, rnk was able to restore growth of the algR2 mutant in the presence of Tween 20, which inhibits other Ndk-like activities.

Mol Microbiol, 1995 Apr, 16(2), 263 - 70
Purification and characterization of LasD: a second staphylolytic proteinase produced by Pseudomonas aeruginosa; Park S et al.; We have previously described studies of a 22 kDa active fragment of the LasA proteinase . In follow-up studies of LasA, we have discovered the separate existence of a 23 kDa proteinase which shares many of the enzymatic properties of LasA, including the ability to lyse heat-killed staphylococci . However, this apparent serine proteinase, which we designate LasD, is distinct from the 22 kDa active LasA protein for the following reasons: (i) the N-terminal sequence of LasD shares no homology with LasA or the LasA precursor sequence; (ii) Pseudomonas aeruginosa LasA mutant strains AD1825 and FRD2128 do not produce LasA yet produce LasD; and (iii) specific antibodies to each proteinase do not show any cross-reactivity . LasD appears to be produced as a 30 kDa protein, which is possibly cleaved to produce a 23 kDa active fragment . The purified LasD fragment (23 kDa) shows strong staphylolytic activity only at higher pH conditions, while LasA exhibits staphylolytic activity over a broad pH range . In addition to their ability to cleave at internal diglycine sites, both the LasD and LasA endoproteinases efficiently cleave beta-casein.

J Comp Pathol, 1995 Apr, 112(3), 273 - 82
Granulomatous lesions caused by Pseudomonas aeruginosa in the ostrich (Struthio camelus); Momotani E et al.; Granulomatous lesions were observed in imported ostriches aged 3 months . Clinically, the birds showed lassitude, incoordination, and inappetence . At necropsy, yellowish white nodules often accompanied by a pseudodiphtheritic membrane were found in the oral, pharyngeal, tracheal and air sac mucosae, the lungs, oesophageal serosa, and abdominal peritoneum . Histopathological examination revealed purulent granulomatous lesions containing central bacterial colonies with an outer shell and club formation . The bacteria were small Gram-negative bacilli, which showed positive immunohistochemical staining for Pseudomonas aeruginosa . The bacterial colonies were positive for chicken IgM . Clubs around the colonies were negative for P . aeruginosa and chicken IgM . Such findings have not previously been reported in the ostrich.

Zentralbl Bakteriol, 1995 Apr, 282(3), 287 - 95
Blood group phenotype determines lectin-mediated adhesion of Pseudomonas aeruginosa to human outer ear canal epithelium; Steuer MK et al.; Pseudomonas aeruginosa is the most frequent bacterial pathogen causing acute diffuse otitis externa . In a recent prospective phase II study we demonstrated that lectin-mediated bacterial adhesion can be blocked by receptor-analogue carbohydrates in patients suffering from Pseudomonas aeruginosa-induced acute otitis externa . In this investigation, human ABO blood group antigens were analysed on outer ear canal epithelial cells with standard routine histological procedures by monoclonal antibodies for the blood groups A and B, and with Ulex europaeus I lectin for the blood group O, respectively . In all cases (n = 20) the blood groups could be shown immunohistologically . P . aeruginosa-specific adhesion and inhibition assays were performed in the presence of N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc), D-mannose and A-like substance . Outer ear canal tissue sections were incubated with P . aeruginosa (strain PA 60), presenting lectin-specificity for GalNAc . Sections from patients presenting with blood group A were closely settled with bacteria in the presence of non-specific GlcNAc, D-mannose and PBS however, GalNAc and A-like substance inhibited the microbial adhesion . Amongst others, P . aeruginosa present adhesion molecules (lectins) with specificity for GalNAc . Thus, the correlation between blood group A phenotype and P . aeruginosa-induced acute diffuse otitis externa was investigated . Statistical evaluation proved a highly significant association . These data support the hypothesis that P . aeruginosa lectins with GalNAc specificity apparently adhere to GalNAc moieties, representing the terminal blood group A-determinant and further indicate that patients presenting with blood group A may have a genetic disposition for this form of otitis externa.

Infect Immun, 1995 Apr, 63(4), 1311 - 7
Ability of bacteria associated with chronic inflammatory disease to stimulate E-selectin expression and promote neutrophil adhesion; Darveau RP et al.; Porphyromonas gingivalis, Pseudomonas aeruginosa, and Helicobacter pylori have been shown to be associated with adult periodontal disease, chronic lung infections, and peptic ulcers, respectively . The ability of these bacteria to stimulate E-selectin expression and promote neutrophil adhesion, two components necessary for the recruitment of leukocytes in response to infection, was investigated . Little or no stimulation of E-selectin expression was observed with either P . gingivalis or H . pylori when whole cells, lipopolysaccharide (LPS), or cell wall preparations added to human umbilical cord vein endothelial cells were examined . P . aeruginosa was able to induce E-selectin to near-maximal levels; however, it required approximately 100 to 1,000 times more whole cells or LPS than that required by E . coli . Neutrophil-binding assays revealed that LPS and cell wall preparations obtained from these bacteria did not promote endothelial cell adhesiveness by E-selectin-independent mechanisms . In addition, P . gingivalis LPS blocked E-selectin expression by LPS obtained from other bacteria . We propose that lack of E-selectin stimulation and the inability to promote endothelial cell adhesiveness are two additional indications of low biologically reactive LPS . We suggest that this property of LPS may contribute to host tissue colonization . In addition, the ability of P . gingivalis to inhibit E-selectin expression may represent a new virulence factor for this organism.

Mol Gen Mikrobiol Virusol, 1995 Apr-Jun, (2), 36 - 40
{Determinant of plasmid pBS221 resistance to potassium tellurite}; Anisimova LA et al.; The genes regulating resistance to potassium tellurite were cloned from plasmid pS221 with a broad host range . The studied DNA fragment encodes for the synthesis of two polypeptides with molecular mass of 38 and 43 kD . Metal tellurium crystals were localized in the periplasm of Pseudomonas aeruginosa ML4262 (pBS221) cells by electron microscopy . Results of colony hybridization permit us to propose that the cloned genes of potassium tellurite resistance have sites of homology with plasmids RP4(IncP), R27 (IncH), pBS38, pBS79, and R931(IncP-2).

Proc Natl Acad Sci U S A, 1995 Mar 28, 92(7), 3019 - 23
Cystic fibrosis epithelial cells have a receptor for pathogenic bacteria on their apical surface; Imundo L et al.; Chronic colonization and infection of the lung with Pseudomonas aeruginosa is the major cause of morbidity and mortality in cystic fibrosis (CF) patients . We found that polarized CF bronchial and pancreatic epithelia bound P . aeruginosa in a reversible and dose-dependent manner . There was significantly greater binding to CF bronchial and pancreatic cells than to their matched pairs rescued with the wild-type CF transmembrane conductance regulator . Bound P . aeruginosa were easily displaced by unlabeled P . aeruginosa but not by Escherichia coli, an organism that does not cause significant pulmonary disease in CF . In contrast, Staphylococcus aureus, a frequent pathogen in CF, could effectively displace bound P . aeruginosa from its receptor . We found undersialylation of apical proteins and a higher concentration of asialoganglioside 1 (aGM1) in apical membranes of CF compared with rescued epithelia . Incubation of P . aeruginosa with aGM1 reduced its binding, as did treatment of the epithelia with the tetrasaccharide moiety of this ganglioside (Gal beta 1-3GalNAc beta 1-4Gal beta 1-4Glc) . Finally, an antibody to aGM1 effectively displaced P . aeruginosa from its binding site and blocked binding of S . aureus to CF cells but not to rescued cells . These results show that the tetrasaccharide of aGM1 is a receptor for P . aeruginosa and S . aureus and that its increased abundance in the apical membrane of CF epithelia makes it a likely contributor to the pathogenesis of bacterial infections in the CF lung.

Gene, 1995 Mar 21, 155(1), 73 - 6
Cloning and primary sequence of the rpoH gene from Pseudomonas aeruginosa; Benvenisti L et al.; A DNA fragment from Pseudomonas aeruginosa containing the rpoH gene encoding the heat-shock sigma factor sigma 32 has been cloned and sequenced . The gene is expressed in Escherichia coli and complements an rpoH- strain . An open reading frame encoding 284 amino acids shows 61% identity and 78% similarity to the RpoH protein of Escherichia coli.

J Med Chem, 1995 Mar 17, 38(6), 973 - 82
6-Aminoquinolones: a new class of quinolone antibacterials?
Cecchetti V, Clementi S, Cruciani G, Fravolini A, Pagella PG, Savino A, Tabarrini O.
A series of quinolone- and 1,8-naphthyridone-3-carboxylic acids, designed by previous QSAR studies and characterized by an amino group at the C-6 position instead of the usual fluorine atom, were synthesized for the first time and evaluated for in vitro antibacterial activity . All of the synthesized compounds maintain good activity against Gram-negative bacteria (Pseudomonas aeruginosa excluded), and those compounds having a thiomorpholine group as the C-7 substituent also have good activity against Gram-positive bacteria . Some aspects of structure-activity relationships associated with the C-1, C-5, C-7, and C-8 substituents are also discussed . Derivatives 18g and 38g displayed the best activity with geometric mean MICs of 0.45 and 0.66-0.76 micrograms/mL against Gram-negative and Gram-positive bacteria, respectively . This antimicrobial activity reflects their ability to inhibit bacterial DNA-gyrase . The results of this study show that, while the C-6 fluorine is still the preferred substituent, good activity can still be obtained by replacing it with an amino group.

Invest Ophthalmol Vis Sci, 1995 Mar, 36(3), 634 - 43
In the immature mouse, Pseudomonas aeruginosa pili bind a 57-kd (alpha 2-6) sialylated corneal epithelial cell surface protein: a first step in infection; Hazlett L et al.; PURPOSE . To test the hypothesis that in the unscarified immature eye, Pseudomonas aeruginosa pili bind glycoprotein receptors, one or more of which are surface associated . METHODS . Several methods--including radioiodination of bacterial pili and surface-associated corneal epithelial proteins (CEPs), solid-phase binding assays, carbohydrate detection, and immunoblotting techniques in which periodate oxidation and preincubation of blots with purified pili, neuraminidase, sialic acid, other sugars, and SNA and MAA lectins--were used to identify and characterize host proteins . Some of these proteins in the immature mouse corneal epithelium interacted with bacterial pili . RESULTS . Seven proteins, with molecular weights from 14 to 66 kd were identified that strongly bound PAK/PR1 pili . To determine if any protein(s) was cell surface localized, corneal epithelial surface membrane proteins were radioiodinated and examined using a pilus overlay assay and lectin analysis . Only one protein of 57 kd was cell surface labeled and bound pili in an overlay assay . This protein was alpha (2-6) sialylated, as shown by SNA binding . Furthermore, SNA lectin was able to block pilus binding to CEPs . 125I labeling of pili and a solid-phase binding assay confirmed that pili bind to CEPs and, further, that binding could be competitively inhibited by excess unlabeled pili and that the receptors appeared saturable . GlycoTrack reagents were used to show that the epithelial proteins of the postnatal day 5 (P5) mouse cornea were glycosylated . Removal of carbohydrates by preincubation of blots with periodate, or combining pili with sialic acid, eliminated pili binding . Pretreatment of blots with either neuraminidase (N'ase) to decrease and/or remove sialic acid residues, or pretreatment with SNA lectin with specificity for alpha (2-6) linked sialic acid to galactose, also diminished pili binding to CEPs . Other sugars or MAA lectin, specific for sialic acid alpha (2-3) linked to galactose, had no inhibitory effect . CONCLUSIONS . These data show that a 57-kd surface membrane protein bound pili in the immature cornea and that for both this protein and the other nonsurface proteins, sialic acid alpha (2-6) linked to galactose was important in receptor recognition by the pilus adhesion . The 57-kd protein is putatively important in the initial interaction of pili with the unwounded ocular epithelium and may be the initial pathogenic event in this model.

J Med Microbiol, 1995 Mar, 42(3), 220 - 4
Mixed morphotype testing of Pseudomonas aeruginosa cultures from cystic fibrosis patients; Wolter JM et al.; Bronchial secretions of patients with cystic fibrosis (CF) inevitably become colonised with Pseudomonas aeruginosa . This organism often exhibits multiple phenotypes with different antibiotic susceptibility profiles . Isolating each colonial morphotype and determining its antibiotic susceptibility profile is labour-intensive and time-consuming . Two disk diffusion methods for mixed morphotype susceptibility testing were examined; 134 morphotypes of P . aeruginosa from 50 respiratory specimens from CF patients were tested . Mixed cultures were prepared by (a) combining morphologically different colonies of P . aeruginosa directly from the sputum specimen primary culture plates from individual patients or (b) mixing colonies of individual morphotypes after isolation and subculture . Agreement with the results for testing morphotypes individually were 85.8% and 91.6%, respectively, for the two methods . These agreement rates rose to 95.6% and 99.4%, respectively, when minor errors (intermediate misclassified as susceptible or resistant or vice versa) were excluded . Mixed morphotype testing of P . aeruginosa directly from sputum specimen culture plates in chronically colonised CF patients is more efficient, reduces turn-around time and provides clinically meaningful information about antibiotic susceptibility.

J Infect Dis, 1995 Mar, 171(3), 570 - 5
Adenovirus-mediated blockade of tumor necrosis factor in mice protects against endotoxic shock yet impairs pulmonary host defense; Kolls JK et al.; A replication-deficient recombinant adenovirus encoding a chimeric protein capable of binding tumor necrosis factor (TNF) and lymphotoxin was given to mice . Administration of this virus (10(9) pfu intravenously) yielded high levels of the recombinant protein in plasma and afforded significant protection to a lethal challenge with lipopolysaccharide with or without D-galactosamine . However, this protein inhibitor was readily detectable in the lung and was associated with decreased neutrophil recruitment and bacterial killing after intratracheal LPS or Pseudomonas aeruginosa, respectively . These data reflect the dual role of many proinflammatory cytokines . This model of TNF inhibition is similar to the homozygous 55-kDa TNF receptor deletion; thus, adenovirus-mediated gene transfer of cytokine inhibitors in vivo is a useful tool to abrogate the function of single or multiple cytokines for investigational or therapeutic purposes.

Am J Respir Cell Mol Biol, 1995 Mar, 12(3), 296 - 306
Mucus glycoconjugate complexes released from feline trachea by a bacterial toxin; Fung DC et al.; This paper describes low-density mucus glycoconjugates released from feline trachea by dirhamnolipid (DRL), a toxin from Pseudomonas aeruginosa . Mucus glycoconjugates in feline tracheas were radiolabeled in vivo with 3H-proline and 14C-glucose . Control mucus and that released by 200 micrograms/ml DRL were dissolved in guanidine hydrochloride buffer (GuHCl) and chromatographed on Sepharose CL-2B . Molecules eluting in the void volume (V0) of the column were isolated by isopycnic density gradient centrifugation in CsCl/GuHCl . All samples gave peaks of radiolabeled and periodic acid/Schiff (PAS)-reactive material at rho = approximately 1.50 and approximately 1.60 g/ml, but DRL-stimulated samples contained low-density material (rho < 1.32 g/ml), also PAS-reactive and radiolabeled . Control secretions incubated with DRL in vitro did not form low-density material . In Triton X-100 (1% vol/vol), a nonionic detergent, low-density material behaved as smaller molecules, running in the partially included volume (Vi) of the column of Sepharose CL-2B, but still in the V0 of Sephacryl S-300 . Incubation with chondroitinase ABC, heparinase II and III, and keratanase failed to change its elution profile on S-300, evidence against glycosaminoglycans; but proteolysis with trypsin or proteinase K gave two peaks, peptide fragments near the totally included volume of the column and glycopeptides in V0 . The V0 glycopeptides banded between 1.50 and 1.55 g/ml in a CsCl gradient and eluted as a single peak in the Vi of Sephacryl S-400, suggesting a distinct homogeneous glycopeptide, smaller than those from normal mucins . The main 14C-labeled sugars in this glycopeptide were fucose, glucosamine, galactosamine, and galactose, consistent with a mucin . Thus, DRL releases stable but noncovalent complexes containing one or more distinct mucinlike glycoconjugates, probably combined with lipids and peptides . We discuss their possible relevance to airway diseases, including cystic fibrosis.

Infect Immun, 1995 Mar, 63(3), 1107 - 12
TH1 cells trigger tumor necrosis factor alpha-mediated hypersensitivity to Pseudomonas aeruginosa after adoptive transfer into SCID mice; Fruh R et al.; Recent experiments have shown that gamma interferon (IFN-gamma), either administered or induced in vivo, e.g., by certain bacteria, is a key mediator in inducing hypersensitivity to bacterial lipopolysaccharides . The source of endogenous IFN-gamma in this context (natural killer versus TH1 cells) has not been investigated yet . In order to investigate the role of antigen-specific, IFN-gamma-producing TH1 cells in murine Pseudomonas aeruginosa infection, a murine TH1 cell line was propagated in vitro by using recombinant P . aeruginosa outer membrane protein I . Adoptive transfer experiments were performed by intravenous injection of various amounts of TH1 cells into P . aeruginosa-challenged SCID mice . Adoptive transfer of 5 x 10(6) T cells into SCID mice followed by an intraperitoneal challenge with 1.4 x 10(6) CFU of live P . aeruginosa resulted in the rapid death of the animals within 12 h postchallenge, whereas transfer of lower T-cell doses and saline as a control did not cause any detrimental effects . After challenge with 2.8 x 10(6) CFU of P . aeruginosa, similar results were obtained 18 h postchallenge; however, at the end of the 72-h observation period, no significant differences in survival rates were obtained between the groups treated with different amounts of T cells . The rapid death of mice treated with 5 x 10(6) T cells was reflected by 860-fold-elevated levels of tumor necrosis factor alpha (TNF-alpha) present in serum 2 h postchallenge, whereas no significant differences in TNF-alpha serum levels were detectable in mice treated with lower doses of T cells or with saline . Pretreatment of T-cell-reconstituted SCID mice with neutralizing anti-IFN-gamma monoclonal antibodies completely protected mice from bacterial challenge and reduced TNF-alpha levels in serum . We conclude that under the experimental conditions described here, IFN-gamma- and interleukin-2-producing TH1 cells represent an important trigger mechanism inducing TNF-alpha-mediated hypersensitivity to bacterial endotoxin.

Curr Eye Res, 1995 Mar, 14(3), 229 - 34
Effectiveness of specific antibiotic/steroid combinations for therapy of experimental Pseudomonas aeruginosa keratitis; Engel LS et al.; Ciprofloxacin and prednisolone, but not an aminoglycoside and dexamethasone, were previously found to be effective in killing bacteria and reducing inflammation for the treatment of Pseudomonas keratitis . We investigated the therapeutic effectiveness of tobramycin/prednisolone and ciprofloxacin/dexamethasone in a rabbit model of experimental keratitis to increase our understanding of the effectiveness of antibiotic/steroid combinations . To our knowledge, this is the first analysis of the effectiveness of a combination of ciprofloxacin and dexamethasone for experimental keratitis . Two experiments were conducted . In the first experiment, 36 rabbits were divided into six groups: 1) untreated; 2) prednisolone acetate, 1.0%; 3) prednisolone phosphate, 1.0%; 4) tobramycin, 1.36%; 5) tobramycin plus prednisolone acetate; 6) tobramycin plus prednisolone phosphate . In the second experiment, 23 rabbits were divided into four groups: 1) untreated; 2) ciprofloxacin, 0.3%, plus dexamethasone alcohol, 0.1%; 3) ciprofloxacin; 4) dexamethasone alcohol . Topical antibiotic and/or steroid was given for 10 h, from 16 to 26 h postinfection, one drop every 15 min for the first hour and then every 30 min for the remaining 9 h . At 27 h postinfection, eyes were evaluated by slit lamp examination (SLE) and assayed for the presence of bacteria in terms of colony forming units (CFU) per cornea . Both prednisolone acetate and prednisolone phosphate reduced ocular inflammation (as determined by SLE), compared with no treatment (P < or = 0.036); the phosphate was more effective (P = 0.005) . Tobramycin alone and in combination with prednisolone also significantly reduced SLE, compared with no treatment (P < or = 0.006) . The bactericidal activity of tobramycin was not affected by either steroid formulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Antimicrob Agents Chemother, 1995 Mar, 39(3), 790 - 2
nfxC-type quinolone resistance in a clinical isolate of Pseudomonas aeruginosa; Fukuda H et al.; Quinolone resistance gene nqr-T91 in a clinical isolate of Pseudomonas aeruginosa P1481 was cotransducible with catA1 in P . aeruginosa PAO . The nqr-T91 transductant, PKH-T91, was resistant to norfloxacin, imipenem, and chloramphenicol and showed less norfloxacin accumulation than the parent strain did . Loss of the 46-kDa outer membrane protein (D2) and an increase in the 50-kDa outer membrane protein in PKH-T91 were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Lipopolysaccharides in the transductant were also changed . These alterations were considered to be related to lower levels of norfloxacin accumulation in PKH-T91 . These genetic and biochemical properties suggested that an nfxC type of quinolone-resistant mutation occurred in a clinical isolate of P . aeruginosa P1481.

Antimicrob Agents Chemother, 1995 Mar, 39(3), 731 - 4
In vitro activity and killing effect of DX-8739, a new carbapenem, compared with those of meropenem and imipenem against multiresistant Pseudomonas aeruginosa; Giamarellos-Bourboulis EJ et al.; DX-8739 is a new dehydropeptidase I-stable carbapenem . In order to evaluate its activity in comparison with those of meropenem and imipenem, 147 multiresistant Pseudomonas aeruginosa isolates acquired nosocomially were simultaneously exposed to the actions of the three carbapenems in vitro, whereas to compare their killing effects on 14 strains, 56 killing curve studies were performed . Overall DX-8739 was found to possess inhibitory activity as well as bactericidal activity statistically superior to those of meropenem and imipenem . At a concentration of 4 micrograms/ml, 106 strains (72.1%) were found to be imipenem resistant; 33 and 27.4% of these strains were inhibited by DX-8739 and meropenem, respectively, a statistically significant difference (P < 0.05) . DX-8739 was also shown to possess intrinsic activity in vitro superior to those of meropenem and imipenem against the imipenem-susceptible population of strains . However, no statistically significant difference regarding the comparative killing activities of the three studied carbapenems was observed . Following exposure to carbapenem for 24 h, 33.3, 44.4, and 70% of the strains which survived became resistant to DX-8739, meropenem, and imipenem, respectively . The reported results demonstrate the significant activity of DX-8739 against multiresistant P . aeruginosa strains acquired nosocomially . The mechanism of action of DX-8739 on P . aeruginosa is unknown, and various hypotheses that might explain its in vitro superiority over meropenem and imipenem are proposed.

Antimicrob Agents Chemother, 1995 Mar, 39(3), 702 - 6
Mechanism of enhanced antipseudomonal activity of BO-2727, a new injectable 1-beta-methyl carbapenem; Hazumi N et al.; The mechanism of the enhanced activity of BO-2727 against imipenem-resistant Pseudomonas aeruginosa was studied by using a set of four isogenic strains derived from beta-lactamase-deficient P . aeruginosa PAO4089 (blaJ blaP) . Complementation of the blaJ and blaP mutations conferred greater resistance to biapenem, panipenem, and imipenem than to BO-2727 and meropenem, most notably in the outer membrane protein D2-deficient strain . The higher levels of resistance to biapenem, panipenem, and imipenem can be explained by the slow but significant hydrolysis by beta-lactamase, whereas the reduced levels of resistance to BO-2727 and meropenem would be attributable to their stability in the presence of high levels of beta-lactamase and the fact that they cause only low induction of beta-lactamase . It is also noted that the activity of BO-2727 against the beta-lactamase-deficient strain was less affected by the loss of the D2 porin than was that of meropenem, indicating that BO-2727 in comparison with meropenem can overcome an intrinsic resistance caused by the loss of D2 . Moreover, comparative in vitro resistance studies have shown that BO-2727 and meropenem selected fewer resistant cells than other carbapenems . In conclusion, BO-2727 exhibited improved activity against imipenem-resistant P . aeruginosa, probably because of its ability to overcome loss of the D2 porin and beta-lactamase hydrolysis.

Antimicrob Agents Chemother, 1995 Mar, 39(3), 686 - 93
New mobile gene cassettes containing an aminoglycoside resistance gene, aacA7, and a chloramphenicol resistance gene, catB3, in an integron in pBWH301; Bunny KL et al.; The multidrug resistance plasmid pBWH301 was shown to contain a sull-associated integron with five inserted gene cassettes, aacA7-catB3-aadB-oxa2-orfD, all of which can be mobilized by the integron-encoded DNA integrase . The aadB, oxa2, and orfD cassettes are identical to known cassettes . The aacA7 gene encodes a protein that is a member of one of the three known families of aminoglycoside acetyltransferases classified as AAC(6')-I . The chloramphenicol acetyltransferase encoded by the catB3 gene is closely related to members of a recently identified family of chloramphenicol acetyltransferases . The catB3 gene displays a relatively high degree of sequence identity to a chromosomally located open reading frame in Pseudomonas aeruginosa, and this may represent evidence for the acquisition by a cassette of a chromosomal gene.

Antimicrob Agents Chemother, 1995 Mar, 39(3), 677 - 9
Should Pseudomonas aeruginosa isolates resistant to one of the fluorinated quinolones be tested for the others? Studies with an experimental model of pneumonia; Chidiac C et al.; A clinical isolate of Pseudomonas aeruginosa resistant to pefloxacin (Pef) but susceptible to ciprofloxacin (Cip) was studied to compare the in vitro and in vivo activities of Pef, ofloxacin (Ofl), and Cip . The time-kill curve method showed no bactericidal activity for Pef and Ofl, but a reduction of 4 log10 CFU/ml was achieved with Cip at 1 h . A model of experimental P . aeruginosa pneumonia was used to evaluate in vivo the relevance of the difference in susceptibility observed in vitro . At 36 h, a 100% cumulative survival rate was observed in Cip-treated rats, which was far higher than the survival rate obtained with Pef (53%) or Ofl (46%) (P < 0.001) . At 4 h, no bacteremia was observed in Cip-treated rats, whereas 93% of the Pef-treated rats and 80% of the Ofl-treated rats were bacteremic (P < 0.001) . The best pulmonary bacterial clearance was observed with Cip . Interestingly, Pef and Ofl, to which the strain was resistant in vitro, showed a fairly good in vivo activity despite sub-MIC concentrations . Cip was more effective than Pef and Ofl in terms of pulmonary and systemic bactericidal activity and provided the best survival rate in animals . We conclude that differences between the different quinolones in terms of the organism's sensitivity assessed in vitro may be relevant and that it might be useful to reconsider the use of a quinolone to which P . aeruginosa shows resistance if the organism shows sensitivity to no other agent.

Antimicrob Agents Chemother, 1995 Mar, 39(3), 645 - 9
Outer membrane proteins responsible for multiple drug resistance in Pseudomonas aeruginosa; Masuda N et al.; Three types of multiple-drug-resistant mutants which were phenotypically similar to previously described nalB, nfxB, and nfxC mutants were isolated from Pseudomonas aeruginosa PAO1 and two clinical isolates . Type 1 (nalB-type) mutants showed cross-resistance to meropenem, cephems, and quinolones . They overproduced an outer membrane protein with an apparent molecular mass of 50 kDa (OprM) . Type 2 (nfxB-type) mutants showed cross-resistance to quinolones and new cephems, i.e., cefpirome and cefozopran, concomitant with overproduction of an outer membrane protein with an apparent molecular mass of 54 kDa (OprJ) . Type 3 (nfxC-type) mutants showed cross-resistance to carbapenems and quinolones . They produced decreased amounts of OprD and increased amounts of a 50-kDa protein (OprN), which was almost the same molecular weight as that of OprM, but it was distinguishable from OprM by its heat modifiability on sodium dodecyl sulfate-polyacrylamide gel electrophoresis . In the presence of salicylate, the parent strains showed an increased level of resistance to carbapenems and quinolones and produced decreased amounts of OprD and increased amounts of OprN . Salicylate caused the repression of OprJ production and the loss of resistance to cefpirome and cefozopran in two of the three OprJ-overproducing mutants, although salicylate slightly increased the level of resistance in the parent strains . The changes in susceptibilities were transient in the presence of salicylate . These data suggest that at least three different outer membrane proteins, OprM, OprJ, and OprN, are associated with multiple drug resistance in P . aeruginosa.

Intensive Care Med, 1995 Mar, 21(3), 226 - 8
Septic shock due to Pseudomonas aeruginosa in a previously healthy woman; Ishihara S et al.; Pseudomonas aeruginosa septicemia rarely occurs in non-immunocompromised adults . We present a case of septic shock following Pseudomonas aeruginosa pneumonia in a previously healthy 48-year-old woman . The onset was sudden, with back pain, pyrexia and shock . Chest radiographs revealed pneumonia, and Pseudomonas aeruginosa was identified from blood and sputum cultures . Therapy with dopamine, piperacillin and fluid replacement led to a prompt recovery . Laboratory tests failed to reveal any immunological deficits . Including this case, only five cases of Pseudomonas aeruginosa septicemia in patients though to be non-immunocompromised have been reported . Two remarkable features of this type of Pseudomonas infection are apparent: i) it commonly develops from pneumonia and ii) it has a better prognosis than that in immunocompromised hosts.

Rinsho Ketsueki, 1995 Mar, 36(3), 206 - 11
{Aspergillus lumbar discitis in a patient with acute lymphoblastic leukemia following induction therapy}; Kawamura M et al.; A 47-year-old female was admitted in October 1988 because of anemia and lymphoblastic cells in peripheral blood . A bone marrow aspirate was hypercellular with 93.9% lymphoblasts negative for peroxidase staining . The case was diagnosed as ALL (L2), and treated with JALSG ALL-87 regimen . She developed spiky fever and endotoxin shock due to bacteremia caused by pseudomonas aeruginosa, then was treated with several antibiotics . With the recovery of leukocytes, the chest X-ray showed an infiltrative shadow and a cavity forming lung abscess resembling aspergilloma in her left lung . The cavity improved of transbronchial infusion following amphotericin B (AMPH-B) . Although she achieved complete remission, she felt severe lumbago accompanied by a marked erosion of the vertebral body with disc space narrowing on her X-ray . Then she underwent surgery to remove a disc abscess, and 1 colony of the aspergillus species was cultured from the specimen . She was treated with intravenous AMPH-B, and post remission therapies were performed under the injection of anti-fungal agents . No remarkable symptoms of complications were recognized during the chemotherapy . AMPH-B is useful and safe for the management of aspergillus discitis.

J Antimicrob Chemother, 1995 Mar, 35(3), 425 - 7
Comparing antimicrobial activity against resistant Pseudomonas aeruginosa using an index for the absence of cross-resistance; Takigawa K et al.; We devised an index to estimate the degree cross-resistance between piperacillin, ceftazidime, sulbactam/cefoperazone, amikacin, tobramycin, carumonam and imipenem against 139 separate clinical isolates of Pseudomonas aeruginosa . A negative value of the index indicated the cross-resistance whereas a positive value suggested the converse making the device an index for the absence of cross resistance (ACR) . Using the ACR index, we identified pairs of antibiotics exhibiting the least degree of cross-resistance and therefore the highest potential for treating infections due to P . aeruginosa.

Int J Pediatr Otorhinolaryngol, 1995 Mar, 31(2-3), 153 - 7
Role of anaerobic bacteria in chronic otitis media and cholesteatoma; Brook I; Otitis media (OM), a common infection in children, can cause significant morbidity . Selection of the most appropriate treatment regimen directed against the pathogens responsible for the OM can minimize complications . The most frequently isolated bacteria from chronic OM are Staphylococcus aureus, Pseudomonas aeruginosa and anaerobic bacteria . The predominant anaerobes are Peptostreptococcus spp., pigmented Prevotella and Porphyromonas spp., Bacteroides spp . and Fusobacterium spp . Many of the organisms causing OM can produce beta-lactamase, which can contribute to the failure of penicillins therapy . The appropriate surgical and medical therapy for chronic OM is reviewed.

Arch Microbiol, 1995 Mar, 163(3), 217 - 22
Growth-phase-dependent alginate synthesis, activity of biosynthetic enzymes and transcription of alginate genes in Pseudomonas aeruginosa; Leitao JH et al.; Alginate synthesis by the highly mucoid Pseudomonas aeruginosa 8821 M is growth-phase-dependent, and the alginate produced per unit of biomass reaches maximum values in the deceleration phase of growth . However, the degree of polymerization increases as batch growth proceeds, reaching maximum values at the stationary phase of growth . The activity of the four enzymes leading to GDP-mannuronic acid formation, phosphomannose isomerase, phosphomannomutase, GDP-mannose pyrophosphorylase and GDP-mannose dehydrogenase peaked earlier at the late exponential phase . Growth-phase-dependent activity of alginate biosynthetic enzymes correlates with the level of transcription of the encoding alginate genes algA, algC and algD during growth, as indicated by Northern blot hybridization experiments . The pattern of coordinate transcriptional growth-phase regulation of these alginate structural genes concurs with the growth-dependent transcription of the regulatory gene algR1.

Infect Dis Clin North Am, 1995 Mar, 9(1), 195 - 216
Infections of the head and neck in diabetes mellitus; Tierney MR et al.; Patients with diabetes mellitus exhibit particular susceptibility to three severe infections of the head and neck: rhinocerebral mucormycosis, postoperative endophthalmitis, and malignant otitis externa . Rhinocerebral mucormycosis is an extensive life-threatening infection beginning in the nasal passages and sinuses and extending often into the orbit and the cerebrum . Endophthalmitis, which is infection of the vitreal contents, can occur secondary to bacteremia, trauma, or postoperatively . Invasive external otitis or malignant otitis externa is an invasive infection beginning in the adjacent soft tissue and into bone . It is usually secondary to Pseudomonas aeruginosa and occurs almost exclusively in diabetics . These will all be discussed in this article.

Curr Microbiol, 1995 Mar, 30(3), 123 - 6
Reactivity of the co-type and baa3-type cytochrome c oxidases from Pseudomonas aeruginosa with different endogenous cytochromes c; Okamoto A et al.; The reactivity between different cytochromes c purified from Pseudomonas aeruginosa cells grown aerobically in the absence of nitrate and isolated cytochromes co and baa3 was determined . The P . aeruginosa cytochrome co reacted most rapidly with the membrane-bound cytochrome c-551 among three c-type cytochromes analyzed, whereas the cytochrome baa3 reacted best with the membrane-bound cytochrome c-555 . The results indicated that two terminal electron transfer systems are present in aerobic P . aeruginosa: one contains the cytochrome c-551 and cytochrome co, and the other contains the cytochrome c-555 and cytochrome baa3.

Chemotherapy, 1995 Mar-Apr, 41(2), 92 - 9
Effect of quinolones against slowly growing bacteria; Dalhoff A et al.; Bacteria growing in vivo multiply much more slowly than in vitro . If the bactericidal activity of quinolones may be affected by an increase in generation time (g) was studied in batch cultures as well as under the well-controlled conditions of a continuous-flow culture . By limiting the nutrient supply, generation times were lengthened from approximately 0.45 to 1.5 h up to 3.9 h . Three recent clinical isolates each of Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa were exposed to twice the MIC of ciprofloxacin, norfloxacin, fleroxacin and ofloxacin . The 'killing rates' were calculated in analogy to the growth rate . The bactericidal activity of the quinolones tested against E . coli was minimally influenced by the reduced generation time and the effect against S . aureus was moderate . As compared to their rapidly growing counterparts (g = 0.4 h) slowly growing (g = 1.3 h) P . aeruginosa were killed even more effectively by ciprofloxacin (176% increase) fleroxacin (48% increase) norfloxacin (36% increase) and ofloxacin (86% increase) . These changes may likely be due to adaptive responses of the outer membranes of the bacteria to the limited nutrient supply thereby sensitizing the bacteria to the bactericidal activity of quinolones.

J Clin Microbiol, 1995 Mar, 33(3), 572 - 5
Risk of cross-colonization and infection by Pseudomonas aeruginosa in a holiday camp for cystic fibrosis patients; Hoogkamp-Korstanje JA et al.; The risk of cross-colonization and subsequent infection by Pseudomonas aeruginosa in holiday camps for cystic fibrosis patients was studied in 91 children by culturing sputum at their arrival, at their departure, 2 months later, and at regular intervals thereafter . The isolated strains were subjected to serotyping, phage typing, pyocin typing, and genotyping by random amplified polymorphic DNA fingerprinting-PCR . It was concluded from random amplified polymorphic DNA fingerprinting-PCR typing that the Pseudomonas flora was not constant in most children . Some children harbored one genotype, whereas some harbored two or more different genotypes simultaneously . Most culture-positive children easily acquired a strain of another genotype which replaced the former one or coexisted with the original one . The incidence of sputum conversion was 7.7% in previously negative children; the incidence of permanent colonization and infection was 1.9% . This risk was comparable with that observed in the community . We conclude that the risk of cross-infection is trivial compared with the obvious joy and social benefit derived from a holiday camp.

J Clin Microbiol, 1995 Mar, 33(3), 528 - 34
Discriminatory power of three DNA-based typing techniques for Pseudomonas aeruginosa; Grundmann H et al.; We assessed the capacity of three DNA typing techniques to discriminate between 81 geographically, temporally, and epidemiologically unrelated strains of Pseudomonas aeruginosa . The methods, representing powerful tools for hospital molecular epidemiology, included hybridization of restricted chromosomal DNA with toxA and genes coding for rRNA (rDNA) used as probes and macrorestriction analysis of SpeI-digested DNA by pulsed-field gel electrophoresis . The probe typing techniques were able to classify all strains into a limited number of types, and the discriminatory powers were 97.7 and 95.6% for toxA and rDNA typing, respectively . Strains that were indistinguishable on the basis of both toxA and rDNA types defined 12 probe type homology groups . Of these, one contained five strains, three contained three strains each, and eight groups were represented by two strains each . Strains in 10 of the homology groups had the same O serotype . SpeI macrorestriction patterns discriminated between all strains with at least four band differences, which corresponded to a similarity level of 85% . Fifteen pairs of strains were similar at a level of > 75% and differed by only four to seven bands . Of these pairs, 11 belonged to the same probe type homology group, indicating their clonal relatedness . We conclude that macrorestriction analysis of P . aeruginosa with SpeI provides the best means of discrimination between epidemiologically unrelated strains . However, DNA probe typing with either toxA or rDNA reveals information on the strain population structure and evolutionary relationships.

Kansenshogaku Zasshi, 1995 Mar, 69(3), 272 - 9
Detection of Mycobacterium tuberculosis in clinical specimens by polymerase chain reaction method; Bi HG et al.; An insertion sequence repeated multiple times in the chromosome of Mycobacterium tuberculosis was used as a target for amplification using the polymerase chain reaction (PCR) assay for detecting Mycobacterium tuberculosis in clinical specimens . The sequences of primers were 5'-CCTGCGAGCGTAGGCGTCGG-3' (primer 1) and 5'-CTCGTCCAGCGCCGCTTCGG-3' (primer 2) . One cycle of amplification consisted of denaturing at 94 degrees C for 2 min, primer annealing at 68 degrees C for 2 min, and extension at 72 degrees C for 2 min . DNA (5 fg) extracted from M . tuberculosis was detected by gel electrophoresis and Southern blot hybridization after 40 cycles of amplification . The amplification products were not obtained by DNA extracted from M . kansasii, M . intracellulare, M . avium, M . fortuitum, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Legionella pneumophila and Staphylococcus aureus; only from the M . tuberculosis complex . PCR results were compared with conventional cultural, pathological and microscopic findings in the detection of M . tuberculosis in 112 clinical specimens . There were 25 specimens that were positive for M . tuberculosis by cultural or pathological examination, of which 20 (80%) were positive by PCR . PCR detected the organism in 5 (83%) of 6 smear-positive specimens and 15 (79%) of 19 smear-negative specimens in which culture or pathology revealed M . tuberculosis . In addition, 2 smear-negative specimens and 8 smear-negative and culture-negative specimens were positive by PCR . These 10 samples were collected from the patients suspected as having tuberculosis by the clinical diagnosis based on the clinical history, characteristic radiographs, a positive PPD skin test and the effectiveness of anti-tuberculous drugs.(ABSTRACT TRUNCATED AT 250 WORDS)

Arzneimittelforschung, 1995 Mar, 45(3), 327 - 30
Suppression of virulence factors of pseudomonas aeruginosa by subinhibitory concentrations of quaternary ammonium salts; Majtan V et al.; The effect of subinhibitory concentrations (sub-MICs) of two homologous series of "soft" quaternary ammonium salts (QATs) upon the expression of phospholipase C (Pseudomonas aeruginosa strain 72/92), elastase, proteinase and permeability activity (P . aeruginosa 9/92) was studied . Both strains were isolated from the patients suffering from nosocomial infections . Phospholipase C was the most markedly inhibited enzyme . The inhibitory effect was directly proportional to the concentration of substances but the degree of inhibition in the two homologous series was different . In the second strain of P . aeruginosa its production of elastase, permeability factor and mainly its proteinase activity were less affected by the series of tested QATs . A significant inhibition of production of these factors was manifested only in higher concentrations particularly by the substances with longer alkyls . The effect of QATs on the production of virulence factors could be attributed to their influence on the metabolic activity of organisms.

N J Med, 1995 Mar, 92(3), 165 - 6
Case report: fatal Pseudomonas aeruginosa pneumonia and sepsis; Parikh P et al.; The authors report a case of overwhelming Pseudomonas pneumonia with sepsis and death in a 54-year-old male; death resulted from jacuzzi exposure in an immunocompetent host . The patient died 24 hours after hospital admission from gram negative sepsis shock.

J Invest Surg, 1995 Mar-Apr, 8(2), 115 - 22
Hemodynamic effects of bradykinin antagonism in porcine gram-negative sepsis; Ridings PC et al.; Activation of the kallikrein-kinin system in sepsis has long been recognized, but its role, beneficial or pathologic, has not been defined . Recently, however, specific bradykinin (BK) antagonists have become available and this study investigated the effects of a BK antagonist, NPC17731 (Scios-Nova) on systemic and pulmonary hemodynamics in a model of gram-negative sepsis . Anesthetized swine were studied for 5 h receiving a 1-h infusion of saline (controls, group 1, N = 8) or live Pseudomonas aeruginosa (septic, group 2, N = 8) . Group 3 (treatment, N = 6) received NPC17731 (5 mg/kg initial bolus followed by 1 mg/kg hourly) just prior to the onset of sepsis . Group 2 animals showed a rapid decrease in systemic arterial pressure (SAP) from 30 min onward, and sustained significant hypotension from 2 h onward . In group 3, SAP fell similarly until 2 h then progressively rose, returning to baseline levels by 5 h . In contrast, cardiac index fell progressively from 3 h onward in groups 2 and 3 . Systemic vascular resistance index (SVRI) fell significantly by 2 h in group 2 animals, recovering to baseline by 5 h . Group 3 showed a similar initial fall followed by a rebound increase in SVRI, which, at 5 h was significantly raised above the other groups . Group 2 developed significant, persistent pulmonary artery hypertension which was not reduced by NPC17731 . The data imply a significant role for bradykinin in the pathogenesis of hypotension in septic shock in this model . Septic shock was reversed by a BK antagonist which increased peripheral resistance without affecting cardiac output.(ABSTRACT TRUNCATED AT 250 WORDS)

Zentralbl Hyg Umweltmed, 1995 Mar, 196(6), 532 - 44
{Acanthamoeba, naturally intracellularly infected with Pseudomonas aeruginosa, after their isolation from a microbiologically contaminated drinking water system in a hospital}; Michel R et al.; The drinking water system of a new hospital building that was highly contaminated with bacteria before opening was investigated too for the prevalence of small free living amoebae . Germ counts resulted in > 100 CFU/ml in 100% of the cold water samples, that showed also growth of P . aeruginosa, whereas E . coli and coliforme bacteria could not be identified . The investigation of 37 water samples for protozoa revealed growth of small freeliving amoebae in 20 samples (54%) belonging to 10 species of the genus Acanthamoeba, Naegleria, Hartmannella, Echinamoeba among others . In addition 2 Ciliate- and 2 Microflagellate-species could be observed . While all Naegleria strains isolated belonged to the N . gruberi-complex two of 16 Acanthamoeba-isolates proved to be pathogenic for laboratory mice . From 7 watersamples positive with P . aeruginosa 5 Acanthamoeba- and 2 Echinamoeba strains could be isolated which revealed intracellular multiplication of P . aeruginosa . Because of their well known resistances against chlorine, the amoebae and their cysts are considered to be vectors for these intracellular bacteria . A complete sanitation of the incriminated drinking water system was accomplished by combined chemical and thermic disinfection measures.

Naunyn Schmiedebergs Arch Pharmacol, 1995 Mar, 351(3), 315 - 9
Pseudomonas aeruginosa cytotoxin: the Asp197-Gly-Asp-Tyr-His-Tyr-His-Tyr202 containing loop is critical for plasma membrane binding; Struckmeier M et al.; The cytotoxin from Pseudomonas aeruginosa is a pore-forming toxin that binds specifically to water channel-related molecules of the erythrocyte membrane . Here, we have defined a domain, Asp197-Gly-Asp-Tyr-His-Tyr202 of the cytotoxin, to be essential for receptor binding . Cytotoxin point mutants from the recombinant gene carrying substitutions in the domain were characterized in terms of inhibiting the binding of radioiodinated natural cytotoxin to rat erythrocyte and producing cytotoxic effects in human granulocytes . A synthetic peptide representing residues 191-211 of the cytotoxin acted as a competitive inhibitor at a concentration of 10(-5) M . In contrast, two other cytotoxin-specific peptides were inactive . Structure prediction of the binding sequence shows a loop structure with similarities to the sequence around His332 in Aeromonas aerolysin essential to receptor binding.

Eur J Radiol, 1995 Mar, 19(3), 198 - 205
Risk of microbial contamination of iodinated contrast media on multiple use of large-volume bottles; Dominik RH et al.; Large-volume bottles of iodinated contrast material (CM) offer advantages over single-dose vials in respect of handling, radiation protection of personnel, cost and waste . Because non-ionic CM are considered to be susceptible to microbial contamination, the probability of such contamination was investigated under practical conditions with reference to the duration of use . Under conditions identical to those in practice, contamination was found in only about 1% of bottles examined after a single piercing . Under worst-case conditions, a maximum of 9 microorganisms/bottle was found . No pathogen multiplication was observed within 24 h in microbial challenge tests; rather, the nosocomial pathogens examined died quickly in iopromide, the only exception being Pseudomonas aeruginosa . There was not a single case of bacterial multiplication on clinical use of 500-ml infusion bottles in patients undergoing routine computed tomography, using either injection by hand or an automatic infusion device . In conclusion, the microbial count rate did not differ from that of small-volume parenterals used up immediately after piercing . To ensure an adequate safety margin for the avoidance of septicemia or pyrogenic reactions, the CM should be used within 1 examination day.

Proc Natl Acad Sci U S A, 1995 Feb 28, 92(5), 1490 - 4
A second N-acylhomoserine lactone signal produced by Pseudomonas aeruginosa; Pearson JP et al.; Quorum sensing systems are used by a number of Gram-negative bacterial species to regulate specific sets of genes in a cell density-dependent manner . Quorum sensing involves synthesis and detection of extracellular signals termed autoinducers . As shown in recombinant Escherichia coli, the Pseudomonas aeruginosa autoinducer (PAI) N-(3-oxododecanoyl)homoserine lactone, together with the lasR gene product, activate the P . aeruginosa lasB gene . In this study, PAI was shown to activate lasB-lacZ expression in a P . aeruginosa lasR mutant containing a plasmid with lasR under the control of the lac promoter . The concentration of PAI necessary for half-maximal activation of the lasB-lacZ fusion was approximately 1 microM, which is within the range of PAI levels found in P . aeruginosa culture fluids . The effect of PAI on a P . aeruginosa lasR mutant containing a plasmid with lasR under the control of its own promoter and containing the lasB-lacZ fusion was also tested . Although extracts of culture fluid activated the lasB promoter in this construct, concentrations of PAI as high as 10 microM did not . This indicates the presence of a second extracellular factor (factor 2) that is required for lasB activation in P . aeruginosa when lasR is controlled by its own promoter but not when lasR is controlled by a strong foreign promoter . Factor 2 was shown to be N-butyrylhomoserine lactone . Although recombinant E . coli cells containing the PAI synthase gene, lasI, produce PAI, these cells do not produce factor 2 . Furthermore, a P . aeruginosa mutant that produced about 0.1% of the wild-type level of PAI made about 5% of the wild-type level of factor 2 . This indicates that factor 2 synthesis results from the activity of a gene product other than PAI synthase . The role of factor 2 in virulence gene regulation remains to be determined, but this compound may affect the expression of lasR, which in turn activates transcription of numerous virulence genes in the presence of sufficient PAI . Apparently, multiple quorum sensing systems can occur and interact with each other in a single bacterial species.

Gene, 1995 Feb 27, 154(1), 61 - 4
Cloning and sequence analyses of the genes coding for the integration host factor (IHF) and HU proteins of Pseudomonas aeruginosa; Delic-Attree I et al.; Histone-like proteins, such as HU and the integration host factor (IHF), are small, dimeric, DNA-bending proteins which play a role in maintaining constrained DNA structures and hence in regulating gene expression . Two different strategies were used to isolate the genes coding for Pseudomonas aeruginosa (Pa) HU and IHF, two proteins that we have previously isolated from a mucoid strain . By use of a PCR-based technique with oligodeoxyribonucleotides (oligos) designed from the N-terminal amino acid (aa) sequences of HU and the beta-subunit of IHF, and Southern blot analyses, hupB and himD, encoding HU and IHF beta, respectively, have been cloned . The himA gene of Pa, encoding the alpha-subunit of IHF, was isolated using himA of Escherichia coli (Ec) as a probe in Southern blot analyses . The deduced hupB product (90 aa, 9 kDa) is 79% identical to HU beta and 61% to HU alpha of Ec . The predicted products of himA (100 aa, 11.5 kDa) and of himD (94 aa, 10.6 kDa) share 77 and 70% identity with IHF alpha and IHF beta of Ec, respectively . The promoter region of himD contains an IHF consensus sequence, as is the case for Ec himD.

Gene, 1995 Feb 27, 154(1), 15 - 21
ToxR (RegA) activates Escherichia coli RNA polymerase to initiate transcription of Pseudomonas aeruginosa toxA; Walker SL et al.; The Pseudomonas aeruginosa (Pa) structural gene (toxA), which encodes the exotoxin A protein has been shown to be regulated at the transcriptional level by a protein designated ToxR (also known as RegA) . We have previously reported that ToxR directly enhances toxA transcription in vitro; however, in the absence of ToxR, Pa RNA polymerase (RNAP) transcribes toxA with low efficiency . In the present study, we have examined the ability of ToxR to initiate toxA transcription using the heterologous Escherichia coli (Ec) RNAP and found that ToxR can function with Ec RNAP to efficiently transcribe toxA both in vitro and in vivo . Antibodies produced against the sigma 70 subunit of Ec RNAP inhibit ToxR-mediated enhancement of toxA transcription, suggesting that the RNAP holoenzyme (E sigma 70) is required for transcriptional activation of toxA . We further demonstrate that ToxR is required for open-complex formation at the toxA promoter . By selectively deleting toxA upstream sequences, we have localized at 214-bp region containing both the toxA promoter and a putative ToxR-binding site sufficient for ToxR-mediated transcription of toxA.

Mol Gen Genet, 1995 Feb 20, 246(4), 519 - 28
Identification of a DNA sequence motif required for expression of iron-regulated genes in pseudomonads; Rombel IT et al.; Many bacteria respond to a lack of iron in the environment by synthesizing siderophores, which act as iron-scavenging compounds . Fluorescent pseudomonads synthesize strain-specific but chemically related siderophores called pyoverdines or pseudobactins . We have investigated the mechanisms by which iron controls expression of genes involved in pyoverdine metabolism in Pseudomonas aeruginosa . Transcription of these genes is repressed by the presence of iron in the growth medium . Three promoters from these genes were cloned and the activities of the promoters were dependent on the amounts of iron in the growth media . Two of the promoters were sequenced and the transcriptional start site were identified by S1 nuclease analysis . Sequences similar to the consensus binding site for the Fur repressor protein, which controls expression of iron-repressible genes in several gram-negative species, were not present in the promoters, suggesting that they are unlikely to have a high affinity for Fur . However, comparison of the promoter sequences with those of iron-regulated genes from other Pseudomonas species and also the iron-regulated exotoxin gene of P . aeruginosa allowed identification of a shared sequence element, with the consensus sequence (G/C)CTAAAT-CCC, which is likely to act as a binding site for a transcriptional activator protein . Mutations in this sequence greatly reduced the activities of the promoters characterized here as well as those of other iron-regulated promoters . The requirement for this motif in the promoters of iron-regulated genes of different Pseudomonas species indicates that similar mechanisms are likely to be involved in controlling expression of a range of iron-regulated genes in pseudomonads.

J Chromatogr B Biomed Appl, 1995 Feb 17, 664(2), 458 - 62
Rapid identification of 4-hydroxy-2-alkylquinolines produced by Pseudomonas aeruginosa using gas chromatography-electron-capture mass spectrometry; Taylor GW et al.; The 4-hydroxy-2-alkylquinolines and their N-oxides are secondary metabolites produced by Pseudomonas aeruginosa which inhibit the growth of a number of Gram-positive organisms including Staphylococcus aureus . To facilitate the identification of these compounds in biological fluids, we have developed a rapid profiling system based on gas chromatography-electron-capture mass spectrometry of the O-bistrifluoromethylbenzoyl derivatives . Using the technique, over twenty hydroxyalkylquinolines have been identified from a culture obtained from a strain of P . aeruginosa obtained from a patient with severe bronchiectasis.

J Chromatogr A, 1995 Feb 17, 693(1), 7 - 13
High-performance liquid chromatographic determination of the rhamnolipids produced by Pseudomonas aeruginosa; Schenk T et al.; The bacterial biosurfactants 3-{3'-(L-rhamnopyranosyloxy)decanoyloxy}decanoic acid (RL-1) and 3-{3'-(2"-O-alpha-L-rhamnopyranosyl-alpha-L-rhamnopyranosyloxy) decanoyloxy}decanoic acid (RL-2) were isolated from Pseudomonas aeruginosa DSM 2659 cultures . An HPLC method was developed for the p-bromophenacyl esters of the rhamnolipids . Separation was obtained within 25 min on a RP C18 column using a linear gradient of water-acetonitrile (30:70 to 0:100) and UV detection (265 and 320 nm) . Linearity of response existed for 1.2-25 microg of the RL-1 and 1.8-25 microg of the RL-2-p-bromophenacyl ester (0.9-19.2 microg RL-1 and 1.3-18.0 microg RL-2) . The reproducibility of the entire analytical method (extraction, derivatisation) was tested.

Eur J Biochem, 1995 Feb 15, 228(1), 39 - 44
The binding of divalent cations to Escherichia coli alpha-haemolysin; Ostolaza H et al.; alpha-haemolysin, an extracellular protein toxin of Escherichia coli, is known to disrupt eukaryotic cell membranes . In spite of genetic evidence of Ca(2+)-binding motifs in its sequence, conflicting results are found in the literature on the requirement of divalent cations for the membranolytic activity of the toxin . Moreover, Ca(2+)-binding sites have not been characterized to date in the native protein . The results in this paper show that when Ca2+ levels are kept sufficiently low during bacterial growth and toxin purification, membrane lysis does not occur in the absence of added divalent cations . Ca2+ and, at higher concentrations, Sr2+ and Ba2+, support the lytic activity, but Mg2+, Mn2+, Zn2+ and Cd2+ appear to be inactive in this respect . Binding of metal ions can be followed by changes in the intrinsic fluorescence of alpha-haemolysin; ions supporting lytic activity produce changes in the intrinsic fluorescence that are not caused by the inactive ones . Scatchard analysis of 45Ca2+ binding reveals three equivalent, independent sites, with Kd approximately 0.11 mM . No 45Ca2+ binding is observed when the protein is incubated with Zn2+; conversely, incubation with Ca2+ prevents subsequent binding of 65Zn2+ . In the light of three-dimensional data available for a structurally related protein, alkaline protease of Pseudomonas aeruginosa {Baumann, U., Wu, S., Flaherty, K . M . & McKay, D . B . (1993) EMBO J . 12, 3357-3364} it is suggested that alpha-haemolysin may bind a larger number of Ca2+ than the three that are more easily exchangeable and are thus detected in the 45Ca(2+)-binding experiments . In addition, structural similarities and conservation of ion-binding motifs support the hypothesis that His 859 is involved in the mutually exclusive binding of Zn2+ and Ca2+.

J Inorg Biochem, 1995 Feb 15, 57(3), 169 - 81
Monomeric Pseudomonas aeruginosa nitrite reductase: preparation, characterization, and kinetic properties; Silvestrini MC et al.; Monomeric nitrite reductase in an active form has been prepared by controlled succinylation of the dimeric native enzyme of Pseudomonas aeruginosa and subsequent purification . The monomeric enzyme has an optical spectrum indistinguishable from that of the native enzyme . On the other hand, circular dichroic spectra in the heme and peptide absorption regions show differences with respect to the dimer that indicate that the chemical modification and/or the dissociation into monomers somewhat perturb the chromophores' environment and the secondary structure . The (negatively charged) monomer is unable to oxidize its physiological substrates, azurin and cytochrome c551 . This loss of activity is not due to monomerization, but is linked to the total net charge of the succinylated molecule, which interestingly enough acquires the ability to oxidize efficiently eukaryotic cytochrome c (which is not a substrate of the native dimeric enzyme) . Stopped-flow studies show that the reduced monomer reacts with oxygen with a kinetic pattern similar to that shown by the dimeric enzyme . However, a higher reaction rate in the bimolecular binding of oxygen and a much higher oxygen affinity than for the native enzyme are observed . The evidence reported in this paper indicates that the dimeric state of Pseudomonas nitrite reductase is not a prerequisite for the ferrocytochrome c-oxygen oxidoreductase activity of this enzyme.

Eur J Biochem, 1995 Feb 1, 227(3), 829 - 37
Determination of haem electronic structure in His-Met cytochromes c by 13C-NMR . The effect of the axial ligands; Turner DL; The assignment of 13C resonances of nuclei alpha to the haem in horse ferricytochrome c is completed and the Fermi contact shifts are evaluated at 30 degrees C and 50 degrees C using empirical magnetic susceptibility tensors to correct for dipolar interactions . The Fermi contact shifts are fitted to a model of molecular orbitals of eg symmetry, which are subject to a rhombic perturbation . A similar analysis is performed using published data for Pseudomonas aeruginosa cytochrome c551 . The relationship between the orientation of the effective g tensor and that of the rhombic perturbation in these proteins is shown to agree with theoretical predictions . A comparison between the orientation of the rhombic perturbations and the crystal structures of horse cytochrome c and P . aeruginosa cytochrome c551 reveals that the orientation of the histidine and methionine axial ligands dominates the rhombic perturbation and that the two ligands have approximately equal influence . The magnitude of the perturbation shows that the orientation of the axial ligands has little effect on the haem redox potential . However, the relationship that is established between the magnetic susceptibility tensor, the partially filled haem molecular orbitals, and the orientation of the haem ligands offers a new source of precise structural information.

Am J Physiol, 1995 Feb, 268(2 Pt 1), L181 - 6
A type I cell-specific protein is a biochemical marker of epithelial injury in a rat model of pneumonia; McElroy MC et al.; In this study we determined whether the alveolar fluid content of a specific epithelial type I cell protein, rTI40, can be used as a biochemical marker for lung injury . A model of alveolar epithelial injury was developed by instilling Pseudomonas aeruginosa bacteria (PA103) into the airspaces of anesthetized, ventilated rats . After 6 h, the alveolar fluid content of rTI40 from PA103-treated rats was increased over 80-fold in comparison to alveolar fluid from control rats (P < 0.05) . This increase in rTI40 correlated with both morphological evidence of injury to alveolar epithelial type I cells and increased permeability of the alveolar epithelium to protein tracers . In contrast, the lactate dehydrogenase activity of alveolar fluid from PA103-treated rats was elevated only threefold over control values at 6 h (P < 0.05) . In a second study using a less injurious strain of P . aeruginosa (PA103 exsA::omega), the alveolar fluid content of rTI40 was the same as control values . These findings indicate that the alveolar fluid content of a type I cell-specific protein can be used as a sensitive and specific biochemical marker of type I cell injury.

J Bacteriol, 1995 Feb, 177(4), 948 - 52
Isolation and characterization of chemotaxis mutants and genes of Pseudomonas aeruginosa; Masduki A et al.; Two chemotaxis-defective mutants of Pseudomonas aeruginosa, designated PC1 and PC2, were selected by the swarm plate method after N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis . These mutants were fully motile but incapable of swarming, suggesting that they had a defect in the intracellular signalling pathway . Computer-assisted capillary assays confirmed that they failed to show behavioral responses to chemical stimuli, including peptone, methyl thiocyanate, and phosphate . Two chemotaxis genes were cloned by phenotypic complementation of PC1 and PC2 . From nucleotide sequence analysis, one gene was found to encode a putative polypeptide that was homologous to the enteric CheZ protein, while the other gene was cheY, which had been previously reported (M . N . Starnbach and S . Lory, Mol . Microbiol . 6:459-469, 1992) . Deletion and complementation analysis showed that PC1 was a cheY mutant, whereas PC2 had a double mutation in the cheY and cheZ genes . A chromosomal cheZ mutant, constructed by inserting a kanamycin resistance gene cassette into the wild-type gene, changed its swimming direction much more frequently than did wild-type strain PAO1 . In contrast, cheY mutants were found to rarely reverse their swimming directions.

Ann Thorac Surg, 1995 Feb, 59(2), 408 - 11
Pneumonectomy for chronic infection: fraught with danger?
Reed CE.
Pneumonectomy for chronic lung infections has been avoided because of potential intraoperative and postoperative complications . A retrospective review of 13 cases requiring pneumonectomy for aspergillus (8), Mycobacterium tuberculosis (2), actinomycosis, Pseudomonas aeruginosa, and bronchiectasis revealed increased operating time, blood loss, and transfusion requirements . Operative records documented problems with dense adhesions, lack of an extrapleural plane, and distortion of hilar structures . Although mortality was acceptable (8%), early and late morbidity (total, 38%), especially bronchopleural fistula (23%), was significant . It is concluded that when justified, pneumonectomy for complete resection of chronic infection can be performed with acceptable risk . However, specific problems should be anticipated . This review has led to modifications in operative technique.

Chest, 1995 Feb, 107(2 Suppl), 61S - 64S
Aerosol delivery of antibiotics to the lower airways of patients with cystic fibrosis; Fiel SB; Aerosolized tobramycin is well tolerated in children and adults with cystic fibrosis (CF) . It represents a potential alternative to intravenous administration of antibiotics to control respiratory infection with Pseudomonas aeruginosa . Although its exact role in acute therapy for a CF exacerbation is not known, it has been shown to improve lung function and reduce the frequency of exacerbations in the chronic stable outpatient.

Infect Immun, 1995 Feb, 63(2), 640 - 6
Binding of the fibrillar CS3 adhesin of enterotoxigenic Escherichia coli to rabbit intestinal glycoproteins is competitively prevented by GalNAc beta 1-4Gal-containing glycoconjugates; Wenneras C et al.; We have attempted to characterize the binding specificity of the coli surface 3 (CS3) subcomponent of colonization factor antigen II of enterotoxigenic Escherichia coli, by means of an immunoblot method in which the binding of fimbriated bacteria to sodium dodecyl sulfate-polyacrylamide gel electrophoresis-separated rabbit intestinal cell membranes was evaluated . Isolated CS3 fibrillae as well as bacteria expressing CS3 on their surface bound to several intestinal cell membrane structures, i.e., structures present in the electrophoretic front and in the 30- to 35-kDa range and, most prominently, 120- to 140-kDa structures . Delipidization and protein digestion of the rabbit brush borders revealed that CS3 bound to structures of a proteinaceous nature . Sodium meta-periodate oxidation of the intestinal cell membranes abolished all their CS3 binding activity, indicating that CS3 bound to carbohydrate moieties of glycoproteins . The binding of CS3 to the separated intestinal proteins could also be inhibited by preincubation with the lectin derived from Maackia amurensis, indicating that CS3 bound to galactoproteins in the rabbit intestine . Inhibition experiments using equimolar amounts of various gangliosides demonstrated that GM1, asialo-GM1, and GM2 inhibited the binding of CS3 equally well, whereas GM3 was not as effective . These results suggested that the critical CS3 binding epitope consisted of the carbohydrate sequence GalNAc beta 1-4Gal . This was supported by electron microscopic experiments showing that this disaccharide, O linked to bovine serum albumin via a spacer, localized around CS3-positive bacteria but not at all around corresponding CS3-negative mutants . Furthermore, CS3-expressing bacteria recognized this neoglycoprotein when it was immobilized on nitrocellulose . The GalNAc beta 1-4Gal disaccharide has also been implicated as a binding structure for other pathogenic bacteria such as enteropathogenic E . coli and Pseudomonas aeruginosa.

Mol Microbiol, 1995 Feb, 15(4), 703 - 17
The Pseudomonas aeruginosa pilK gene encodes a chemotactic methyltransferase (CheR) homologue that is translationally regulated; Darzins A; A new locus, designated pilK, located immediately adjacent to the previously described Pseudomonas aeruginosa pilG-J gene cluster, has been identified . Sequence analysis of a 1.3 kb region revealed the presence of a single open reading frame of 291 amino acid residues (M(r) 33,338) that contained significant homology to the chemotactic methyltransferase proteins of Escherichia coli, Bacillus subtilis and the gliding bacterium Myxococcus xanthus . The 60 bp pilJ-pilK intergenic region was devoid of promoter consensus sequences, suggesting that pilJ and pilK are contained within the same transcriptional unit . The intergenic region did contain, however, a large, highly GC-rich, inverted repeat that prevented PilK production in expression studies . To investigate the regulatory role of these sequences, pilK-lacZ gene fusions, as well as derivatives containing sequence alterations in the potential stem-loop region, were constructed and analysed in E . coli and P . aeruginosa . Modification of the inverted repeat region in pilK-lacZ protein fusion constructs resulted in as much as a 24-fold increase in beta-galactosidase activity, whereas similar modifications in pilK-lacZ transcriptional fusions had only a marginal effect on beta-galactosidase levels . These results indicated that PilK production may be largely regulated at the level of translation . In stark contrast to pilG-J mutants, which are dramatically impaired in pilus production and/or function, a PAO1 pilK deletion mutant was indistinguishable from the wild type . In addition, complementation studies suggested that the PilK and E . coli CheR proteins are not functionally interchangeable.

Nippon Ronen Igakkai Zasshi, 1995 Feb, 32(2), 101 - 4
{An analysis of 41 elderly patients with urosepsis}; Kaneko H et al.; We performed a clinical evaluation in 41 patients with urosepsis at Tokyo Metropolitan Geriatric Hospital from July 1992 through March 1993 . The most common organism isolated from the patients was Escherichia coli (46.3%), followed by Pseudomonas aeruginosa (9.8%), Methicillin-resistant staphylococcus aureus (7.3%), and mycetes (7.3%) . The most frequent underlying disease was cerebrovascular disease (34.1%) and malignancies were observed 29.2% of all cases . Twenty-six patients (63.4%) had indwelling urethral catheters . Indwelling catheters were suspected to be related to the onset of urosepsis in 16 cases . Total mortality of urosepsis was 4.9% (2/41) in this study . We speculate that the main cause of urosepsis is a long-term use of urethral catheterization, especially in elderly patients with severe complications who are vulnerable to infections . It is important to assess and correct the conditions of dysuria of individual patients before placing indwelling urethral catheters.

Glycobiology, 1995 Feb, 5(1), 39 - 44
A beta-linked mannan inhibits adherence of Pseudomonas aeruginosa to human lung epithelial cells; Azghani AO et al.; Adherence through carbohydrate-binding adhesins is an early step in colonization of the lung by gram-negative organisms, and because published data indicate that binding involves mannose groups, we tested the ability of a beta-linked acetyl-mannan (acemannan) to inhibit adherence of Pseudomonas aeruginosa to cultures of human lung epithelial cells . Adherence of radiolabelled P.aeruginosa to A549 cells (a type II-like pneumocyte line) increased linearly with the duration of the incubation . Acemannan inhibited adherence of bacteria, and the extent of inhibition was related to the concentration of the mannan . Inhibition required continued contact between acemannan and the target epithelial cells; cells washed free of acemannan no longer discouraged bacterial binding . Comparison of binding between seven different strains of P.aeruginosa indicated that fewer mucoid than non-mucoid bacteria adhered, but binding of either phenotype was inhibited by acemannan . Mannose, methyl alpha-D-mannopyranoside, methyl beta-D-mannopyranoside and dextran did not affect adherence of any of the non-mucoid strains . Mannose inhibited adherence by one mucoid strain, but not the other, indicating differences between strains of the same phenotype . Since prior treatment of epithelial cells with concanavalin A did not affect acemannan-induced inhibition of bacterial adherence, we concluded that the inhibitory effect of acemannan probably does not involve mannose-containing receptors.

Farmaco, 1995 Feb, 50(2), 143 - 8
Synthesis and antimicrobial activity of 1-(p-substituted)phenyl-3-(p-alkoxy)phenyl-4-phenyl-azetidin-2-ones; Diurno MV et al.; A series of 1-(p-substituted)phenyl-3-(p-alkoxy)phenyl-4-phenyl-azetidin-2-one s 1a-20a, was synthesized and characterized . Their antimicrobial activity, against Gram+ and Gram- bacteria and Fungi, was tested . The compounds 8a, 13a, 14a, 18a and 6a, 9a, 10a showed remarkable activity respectively against Pseudomonas aeruginosa and against Fungi.

Biosci Biotechnol Biochem, 1995 Feb, 59(2), 314 - 5
A catechol 2,3-dioxygenase gene as a reporter; Shindo T et al.; A catechol 2,3-dioxygenase (C23O) gene of Pseudomonas aeruginosa was expressed under the Simian virus 40 or Rous sarcoma virus promoter in mammalian cells; it was found that the gene could be used as a reporter for the study of gene expression . The C23O gene was a more sensitive reporter than the generally used beta-galactosidase gene.

J Hosp Infect, 1995 Feb, 29(2), 143 - 51
Evaluation of the Steris System 1 Peracetic Acid Endoscope Processor; Bradley CR et al.; An automated endoscope sterilizing machine, the Steris System 1 Processor, was tested for bactericidal and sporicidal efficacy . The disinfectant, peracetic acid, was diluted to 0.2% within an enclosed system . The exposure time to the disinfectant was 12 min and the overall cycle time ranged from 25-38 min, mean 29 min . Preliminary suspension tests, with and without yeast or serum, showed a log10 reduction of > 5 with Pseudomonas aeruginosa, Staphylococcus aureus and Bacillus subtilis in 5 min with 0.2% peracetic acid . After a routine cycle in the machine, endoscopes contaminated with the same organisms showed no growth . Two of 24 spore strips, containing 10(6) B . subtilis showed a small number of survivors (less than 10 per strip) . No significant damage to the endoscope was observed although the number of cycles tested was small (i.e . 31) . The advantage of the system is that staff are not directly exposed to the agent, but the costs per cycle are higher than glutaraldehyde, since peracetic acid is not renewed . Unlike other automated processors the Steris machine has no cleaning cycle.

J Antimicrob Chemother, 1995 Feb, 35(2), 339 - 43
The in-vitro effect of temperature on MICs, bactericidal rates and postantibiotic effects in Staphylococcus aureus, Klebsiella pneumoniae and Pseudomonas aeruginosa; Magnusson V et al.; Varying temperatures (35.5 degrees C, 38.5 degrees C, 41 degrees C) only minimally affected growth rates in vitro of Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa, as well as bactericidal rates and postantibiotic effects of several antibiotics . However, MICs were reduced at least four-fold by increasing temperature in 25% of the drug-organism combinations tested.

J Antimicrob Chemother, 1995 Feb, 35(2), 295 - 304
Antibodies against Pseudomonas aeruginosa chromosomal beta-lactamase inpatients with cystic fibrosis are markers of the development of resistance of P . aeruginosa to beta-lactams; Ciofu O et al.; Chromosomal beta-lactamase production is considered to be the most important resistance mechanism of Pseudomonas aeruginosa against beta-lactams . Recently we have detected serum and sputum antibodies against P . aeruginosa chromosomal beta-lactamase (a beta ab), using immunoblotting techniques . In this study we have developed an enzyme-linked-immunosorbent assay to measure serum a beta ab response in 124 cystic fibrosis patients in a cross-sectional study and in 54 cystic fibrosis patients in a longitudinal study . The a beta ab response occurred after a median of 3 years following onset of chronic infection and was significantly higher (P < 0.0002) in patients chronically infected with resistant strains than in those from whom resistant strains were occasionally isolated . The a beta ab levels correlated (r = 0.51, P = 0.0001) with the number of beta-lactam courses . A 14 fold increase in a beta ab levels occurred during the 14 year period covered by the longitudinal study . The results of this study show that a beta ab to P . aeruginosa is a specific marker for resistance development of P . aeruginosa to beta-lactams.

J Antimicrob Chemother, 1995 Feb, 35(2), 281 - 94
Transferable production of PER-1 beta-lactamase in Pseudomonas aeruginosa; Danel F et al.; PER-1, an extended-spectrum class A beta-lactamase, has been described only from Pseudomonas aeruginosa RNL-1, which was obtained in France in 1991 . During studies on ceftazidime-resistant P . aeruginosa collected from December 1991 to July 1993 in Ankara, Turkey, we found 14 further isolates with PER-1 enzyme, as recognised by isoelectric point (pI 5.4), hydrolytic activity, gene hybridization and DNA sequence . Five of these isolates also had OXA-10 (PSE-2)-related enzymes and one hyper-produced ampC beta-lactamase, whereas eight had PER-1 enzyme alone . The last group, from four wards, appeared to be identical, giving the same DNA restriction patterns and carrying the PER-1 gene on an 8.5 kb HincII fragment . Two more producers were related but the other four were unique . In several representatives of the group of eight replicates, the PER-1 gene was shown to be encoded on a plasmid, larger than 154 kb in size, which transferred to P . aeruginosa PU21 . A further isolate had the gene on an even larger conjugative plasmid . By contrast, the PER-1 gene reportedly was chromosomally-inserted in strain RNL-1 . The PER-1 producers and their transconjugants were highly resistant to ceftazidime and aztreonam (MIC > or = 128 mg/L) but not to carbapenems or latamoxet . Piperacillin insusceptibility was marginal (MIC 8 mg/L) . Clavulanate 4 mg/L, but not tazobactam 4 mg/L, reversed resistance to ceftazidime and carbenicillin . Purification of the enzyme to homogeneity was achieved by three ion exchanges and one gel filtration . We found much lower Vmax rates for aminothiazolyl cephalosporins than reported previously for PER-1 enzyme . This reflected the present assays being in 0.1 M phosphate buffer pH 7.0, whereas the previous were pH-stat-regulated; concentrated phosphate reduced enzyme activity against ceftazidime, but not against cephaloridine.

Pediatr Infect Dis J, 1995 Feb, 14(2), 140 - 3
The changing epidemiology of bacteremia in neutropenic children with cancer; Aquino VM et al.; Gram-positive bacteria have been the predominant organisms causing bacteremia in febrile neutropenic cancer patients during the past decade . Recently we have noted an increase in Gram-negative bacteremia in children and adolescents with cancer . Therefore we retrospectively reviewed 153 episodes of bacteremia during a 6-year period to investigate changes in the etiology of bacteremia in pediatric oncology patients . In the early 3-year period (January, 1988, to December, 1990) Gram-positive organisms comprised 73 (74%) of the 99 isolates, and Staphylococcus epidermidis was the most common isolate . In the later 3-year period (January, 1991, to December, 1993) Gram-negative organisms were seen with greater frequency and represented 50% of isolates (P = 0.004) . Pseudomonas aeruginosa was the most commonly isolated organism during this period (22% of all isolates) . We speculate that the recent utilization of more intensive chemotherapy regimens has caused an alteration in the epidemiology of bacteremia in children and adolescents with cancer which could influence the initial empiric antibiotic regimens and the outcome of such infections.

Kansenshogaku Zasshi, 1995 Feb, 69(2), 199 - 201
{A report to 2 cases where Stenotrophomonas maltophilia with a mucoid phenotype was isolated from the sputum}; Irifune K et al.; Mucoid Stenotrophomonas maltophilia was isolated from the sputum of 2 women . Case 1 which we reported recently was primary pneumonia caused by S . maltophilia . However, in Case 2, mucoid S . maltophilia represented part of the transient flora . Interestingly, colonization of mucoid Pseudomonas aeruginosa on respiratory tract occurred in both cases after mucoid S . maltophilia was isolated from their sputum.

Clin Infect Dis, 1995 Feb, 20(2), 302 - 8
Pseudomonas aeruginosa infections of intact skin; Agger WA et al.; Pseudomonas aeruginosa infections of healthy skin are uncommon . We report four cases of P . aeruginosa infections of intact skin . These cases illustrate the clinical spectrum of these cutaneous infections: localized, mild epidermal infections (the green nail syndrome and webbed space infections), moderately serious infections (cutaneous folliculitis and otitis externa), and, in immunocompromised patients, extremely serious infections (malignant otitis externa, perirectal infection, and ecthyma gangrenosum).

Intern Med, 1995 Feb, 34(2), 100 - 3
Jaundice associated with Pseudomonas aeruginosa bacteremia complicating acute leukemia; Funada H et al.; Two adult patients with acute leukemia developed marked jaundice during Pseudomonas aeruginosa bacteremia . The progressive increase in serum conjugated bilirubin levels was disproportionate to the gradual decrease in serum alkaline phosphatase and transaminase activity to or below low normal . The production of coagulation factors decreased . Autopsy revealed periportal cholestasis with minimal liver-cell damage . These findings suggested decreased metabolic activity of liver cells associated with bacteremia, probably leading to impaired bilirubin excretion . Both patients died despite appropriate antibiotic therapy . Isolated hyperbilirubinemia, thus, seemed to be an ominous prognostic sign in severe infection.

Antimicrob Agents Chemother, 1995 Feb, 39(2), 496 - 9
Comparative efficacies of ciprofloxacin and pefloxacin alone or in combination with fosfomycin in experimental endocarditis induced by multidrug-susceptible and -resistant Pseudomonas aeruginosa; Xiong YQ et al.; The in vivo efficacy of ciprofloxacin or pefloxacin alone or in combination with fosfomycin was evaluated in experimental aortic valve endocarditis induced in 133 rabbits by a multidrug-susceptible or multidrug-resistant strain of Pseudomonas aeruginosa . Therapy was initiated early (12 h after infection), when bacterial counts in aortic valve vegetations were relatively low, or late (48 h after infection), when vegetations contained a larger inoculum . Antibodies were administered as a continuous 24-h intravenous infusion . Mean steady-state levels of ciprofloxacin (64 mg/kg), pefloxacin (64 mg/kg), and fosfomycin (300 mg/kg) in serum were 2.5, 4.2, and 63.9 mg/liter, respectively . For the multidrug-susceptible strain, all regimens except pefloxacin alone significantly reduced the number of CFU per gram of vegetation versus controls, whether treatment was performed early or late . For the multidrug-resistant strain, none of the regimens showed differences from untreated controls, except ciprofloxacin-fosfomycin, which significantly reduced bacterial counts in vegetations compared with controls when therapy was begun early (4.1 +/- 1.1 log10 CFU/g of vegetation; P < 0.001 versus the control) . These data suggest that combination of fosfomycin with ciprofloxacin or pefloxacin is more effective than ciprofloxacin or pefloxacin alone for the therapy of severe infections caused by multidrug-susceptible P . aeruginosa.

Antimicrob Agents Chemother, 1995 Feb, 39(2), 489 - 95
Development of multiple-antibiotic-resistant (Mar) mutants of Pseudomonas aeruginosa after serial exposure to fluoroquinolones; Zhanel GG et al.; Laboratory-derived fluoroquinolone-resistant mutants were created by serially passaging wild-type Pseudomonas aeruginosa on fluoroquinolone-containing agar to obtain high-level fluoroquinolone resistance (e.g., ciprofloxacin MIC of 1,024 micrograms/ml) . With increases of 4- to 32-fold in MICs of fluoroquinolones, these organisms demonstrated (relative to wild-type) normal morphology, resistance to fluoroquinolones only, no change in fluoroquinolone uptake, and no change in lipopolysaccharide profiles or outer membrane protein profiles . Complementation with wild-type Escherichia coli gyrA restored fluoroquinolone susceptibility, suggesting that these were gyrA mutants . After 4- to 32-fold increases in fluoroquinolone MICs (with continued passage on fluoroquinolone-containing agar) isolates demonstrated altered morphology, a multiple-antibiotic-resistant (Mar) phenotype (including cross-resistance to beta-lactams, chloramphenicol, and tetracycline), reduced fluoroquinolone uptake and altered outer membrane proteins (reductions in the 25- and 38-kDa bands as well as several bands in the 43- to 66-kDa region) . Complementation with wild-type E . coli gyrA partially reduced the level of fluoroquinolone resistance by approximately 8- to 32-fold, suggesting that these mutants displayed both gyrA and non-gyrA mutations.

Antimicrob Agents Chemother, 1995 Feb, 39(2), 394 - 9
Structure-activity relationships of carbapenems that determine their dependence on porin protein D2 for activity against Pseudomonas aeruginosa; Fung-Tomc JC et al.; A number of carbapenem derivatives were examined to determine the structure-activity relationships required for dependence on porin protein D2 for activity against Pseudomonas aeruginosa . As suggested by J . Trias and H . Nikaido (Antimicrob . Agents Chemother . 34:52-57, 1990), carbapenem derivatives, such as imipenem and meropenem, containing a sole basic group at position 2 of the molecule utilize the D2 channel for permeation through the outer membrane of pseudomonads; they are more active against D2-sufficient strains of P . aeruginosa . Our results indicated that carbapenems with a basic group at position 1 or 6 of the molecule did not depend on the D2 channel for activity; i.e . they were equally active against D2-sufficient and D2-deficient pseudomonal strains . However, addition of a basic group at position 1 or 6 of a carbapenem derivative already containing a basic group at position 2 resulted in its lack of dependency on the D2 pathway . Comparison between meropenem and its 1-guanidinoethyl derivative, BMY 45047, indicated that they differed in their dependence on D2; while meropenem required the D2 channel for uptake, BMY 45047 activity was independent of D2 . Meropenem and BMY 45047 had similar affinities for the penicillin-binding proteins of P . aeruginosa . However, BMY 45047 and meropenem differed in the morphological changes that they induced in pseudomonal cells . While meropenem induced filamentation, BMY 45047 induced filaments only in BMS-181139-resistant mutants and not in imipenem-resistant mutants or in carbapenem-susceptible P . aeruginosa strains . These results suggested that in Mueller-Hinton medium the uptake of BMY 45047 through the non-D2 pathway is more rapid than that of meropenem through the D2 porin . In summary, the presence of a basic group at position 2 of a carbapenem is important for its preferential uptake by the D2 channel . However the addition of a basic group at position 1 or 6 of a carbapenem already containing a basic group at position 2 dissociates its necessity for porin protein D2 for activity.

Antimicrob Agents Chemother, 1995 Feb, 39(2), 386 - 93
Activity of carbapenem BMS-181139 against Pseudomonas aeruginosa is not dependent on porin protein D2; Fung-Tomc JC et al.; The broad antipseudomonal spectrum of the carbapenem BMS-181139 includes clinical strains and laboratory mutants of Pseudomonas aeruginosa that are resistant to imipenem . Unlike other known carbapenems (meropenem, panipenem, biapenem, and BO-2727), which have reduced activity against imipenem-resistant strains of P . aeruginosa, BMS-181139 was equally active against imipenem-susceptible (D2-sufficient) and imipenem-resistant (D2-deficient) strains . Conversely, imipenem and meropenem activities were the same against the susceptible parental strains and their BMS-181139-resistant mutants . Whereas basic amino acids antagonized the antipseudomonal activities of imipenem and meropenem, they had no effect on the activity of BMS-181139 . These results suggest that the uptake of BMS-181139 into pseudomonal cells occurs by a non-D2 pathway . Compared with imipenem and meropenem, BMS-181139 may have a slightly higher affinity for penicillin-binding protein 2 (PBP-2) of P . aeruginosa . The rates of resistance development to imipenem, meropenem, and BMS-181139 in P . aeruginosa strains were similar; resistance occurred at frequencies of approximately 10(-7) to 10(-8) . Resistance to BMS-181139 in P . aeruginosa is presumed to be caused by its diminished permeability since no change in their penicillin-binding protein affinities or beta-lactamase levels could be detected . In summary, BMS-181139 is a new carbapenem which differs from other known carbapenems in its lack of cross-resistance with imipenem . This difference could be explained by the permeation of BMS-181139 through a non-D2 channel, compared to the preferential uptake of other carbapenems by the D2 porin.

Eur J Pediatr, 1995 Feb, 154(2), 157 - 60
Specific decrease of anti-pseudomonal IgA after anti-pseudomonal therapy in cystic fibrosis; de Boeck K et al.; In patients with cystic fibrosis (CF) and chronic colonisation with Pseudomonas aeruginosa, specific anti-pseudomonal IgG and IgA, as well as serum immunoreactive protein C, WBC and differential count, ESR, pulmonary function and chest radiograph score were determined before and after a 2 week intravenous course of anti-pseudomonal antibiotics in 32 cases of acute exacerbation of pulmonary infection . Specific anti-pseudomonal IgA but not specific anti-pseudomonal IgG decreased significantly after treatment . Log of anti-pseudomonal IgA but not log anti-pseudomonal IgG correlated well with disease severity as assessed by the Brasfield chest radiograph score (r 0.57), forced expiratory volume in 1 s (r 0.6) as well as C-reactive protein (r 0.62) . CONCLUSION: Specific anti-pseudomonal IgA may be a better parameter than specific IgG in the follow up of lung infection in patients with CF, probably because it more closely reflects ongoing endobronchial infection, the major pathology in CF lungs.

Gesundheitswesen, 1995 Feb, 57(2), 97 - 100
{Microbial contamination of immersed massage devices}; Bethe M et al.; Investigations in the District of Schaumburg . On the occasion of the inspection of a hospital we became aware of the problem of contamination of underwater-massage tubs with microorganisms . Hence, investigations according to the German DIN standard 19,643 were performed on 41 tubs . Very often we found a high count of microorganisms . If Pseudomonas aeruginosa was detected, the use of the respective tub was prohibited because of the risk of infection (about 70% of all tubs) . The reasons are the design of the pumps and different procedures of cleaning and disinfection . During the investigations the suppliers of the apparatus were queried . In some cases, technical improvements have already been effected . Reliable standard disinfection procedures must be developed.

Microbiology, 1995 Feb, 141 ( Pt 2), 431 - 9
A direct sulfhydrylation pathway is used for methionine biosynthesis in Pseudomonas aeruginosa; Foglino M et al.; The relationship between genes and enzymes in the methionine biosynthetic pathway has been studied in Pseudomonas aeruginosa . The first step is catalysed by an O-succinylhomoserine synthase, the product of the metA gene mapped at 20 min on the chromosome . The second step is achieved by direct sulfhydrylation, involving the enzyme encoded by a metZ gene that we have identified and sequenced, located at 40 min . Thus Pseudomonas appears to be the only organism so far described that uses O-succinylhomoserine as substrate for a direct sulfhydrylation . As in yeast, the two transsulfuration pathways between cysteine and homocysteine, with cystathionine as an intermediate, probably exist in parallel in this organism.

Environ Health Perspect, 1995 Feb, 103 Suppl 1, 59 - 62
Biosurfactant-facilitated remediation of metal-contaminated soils; Miller RM; Bioremediation of metal-contaminated wastestreams has been successfully demonstrated . Normally, whole cells or microbial exopolymers are used to concentrate and/or precipitate metals in the wastestream to aid in metal removal . Analogous remediation of metal-contaminated soils is more complex because microbial cells or large exopolymers do not move freely through the soil . The use of microbially produced surfactants (biosurfactants) is an alternative with potential for remediation of metal-contaminated soils . The distinct advantage of biosurfactants over whole cells or exopolymers is their small size, generally biosurfactant molecular weights are less than 1500 . A second advantage is that biosurfactants have a wide variety of chemical structures that may show different metal selectivities and thus, metal removal efficiencies . A review of the literature shows that complexation capacities of several bacterial exopolymers was similar to the complexation capacity of a rhamnolipid biosurfactant produced by Pseudomonas aeruginosa ATCC 9027.

Int J Exp Pathol, 1995 Feb, 76(1), 21 - 8
Role of alpha-2-macroglobulin and bacterial elastase in guinea-pig pseudomonal septic shock; Khan MM et al.; An essential role of alpha-2-macroglobulin (alpha 2M) was revealed in the prevention of septic shock induced in guinea-pigs by an elastase producing strain (IFO-3455) of Pseudomonas aeruginosa . When bacterial peritonitis was induced by inoculating fibrin-thrombin clot containing viable bacteria at a dose of 10(9) c.f.u./kg body weight, the guinea-pigs (n = 6) died within 7-8 hours due to septic shock . Prior to the shock, consumption of two-thirds of the circulating alpha 2M was observed . When circulating alpha 2M was depleted 4 hours after the bacterial inoculation, the guinea-pigs immediately developed shock and died within one hour . This shock was prevented either with a specific elastase inhibitor, HONHCOCH(CH2C6H5)CO-Ala-Gly-NH2, zincov (6 microM), or with human alpha 2M . Simultaneous depletion of circulating Hageman factor also prevented shock in the alpha 2M-depleted animals . These results indicate that septic shock was induced through activation of the Hageman factor dependent system by the bacteria-produced elastase which survived alpha 2M in the circulation.

J Pediatr, 1995 Feb, 126(2), 230 - 3
Binding of Pseudomonas aeruginosa to respiratory epithelial cells from patients with various mutations in the cystic fibrosis transmembrane regulator; Zar H et al.; OBJECTIVE: To determine whether there is an association between mutations of the cystic fibrosis transmembrane regulator (CFTR) and the predilection of patients with cystic fibrosis (CF) for Pseudomonas aeruginosa infection . METHOD: We quantified the adherence of P . aeruginosa PA01, labeled with sulfur 35-methionine, to epithelial monolayers derived from nasal scrapings of patients with specific CFTR mutations, and of carriers and normal subjects . RESULTS: Adherence of P . aeruginosa to epithelial cells from patients with CF was significantly greater than to cells from either carriers (t = 2.94; p = 0.009) or normal subjects (t = 3.32; p = 0.004) . Adherence to epithelial cells from patients with CF who were homozygous for the delta F508 mutation ranged from 12% to 35% (mean, 23.7%) of the added inoculum, which was significantly greater than the binding to cells from patients with other mutations, which ranged from 3% to 18% (mean, 9.4%; t = 3.71; p = 0.002), from heterozygote carriers (3% to 11%; mean, 7.9%; t = 4.87; p = 0.002), or from normal subjects (2% to 10%: mean, 7.0%; t = 5.21; p = 0.002) . CONCLUSION: Adherence to P . aeruginosa can be correlated with homozygosity for the delta 508 mutation; CFTR dysfunction may be one of the factors involved in the pathogenesis of pulmonary infection in CF.

FEMS Microbiol Lett, 1995 Jan 15, 125(2-3), 265 - 73
Comparison of type IV-pilin genes of Pseudomonas aeruginosa of various habitats has uncovered a novel unusual sequence; Spangenberg C et al.; All known pilin sequences in Pseudomonas aeruginosa were amplified by a set of consensus primers located in the 5"-conserved region of pilA and the threonine-specific t-RNA following pilA . This also enabled the discovery of a novel pilin gene in a strain pair of clonal variants, which differs from known pilin genes in its increased GC-content . The mature protein has 173 amino acids making it the longest pilin known to date in P . aeruginosa . Different inserted sequences detected between the 3"-end of the pilin gene and the t-RNA in this strain and in strains with group I pilin genes seemed to be specific for each pilin group indicating a horizontal cotransfer of sequences.

J Biol Chem, 1995 Jan 13, 270(2), 679 - 84
Active site mutations of Pseudomonas aeruginosa exotoxin A . Analysis of the His440 residue; Han XY et al.; Pseudomonas aeruginosa exotoxin A (ETA) is a member of the family of bacterial ADP-ribosylating toxins which use NAD+ as the ADP-ribose donor . By analogy to diphtheria and pertussis toxins, the His440 residue of ETA has been proposed to be one of the critical residues within the active site of the toxin . In this study the role of the His440 residue was explored through site-directed mutagenesis which resulted in the production of ETA proteins containing Ala, Asn, and Phe substitutions at the 440 position . The His440-substituted ETA proteins were purified and analyzed . All substitutions at the 440 site displayed severely reduced ADP-ribosylation activity (> 1000-fold) . However, NAD glycohydrolase activity remained intact and in the case of ETAH440N actually increased 10-fold . NAD+ binding is not affected by substitutions at the 440 site as indicated by similar Km values for the ETA variants tested . Conformational integrity of the mutant toxins appears to be largely unaffected as assessed by analysis with a conformation-sensitive monoclonal antibody as well as sensitivity to proteinase digestion . In view of the location of His440 residue within or close to the proposed NAD(+)-binding site, these results suggest that His440 may be a catalytic residue involved in the transfer of the ADP-ribose moiety to the EF-2 substrate.

Virology, 1995 Jan 10, 206(1), 611 - 25
Nucleotide sequence of a single-stranded RNA phage from Pseudomonas aeruginosa: kinship to coliphages and conservation of regulatory RNA structures; Olsthoorn RC et al.; We report the complete nucleotide sequence of the single-stranded RNA phage PP7 from Pseudomonas aeruginosa . There are three open reading frames which code for apparent protein homologues of the single-stranded RNA coliphages, i.e., maturation protein, coat protein, and replicase . A fourth overlapping reading frame exists that probably encodes a lysis protein, similar to what has been found in the group A coliphages such as MS2 . The genetic map of PP7 is colinear with group A coliphages and we accordingly classify the phage as a levivirus . There is, generally speaking, no significant nucleotide sequence identity between PP7 and the coliphages except for a few regions where homologous parts of proteins are encoded, most notable in the replicase gene . In these regions the nucleotide sequence similarity between PP7 and MS2 is no greater than between PP7 and the group B coliphages such as Q beta . Surprisingly, Q beta and MS2 are no closer to each other than they are to PP7 . Several regulatory RNA secondary structure features that are present in the coliphages were identified also in PP7 RNA although the sequences involved cannot be aligned . Among these are the coat protein binding helix at the start of the replicase gene, structures at the 5' and 3' terminus of the RNA, a replicase binding site, and the structure of the coat protein cistron start . Some of these features resemble MS2 type coliphages but others the Q beta type . These findings suggest that PP7 is related to the coliphages but branched off before the coliphages diverged into separate groups.

Biochemistry, 1995 Jan 10, 34(1), 257 - 63
Azotobacter vinelandii citrate synthase; Rault-Leonardon M et al.; We have purified the citrate synthase from Azotobacter vinelandii and have determined that the size of the subunit is 48,000 Da and the structure of the holoenzyme is a hexamer . This contrasts with earlier estimates that indicate a 58,000 Da subunit and a tetrameric structure . In addition, the enzyme is allosteric with a Hill coefficient of 1.5 and is inhibited by NADH . The Hill coefficient is changed to about 1 by high ionic strength and AMP . The enzyme is thus similar to the citrate synthases of many other Gram-negative, facultative, anaerobic organisms . In addition, the amino acid sequence of about 100 residues has been determined and found to be highly similar to the sequence of Pseudomonas aeruginosa citrate synthase.

Mol Gen Genet, 1995 Jan 6, 246(1), 72 - 9
Site-specific integration of the phage phi CTX genome into the Pseudomonas aeruginosa chromosome: characterization of the functional integrase gene located close to and upstream of attP; Wang Z et al.; The site-specific integration of the phage phi CTX genome, which carries the gene for a pore-forming cytotoxin, into the Pseudomonas aeruginosa chromosome was analysed . The 1,167 bp integrase gene, int, located immediately upstream of the attachment site, attP, was characterized using plasmid constructs, harbouring the integration functions, and serving as an integration probe in both P . aeruginosa and Escherichia coli . The attP plasmids p1000/p400 in the presence of the int plasmid pIBH and attP-int plasmids pINT/pINTS can be stably integrated into the P . aeruginosa chromosome . Successful recombination between the attP plasmid p1000 and the attB plasmid p5.1, in the presence of the int plasmid pIBH in E . coli HB101 showed that the int gene is active in trans in E . coli . The int gene product was detected as a 43 kDa protein in E . coli maxicells harbouring pINT . Proposed integration arm regions downstream of attP are not necessary for the integration process . pINT and phage phi CTX could be integrated together into P . aeruginosa chromosomal DNA, yielding double integrates.

Ophthalmic Res, 1995, 27(6), 322 - 9
Bacterial keratitis therapy in guinea pigs with lomefloxacin by initially high-followed by low-dosage regimen; Malet F et al.; Lomefloxacin is known to have an excellent corneal penetration . The therapeutic effect of two specific treatment regimens with lomefloxacin 0.3% eye drops was studied in a Pseudomonas aeruginosa-induced keratitis model in guinea pigs . An initial loading dose of 1 drop every 5 min for 5 times was used in all groups . Its purpose was to obtain high bactericidal corneal levels in the two actively treated groups . The follow-up treatment for 7 days was by a twice daily regimen in one group and 3 times daily in the other treated group . There were two control groups - one with no treatment and the other with vehicle treatment (initial loading dose followed by twice daily regimen) . Clinical signs of cornea, conjunctiva and eye adnexae improved significantly within 2 days in both lomefloxacin regimens, whereas clinical signs deteriorated with vehicle treatment . Fifty percent of the animals treated with lomefloxacin showed no colonies in the swab culture after 2 days of therapy, while all vehicle-treated animals were positive even at day 6, with 5 out of 9 animals continuing to be positive at day 8 . Pseudomonas in the range of 100-2,300 colonies was isolated from the grounded and cultured cornea at the end of the study in 4 out of 9 vehicle-treated animals but in none of the lomefloxacin-treated guinea pigs . The biggest difference in the degree of secondary inflammation between lomefloxacin and vehicle-treated groups was observed in the cornea which was the target tissue of infection . An unexpected observation was the lower degree of corneal inflammation in the twice daily treated animals when compared to the 3 times daily treatment group . This finding may be due to the somewhat lower initial degree of corneal inflammation in this group and suggests that the course of corneal recovery is predominately dependent on the initial degree of infection with both dosage regimens of lomefloxacin capable of eradicating corneal organisms.

Folia Microbiol (Praha), 1995, 40(3), 283 - 7
Antimicrobial efficacy of quaternary bisammonium salts and the effect of their sub-MICs on Pseudomonas aeruginosa virulence factors; Hostacka A et al.; Antipseudomonadal activity of homologous series of six quaternary bisammonium salts (QBAS) (4,7-dioxo-3,8-dioxadekan-1,1-{bis(alkyldimethyldiammonium dibromide)} as well as the effect of their subinhibitory concentrations (sub-MICs) on Pseudomonas aeruginosa virulence factors was studied . Antibacterial activity of QBAS increased up to a certain length of the chain and then decreased with further elongation . All the tested sub-MICs of QBAS caused a significant suppression of phospholipase C activity (to 0-41%) . Elastase and proteinase activity were less efficiently reduced . A more effective decrease of these activities was only found after treatment with one-fourth of the MICs of the tested substances . QBAS caused only an erratic decrease of alginate production.

Acta Microbiol Pol, 1995, 44(2), 111 - 7
Identification of Pseudomonas aeruginosa on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms; Kur J et al.; In the present study, 40 clinical strains of Pseudomonas aeruginosa were investigated by using the polymerase chain reaction (PCR) . We used primers to amplify spacer regions between the 16S and 23S genes in the prokaryotic rRNA genetic loci . When the spacer amplification products were resolved by electrophoresis, the resulting patterns were characteristic for all tested strains . Only one specific PCR fragment of 580 bp was formed . This product was digested with HaeIII, HinfI and AluI restriction endonucleases . Restriction fragments produced by all three restriction endonucleases were characteristic and have the same sizes for all strains tested . The amplification product contains a conserved, internal single HaeIII restriction site . On the basis of our results, PCR amplifications of the 16S-23S spacer region for Pseudomonas aeruginosa and subsequent RFPL analysis show significant promise as a tool for the simple identification of this bacteria.

Eye, 1995, 9 ( Pt 6), 679 - 83
The use of epidemiological techniques to assess risk: the epidemiology of microbial keratitis; Dart JK; The role of the three principal epidemiological study designs-- descriptive, cohort and case-control studies--for the evaluation of risk, is illustrated by describing their use for the investigation of microbial keratitis . Descriptive studies have identified potential risk factors and causes of microbial keratitis by both case reports and case series; trauma, ocular surface diseases and, latterly, contact lenses have been identified as potential risk factors . These studies have also shown that Pseudomonas aeruginosa and Acanthamoeba are particularly associated with contact lens wear . However, these studies are limited because they cannot be used to quantify risk . Cohort studies, in which the number of cases of a disease developing in a defined and initially unaffected population are identified, are usually inappropriate for assessing rare conditions because the size of the study often has to be too large, and follow-up too long, to generate sufficient numbers of cases . Some of the disadvantages of this study type can be overcome by sampling techniques and have been successfully carried out to obtain incidence figures for microbial keratitis . Case-control studies use multivariable analyses to examine the risk of microbial keratitis associated with various factors . This is an economical study design for investigating rare diseases because a group of subjects with the disease is compared with a control group from the same population who are unaffected . The selection of an appropriate control group is a difficult problem in epidemiology but this study design has been crucial in identifying risk factors and potential causes of microbial keratitis.

Scand J Infect Dis, 1995, 27(6), 619 - 22
Pneumonia in mechanically ventilated children; Brook I; The quantitative aerobic and anaerobic microbiology of bronchial aspirates, obtained using protective brush catheters, from 10 children with ventilator-associated pneumonia, is presented . Aerobic or faculative organisms only were isolated in 1 child, anaerobic bacteria only in 3, and aerobic mixed with anaerobic bacteria in 6 . There were 10 aerobic or faculative and 17 anaerobic isolates . The predominant aerobes were Pseudomonas aeruginosa (2 isolates) and Klebsiella sp . (2) . The predominant anaerobes were pigmented Prevotella and Porphyromonas species (5), Peptostreptococcus sp . (4), Fusobacterium sp . and B . fragilis group (2) . A total of 10 beta-lactamase-producing aerobic and anaerobic bacteria were isolated in 8 patients . All patients except 1 responded to antimicrobial therapy directed against the recovered isolates . These data highlight the polymicrobial aerobic-anaerobic flora isolated from pulmonary specimens of patients with ventilator-associated pneumonia.

Microbios, 1995, 84(339), 87 - 90
In vitro activity of imipenem and six other beta-lactam antibiotics against aminoglycoside resistant gram-negative bacilli; Bujdakova H et al.; The in vitro activity of imipenem and six other beta-lactams (ampicillin, cefamandole, cefoxitin, ceftriaxone, ceftazidime, cefotaxime) against twenty Gram-negative bacilli resistant to clinically important aminoglycosides was studied . The bacterial strains showed high resistance (R) to gentamicin, tobramycin (R 100%), netilmicin (R 75%) and to the beta-lactams ampicillin (R 100%), cephamandole (R 90%) and cefoxitin (R 75%) . The strains were susceptible to isepamicin (R 10%) and imipenem (R 10%) as well as susceptible to at least third generation cephalosporin . Three Pseudomonas aeruginosa strains were resistant to the majority of antibiotics, except for imipenem.

Res Exp Med (Berl), 1995, 195(3), 163 - 9
The effects of experimental splenic autotransplantation and imipenem-cilastatin treatment in postsplenectomy Pseudomonas aeruginosa sepsis; Eskiturk A et al.; Male Wistar albino rats were allocated to three groups-sham operated (n: 10), splenectomized (n: 20) and splenic autotransplanted (n: 10) . Twelve weeks after operation, all were challenged with 1.8 x 10(8) cfu/ml Pseudomonas aeruginosa intranasally . Half of the splenectomized rats received imipenem-cilastatin after 2 h of bacterial challenge . Mortality was then observed for the next 12 days . All animals were autopsied and liver, kidney, spleen and lung specimens were processed for microbiological culture and histopathological examination . In 80% of autotransplanted rats, splenic tissue regeneration was histopathologically verified . Hemoglobin oxidation of erythrocytes increased in splenectomized rats and remained close to control levels in the autotransplanted group . No significant difference was detected between IgM levels of splenectomized and autotransplanted groups . Mortality rates were the same for all groups, except that splenectomized animals given antimicrobial therapy had increased survival rates . In conclusion, it is likely that the spleen has no role in protection against pulmonary sepsis and that only appropriate antimicrobial therapy can protect the splenectomized host from Pseudomonas sepsis.

Microbios, 1995, 84(338), 41 - 51
Effect of quaternary ammonium salts and amine oxides on Pseudomonas aeruginosa; Majtan V et al.; Two quaternary ammonium salts, viz (1-methyldodecyl) trimethylammonium bromide (ATDBr) and tetramethylammonium bromide (TMABr), as well as two amine oxides, (1-methyldodecyl)dimethylamine oxide (ATDNO) and trimethylamine oxide (TMANO), were tested for their inhibitory activity on a Pseudomonas aeruginosa strain isolated from nosocomial infections . Only those compounds with long alkyl chains in their molecules (ATDBr and ATDNO) showed antimicrobial efficacy . In subinhibitory concentrations both compounds inhibited incorporation of {14C}-adenine and {14C}-leucine as precursors of macromolecular biosynthesis . Endogenous respiration of the cells was more sensitive to both agents than the respiration of various substrates . Among the virulence factors only the production of phospholipase C was inhibited by sub-MICs, while the activities of elastase and proteinase were stimulated until the inhibitory concentrations were reached . The short-chain analogues TMABr and TMANO did not show these effects . It is suggested that the production of virulence factors is affected by amphiphilic compounds due to their antimicrobial activity.

Trop Geogr Med, 1995, 47(5), 193 - 6
Pseudomonas aeruginosa bacteraemia in Enugu, Nigeria . A review of 24 cases; Ozumba UC; A 4-year study of Pseudomonas aeruginosa bacteraemia was carried out at the University of Nigeria Teaching Hospital (UNTH) in Enugu, Nigeria . The average age of the patients was 14.7 years . Thirty-three per cent of the patients were between 0-6 months of age, with males being in the majority . Underlying factors/diseases were present in 70.8% of patients, with prematurity, chronic suppurative otitis media and leukaemia being the major ones in infancy and childhood and diabetes mellitus, urogenital disorders and head injuries in adults . Overall mortality was 50% and highest (16.6%) in the 0-6 months age group . Antipseudomonas antibiotics are extremely costly and therefore beyond the financial scope of many people in the developing world . In Nigeria aminoglycosides are the best affordable antibiotics.

Adv Perit Dial, 1995, 11, 152 - 6
The effect of ultraviolet rays on the prevention of exit-site infections; Shimomura A et al.; The aim of this study was to show that ultraviolet (UV) irradiation to the skin around the catheter exit site (ES) could inhibit its infection . First, bacterial cultures of swabbed fluid from the ES were obtained from 68 continuous ambulatory peritoneal dialysis (CAPD) outpatients six times during the 24-month observation period . Second, the bactericidal effects of UV irradiation on the catheter ES were examined . The results were as follows: (1) In spite of disinfection of the catheter ES by the strict application of povidone-iodine once or twice a day, 23%-45% of the cases were found to be micro-organism positive . The most prevalent micro-organisms from the catheter ES were, in order of highest to lowest prevalence, Staphylococcus epidermidis (SE), Staphylococcus aureus (SA), and Pseudomonas aeruginosa (PA) . (2) In the nasal cavity SA was detected in 20%-25% of patients . There was a high incidence of ES infection among the SA nasal carriers . (3) UV irradiation was performed in 18 cases that constantly revealed bacteria on culture at the catheter ES . Ten cases (55%) became culture-negative, 3 cases showed a microbial decrease, and 5 cases remained unchanged . These results suggest that UV irradiation can eliminate bacteria and can be of prophylactic use for ES infections.

Adv Perit Dial, 1995, 11, 149 - 51
Frequency of various types of peritoneal catheter infections and therapeutic outcome of treatment; Gucek A et al.; To analyze peritoneal catheter infections (PCIs), primarily the type (acute or chronic), frequency, and therapeutic outcome, we assessed 113 patients treated between January 1992 and December 1994 . The average age was 56.3 +/- 15.3 years, and 38% were diabetics . One hundred and thirty peritoneal catheters (PCs) were placed surgically in the lateral abdominal wall . The peritonitis rate fell from 0.61 episodes/year to 0.33 episodes/year, but the exit-site and/or tunnel infection (ESI/TI) rate increased (from 0.48 episodes/year to 0.61 episodes/year) . Seventy-nine cases of PCI were observed; 58 (73.4%) were acute ESI/TI and 21 (26.6%) were exacerbations of chronic ESI/TI . Thirty-one (53.4%) acute PCIs were cured, 17 (29.3%) became persistent, and in 10 (17.2%) cases the PC was removed . In chronic ESI/TI, of the 21 exacerbations registered, in 12 cases (57.1%) conservative treatment was effective, while in 9 cases (42.9%) the PC was removed . We conclude that ESI/TIs are the most frequent type of continuous ambulatory peritoneal dialysis (CAPD) infection and the more frequent cause of PC removal compared to peritonitis (p < 0.001) . PC removal is more frequent in chronic than in acute ESI/TI (p < 0.005) . The progression of infection towards the external and even the internal cuff is a poor prognostic sign . Staphylococcus aureus and Pseudomonas aeruginosa were the most common causes of infection and the most serious infective agents, causing chronic infection or catheter removal . Clinical evaluation of ESI/TI can be helped significantly by ultrasound examination, which is 100% positive in chronic ESI/TI and not more than 52.1% positive in acute ESI/TI (p < 0.005).

Eur J Pediatr, 1995, 154(9 Suppl 4), S69 - 73
The age at onset of chronic Pseudomonas aeruginosa colonization in cystic fibrosis--prognostic significance; Aebi C et al.; To evaluate the prognostic significance of the age at onset of chronic Pseudomonas aeruginosa colonization (OPCP) with respect to pulmonary disease progression in patients with cystic fibrosis (CF), a retrospective long-term analysis using annual chest radiographs was performed on 54 CF patients . Thirty-seven patients (68%) were chronically colonized before the age of 12 years (group 1), 17 patients (32%) thereafter (group 2) . These two groups did not significantly differ in terms of mean duration of follow up (16.2 +/- 5.9 years), sex, CF genotypes, colonization with other respiratory pathogens, supportive medical treatment and death rate during the study period . Chest radiographs were evaluated according to the Chrispin-Norman score, increasing scores representing increasing severity of respiratory disease . In both groups, progression of score means was not accelerated of score means was not accelerated up to 6 years after OCPC (Scores at OCPC set 0; mean score +/- SEM 6 years prior to OCPC -5.6 +/- 2.0; 10 years after OCPC +3.6 +/- 0.7 points) . Patients chronically colonized prior to age 12 years (group 1) scored significantly higher between age 2 and 11 years (maximum difference at age 8 years {mean +/- SEM}: 9.4 +/- 0.7 vs . 4.3 +/- 1.3 points; P = 0.002) as compared to group 2 . After age 11 years, mean scores were similar in both groups, since in group 2 scores increased rapidly after age 8 years . We conclude that OCPC did not cause an immediate acceleration of CF lung disease judged by serial chest radiographs . Rapid progression in group 2 (OCPC after age 12 years) was independent of OCPC since it occurred earlier . These data indicate that OCPC may be a marker rather than the cause of respiratory disease progression.

Drugs Exp Clin Res, 1995, 21(4), 139 - 44
Outer membrane alterations in Pseudomonas aeruginosa after five-day exposure to quinolones and carbapenems; Cipriani P et al.; A Pseudomonas aeruginosa strain (P.aeruginosa), recently isolated in clinical practice, was tested to evaluate the changes induced in the minimal inhibitory concentration (MIC) values and in the outer membrane (OM) components by a five-day exposure to sub-MIC concentrations of the quinolones ciprofloxacin and PD-131.628 and the carbapenems imipenem and meropenem . The treated strain showed a thirty two-fold increase in MIC values for quinolones and a sixteen-fold increase for carbapenems . The electrophoretic profile of the OM proteins of the strain treated with quinolones showed that ciprofloxacin induces loss of the 47 Kd protein band, whereas PD-131.628 modifies the protein pattern of the strain only after five days of exposure . The carbapenems engendered disappearance of the same protein band . Qualitative lipopolysaccaride (LPS) analysis did not reveal any change after antibiotic treatment of the strain, whereas the 2-keto-3-deoxyoctonic acid (KDO) assay showed considerable reduction in the strain treated with sub-MIC doses of meropenem . It can therefore be safely stated that the D2 protein plays an important though not exclusive role in enhancing strain resistance against the two classes of antibiotics tested in our study.

Surg Today, 1995, 25(8), 672 - 8
Severity and predicted outcome of postoperative Pseudomonas aeruginosa infections; Kodama T et al.; The severity and predicted outcome of postoperative Pseudomonas aeruginosa (P . aeruginosa) infections (PPAI) was evaluated using a severity scoring system based on a simplification and modification of the APACHE II system . A total of 86 patients in whom P . aeruginosa was isolated from various sources were examined . PPAI developed in 50 patients, resulting in an overall mortality rate of 24% . An increased severity score (SS) correlated with an increased risk of developing PPAI . Thus, PPAI developed in 33% of the patients with an SS of 0-1, in 66.7% of those with an SS of 2-3, and in 100% of those with an SS of 6 or higher . Moreover, the mortality rate of the patients with an initial score of 6 or higher was 50% . The mean (+/- SD) initial severity score was 5.4 +/- 2.9 for survivors and 2.9 +/- 2.6 for nonsurvivors (P < 0.01) . In the patients who subsequently died, the SS remained high throughout the clinical course despite therapy, whereas in the survivors the SS decreased progressively, reflecting a favorable clinical course . These results suggest that our severity scoring system was useful for predicting outcome and monitoring the response of PPAI to therapy.

Thorax, 1995 Jan, 50(1), 14 - 7
Use of elastin fibre detection in the diagnosis of ventilator associated pneumonia; el-Ebiary M et al.; BACKGROUND--Elastin fibre detection could be a simple and reliable marker of ventilator associated pneumonia . To confirm this, a prospective study was undertaken to evaluate the diagnostic yield of elastin fibre detection in the diagnosis of ventilator associated pneumonia . METHODS--Seventy eight mechanically ventilated patients were evaluated by examining endotracheal aspirates for the presence of elastin fibres . All patients were previously treated with antibiotics . Quantitative bacterial cultures of endotracheal aspirates and protected specimen brush samples were also performed . Patients were classified into three diagnostic categories: group 1, definite pneumonia (n = 25); group 2, probable pneumonia (n = 35); and group 3, controls (n = 18) . RESULTS--Patients with definite and probable pneumonia were grouped together . The presence of elastin fibres in endotracheal aspirate samples was more frequent in groups 1 and 2, being found in 19 of the 60 patients compared with five of the control group . Although the presence of elastin fibres had a low sensitivity (32%), it was a reasonably specific marker (72%) of pneumonia . This specificity increased to 86% and 81% respectively when only Gram negative bacilli and Pseudomonas aeruginosa pneumonia were considered . Again, calculated sensitivity was 43% and 44% when analysing cases infected by Gram negative bacilli and Ps aeruginosa, respectively . The negative predictive value of the detection of elastin fibres in pneumonia caused by Ps aeruginosa was 81% . Detection was more frequent with infection by Gram negative bacilli (14/19), particularly with Ps aeruginosa (8/14) . By contrast, pneumonia due to Gram positive cocci or non-bacterial agents uncommonly resulted in positive elastin fibre preparations (4/19, 21%) . When analysing patients with and without chronic obstructive pulmonary disease, the diagnostic value of elastin fibre detection did not change . CONCLUSIONS--Potassium hydroxide preparation of elastin fibres is a rapid and simple specific marker of ventilator associated pneumonia and may be a useful technique to help diagnose pulmonary infections in mechanically ventilated patients, although this assessment is at present limited to patients without adult respiratory distress syndrome.

Invest Ophthalmol Vis Sci, 1995 Jan, 36(1), 16 - 23
Characterization of arachidonic acid metabolism and the polymorphonuclear leukocyte response in mice infected intracorneally with Pseudomonas aeruginosa; Kernacki KA et al.; PURPOSE . To examine the activity of myeloperoxidase (MPO) and the concentrations of the proinflammatory metabolites of arachidonic acid (AA) in ocular tissue of mice that are either capable or incapable of restoring corneal clarity during an intraocular Pseudomonas aeruginosa infection . METHODS . For a period of 11 days after infection, whole eyes were enucleated and homogenized in buffer from mice given only an initial infection as well as from mice given a subsequent infection in the previously uninfected eye either 4 or 8 weeks after the initial infection . Tissue-free supernatants from the ocular homogenates were used for the determination of MPO activity by quantitating the conversion of specific substrate by spectrophotometric methods and for the quantitation of AA metabolites by ELISA . RESULTS . Overall, animals reinfected at 4 and 8 weeks had a lower inflammatory response when compared to the mice given only the initial infection . The lowest levels of LTB4 and MPO activity, indicators of PMN involvement, were observed in the the 8-week reinfected mice, which restored corneal clarity in an enhanced manner . CONCLUSIONS . These results suggest that induced ocular PMN responses may play a role, in part, in the inflammatory response leading to the tissue destruction observed during ocular P . aeruginosa infection.

J Bacteriol, 1995 Jan, 177(2), 432 - 8
Isolation and characterization of mutants defective in the cyanide-insensitive respiratory pathway of Pseudomonas aeruginosa; Cunningham L et al.; The branched respiratory chain of Pseudomonas aeruginosa contains at least two terminal oxidases which are active under normal physiological conditions . One of these, cytochrome co, is a cytochrome c oxidase which is completely inhibited by concentrations of the respiratory inhibitor potassium cyanide as low as 100 microM . The second oxidase, the cyanide-insensitive oxidase, is resistant to cyanide concentrations in excess of 1 mM as well as to sodium azide . In this work, we describe the isolation and characterization of a mutant of P . aeruginosa defective in cyanide-insensitive respiration . This insertion mutant was isolated with mini-D171 (a replication-defective derivative of the P . aeruginosa phage D3112) as a mutagen and by screening the resulting tetracycline-resistant transductants for the loss of ability to grow in the presence of 1 mM sodium azide . Polarographic studies on the NADH-mediated respiration rate of the mutant indicated an approximate 50% loss of activity, and titration of this activity against increasing cyanide concentrations gave a monophasic curve clearly showing the complete loss of cyanide-insensitive respiration . The mutated gene for a mutant affected in the cyanide-insensitive, oxidase-terminated respiratory pathway has been designated cio . We have complemented the azide-sensitive phenotype of this mutant with a wild-type copy of the gene by in vivo cloning with another mini-D element, mini-D386, carried on plasmid pADD386 . The complemented cio mutant regained the ability to grow on medium containing 1 mM azide, titration of its NADH oxidase activity with cyanide gave a biphasic curve similar to that of the wild-type organism, and the respiration rate returned to normal levels . Spectral analysis of the cytochrome contents of the membranes of the wild type, the cio mutant, and the complemented mutant suggests that the cio mutant is not defective in any membrane-bound cytochromes and that the complementing gene does not encode a heme protein.

J Bacteriol, 1995 Jan, 177(2), 423 - 31
Genetic and physical mapping of genes involved in pyoverdin production in Pseudomonas aeruginosa PAO; Tsuda M et al.; Pseudomonas aeruginosa PAO was mutagenized with Tn1737KH, a type I transcription probe transposon containing a promoterless lacZ (beta-galactosidase) gene, and 24 insertion mutants that did not grow under iron-deficient conditions were isolated . None of the culture supernatants from any mutants contained pyoverdin, a low-molecular-weight siderophore able to sequester ferric iron at very high affinity, and the growth defects of the mutants were all phenotypically recovered by the addition of the culture supernatant from the wild-type strain . These phenotypes led to the inference that all the mutants had defects in the genes (pvd genes) for production of pyoverdin . In some pvd::Tn1737KH mutants, high levels of beta-galactosidase activities were observed, and such activities were drastically reduced by the addition of ferric ion in the culture media, indicating that the expression of at least some pvd genes is regulated at the transcriptional level . Molecular cloning and physical analysis of the chromosomal fragments with Tn1737KH insertions allowed us to allocate all the mutations within a 103-kb region, referred to as the pvd region, that was found to locate at 47 min on the genetic map of PAO . Further physical mapping and Southern analysis showed that there is a 10-kb overlap between the pvd region and the 125-kb catA region described by Zhang and Holloway (C . Zhang and B . W . Holloway, J . Gen . Microbiol . 138:1097-1107, 1992) . We could hence illustrate the physical map of the P . aeruginosa chromosome with a size of 218 kb.

Clin Exp Immunol, 1995 Jan, 99(1), 98 - 105
In vitro and in vivo T cell responses in mice during bronchopulmonary infection with mucoid Pseudomonas aeruginosa; Stevenson MM et al.; In vitro and in vivo T cell responses were determined during the course of bronchopulmonary infection with mucoid Pseudomonas aeruginosa . T cell responses were compared in two inbred mouse strains, namely BALB/c mice, which are resistant to the establishment of chronic bronchopulmonary Ps . aeruginosa infection, and C57Bl/6 mice, which have high numbers of bacteria in the lungs through 14 days post-infection . Unseparated lung cells and lung T cells from BALB/c mice exhibited significantly higher in vitro proliferative responses to both heat-killed Ps . aeruginosa and concanavalin A (Con A) than cells from C57Bl/6 mice through 20 days post-intratracheal infection with 10(4) colony-forming units (CFU) Ps . aeruginosa . Proliferation of unseparated lung cells but not lung T cells from BALB/c mice infected 6 days previously with 10(5) CFU Ps . aeruginosa was suppressed in response to Con A; these cells were unresponsive to specific antigen . Suppression of lymphocyte proliferation in the lungs of C57Bl/6 mice infected with 10(4) CFU Ps . aeruginosa and in BALB/c mice infected with 10(5) CFU was found to be mediated by adherent lung cells via the production of nitric oxide and prostaglandins . Determination of in vivo T cell-mediated responses in infected mice demonstrated that resistant BALB/c mice had high DTH and low Pseudomonas-specific antibody responses, while C57Bl/6 mice had low DTH and high antibody levels, in particular, IgG2b and IgM.

Infect Immun, 1995 Jan, 63(1), 38 - 42
Epitope mapping of the Pseudomonas aeruginosa major outer membrane porin protein OprF; Rawling EG et al.; The Pseudomonas aeruginosa major outer membrane protein OprF has been proposed for use as a vaccine and as a target for immunotherapeutic and diagnostic monoclonal antibodies . The well-conserved epitopes for 10 surface-reactive, OprF-specific monoclonal antibodies were localized by both overlapping peptide analysis and immunodetection of OprF peptides generated by cyanogen bromide and the protease papain . Three of the monoclonal antibodies bound to specific overlapping octapeptides, which had been synthesized on 160 pins to cover the entire 326 amino acids of OprF . The highest reactivities were as follows: MA7-1 to the pin with attached peptide GTYETGNK (amino acids 55 to 62), MA7-2 to NLADFMKQ (amino acids 237 to 244), and MA5-8 to TAEGRAIN (amino acids 307 to 314) . The other monoclonal antibodies showed no reactivity, indicating that they do not recognize linear epitopes . Two polyclonal sera were also tested and demonstrated weak reactivity with discrete regions of OprF, suggesting that the majority of antibodies produced might recognize conformational epitopes . Utilizing defined peptides generated with cyanogen bromide and papain, the conformational epitopes recognized by the seven monoclonal antibodies were localized to regions that were 42 to 90 amino acids long . These regions were located on two adjacent loops in the middle of an amended structural model of OprF.

Infect Immun, 1995 Jan, 63(1), 353 - 5
Pseudomonas aeruginosa infection of the cornea and asialo GM1; Zhao Z et al.; Extensive immunohistochemical and thin-layer chromatogram-immunostain analyses were carried out to establish whether asialo GM1, a glycolipid which contains binding sites for Pseudomonas aeruginosa, is present in corneal epithelium . The data suggest that rabbit corneal epithelium does not contain detectable levels of asialo GM1 even after corneas are scarified and incubated with trypsin, P . aeruginosa, or P . aeruginosa exoproducts to expose potential cryptic sites . Preliminary immunohistochemical analyses indicated that asialo GM1 is also not found in human corneas.

Infect Immun, 1995 Jan, 63(1), 333 - 9
Role of endotoxin in acute inflammation induced by gram-negative bacteria: specific inhibition of lipopolysaccharide-mediated responses with an amino-terminal fragment of bactericidal/permeability-increasing protein; Kohn FR et al.; A recombinant 23-kDa amino-terminal fragment of human bactericidal/permeability-increasing protein (rBPI23), a potent lipopolysaccharide (LPS)-binding/neutralizing protein, was used as a probe to assess the role of endotoxin in the acute inflammatory responses elicited by gram-negative bacteria in rat subcutaneous air pouches . In initial experiments, rBPI23 prevented the Escherichia coli O111:B4 LPS-induced accumulation of polymorphonuclear leukocytes (PMN), tumor necrosis factor alpha (TNF-alpha), and nitrite (a stable end product of nitric oxide formation) in exudate fluids . Significant inhibition of TNF-alpha production was still evident when rBPI23 treatment was delayed for 30 min after LPS instillation . In subsequent experiments, rBPI23 also prevented the nitrite and early (2-h) TNF-alpha accumulation induced by three different strains of formaldehyde-killed gram-negative bacteria (E . coli O7:K1, E . coli O111:B4, and Pseudomonas aeruginosa 12.4.4) but did not inhibit the PMN or late (6-h) TNF-alpha accumulation induced by these bacteria . As with LPS challenge, a significant inhibition of early TNF-alpha production was still evident when rBPI23 treatment was delayed for 30 to 60 min after instillation of killed bacteria . The results indicate that in this experimental model the NO and early TNF-alpha responses to gram-negative bacterial challenge are mediated predominantly by endotoxin, whereas the PMN and late TNF-alpha responses may be mediated by other bacterial components . Moreover, the results indicate that rBPI23 can inhibit the bacterially induced production of certain potentially harmful mediators (TNF-alpha and NO) without entirely blocking the host defense, i.e., PMN response, against the bacteria.

Infect Immun, 1995 Jan, 63(1), 1 - 8
Expression of recombinant exoenzyme S of Pseudomonas aeruginosa; Kulich SM et al.; The structural gene for the 49-kDa form of exoenzyme S (exoS) isolated from Pseudomonas aeruginosa 388 was expressed in both Escherichia coli and P . aeruginosa PA103 . Expression of exoS in E . coli under the transcriptional regulation of the T7 promoter yielded a soluble cytosolic protein with an apparent molecular mass of 49 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Expression of exoS in P . aeruginosa PA103 under the transcriptional regulation of the 0.9 kbp of Pseudomonas chromosomal DNA flanking the 5' end of exoS yielded a nitrilotriacetic acid-inducible extracellular protein with an apparent molecular mass of 49 kDa . Recombinant ExoS (rExoS) reacted with the anti-49-kDa form of exoenzyme S immunoglobulin G, existed as an aggregate as determined by gel filtration chromatography, and ADP-ribosylated soybean trypsin inhibitor at a specific activity that was similar (within twofold) to that of native exoenzyme S . Allelic exchange of exoS with a tetracycline gene cartridge yielded a strain of P . aeruginosa 388 that did not express detectable amounts of either ExoS in an immunoblot analysis using the anti-49-kDa form of exoenzyme S immunoglobulin G or ADP-ribosyltransferase activity under standard enzyme assay conditions . Expression of catalytically active rExoS in E . coli demonstrated that exoS was necessary and sufficient for the factor-activating exoenzyme S-dependent ADP-ribosyltransferase activity of exoenzyme S . Expression of nitrilotriacetic acid-inducible rExoS in P . aeruginosa PA103 demonstrated that the 0.9 kbp of Pseudomonas chromosomal DNA flanking the 5' end of exoS encoded a functional exoenzyme S promoter . Expression analysis and allelic exchange experiments suggest that the 49- and 53-kDa forms of exoenzyme S are encoded by separate genes.

J Bacteriol, 1995 Jan, 177(1), 252 - 8
Nucleotide sequence of pvdD, a pyoverdine biosynthetic gene from Pseudomonas aeruginosa: PvdD has similarity to peptide synthetases; Merriman TR et al.; Pseudomonas aeruginosa secretes a fluorescent siderophore, pyoverdine, when grown under iron-deficient conditions . Pyoverdine consists of a chromophoric group bound to a partly cyclic octapeptide . As a step toward understanding the molecular events involved in pyoverdine synthesis, we have sequenced a gene, pvdD, required for this process . The gene encodes a 2,448-residue protein, PvdD, which has a predicted molecular mass of 273,061 Da and contains two highly similar domains of about 1,000 amino acids each . The protein is similar to peptide synthetases from a range of bacterial and fungal species, indicating that synthesis of the peptide moiety of pyoverdine proceeds by a nonribosomal mechanism . The pvdD gene is adjacent to a gene, fpvA, which encodes an outer membrane receptor protein required for uptake of ferripyoverdine.

Zh Mikrobiol Epidemiol Immunobiol, 1995 Jan-Feb, (1), 6 - 10
{The effect of the level of lipid peroxidation and the pH value of a liposomal suspension on its antibacterial activity}; Mel'iantseva LP et al.; Multilamellar phosphatidylcholine liposomes have been shown to possess antibacterial activity with respect to Pseudomonas aeruginosa and Staphylococcus aureus . An increase in the level of lipid peroxidation products in liposomal suspension as the consequence of the earlier oxidation of liposomal lipid leads to a considerable increase in the antibacterial activity of liposomes . The pH of the liposomal suspension has also an essential influence on its activity . If the pH of the suspension has acidic values, its antibacterial activity increases . The bactericidal action of liposomal suspension is more pronounced with respect to S . aureus, than to P . aeruginosa.

Zh Mikrobiol Epidemiol Immunobiol, 1995 Jan-Feb, (1), 59 - 61
{The determination of specific Pseudomonas aeruginosa antitoxins in commercial preparations of human immunoglobulin}; Vaneeva NP et al.; The comparative study of the activity of P . aeruginosa specific antitoxins in 134 lots of commercial preparations of human immunoglobulin in parallel mouse experiments on the neutralization of the lethal effect of exotoxin and in ELISA made it possible to establish a high degree of correlation between their results and the possibility of using immunochemical assays instead of biological tests in the large-scale production of anti-P . aeruginosa immunoglobulin.

Protein Eng, 1995 Jan, 8(1), 63 - 70
Interactions of (Ala*Ala*Lys*Pro)n and (Lys*Lys*Ser*Pro)n with DNA . Proposed coiled-coil structure of AlgR3 and AlgP from Pseudomonas aeruginosa; Medvedkin VN et al.; The proteins, AlgR3 and AlgP, are involved in the regulation of alginate synthesis in Pseudomonas . They contain multiple repeats of Ala*Ala*Lys*Pro as do several other proteins that resemble histones . The interactions of synthesis oligopeptides composed of repeated Ala*Ala*Lys*Pro or Lys*Lys*Ser*Pro units with DNA were studied by fluorescence of the Fmoc (9-fluorenylmethyloxycarbonyl) group attached to the N-termini of the peptides . DNA quenching of the Fmoc fluorescence of the peptides was used to estimate the apparent association constants for the interaction of Fmoc(AAKP)nOH (n = 2, 4, 8, 18, 32) and of Fmoc (KKSP)nOH (n = 2, 4, 8, 16, 20, 32) with DNA . The Fmoc(AAKP)nOH peptides bind to DNA only at low ionic strength; the Fmoc(KKSP)n OH peptides interact with DNA at both low (0.05 M KCl) and high (0.2 M KCl) salt . At low ionic strength an increase in the number of the repeat units causes an increase in the apparent association constant up to approximately 2 x 10(6) M-1 for both types of peptides at N congruent to 24 . The insertion of an AAKTA unit into the middle of the Fmoc(AAKP)8OH peptide increases its affinity to DNA . We propose a model of (AAKP)n and of its interaction with DNA . The repeat unit consists of a single turn of alpha-helix followed by a bend necessitated by Pro . The resultant coiled-coil forms a right-handed superhelix with 10 AAKPs per repeat distance of approximately 33 A.(ABSTRACT TRUNCATED AT 250 WORDS)

Biodegradation, 1995 Jan, 6(1), 39 - 46
Degradation of 2,5-dichlorobenzoic acid by Pseudomonas aeruginosa JB2 at low oxygen tensions; van der Woude BJ et al.; From long-term chemostat experiments, variants of Pseudomonas aeruginosa JB2 were obtained which exhibited altered properties with respect to the metabolism of 2,5-dichlorobenzoic acid (2,5-DBA) . Thus, unlike the original strain JB2-WT, strain JB2-var1 is able to grow in continuous culture on 2,5-DBA as the sole limiting carbon and energy source . Yet, at a dilution rate of 0.07 h-1 and a dissolved oxygen concentration of < or = 12 microM, even with this strain no steady states with 2,5-DBA alone could be established in continuous cultures . Yet another strain was obtained after prolonged continuous growth of JB2-var1 in the chemostat . It has improved 2,5-DBA degrading capabilities which become apparent only during growth in continuous culture: a lower apparent Km for 2,5-DBA and lowered steady-state residual concentrations of 2,5 DBA . Although with this strain steady states were obtained at oxygen concentrations as low as 11 microM, at further lowered concentrations this was no longer possible . In C-limited continuous cultures of JB2-var1 or JB2-var2, addition of benzoic acid (BA) to the feed reduced the amounts of 2,5-DBA degraded, which was most apparent at low oxygen concentrations (< 30 microM) . At higher dissolved oxygen concentrations the addition of BA resulted in increasing cell-densities but did not affect the residual steady state concentration of 2,5-DBA . Indeed, whole cell suspensions from chemostat cultures grown on BA plus 2,5-DBA did show a lower apparent affinity for 2,5-DBA than those from cultures grown on 2,5-DBA alone.(ABSTRACT TRUNCATED AT 250 WORDS)

Curr Microbiol, 1995 Jan, 30(1), 55 - 60
Carnitine resembles choline in the induction of cholinesterase, acid phosphatase, and phospholipase C and in its action as an osmoprotectant in Pseudomonas aeruginosa; Lucchesi GI et al.; The present study demonstrates that under conditions of iso or hyperosmolarity, P . aeruginosa utilized carnitine as the carbon, nitrogen or carbon and nitrogen sources . As occurred in the case of choline, the bacteria synthesized cholinesterase (ChE), acid phosphatase (Ac.Pase) and phospholipase C (PLC) under any of these conditions and in the presence of high or low Pi concentrations . Carnitine acted as an osmoprotectant when the cells were grown in the presence of preferred carbon and nitrogen sources and high NaCl concentrations . Under these conditions the three enzyme activities were not produced . The osmotically stressed bacteria grown under any of the above conditions accumulated betaine . Its presence indicated that carnitine may be metabolized by P . aeruginosa to produce betaine which could account for the induction of the three enzyme activities or its action as an osmoprotectant . The phosphatidylcholine encountered in the host cell membranes allows the bacteria to obtain free choline by the coordinated action of PLC and Ac.Pase . Since the consequence of this action may be cell disruption, the increase of free carnitine in the natural environment of the bacteria is also possible . These two compounds, choline and carnitine, acting in conjunction or separately, may increase the production of PLC and Ac.Pase activities by P . aeruginosa and thus enhance the degradative effect upon the host cells.

Vaccine, 1995 Jan, 13(1), 67 - 71
Human immunodeficiency virus-1 principal neutralizing domain peptide-toxin A conjugate vaccine; Cryz SJ Jr et al.; To enhance the potential efficacy of peptide-based vaccines for human immunodeficiency virus-1 (HIV-1), a principal neutralizing domain (PND) peptide (KRIHIGPGRAFYT) (HIV-1MN) was covalently coupled to Pseudomonas aeruginosa toxin A (TA) . Immunization of guinea-pigs with this conjugate vaccine, in the absence of an adjuvant, engendered a high-affinity antibody response to the homologous HIV-1MN PND peptide and to analogous peptides from variant strains of HIV-1 . A substantial proportion of such antibodies was directed to the conserved GPGRAF motif . Anti-PND peptide antibodies were capable of neutralizing the homologous strain, HIV-1MN, in addition to two heterologous (RF, IIIB) variants, as determined either by inhibition of syncytia formation or by suppression of p24 antigen production in cultured cells . Therefore, the method of conjugation used preserved critical neutralizing epitopes expressed by the PND peptide . Monovalent or polyvalent PND-TA conjugates, which meet all safety criteria for human use, are a promising approach towards the development of an acquired immunodeficiency syndrome (AIDS) vaccine.

J Nat Prod, 1995 Jan, 58(1), 74 - 81
Microbial metabolites of ophiobolin A and antimicrobial evaluation of ophiobolins; Li E et al.; Ophiobolin A {1}, 3-anhydroophiobolin A {2}, ophiobolin B {3}, and ophiobolin L {4} were isolated from fermentation broths of Cochliobolus heterostrophus . Preliminary screening showed that a number of organisms were capable of metabolizing the sesterterpene ophiobolin A {1} . Large-scale transformations of ophiobolin A {1} with Polyangium cellulosum produced 6 and 7 while Pseudomonas aeruginosa produced 8 . Resting-cell preparations of Penicillium patulum afforded 9 and 10 . The structures of these metabolites were established by spectroscopic methods and by comparison of the spectral data with those of the starting material . The antimicrobial activity of the ophiobolins was also evaluated.

APMIS Suppl, 1995, 50, 1 - 30
Lipopolysaccharide (LPS), LPS-immune complexes and cytokines as inducers of pulmonary inflammation in patients with cystic fibrosis and chronic Pseudomonas aeruginosa lung infection; Kronborg G; The aim of the present thesis was to summarize some important immunological mechanisms in the pathogenesis of chronic Pseudomonas aeruginosa lung infection in patients with cystic fibrosis (CF) . The continuous presence of bacteria in the lungs induce a strong immunological response in the patients both locally in the lungs and systemically with high amounts of circulating specific anti-P . aeruginosa antibodies . Our work has been concentrating on anti-lipopolysaccharide (LPS) antibodies . We have shown an increasing antibody response in CF patients, to all three parts of the LPS molecule; lipid A, core, and O-sugars, during the course of chronic P . aeruginosa infection . The antibodies belonged to both IgA, IgM, and all four subclasses of IgG, and were detected in serum and sputum . We detected immune complexes (IC)s in sputum from chronically infected CF patients . The ICs were composed of P . aeruginosa LPS and immunoglobulins of both IgG1-4, IgA and IgM . The concentration of circulating ICs were significantly higher in chronically infected patients compared to non-infected CF patients . The presence of ICs containing LPS in sputum were positively correlated to the amount of tumor necrosis factor alpha (TNF alpha) in the same sputum sample . TNF alpha is a very potent inflammatory mediator, stimulating cells for release of several cytokines attracting polymorphonuclear neutrophil granulocytes (PMNs), which release proteolytic enzymes and toxic oxygen radicals . We detected high concentrations of both TNF alpha, interleukin (IL)-1 alpha, IL-1 beta, IL-6, and the IL-1 receptor antagonist (IRAP) in sputum from chronically infected CF patients . The corresponding concentrations of the cytokines in serum were low or undetectable . Relatively high concentrations of serum-IRAP before the diagnosis of chronic P . aeruginosa infection were correlated to development of poor pulmonary function . We made ICs in vitro of purified P . aeruginosa LPS and hyperimmune serum from chronically infected CF patients . The biological activity of these ICs was investigated in two different assays . LPS by itself induced TNF alpha liberation in vitro, but the ICs made in vitro were also able to stimulate TNF alpha release from mononuclear cells, and they were a more potent stimuli compared to the corresponding amount of LPS alone . The IC preparation did also induce an oxidative burst response in PMNs . We conclude P . aeruginosa LPS is biological active and formation of ICs involving P . aeruginosa LPS and anti-LPS antibodies takes place in the lungs of chronically infected CF patients.(ABSTRACT TRUNCATED AT 400 WORDS)

Nephrol Dial Transplant, 1995, 10(2), 207 - 11
The role of plasma coating on the permeation of cytokine-inducing substances through dialyser membranes; Lonnemann G et al.; We studied the effects of coating of dialyser membranes with plasma proteins on the permeation of bacteria-derived cytokine-inducing substances (CIS) . An in vitro dialysis circuit using polysulphone (PS) or modified cellulose triacetate (mCT) dialysers was used . Precoating of the dialysers was performed by recirculation of 10% normal human plasma for 30 min in the blood compartment and subsequent rinse with pyrogen-free saline . Samples from the blood compartment were tested for induction of interleukin-1 alpha (IL-1 alpha), interleukin-1 beta (IL-1 beta) and tumour necrosis factor (TNF alpha) at various time points after challenging the dialysate with sterile culture supernatants from Pseudomonas aeruginosa . Contamination of the dialysate resulted in the appearance of CIS in the blood compartment of both polysuphone modified cellulose triacetate (IL-1 alpha: PS, time 0: 81 +/- 11 pg/ml, time 60 min: 4747 +/- 1822 pg/ml, P < 0.05; mCT, time 0: 235 +/- 141 pg/ml, time 60 min: 1632 +/- 531 pg/ml, P < 0.05) . The plasma protein layer reduced the penetration of CIS significantly only for polysulphone (IL-1 alpha: PS, time 60: 4747 +/- 1822 versus 880 +/- 525 pg/ml, P < 0.05; modified cellulose triacetate, time 60 min: 1632 +/- 531 pg/ml versus 930 +/- 326 pg/ml) . Samples from the blood compartment contained < 6 pg/ml LAL-reactive material at all time points . We conclude that plasma coating of polysulphone dialysers reduces the permeability for CIS derived from Pseudomonas, either by reducing the effective pore size or by adsorption of proteins that bind CIS.(ABSTRACT TRUNCATED AT 250 WORDS)

Eur Respir J, 1995 Jan, 8(1), 10 - 4
Cystic fibrosis sputum induces a secretory response from airway gland serous cells that can be prevented by neutrophil protease inhibitors; Schuster A et al.; High activities of the neutrophil proteases, elastase and cathepsin G, are found in the sputum of patients with cystic fibrosis (CF) . Because both proteases have been shown to be potent secretagogues for airway submucosal glands, and because hypersecretion is a characteristic feature of CF, the objective of the present study was to examine whether there is secretagogue activity in CF sputum, and to determine the contribution of neutrophil proteases to the secretagogue activity . Confluent monolayers of cultured bovine tracheal serous cells were pulse-labelled with Na2(35)SO4, incubated with diluted CF sputum supernatants in the presence or absence of different protease inhibitors, and the subsequent release of the radio-labelled macromolecules was measured . CF sputum potently induced secretion concentration-dependently . Addition of the selective neutrophil elastase inhibitor ICI 200,355 inhibited the secretory response to CF sputum supernatant by 89% . Addition of a cathepsin G-inhibitor resulted in further inhibition of the secretory response . Addition of phosphoramidon, a drug known to inhibit Pseudomonas aeruginosa elastase, had no effect . We conclude that CF sputum potently stimulates airway submucosal gland cell secretion . These studies with protease inhibitors suggest that neutrophil proteases account substantially for the secretagogue activity present in CF sputum.

Pediatr Med Chir, 1995 Jan-Feb, 17(1), 53 - 5
{Use of quinolones in the treatment of Pseudomonas aeruginosa infections in children with cystic fibrosis}; Milio MC et al.; Quinolones, elective drugs for Pseudomonas aeruginosa (P . aeruginosa) pulmonary infections in Cystic Fibrosis (C.F.) patients, are controversially administered in prepuberal age for their arthropathic toxicity . We report the result of retrospective study on the use of quinolones in a group of 43 CF patients . The patients were divided into two groups: below 18 years (22 pts.) and over 18 (21 pts.) . All patients were evaluated clinically with the scoring system Shwachman and Kulczyk . Ciprofloxacin and Ofloxacin (15/20 mg/kg/die) were administered . In 11.6% of the patients, all belonging to the second group, side effects, such as urticaria, tongue oedema, foreskin erythema, generalised erythema and itch, were described . No side effect has been reported in the patients below 18 years . Two patients complained knee arthralgias not related to the quinolones administration: in fact in the first case the demineralisation seemed to be responsible of the arthralgia, while in the second case an immunological disorder (ANA+, ICC+) should be involved in the pathogenesis of arthritis . Height velocity evaluation showed the same slowering of the CF untreated patients . In conclusion, our study confirms that quinolones use in indicated in severe and infective complications of CF on the basis of their efficacy, safety and their slight adverse effects similar to those of other potent antibiotic . Moreover our results confirm that no quinolone-induced cartilage toxicity is present in CF patients.

J Hosp Infect, 1995 Jan, 29(1), 1 - 7
Epidemiology of Pseudomonas aeruginosa infection and the role of contamination of the environment in the Danish Cystic Fibrosis Centre; Zembrzuska-Sadkowska E et al.; In order to identify the possible reservoirs and routes of cross-infection with Pseudomonas aeruginosa, samples were collected during a six-week period in autumn 1992 from patients, their visiting parents, staff and the inanimate environment of the Danish Cystic Fibrosis (CF) Centre and from a control ward with common paediatric diseases . All the P . aeruginosa strains were phage typed and serotyped . From 240 CF patients, 310 strains of P . aeruginosa were isolated, and of these 283 (91.3%) belonged to the polyagglutinable phenotype, most often with a short phage type (31/188 or 109) . P . aeruginosa was isolated from only six (0.6%) of 1000 swabs taken from the environment . These six environmental strains and 20 P . aeruginosa strains from CF patients with identical serotype and phage type were examined with pulsed field gel electrophoresis . None of the patients harboured strains similar to the environmental strains, indicating the present isolation procedure and hygienic precautions were effective in our CF centre, and prevented contamination of the environment with P . aeruginosa.

Zentralbl Bakteriol, 1995 Jan, 282(1), 54 - 66
Phospholipase A activity in Pseudomonas aeruginosa; Steinbrueckner BE et al.; Our study describes the production, purification and properties of an enzyme from Pseudomonas aeruginosa displaying the properties of phospholipase A . Maximal amounts of enzyme could be detected in the culture supernatant when the bacterium was grown for 3 to 5 days at 37 degrees C in stirred flask cultures containing brain heart infusion . The enzyme was purified by polyethylenimine precipitation and ammonium sulfate precipitation followed by gel filtration . In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the enzyme preparation exhibited two bands with molecular weights of 13.5 and 60 kD, respectively . Correspondingly, two peaks of the same molecular weight could be demonstrated by high performance size exclusion chromatography . The activity toward the sn-2 ester binding of phospholipids was characterized and found to be highest towards phosphatidylcholine . Enzymatic activity was not influenced by the addition of calcium or EDTA while magnesium and strontium caused a decrease of activity . The lyophilized enzyme was found to be stable when stored at -70 degrees C and most active at pH 8.0.

Microbios, 1995, 81(326), 29 - 31
Characterization of resistance to essential oils in a strain of Pseudomonas aeruginosa (VR-6); Pattnaik S et al.; VR-6, a strain of Pseudomonas aeruginosa, harboured a plasmid and was not inhibited by 20 microliters ml-1 of essential oils (eucalyptus, lemongrass, palmarosa, and peppermint) . On treatment with acridine orange, a clone VR-6-AO-1 was obtained which was susceptible to 16.6 microliters ml-1 of eucalyptus or palmarosa oil . The plasmid DNA content of this clone was similar to the parent strain.

Int J Pediatr Otorhinolaryngol, 1995 Jan, 31(1), 23 - 8
Microbiology of chronic suppurative otitis media in children in Surabaya, Indonesia; Brook I et al.; The aerobic and anaerobic microbiology of 38 children from Surabaya, Indonesia, who suffered from chronic suppurative otitis media (CSOM) was studied using strict microbiological methodology . A total of 106 isolates (49 anaerobic and 57 aerobic) were recovered . Aerobic organisms alone were isolated from 11 (29%), anaerobic bacteria only in 4 (11%) and mixed aerobic and anaerobic flora were present in 23 (60%) . The predominant organisms were Peptostreptococcus sp., Prevotella sp., Bacteroides sp., Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae and Fusobacterium sp . Thirty-nine beta-lactamase-producing bacteria were recovered from 22 (58%) patients . These findings demonstrate the role of penicillin resistant aerobic and anaerobic bacteria in the polymicrobial etiology of CSOM in children from Indonesia . Judicious use of antimicrobial therapy may prevent the development of antimicrobial resistance.

Can J Microbiol, 1995 Jan, 41(1), 75 - 87
Cloning the gene for the heat shock response positive regulator (sigma 32 homolog) from Pseudomonas aeruginosa; Naczynski ZM et al.; A 31 base pair synthetic oligonucleotide based on the genes for the Escherichia coli heat shock sigma factor (rpoH) and the Pseudomonas aeruginosa housekeeping sigma factor (rpoD) was employed in conjunction with the Tanaka et al . (K . Tanaka, T . Shiina, and H . Takahashi, 1988 . Science (Washington, D.C.), 242: 1040-1042) RpoD box probe to identify the location of the rpoH gene in P . aeruginosa genomic digests . This gene was cloned into plasmid pGEM3Z(f+), sequenced, and found to share 67% nucleotide identity and 77% amino acid homology with the rpoH gene and its product (sigma 32) of E . coli . The plasmid containing the rpoH gene complemented the function of sigma 32 in an E . coli rpoH deletion mutant . Furthermore, this plasmid directed the synthesis of a 32-kDa protein in an E . coli S-30 in vitro transcription-translation system . Primer extension studies were used to identify the transcriptional start sites under control and heat-stressed (45 and 50 degrees C) conditions . Two promoter sites were identified having sequence homology to the E . coli sigma 70 and sigma 24 consensus sequences.

FEMS Immunol Med Microbiol, 1995 Jan, 10(2), 139 - 44
Modulation of human polymorphonuclear leukocyte chemotaxis and superoxide anion production by Pseudomonas aeruginosa exoproducts, IL-1 beta and piroxicam; Fontan PA et al.; Whereas addition of 200 ng ml-1 exotoxin A (exoA) did not modify PMNL chemotaxis, 20 U ml-1 human recombinant interleukin-1 beta (hrIL-1 beta) primed polymorphonuclear leukocytes (PMNL) for migration towards Pseudomonas aeruginosa peptide chemotactins (PAPCs) . Piroxicam (100 micrograms ml-1), a non-steroidal anti-inflammatory agent (NSAIA), inhibited PMNL chemotaxis and abolished the priming effect of hrIL-1 beta . Both PAPCs and exoA induced PMNL superoxide anion production, but neither hrIL-1 beta nor piroxicam modified significantly PMNL superoxide anion production induced by PAPCs . The fact that hrIL-1 beta can prime PMNL for chemotaxis towards PAPCs and that piroxicam can abolish activation by primed PMNL are findings relevant to the pharmacological control of lung tissue damage during P . aeruginosa pneumonia.

Vet Microbiol, 1995 Jan, 43(1), 21 - 32
Antibody response against Pseudomonas aeruginosa membrane proteins in experimentally infected sheep; Chin JC et al.; Shedded sheep inoculated epicutaneously with P . aeruginosa and then wetted experimentally by a sprinkler system, rapidly develop a green bacterial stain . This was associated with an outpouring of serious exudates onto the skin surface in the fleecerot lesion site . Histopathological analysis of dermatitic lesions revealed an infiltration of polymorphonuclear leucocytes into the dermis and the formation of a mosaic of microabscesses beneath the sloughed sheets of cornified epithelium . P . aeruginosa if present, was always localized as aggregates at the leading front of the seropurulent exudate and was never observed to invade the dermis . Animals that had been inoculated with P . aeruginosa but kept dry, showed no signs of dermatitis or serological reactivity against the inoculated bacterium . In contrast, sheep that had been inoculated and wetted, reacted serologically against P . aeruginosa whole cells in an enzyme-linked immunosorbent assay (ELISA) . Eleven of 18 sheep were considered to be high-antibody responders and registered an ELISA ratio > 2.5 at one or more time points over the duration of the experiment (14 weeks) . Analysis of ELISA reactivity of fleecerot sheep against fractionated cell envelope proteins of P . aeruginosa showed a preferential antibody response to outer (OMP) rather than inner (IMP) membrane proteins . Immunoblots revealed strong antibody activity against 2 major OMPs-Opr F and Opr H with apparent molecular masses of 39 and 21 kDa respectively . OMPs prepared from sarkosyl-resistant outer membrane vesicles were electrophoretically identical to OMPs prepared by a more rapid and efficient organic phase partitioning procedure (Chin and Dai, 1990) . Although two other OMPs-Opr E (44 kDa) and Opr G (25 kDa) were seen in Coomassie blue-stained SDS-PAGE gels of P . aeruginosa OMPs, they were not reactive with sera from fleecerot affected sheep . It is likely that sheep with high levels of circulating serum antibody against major outer membrane proteins of P . aeruginosa may, in the event of a fleecerot episode, exude such antibodies onto the skin surface . This could provide a strategy for the control of ovine fleecerot by vaccination if highly conserved outer membrane proteins of P . aeruginosa were found to be protective.

Cornea, 1995 Jan, 14(1), 56 - 61
Effect of poloxamer 407 on the adherence of Pseudomonas aeruginosa to corneal epithelial cells; Portoles M et al.; The effect of poloxamer 407 on Pseudomonas aeruginosa adherence to cultured epithelial cells from rabbit corneas was investigated . Three methods of bacterial quantification were used to assess P . aeruginosa adherence: scanning electron microscopy (SEM) counts, radioactivity counts, and viable bacteria counts . Confluent monolayers of rabbit corneal epithelial cells were incubated in agitation for 30 min at room temperature with H3-labeled or nonradiolabeled P . aeruginosa (10(10) bacteria/ml) in a solution of poloxamer 407 {2% or 4% in phosphate-buffered saline (PBS)} or PBS as control . Cell monolayers were washed to remove nonadherent bacteria and fixed with 2.5% glutaraldehyde and processed for SEM or processed for radioactivity counting or for culture on agar plates . The results showed that both solutions of poloxamer 407 inhibited approximately 75% of the bacterial adherence to epithelial cells (p < 0.05) . Similar percentages of bacterial inhibition were obtained using the three methods of bacterial quantification . The use of an antiadherent agent such as poloxamer 407 in eye drops could possibly be a prophylactic approach to P . aeruginosa keratitis.

Am J Perinatol, 1995 Jan, 12(1), 25 - 6
Perinatally acquired Pseudomonas infection: a newly recognized maternal risk factor; Insoft RM et al.; Pseudomonas aeruginosa is a less common but, nonetheless, serious bacterial pathogen implicated in early-onset neonatal sepsis . This report demonstrates perinatal transmission from a mother with a long smoking history who had an oral abscess surgically drained prior to delivery and never received antibiotic therapy . This case emphasizes the need to consider prophylactic antibiotic coverage in similar patients to prevent the morbidity and mortality associated with this type of perinatally acquired infection.

Can J Vet Res, 1995 Jan, 59(1), 76 - 8
A study on the effect of Pseudomonas aeruginosa in semen on bovine fertility; Eaglesome MD et al.; Two experiments were done to demonstrate whether the presence of Pseudomonas aeruginosa in bovine semen could affect fertilization and/or early embryonic development . In the first experiment, superovulated heifers were inseminated with semen naturally contaminated with P . aeruginosa (ADRI 102) or clean semen and seven day-old embryos were collected nonsurgically . The endometrium of treated heifers appeared more sensitive to the flush procedures . In experiment 2, heifers were inseminated at synchronized estrus with semen experimentally contaminated with P . aeruginosa (ADRI 102) and processed in the same way as commercial semen with antibiotics (gentamicin, lincomycin, spectinomycin and tylosin) or processed without antibiotics added . Embryos were recovered at slaughter seven days later . In general, there was no significant reduction in fertility or development of embryos in vitro as a result of relatively high numbers of P . aeruginosa in bovine semen.

Can J Vet Res, 1995 Jan, 59(1), 73 - 5
Comparisons of antibiotic combinations to control Pseudomonas aeruginosa in bovine semen; Eaglesome MD et al.; Raw semen experimentally contaminated with 10(6) Pseudomonas aeruginosa cells per milliliter was processed for use in artificial insemination (AI) using three different antibiotic combinations: a) gentamicin, lincomycin, spectinomycin and tylosin (GLST) directly added to contaminated raw semen followed by dilution with whole milk or egg yolk Tris containing GLST; b) penicillin, streptomycin, lincomycin, spectinomycin and minocycline (PSLSM) in whole milk used to dilute the contaminated raw semen followed by further dilution with glycerolated milk containing PSLSM; and c) penicillin, streptomycin, lincomycin and spectinomycin (PSLS) used with egg yolk Tris diluent in the same way as PSLSM and milk . Diluted semen was incubated at 35 degrees C for 5 or 40 min before cooling commenced . To assess the efficacy of the antibiotics in controlling P . aeruginosa, diluted semen samples were cultured for the organism before and after freezing . The GLST antibiotics added to raw semen and milk reduced the counts of P . aeruginosa before or after freezing . When egg yolk Tris was used, GLST inhibited the organism as indicated by its low growth in culture before freezing and absence of growth from samples after freezing . With PSLSM and PSLS treatments, the organism was recovered in milk and egg yolk Tris processed semen both before and after freezing . However, incubation at 35 degrees C for 40 min prior to cooling, compared to incubation of 5 min, appeared to reduce the bacterial counts after freezing.

J Clin Microbiol, 1995 Jan, 33(1), 37 - 40
Auxotrophic variants of Pseudomonas aeruginosa are selected from prototrophic wild-type strains in respiratory infections in patients with cystic fibrosis; Barth AL et al.; Twenty-four nutritionally dependent (auxotrophic) Pseudomonas aeruginosa strains were isolated from 20 cystic fibrosis (CF) patients and tested for their amino acid requirements . Two different methods were necessary to identify the nutritional status of all isolates . Methionine was the most common single amino acid required (9 of 24 isolates), followed by leucine and arginine or ornithine . In total, a requirement for 12 different compounds or combination of compounds was demonstrated . Auxotrophic and prototrophic pairs of isolates from the same patient were compared by macrorestriction analysis of DNA in pulsed-field gel electrophoresis . Thirteen of 18 pairs analyzed presented identical restriction fragment length polymorphism profiles following digestion of DNA with XbaI . Three of the remaining pairs showed percentage similarities of 77, 91, and 98%, and the profiles of two pairs could not be compared because of the excessive degradation of their DNA . These results suggest that auxotrophic and prototrophic P . aeruginosa isolates colonizing the same CF patient constitute an isogenic group and raise the possibility that auxotrophs are selected from the prototrophic population during the course of pulmonary infection in CF patients.

Antimicrob Agents Chemother, 1995 Jan, 39(1), 132 - 6
Improved efficacy with nonsimultaneous administration of first doses of gentamicin and ceftazidime in vitro; Barclay ML et al.; First doses of aminoglycoside and beta-lactam antibiotics, when used in combination, are usually given simultaneously; however, nonsimultaneous administration may be more efficacious . We used a dynamic in vitro model, which simulates in vivo serum kinetics, to assess the effect of spacing the first doses of gentamicin and ceftazidime used against Pseudomonas aeruginosa ATCC 27853 and two clinical isolates of P . aeruginosa, PA1 and PA2 . The following dose regimens against P . aeruginosa ATCC 27853 were compared: (i) gentamicin given alone, (ii) ceftazidime given alone, (iii) gentamicin and ceftazidime given simultaneously, (iv) gentamicin followed by ceftazidime at 15 or 50 min or at 2, 4, or 8 h, and (v) ceftazidime which was followed by gentamicin at 4 h . The effects of regimen iii and the 4-h interval in regimen iv against PA1 and PA2 were also compared . Initial peak concentrations used were 8 mg/liter for gentamicin and 80 mg/liter for ceftazidime, with drug half-lives of 2.5 and 1.8 h, respectively . Compared with simultaneous administration, nonsimultaneous administration (regimens iv and v) produced greater overall bacterial killing and was associated with a delay in bacterial regrowth (p < 0.005) of up to 6.6 to 8.3 h, regardless of the order in which the drugs were given . The optimal interval between gentamicin and ceftazidime doses, which maximized initial bactericidal effect and the time before regrowth, appeared to be 2 to 4 h.

Pediatr Pulmonol, 1995 Jan, 19(1), 10 - 5
Effects of pseudomonas aeruginosa colonization on lung function and anthropometric variables in children with cystic fibrosis; Pamukcu A et al.; The aim of this study was to evaluate how lung function and growth changed over time in children with cystic fibrosis (CF) colonized with pseudomonas aeruginosa (Pa) compared with those free of the organism . A total of 192 children attended our cystic fibrosis clinic between 1982 and 1992 . Sixty-two of these had three or more annual assessments for lung function, and 117 had three or more annual assessments for height and weight . When lung function was expressed as a standard deviation score (SDS), forced expiratory volume in 1 second (FEV1) and forced expiratory flow at 25% of vital capacity (FEF25) decreased significantly more with respect to height in colonized compared with noncolonized children: FEV1, -0.052 verses -0.015 SDS/cm (P < 0.05); FEF25, -0.060 verses -0.007 SDS/cm (P < 0.05); forced vital capacity (FVC), -0.034 versus -0.012 (NS) . In actual values those patients colonized with Pa increased their FEV1 by 16.4 versus 31.6 mL/cm (P < 0.01); FVC by 28.8 versus 41.4 mL/cm, P < 0.01; and FEF25 by -0.001 versus 0.015 L/s/cm, P < 0.01 . In terms of height, colonized children grew at 5.63 versus 6.96 cm/yr, P < 0.001, and height SDS decreased in colonized compared with noncolonized children at -0.031 verses 0.08 SDS/yr, P < 0.05 . Clinically, most children with CF, with or without Pa, grew within +/- 1 SD of the norm for weight and height . However, in terms of lung function despite optimum pulmonary management colonized children deteriorated significantly faster.

Acta Microbiol Pol, 1995, 44(1), 39 - 46
Mouse model of infected wound; Stepinska M et al.; Local Pseudomonas aeruginosa infection of experimental wound in mice was performed . Suspension of living bacteria was injected beneath skin muscle after skin window had been made on the back of animals . On day 1, 2, 3 and 8 mice were sacrificed, biopsies were taken, number of colony forming units in muscle specimens was estimated and histological examination of skin muscle flaps was performed . The number of bacteria dropped from the very beginning of experiment to the eighth day after infection, still remaining higher than critical value of clinical infection (10(5) CFU per 1 g) . Histologically, healing processes beneath skin started from the third day and on the eight day the wounds were practically healed . Developed model of infected wound may be useful to study the antibacterial agents' effectiveness.

Scand J Infect Dis, 1995, 27(2), 153 - 5
Capnocytophaga (Capnocytophaga ochracea group) bacteremia in hematological patients with profound granulocytopenia; Kristensen B et al.; The clinical and microbiological features of 7 cases of bacteremia due to Capnocytophaga (Capnocytophaga ochracea group) are reported . They were diagnosed during 1991-93 at three hospital clinics . Five patients were < 10 years old and all had hematological disorders, 4 acute lymphoblastic leukemia and 1 each had aplastic anemia, non-Hodgkin lymphoma, and myelodysplastic syndrome . All were profoundly granulocytopenic with an absolute granulocyte count < 0.13 x 10(9)/l, and all but 1 had oral lesions as a possible portal of entry . A favourable response to antibiotic therapy was recorded in all patients but one who, being profoundly granulocytopenic, rapidly succumbed to Pseudomonas aeruginosa septicemia . None of the isolates were beta-lactamase producers . In addition to penicillin the isolates were susceptible to broad-spectrum cephalosporins and ciprofloxacin, but resistant to aminoglycosides.

Scand J Infect Dis Suppl, 1995, 96, 45 - 8
Carbapenem treatment of meningitis; Klugman KP et al.; The carbapenem class of antibacterial agents is highly effective in vitro against the bacterial pathogens causing meningitis . Both imipenem and meropenem penetrate into the cerebrospinal fluid of children with inflamed meninges in the acute phase of meningitis . The wider clinical use of imipenem/cilastatin for the treatment of meningitis has been limited by the high incidence of seizures (up to 33%) in patients not having seizures prior to drug therapy . However, imipenem/cilastatin has been successfully used for the treatment of pneumococcal meningitis following failure of treatment with third-generation cephalosporins . The bacteriological and clinical efficacy of meropenem appears similar to that of cefotaxime in the management of meningitis in children and there was no significant propensity of either meropenem or cefotaxime to cause seizures . Meropenem has also been usefully employed to treat multi-resistant Pseudomonas aeruginosa meningitis . These data suggest that meropenem should be further investigated for use in the treatment of meningitis.

Scand J Infect Dis Suppl, 1995, 96, 24 - 7
The treatment of pulmonary infection in cystic fibrosis; Webb AK; The survival of cystic fibrosis patients has improved through an aggressive, multidisciplinary approach to the therapy of pulmonary sepsis . Intravenous antibiotics play a major role in the care of cystic fibrosis patients, even though it is not possible to achieve persistent bacterial eradication due to the complex microbiology and pathology of these patients . The most important pathogen in older patients is Pseudomonas aeruginosa . The increasing incidence of Pseudomonas cepacia, strains of which can be highly resistant to many antibiotics, also represents an important challenge to the efficacy of antibiotic therapy . Choice of appropriate antimicrobial therapy is hampered by the fact that a single patient may harbour several different pseudomonas phenotypes with variable resistance patterns . Carbapenem antibiotics possess a wide range of activity against most Gram-negative and Gram-positive bacteria and are therefore a useful addition to the antimicrobial armamentarium available to the clinician . The new carbapenem meropenem is well tolerated at high doses by both children and adults . Results from a comparative trial against ceftazidime suggests that meropenem has a place in the management of cystic fibrosis.

Ophthalmologica, 1995, 209(3), 155 - 9
Comparative study on topical and oral administration of ofloxacin for Pseudomonas corneal ulcer in rabbits; Shiao S et al.; Pseudomonas aeruginosa is one of the most frequently isolated microorganisms in bacterial corneal ulcers in Japan and other countries . We evaluated a topical application of 0.3% ofloxacin (OFLX) for Pseudomonas corneal ulcer in rabbits and compared it to the clinical effects of OFLX in topical and oral applications . All corneas in the control groups of both experiments showed total leukoma with a score of 4 on the seventh day . On the other hand, in the preventive experiment, all the scores in the OFLX-treated group were 0, and those in the therapeutic experiment were smaller than those of both the control and oral groups . The results indicate that topical OFLX is effective on Pseudomonas corneal ulcer in rabbits.

Microbios, 1995, 82(331), 87 - 93
Antimicrobial activity of some heterocyclic derivatives; Alsaleh FS et al.; A group of newly synthesized heterocyclic compounds was tested for reactivity against Gram-negative Escherichia coli and Pseudomonas aeruginosa and Gram-positive Staphylococcus aureus and Bacillus subtilis . An attempt was made to correlate the structure of the compounds with their antimicrobial activity.

Can J Microbiol, 1995, 41 Suppl 1, 14 - 21
Formation of novel poly(hydroxyalkanoates) from long-chain fatty acids; Eggink G et al.; Poly(hydroxyalkanoates) (PHAs) were isolated from Pseudomonas aeruginosa 44T1 cultivated on euphorbia oil and castor oil . With the aid of 2-D proton NMR spectra and proton-detected multiple bond coherence NMR spectra the structures of the PHAs were determined . In addition to the usual PHA constituents (C6-C14 3-hydroxy fatty acids), PHAs formed from euphorbia oil contained delta 8,9-epoxy-3-hydroxy-5c-tetradecenoate, and probably delta 6,7-epoxy-3-hydroxydodecanoate and delta 4,5-epoxy-3-hydroxydecanoate . These novel constituents account for approximately 15% of the total amount of monomers and are clearly generated via beta-oxidation of vernolic acid (delta 12,13-epoxy-9c-octadecenoic acid), the main component of euphorbia oil . In PHAs formed from castor oil, 7% of the monomers found were derived from ricinoleic acid (12-hydroxy-9c-octadecenoic acid) . The presence of 3,8-dihydroxy-5c-tetradecenoate was clearly demonstrated . Furthermore, NMR analysis strongly suggested the presence of 3,6-dihydroxydodecanoate, 6-hydroxy-3c-dodecenoate, and 4-hydroxydecanoate.

Zhonghua Zheng Xing Shao Shang Wai Ke Za Zhi, 1995 Jan, 11(1), 29 - 31
{The character and clinical application of glutaraldehyde cross-linked collagen artificial skin}; Wang W et al.; The glutaraldehyde cross-linked collagen artificial skin was made with the modified Burke's method . It is an ideal artificial skin in that animal tests proved that it has the effect of bacteriostasis with strong adhesiveness and good porosity, is capable of decreasing vaporization and the loss of protein and electrolyte, and without rejection reaction . Clinical use of the artificial skin in 30 patients resulted in an average healing time of 24 +/- 3 days of the eschar-excised wound covered with the artificial skin and autograft of skin seeds . It is especially effective in Pseudomonas aeruginosa infected wound and chronic ulcer.

Pneumonol Alergol Pol, 1995, 63(5-6), 259 - 63
{Histology of the respiratory system after exposure to bacterial infection and formalin vapors used to induce experimental bronchospasm}; Sachs K et al.; The purpose of the research was to observe the influence of the bacterial infection and inhalation vapours on the histologic picture of bronchi and lungs in the course on the experimental asthma induced in guinea pigs . The animals were divided into 6 groups . The animals were immunized by ovalbumin . Group I was control and was subjected to inhalations of physiologic salt solutions . Group II was immunized by the soluble of ovalbumin intraperitoneally and was inhaled with the solution of ovalbumin . Group III was subjected only to inhalation of the ovalbumin . Group IV was inhaled with the solution of formalin alternately . Group V experienced only formalin inhalations . Group VI was infected with Pseudomonas aeruginosa strain and inhaled with the solution of ovalbumin . On the histologic examination of the lung tissue the authors found the atrophy of the lymphatic system, the hypertrophy of the mucous membrane and muscular coat of the bronchi, the accumulation of large amount of mucus in their lumen and the exfoliation of the bronchial epithelium.

Vestn Ross Akad Med Nauk, 1995, (9), 57 - 60
{Etiology of the infectious process in pulmonary and mixed forms of mucoviscidosis in children}; Vishniakova LA et al.; The specific features of an infectious process were studied in 150 children treated for mucoviscidosis at the State Pulmonology Research Center, Ministry of Health and Medical Industry of the Russian Federation . Hemophilic bacilli, Staphylococcus aureus, and Pseudomonas aeruginosa were fond to play the leading role in the etiology of the infectious process in the bronchopulmonary system . Pneumococcal infection was first ascertained to be important in the course of the disease . Developmental stages of a pyoinfectious process from the onset of its contamination, acute infection to chronic one were followed up . The most severe, prognostically unfavourable course of the disease was demonstrated to be associated with Pseudomonas aeruginosa infection.

Verh K Acad Geneeskd Belg, 1995, 57(2), 109 - 22
New approaches to Pseudomonas aeruginosa lower respiratory tract infections; Matsumoto K; Since 1973, the occurrence of respiratory tract infections due to P . aeruginosa has increased associated with the development of broad-spectrum penicillins . A clinical entity, diffuse panbronchiolitis (DPB) is a representative disease of chronic P . aeruginosa infections in Japan . In this paper, recent advances of research on pathogenesis and treatments of chronic P . aeruginosa lower respiratory tract infections in our department are reported . We examined sputum from patients with chronic P . aeruginosa infections under the electron microscope . Mucoid type of microcolonies were observed with fibrous matrix of exopolysaccharide . Neutrophils were found to be partially surrounding the microcolony in an attempt to defense . Debris was formed mainly by the destruction of the neutrophils . Most neutrophils were found full of phagocytized debris . These data support that instead of phagocytizing bacteria, neutrophils phagocytized debris and bacteria were not completely eradicated . This might be a factor in the pathogenesis of persistent colonization of P . aeruginosa . In the airways of patients with chronic airway diseases (CAD), neutrophils enhance the recruitment of more neutrophils through the production of neutrophil chemotactic factors such as interleukin-8 (IL-8) and LTB4, perpetuating a cycle of inflammation in the lung . We demonstrated increased levels of IL-8, a chemotactic cytokine, in bronchoalveolar lavage (BAL) fluid from patients with CAD associated with P . aeruginosa infections . We also documented a significant correlation between neutrophil numbers and IL-8 levels or IL-1 beta levels or neutrophil elastase levels in BAL fluids from patients with CAD . By immunohistochemical studies and in vitro data, three major sources of IL-8 in the airways of CAD patients were found to be alveolar macrophages, bronchial epithelial cells, and migrated neutrophils . In Japan, the clinical effectiveness of oral erythromycin (EM) for CAD, including DPB seems to be established, but its pharmacological mechanism remains unclear . In addition, we found a marked decrease of IL-8 levels in BAL fluid from two patients with CAD after treatment with EM . Therefore, we postulated that EM inhibited IL-8 production by stimulated respiratory cells . EM and Roxythromycin, suppressed IL-8 production in Pseudomonas-stimulated neutrophils in a dose-dependent manner . 1 alpha, 25-dihydroxy vitamin D3 also inhibited neutrophil-derived IL-8 . Our data encourage the development of new anti-IL-8 agents against persistent P . aeruginosa lower respiratory tract infections.

Rev Pneumol Clin, 1995, 51(3), 151 - 9
{Pseudomonas aeruginosa and mucoviscidosis}; Lenoir G; Pseudomonas aeruginosa causes episodes of infection which always have a deteriorating effect on the clinical course of cystic fibrosis . The factors and mechanisms involved were discussed on the basis of data in the literature and the experience at the Necker-Enfants-Malades hospital . Preventive measures and indications and modalities for treatment are also presented.

Eur Arch Otorhinolaryngol, 1995, 252 Suppl 1, S59 - 60
Cytokines in neutrophil-dominated airway inflammation in patients with cystic fibrosis; Schuster A et al.; Bronchopulmonary disease in patients with cystic fibrosis (CF) is a paradigm of neutrophil-dominated airway inflammation . We hypothesized that proinflammatory cytokines contribute to a localized neutrophil-dominated inflammatory state as present in CF airways . In a cross-sectional study, we analyzed 63 sputum samples from 33 CF patients for concentrations of the cytokines interleukin-1 alpha (IL-1 alpha), IL-1 beta, IL-8, tumor necrosis factor-alpha (TNF-alpha), and granulocyte-colony stimulating factor (G-CSF) by means of enzyme-linked immunosorbent assay . Furthermore, the activity of neutrophil elastase (NE) in the sputum samples was determined using a specific chromogenic substrate . Compared to sputum samples from 10 healthy controls, there were significantly increased concentrations of IL-1 beta, IL-8 and TNF-alpha in the CF sputum samples . The concentration of IL-8 correlated significantly with NE activity in the CF sputum samples . In CF patients with airways chronically colonized with Pseudomonas aeruginosa, IL-8 concentrations in sputum were significantly enhanced . In glucocorticoid-treated patients, IL-1 alpha and G-CSF sputum concentrations were significantly lower when compared to levels in the other patients . These results show that there are high concentrations of proinflammatory cytokines in CF airways which may contribute to the localized neutrophil-dominated inflammatory state found clinically.

Antimicrob Agents Chemother, 1995 Jan, 39(1), 34 - 9
Macromolecular mechanisms of sputum inhibition of tobramycin activity; Hunt BE et al.; Tobramycin, an aminoglycoside antibiotic, is used in the treatment of Pseudomonas aeruginosa infections in cystic fibrosis patients . Tobramycin bioactivity, however, is antagonized by sputum . Glycoproteins (mucins) and high-molecular-weight DNA make up 2 to 3% (P . L . Masson and J . F . Heremans, p . 412-475, In M . J . Dulfano, ed., Sputum: Fundamentals and Clinical Pathology, 1973) and 3 to 10% (W . S . Chernick and G . J . Barbero, Pediatrics 24:739-745, 1959, and R . Picot, I . Das, and L . Reid, Thorax 33:235-242, 1978) of the dry weight of sputum, respectively . tobramycin binds to both mucins and DNA obtained from sputum (R . Ramphal, M . Lhermitte, M . Filliat, and P . Roussel, J . Antimicrob . Chemother . 22:483-490, 1988) . In vitro, recombinant human DNase (rhDNase) hydrolyzes high-molecular-weight DNA of > 50 kb within sputum to fragments of 2 to 4 kb . Studying dialyzable tobramycin, we examined drug binding to whole sputum and to "mock sputum," which consisted of porcine gastric mucin and calf thymus DNA . We also studied the effects of rhDNase treatments of sputum, mock sputum, and calf thymus DNA on tobramycin binding . We found that treatments of sputum, mock sputum, and calf thymus DNA with rhDNase did not significantly increase the tobramycin bioactivity within the dialysates; surprisingly, sputum binding of tobramycin was increased by rhDNase . We conclude that rhDNase does not increase the bioactivity of tobramycin in sputum.

Infect Immun, 1995 Jan, 63(1), 21 - 6
Biologic activities of antibodies to the neutral-polysaccharide component of the Pseudomonas aeruginosa lipopolysaccharide are blocked by O side chains and mucoid exopolysaccharide (alginate); Hatano K et al.; Virulent strains of Pseudomonas aeruginosa are either of a nonmucoid, lipopolysaccharide (LPS)-smooth or mucoid, LPS-rough phenotype, and immunity to these different variants is efficiently mediated by antibodies specific to O antigens or mucoid exopolysaccharide (also called alginate), respectively . In addition to O side chains and core polysaccharide components, the LPS of P . aeruginosa also contains neutral-polysaccharide components that express antigenic determinants common to many clinical isolates . We evaluated antibodies specific to neutral polysaccharides for the ability to mediate opsonic killing and protective immunity . Antibodies to these antigens mediated opsonic killing of poorly virulent nonmucoid LPS-rough isolates but not of isogenic strains with either a LPS-smooth or a mucoid phenotype . Antibodies to neutral-polysaccharide antigens also failed to protect neutropenic mice from challenge with modest doses of LPS-smooth P . aeruginosa strains (< 10(3) CFU per mouse), whereas O-antigen-specific antibodies were highly protective . Antibodies to neutral polysaccharides deposited significantly (P = 0.002) more C3 onto LPS-rough strains than did antibodies to O side chains, but this situation was reversed when isogenic LPS-smooth strains were tested . Given that protective immunity against P . aeruginosa must be directed against either nonmucoid LPS-smooth strains or mucoid LPS-rough strains, it appears that antibodies specific to neutral-polysaccharide antigens do not protect against P . aeruginosa infection . Lack of protection is likely due to the ability of both O side chains and mucoid exopolysaccharide (alginate) to interfere with the opsonic killing activity of neutral-polysaccharide-specific antibodies.

FEBS Lett, 1994 Dec 19, 356(2-3), 357 - 60
Mechanism of the cytolytic action of Pseudomonas aeruginosa cytotoxin: oligomerization of the cytotoxin on target membranes; Ohnishi M et al.; Pseudomonas aeruginosa cytotoxin (CTX) is thought to be a pore-forming polypeptide of 29 kDa . To study whether CTX assembles into oligomer on target membranes, we solubilized membrane-bound toxin with 1% sodium dodecyl sulfate (SDS) at 25 degrees C and analyzed its molecular size using SDS-polyacrylamide gel electrophoresis and immunoblot analysis . The results indicate that CTX forms a complex of approximately 145 kDa on the surface of erythrocytes and lipid vesicles, and that the complex formation is closely correlated with the toxin-induced permeabilization of target membranes . Thus, CTX may assemble into a pore-forming oligomer on target membranes.

J Mol Biol, 1994 Dec 16, 244(5), 586 - 608
Structure of protocatechuate 3,4-dioxygenase from Pseudomonas aeruginosa at 2.15 A resolution; Ohlendorf DH et al.; Protocatechuate 3,4-dioxygenase catalyzes the aromatic ring cleavage of 3,4-dihydroxybenzoate by incorporating both atoms of molecular oxygen to yield beta-carboxy-cis,cis-muconate . The structure of this metalloenzyme from Pseudomonas aeruginosa (now reclassified as P . putida) has been refined to an R-factor of 0.172 to 2.15 A resolution . The structure is a highly symmetric (alpha beta Fe3+)12 aggregate with a root-mean-square (r.m.s.) difference of 0.18 A among symmetry-related atoms . The tertiary structure of the two polypeptides (alpha and beta) are highly homologous (r.m.s . difference of 1.05 A over 127 C alpha atoms), suggesting that the ancestral enzyme was originally a homodimer with two active sites . Indeed, a non-functional, vestigial active site retains many of the properties of the functional active site but does not bind iron . The coordination geometry of the non-heme iron catalytic cofactor can best be described as trigonal bipyramidal with Tyr447 (147 beta) and His462 (162 beta) serving as axial ligands, and Tyr408 (108 beta), His460 (160 beta) and Wat837 serving as equitorial ligands . The active site environment has a number of basic residues that may promote binding of the acidic substrate . Within the putative active site cavity which is located between alpha and beta chains, five approximately coplanar solvent molecules suggest a position for the planar substrate Trp449 (149 beta), Ile491 (191 beta), defined by Gly14 (14 alpha) and Pro15 (15 alpha) . In this position the guanidino group of Arg457 (157 beta) would be buried by the substrate, suggesting a functional role in catalysis.

J Immunol, 1994 Dec 15, 153(12), 5772 - 80
Differential gene expression for IL-1 receptor antagonist, IL-1, and TNF receptors and IL-1 and TNF synthesis may explain IL-1-induced resistance to infection; Vogels MT et al.; IL-1 pretreatment prolongs survival in lethal infection in normal and in neutropenic mice . We investigated whether this protection occurs by interference with deleterious cytokine effects . The effect of IL-1 pretreatment on concentrations of IL-1 alpha, IL-1 beta, IL-6, and TNF-alpha circulating in vivo and the ex vivo cytokine production capacity of macrophages was assessed in uninfected, non-neutropenic and neutropenic Swiss mice, in Swiss mice infected with Klebsiella pneumoniae (non-neutropenic mice) or Pseudomonas aeruginosa (neutropenic mice), and in neutropenic C3H/HeN and C3H/HeJ mice infected with P . aeruginosa . In Swiss and C3H/HeN mice, IL-1 pretreatment enhanced survival and reduced circulating TNF-alpha and IL-6 as well as LPS-stimulated production of IL-1 alpha and TNF-alpha . In C3H/HeJ mice, a lack of IL-1-induced protection was associated with low cytokine concentrations and production . In contrast, up-regulation of mRNA for the IL-1 receptor antagonist (IL-1Ra) was observed in several organs of IL-1-pretreated mice, suggesting that IL-1Ra could attenuate deleterious IL-1 effects . In addition, IL-1 pretreatment down-regulated steady state mRNA for the type I IL-1R and the type I TNFR in several organs at the time of infection, suggesting desensitization of target cells as an additional mechanism of IL-1-induced protection . We conclude that the IL-1-induced protection is at least partially mediated by down-regulating cytokine production, and that the induction of IL-1Ra and the desensitization of target cells by receptor down-modulation may also contribute to this phenomenon.

FEMS Microbiol Lett, 1994 Dec 15, 124(3), 411 - 7
Methionine-321 in the C-terminal alpha-helix of catabolic ornithine carbamoyltransferase from Pseudomonas aeruginosa is important for positive homotropic cooperativity; Nguyen VT et al.; Pseudomonas aeruginosa has a pair of distinct ornithine carbamoyltransferases . The anabolic ornithine carbamoyltransferase encoded by the argF gene catalyzes the formation of citrulline from ornithine and carbamoylphosphate . The catabolic ornithine carbamoyltransferase encoded by the arcB gene promotes the reverse reaction in vivo; although this enzyme can be assayed in vitro for citrulline synthesis, its unidirectionality in vivo is determined by its high concentration at half maximum velocity for carbamoylphosphate ({S}0.5) and high cooperativity toward this substrate . We have isolated mutant forms of catabolic ornithine carbamoyltransferase catalyzing the anabolic reaction in vivo . The corresponding arcB mutant alleles on a multicopy plasmid specifically suppressed an argF mutation of P . aeruginosa . Two new mutant enzymes were obtained . When methionine 321 was replaced by isoleucine, the mutant enzyme showed loss of homotropic cooperativity at physiological carbamoylphosphate concentrations . Substitution of glutamate 105 by lysine resulted in a partial loss of the sigmoidal response to increasing carbamoylphosphate concentrations . However, both mutant enzymes were still sensitive to the allosteric activator AMP and to the inhibitor spermidine . These results indicate that at least two residues of catabolic ornithine carbamoyltransferase are critically involved in positive carbamoylphosphate cooperativity: glutamate 105 (previously known to be important) and methionine 321 . Mutational changes in either amino acid will affect the geometry of helix H2, which contains several residues required for carbamoylphosphate binding.

EMBO J, 1994 Dec 15, 13(24), 5810 - 7
Crystal structure of AmiC: the controller of transcription antitermination in the amidase operon of Pseudomonas aeruginosa; Pearl L et al.; The crystal structure for the negative regulator (AmiC) of the amidase operon from Pseudomonas aeruginosa has been solved at a resolution of 2.1 A . AmiC is the amide sensor protein in the amidase operon and regulates the activity of the transcription antitermination factor AmiR, which in turn regulates amidase expression . The AmiC structure consists of two domains with an alternating beta-alpha-beta topology . The two domains are separated by a central cleft and the amide binding site is positioned in this cleft at the interface of the domains . The overall fold for AmiC is extremely similar to that for the leucine-isoleucine-valine binding protein (LivJ) of Escherichia coli despite only 17% sequence identity, however, the two domains of AmiC are substantially closed compared with LivJ . The closed structure of AmiC is stabilized significantly by the bound acetamide, suggesting a molecular mechanism for the process of amide induction . The amide binding site is extremely specific for acetamide and would not allow a closed conformation in the presence of the anti-inducer molecule butyramide.

Gene, 1994 Dec 2, 150(1), 87 - 92
ToxR (RegA)-mediated in vitro transcription of Pseudomonas aeruginosa toxA; Walker SL et al.; Exotoxin A (ETA) has been described as a major virulence factor produced by the opportunistic pathogen Pseudomonas aeruginosa . The transcription of the ETA structural gene (toxA) has been shown to be positively regulated by the product of the toxR gene (also called regA) . However, the mechanism by which ToxR regulates toxA transcription is still under investigation . We have expressed toxR in Escherichia coli under the control of the T7 promoter and purified the wild-type ToxR protein . We have also produced ToxR as a fusion protein consisting of the first 12 amino acids of the T7 capsid protein attached to the N terminus of the intact ToxR protein . In the present study we have developed and used an in vitro transcription assay in order to investigate the mechanism of ToxR-mediated transcriptional regulation of toxA . Under the conditions of this in vitro assay toxA transcription requires the toxR product in addition to P . aeruginosa RNA polymerase (RNAP) . Both the native and the T7::ToxR fusion proteins facilitate initiation of toxA transcription in vitro in the presence of Pseudomonas RNAP . Additional studies using (i) specific enzyme-linked immunosorbent assay; (ii) indirect immunoprecipitation; and (iii) gel-filtration chromatography, indicate that ToxR binds to the purified Pseudomonas RNAP and strengthens the possibility that ToxR may be an alternative sigma factor . Furthermore, the ToxR-mediated transcription of toxA is increased approx . threefold in the presence of crude cytoplasmic extracts from P . aeruginosa ToxR+ or ToxR-RegB- strains, indicating that additional factors play a role in the efficient and optimal transcription of toxA.

What Is Bioassay?, What Is Amino Acid?, What Is Bioremediation?, What Is Growth Medium?, What Is Molecular Biology?, r, Microorganism, a, Bacteria, r, Bacteriology, i, Microbe, a, Microbiology, i, Biofilms, c, Haemophilus, o, Yeasts, o, Microorganism, r, Microorganisms, a, Microorganisms, i, Campylobacter, n, Functional genomics, a, Microorganisms, c, Cell suspensions, a, Enterobacteriacea, o, Staphylococcus aureus, i, Microbial, a, Escherichia coli, a, Microorganisms, o, Prokaryotes, n, Bacillus anthracis, i, Streptococcal, a, Escherichia coli, s, Antibiotics, e, Bactericidal

Scientific Publications - Work Done by Microbiology Reader Bioscreen C
Agricultural Microbiology Anaerobic Microbiology Antimicrobial Susceptibility Artificial Atmosphere Bioassay of Antibiotics Biofilm Microbiology Bioreactor Technology Biotechnology Cell Biology Clinical Microbiology Environmental Microbiology Experiments with Yeast	Fermentation Food Microbiology Functional Genomics Gene Technology Growth Media Development Growth Rate and Lag Time Industrial Microbiology Medical/Pharmaceutical Field Microbiological Assay Microbiological Research Microbiology of Cosmetics go to a specific theme...	Military Microbiology Molecular Microbiology Mutagenicity and Genotoxicity Oral Microbiology Patents Postantibiotic Studies Soil Microbiology Spore Microbiology Veterinary Microbiology Waste/Wastewater Treatment Water Microbiology Wine Microbiology

� 2005 Transgalactic Ltd (manufacturer of Bioscreen C software) | Privacy Statement | P.O. Box 1393, 00101 Helsinki, Finland, phone: +358 9 85172920, fax: +358 9 8749481, e-mail: microbiology@bionewsonline.com

Last modified: May 25, 2005