Antimicrob Agents Chemother, 1996 Jan, 40(1), 247 - 52 Influence of antibiotic and E5 monoclonal immunoglobulin M interactions on endotoxin release from Escherichia coli and Pseudomonas aeruginosa; Lamp KC et al.; Recent controversy surrounding the activity of monoclonal antibodies against endotoxin highlights the necessity of identifying all factors associated with increased mortality, one of which is endotoxin concentrations . Antibiotics may induce different patterns of endotoxin release . We compared the release of free endotoxin (in endotoxin units per milliliter) over 6 h and changes in numbers of CFU of exponentially growing Escherichia coli and Pseudomonas aeruginosa (10(6) to 10(7) CFU/ml) cultured in chemically defined endotoxin-free broth combined with pooled human serum and/or 10 micrograms of E5 immunoglobulin M monoclonal antibody per ml . MICs and MBCs were tested in each medium at the same inoculum . The inoculum was exposed to antibiotics at a single fixed multiple of the MIC for each medium (range, two to eight times the MIC) . E5 antibody had no effect on MICs, MBCs, bactericidal activity, or endotoxin release . In the presence of 50% serum, amikacin, ceftazidime, imipenem, and ofloxacin each killed equivalent amounts of E . coli over 6 h; however, ceftazidime induced the highest release of endotoxin . Amikacin and ofloxacin produced the most favorable ratio of endotoxin release to amount of bacterial killing . In the presence of 50% serum, ceftazidime and imipenem reduced the P . aeruginosa inoculum to the greatest extent over 6 h . Although its bactericidal activity was diminished, ofloxacin caused the lowest release of free endotoxin . Imipenem and ofloxacin showed similar low ratios of endotoxin release to bacterial killing . In summary, antibiotic class, presence of serum, and type of organism influenced bactericidal activity and endotoxin release. Antimicrob Agents Chemother, 1996 Jan, 40(1), 61 - 4 Serum bactericidal activity of ceftazidime: continuous infusion versus intermittent injections; Nicolau DP et al.; Since beta-lactam antibiotics have concentration-independent killing, bacterial eradication is a function of the time the serum drug concentration remains above the drug's MIC (T > MIC) . We compared the serum bactericidal titers (SBTs) of ceftazidime given by continuous infusion (CI) or by intermittent bolus dosing (BD) against two clinical isolates each of Pseudomonas aeruginosa and Escherichia coli to determine if CI would allow lower daily dosing while still providing equal bactericidal activity compared with BD . This was an open-labeled, randomized, steady-state, four-way crossover study with 12 healthy volunteers . The ceftazidime regimens were 1 g every 8 h (q8h) BD, 1 g q12h BD, 3 g over 24 h CI, and 2 g over 24 h CI . The areas under the bactericidal curves were calculated by the trapezoidal rule using the reciprocal of the SBT . For all organisms the areas under the bactericidal curves for intermittent versus the CI regimens were the same for equal doses (P > 0.05) . For both strains of E . coli all four regimens provided SBTs of > or = 1:2 over the dosing interval and 100% T > MIC . The 1-g q8h BD and q12h BD regimens provided T > MIC of 82 and 52%, respectively, for both P . aeruginosa isolates (MICs, 4 micrograms/ml) . In comparison, the 2- and 3-g CI regimens always maintained SBTs of > or = 1:2 and T > MIC over the 24-h period as serum drug concentrations were 12.8 +/- 3.0 and 18.2 +/- 4.5 micrograms/ml, respectively . CI optimizes the pharmacodynamic and pharmacoeconomic profile of ceftazidime by providing adequate antibacterial activity over the 24-h dosing period with a reduction in the total daily dose of the antimicrobial agent. Antimicrob Agents Chemother, 1996 Jan, 40(1), 35 - 9 Influence of pH on adaptive resistance of Pseudomonas aeruginosa to aminoglycosides and their postantibiotic effects; Xiong YQ et al.; Adaptive resistance to aminoglycosides in Pseudomonas aeruginosa and other gram-negative bacilli is usually induced by the initial exposure to the drug . We investigated the influence of pH on the adaptive resistance of a clinical P . aeruginosa strain to aminoglycosides in vitro and on their postantibiotic effects . For adaptive resistance, the first-exposure concentrations of both amikacin and netilmicin were one, two, four, and eight times the MIC of each drug and the second-exposure concentrations were two times the MIC of each drug . Adaptive resistance was greater and more prolonged with higher initial aminoglycoside concentrations, and the bactericidal effects of the aminoglycosides were concentration dependent at pH 7.4 . At pH 6.5, the killing rates of amikacin and netilmicin were far lower than those observed at pH 7.4 . At pH 5.5, amikacin and netilmicin exerted practically no bactericidal effect on the P . aeruginosa strain used . However, with media at pH 5.5 and 6.5, adaptive resistance of P . aeruginosa preexposed to amikacin and netilmicin was also clearly exhibited, with the degree of adaptive resistance depending on the bactericidal effects of both drugs on nonpreexposed controls . Maximal adaptive resistance occurred between 0 and 4 h after preexposure . The postantibiotic effects of amikacin and netilmicin against the P . aeruginosa strain were shown to be concentration dependent and were reduced at acidic pHs . No changes in outer and inner membrane proteins occurred during the adaptive-resistance interval. Am J Physiol, 1996 Jan, 270(1 Pt 2), R207 - 16 Changes in regional hemodynamics after nitric oxide inhibition during ovine bacteremia; Lingnau W et al.; We studied the action of nitric oxide synthase (NOS) inhibition on changes in regional blood flow during a continuous infusion of live bacteria . Eighteen ewes were chronically instrumented . After a 7-day recovery period, an infusion of 10(6) colony-forming units/min Pseudomonas aeruginosa was begun . At 24 h, cardiac output increased significantly above baseline in all groups (5.9 +/- 0.4 vs . 8.2 +/- 0.6 l.min 1.m-2), systemic vascular resistance decreased (1,362 +/- 120 vs . 821 +/- 145 dyn.g.cm-5.m-2), and cerebral, cephalic mesenteric, and hindlimb blood flows increased . The animals were then equally and randomly assigned to a bolus of a NOS inhibitor, either 25 mg/kg N omega-nitro-L-arginine methyl ester (L-NAME) or 20 mg/kg N omega-monomethyl-L-arginine (L-NMMA), followed by a continuous infusion of 7 mg.kg-1.min-1 L-NMMA or saline . After NOS inhibition, cardiac index decreased {5.6 +/- 0.1 (L-NAME) and 5.5 +/- 0.4 l.min-1.m-2 (L-NMMA)} and remained significantly decreased for 12 h . 1-NAME decreased carotid and mesenteric blood flows to 64% of the preseptic baseline, and they remained below baseline for 20 h . L-NMMA decreased blood flows only to preseptic baseline values . NOS inhibitors may affect blood flows independently of their hemodynamic effects. Ann Dermatol Venereol, 1996, 123(4), 268 - 70 {Pseudomonas aeruginosa folliculitis after depilation}; De La Cuadra J et al.; INTRODUCTION: Pseudomonas aeruginosa is usually the responsible agent for epidemics of folliculitis acquired in swimming pools and whirlpools . Sporadic cases of Pseudomonas folliculitis after epilation of the legs are well known by dermatologists, but unfrequently reported in the literature . CASE REPORT: We report the case of a 18-year-old female affected of folliculitis of the legs caused by Pseudomonas aeruginosa, after depilation . A meticulous investigation of contaminant sources was performed, and the responsible was the sponge used to wash the legs after depilation . DISCUSSION: Pseudomonas aeruginosa folliculitis after depilation is more frequent than usually reported . A meticulous study of the concomitant source of contamination is essential to avoid recurrent episodes. Chemotherapy, 1996 Jan-Feb, 42(1), 47 - 56 Meropenem resistance in Pseudomonas aeruginosa; Sumita Y et al.; Two genetically distinct classes of meropenem-low-susceptibility Pseudomonas aeruginosa PAO2152 mutants, which arose spontaneously, were isolated . Two meropenem resistance genes, mpmA and mpmB, were mapped near ilvB/C and proC, respectively, on the P . aeruginosa PAO chromosome . The mpmA was thought to be identical to oprD2 because of the cross-resistance to carbapenems and the association with the loss of the outer membrane protein D2 (OprD2) . The mpmB mutation conferred a 4-fold increase in resistance to meropenem, and cross-resistance to various types of antimicrobial agents, e.g . carbenicillin, norfloxacin and chloramphenicol . However, the mpmB mutant was susceptible to imipenem . This mutant still possessed OprD2 and showed increased expression of a 48-kD outer membrane protein, although its profiles of beta-lactamase activity and affinities of penicillin-binding proteins for beta-lactams were indistinguishable from those of the parent strain . The resistance gene mpmB was considered to be an allele of nalB (or cfxB or oprK) from the results of the transductional analysis . The mutation frequency of mpmA:mpmB was in the ratio of 4:1 . The same results were obtained in another clinically isolated P . aeruginosa strain . Meropenem resistance caused by both mpmA and mpmB mutations seemed to be due to the reduction in permeability of antibiotics through the outer membrane . These findings suggest a new pathway for the translocation of meropenem other than that mediated by OprD2 across the outer membrane . Thus, meropenem showed about 4- to 8-fold higher activity than imipenem against OprD2-deficient P . aeruginosa. J Clin Microbiol, 1996 Jan, 34(1), 202 - 4 Comparison of ribotyping and genome fingerprinting of Pseudomonas aeruginosa isolates from cystic fibrosis patients; Bennekov T et al.; Forty Pseudomonas aeruginosa strains, previously characterized by pulsed-field gel electrophoresis, were ribotyped with EcoRI, BamHI, ClaI, and PvuII . Ribotyping with PvuII proved to be as discriminatory as pulsed-field gel electrophoresis with XbaI or DraI while EcoRI and BamHI were not . ClaI contributed further ribotypes, some of which might be due to a transposable element. Jpn J Ophthalmol, 1996, 40(1), 123 - 6 Pharmacokinetics of topically applied ciprofloxacin in rabbit tears; Green LC et al.; Fluoroquinolones provide an important single antibiotic therapy for bacterial keratitis caused by Staphylococcus aureus and numerous gram-negative bacteria, including Pseudomonas aeruginosa . The pharmacokinetics of three ocular fluoroquinolones, norfloxacin (CHIBOXIN, Merck, Sharp & Dohme), ciprofloxacin (CILOXAN, Alcon Laboratories), and ofloxacin (OCUFLOX, Allergan Pharmaceuticals), have been studied in the human eye and in both the normal rabbit eye and rabbit models of keratitis . However, the pharmacokinetics of ciprofloxacin have not been previously studied in the tear film of rabbits . This study was done to determine the pharmacokinetics of topical ciprofloxacin in the rabbit tear film . Two drops of CILOXAN were applied to the eyes of normal rabbits . Tear samples were collected at 5, 10 and 30 minutes, and 1, 2, 4 and 6 hours after topical drug application . Tear samples were analyzed for ciprofloxacin concentrations by HPLC . Ciprofloxacin concentrations reached a peak at 5 minutes, then declined in a manner similar to that reported for norfloxacin and ofloxacin . The ciprofloxacin concentrations in tears were substantially higher throughout the length of the study than the MIC90 for most ocular pathogens including Staphylococcus aureus and Pseudomonas aeruginosa. QJM, 1996 Jan, 89(1), 71 - 6 Bacterial meningitis in patients with nasopharyngeal carcinoma; Tang LM et al.; Bacterial meningitis was found in 12 patients with nasopharyngeal carcinoma, accounting for 0.65% of the 1850 patients with the tumour diagnosed between 1981 and 1994 in our hospital . In 11 patients, the time-lag between diagnosis of cancer and the appearance of infection ranged from 9 months to 11 years (mean 57 months) whereas in one patient it was only 5 days . Three patients developed mixed bacterial meningitis . Cerebrospinal fluid culture for bacteria was positive in six patients . Three patients (25%) were bacteraemic . Gram-negative bacilli, especially Pseudomonas aeruginosa, were the most common pathogens . Age, sex and histopathology were not risk factors for infection . Conditions predisposing to meningitis included intracranial invasion of the tumor, neutropenia, otitis media, and neurosurgical procedures . All but two patients had intracranial tumour invasion and erosion of the base of the skull . Local spread of micro-organism to the meninges was more important than haematogenous spread . The overall mortality in our patients was 66.7%, much higher than in patients without cancer. Bioorg Med Chem, 1996 Jan, 4(1), 43 - 8 Synthesis and in vitro antibacterial activity of spermidine-based mixed catechol- and hydroxamate-containing siderophore--vancomycin conjugates; Ghosh M et al.; The first antibiotic conjugates of vancomycin (1) and siderophore analogues containing spermidine-based catechol ligands (conjugate 11) as well as mixed catechol and hydroxamate ligands (conjugate 13) are described . The design of the conjugates was based on the earlier observation that conjugation of siderophore components to beta-lactam antibiotics induced active iron transport-mediated drug delivery . The novel conjugates (11 and 13) were synthesized by selective acylation of the primary amino group of 1 . Preliminary biological studies indicated that siderophore modified vancomycins lost some activity (4- to 16-fold) against Gram-positive bacteria relative to vancomycin itself, and were generally similar to vancomycin in activity against Gram-negative bacteria under iron-sufficient conditions . However, under iron-depleted conditions which mimic human serum, conjugate 11 displayed enhanced antibacterial activity against an antibiotic hypersensitive strain of Pseudomonas aeruginosa. Schweiz Med Wochenschr Suppl, 1996, 76, 49S - 53S {The role of aminoglycosides in HIV-infected patients}; Opravil M; The frequency of serious infections due to Pseudomonas aeruginosa is increasing among patients with advanced HIV infection; most of the cases are community-acquired . Early diagnosis and adequate treatment--usually with the combination of an anti-pseudomonas beta-lactam antibiotic and an aminoglycoside--have a crucial bearing on prognosis . The good in vitro activity of aminoglycosides against mycobacteria contrasts with the need for intravenous administration and toxicity . In patients with Mycobacterium avium complex infection, amikacin should be given only in the event of difficulty with an oral regimen and only on a temporary basis . Similar rules apply to the treatment of multiresistant tuberculosis with streptomycin or amikacin . Paromomycin is not absorbed by the gastrointestinal tract when taken orally, constitutes the treatment of choice against cryptosporidiosis and serves as an alternative against Entamoeba histolytica and Giardia lamblia . Topical treatment with paromomycin ointment can be used in cutaneous leishmaniasis. Rev Mal Respir, 1996, 13(1), 55 - 60 {Pulmonary deposition of colistin aerosols in cystic fibrosis . Comparison of an ultrasonic nebulizer and a pneumatic nebulizer}; Gagnadoux F et al.; The objective of this study was to quantify the deposition in the lung of a Colistine aerosol generated using a pneumatic nebuliser (Pari LL(R) equipped with a Pari Master, Pari, Germany) and to compare this with the results obtained with an ultrasonic nebuliser (DP100, DP Medical, France) in four subjects suffering from cystic fibrosis being colonised with Pseudomonas aeruginosa . To quantify the pulmonary deposition of the aerosols we have used an indirect isotopic method which consists in assimilating the kinetics of the molecules studied with a serum albumin tagged with Technetium 99m (Tc99mm) and added to a preparation of Colistine . We have previously verified that the addition of a radioactive tracer does not change the normal distribution or dynamics of the medication within the aerosol and the radioactive counter linked to the tracer reflects the mass of the medicament . The pulmonary deposition was expressed as a percentage of the nebuliser dose . A regional analysis of the deposition (central, peripheral, superior and inferior) was carried out and in central deposition compared to the periphery (C/P) and superior compared to inferior (S/I) were calculated . With the DP100 nebuliser the pulmonary deposition of the aerosol was very reproducible from one patient to another, varying only between 9.5 to 14 percent of the nebuliser dose . With the Pari LL the fraction deposited varied more from one patient to another from 5.6 to 27% of the nebuliser dose . In three of four patients, the pulmonary deposition was superior or equal to that obtained with the ultrasonic nebuliser . The patients whose pulmonary deposition was inferior, using the pneumatic nebuliser, was the youngest in the group and co-ordinately poorly the triggering of the nebuliser with the beginning of inspiration . With the two nebulisers, the pulmonary deposition of Colisitine was very heterogeneous throughout the pulmonary parenchyma . The mean of the ratio C/P and S/I obtained in all four patients was identical (1.35 an 0.86 respectively), indicating a deposition of the aerosol which was predominantly central and inferior but was distributed equally in the peripheral parts of the lung . Pneumatic nebulisers offer a reliable alternative notably for domiciliary treatment for Colistine aerosols in patients suffering from cystic fibrosis . In younger patients who have not yet acquired good motor co-ordination, nebulisers which function continuously or are triggered by inspiration seem to be the preferred choice. Nephrol Dial Transplant, 1996 Jan, 11(1), 101 - 8 Induction of interleukin-1 and interleukin-1 receptor antagonist during contaminated in-vitro dialysis with whole blood; Schindler R et al.; BACKGROUND . Previous studies on the permeability of cellulosic and synthetic dialysers for bacterial-derived cytokine-inducing substances gave conflicting results . We tried to study this issue as close to the in-vivo situation as possible . METHODS . An in-vitro dialysis circuit with whole human blood present in the blood compartment of cuprophane (Cup), polysulphone (PS), and polyamide (PA) dialysers was employed; sterile filtrates derived from Pseudomonas aeruginosa cultures were added to the dialysate . We studied the induction of interleukin-1 beta (IL-1 beta) by plasma samples taken from the blood compartment as well as the induction of IL-1 beta and interleukin-1 receptor antagonist (IL-1Ra) in mononuclear cells separated from whole blood after circulation by radioimmunoassay and polymerase chain reaction . RESULTS . Plasma samples from the blood side of all dialysers induced IL-1 beta from non-circulated mononuclear cells after addition of pseudomonas filtrates to the dialysate; the maximal amount of IL-1 beta induced by samples from the blood compartment was 4.8 +/- 1.2 ng/ml for Cup, 1.9 +/- 0.5 ng/ml for PS, and 2.0 +/- 0.6 ng/ml for PA . Mononuclear cells separated after contaminated dialysis will all types of dialysers expressed increased mRNA levels for IL-1 beta and IL-1Ra . Production of IL-1Ra by cells separated after contaminated dialysis was determined after Cup and PS dialysis; there was increased production of IL-1Ra by these cells (Cup, 10.3 +/- 4.2; PS, 7.3 +/- 2.5 ng/ml) compared to cells separated after sterile dialysis (Cup, 5.6 +/- 2.1, P < 0.05; PS, 4.5 +/- 1.1 ng/ml, n.s.) or from non-circulated blood (Cup experiments, 4.7 +/- 1.5, P < 0.05; PS experiments, 4.1 +/- 1.2 ng/ml, n.s.) . CONCLUSIONS . These data suggest penetration of cytokine-inducing substances through both cellulosic and synthetic dialysers . Differences between dialysers may exist regarding extent and time course of penetration . The detection of cytokine mRNA as well as the measurement of IL-1Ra synthesis is a more sensitive marker for the transfer of cytokine-inducing substances through dialyser membranes than the measurement of IL-1 beta protein synthesis. Eur J Clin Microbiol Infect Dis, 1996 Jan, 15(1), 82 - 5 Relationship between outer membrane protein profiles and resistance to ceftazidime, imipenem, and ciprofloxacin in Pseudomonas aeruginosa isolates from bacteremic patients; Gimeno C et al.; Outer membrane protein (OMP) profiles of 122 Pseudomonas aeruginosa isolates recovered from the blood of bacteremic patients were analyzed to relate alterations in the expression of OMPs with porin activity to resistance to imipenem, ceftazidime, and ciprofloxacin . Imipenem-resistant isolates lacked or expressed reduced amounts of porin OprD . In contrast, alterations of OMP profiles were absent in most ceftazidime-resistant isolates . Six of 12 ciprofloxacin-resistant isolates had normal OMP profiles . The remaining isolates showed alterations in the expression of either OprC, OprF, or OprD . In addition, imipenem- and ceftazidime-resistant isolates displayed a beta-lactamase activity compatible with that of a group 1 chromosomal cephalosporinase. FEMS Microbiol Lett, 1996 Jan 1, 135(1), 123 - 9 A mutant of Pseudomonas aeruginosa that lacks c-type cytochromes has a functional cyanide-insensitive oxidase; Ray A et al.; Using transposon mutagenesis and screening for the loss of the ability to oxidise the artificial electron donor N,N,N',N'-tetramethyl-p-phenylenediamine, we have isolated a mutant of Pseudomonas aeruginosa that lacks all c-type cytochromes . This mutant is unable to grow anaerobically with nitrate as a terminal electron acceptor . Analysis of its respiratory function indicates that the mutant has lost its cytochrome c oxidase-terminated respiratory pathway but the cyanide-insensitive oxidase-terminated branch remains functional . Complementation of the mutant by in vivo cloning led to recovery of the wild-type characteristics . These data are consistent with the idea that the cyanide-insensitive respiratory pathway does not contain haem c and that the pathway's terminal oxidase is a quinol oxidase. J Surg Res, 1996 Jan, 60(1), 186 - 92 Pulmonary artery endothelial cell function in swine pseudomonas sepsis; Kadletz M et al.; A substantial increase in pulmonary vascular resistance is associated with sepsis and its sequelae (sepsis syndrome and septic shock) . It is postulated that increased resistance may result from sepsis-induced endothelial cell injury or altered vasoreactivity secondary to pulmonary hypertension . We, therefore, tested the hypothesis that sepsis causes endothelial cell injury and that increased pulmonary pressure alters vascular reactivity . Young swine (15-25 kg) were anesthetized and ventilated . Septic animals received a 1-hr infusion of live Pseudomonas aeruginosa (n = 11), and the control cohort received 0.9% NaCl (n = 7) . All animals were studied for 300 min following the infusion . Postmortem branches of peripheral pulmonary arteries were prepared and tested in a vessel myograph . Ring segments were set to 90% of the circumference the vessels would have at pressures of 20, 30, 40, or 50 mmHg (L90), corresponding to varying pulmonary pressures observed in sepsis . A high dose of potassium was used to obtain maximum possible contraction . Prostaglandin was used to precontract the vessels before testing endothelial cell responses to acetylcholine or bradykinin . Sodium nitroprusside was added at the end of each experiment to obtain maximum possible smooth muscle relaxation . No differences in contraction or relaxation were observed when vessels were set to different pressures (i.e., 20 vs 50 mmHg) . Maximum possible contraction to KCl was significantly decreased after 300 min of sepsis compared to control . No differences between groups were found in contractility to prostaglandin . Bradykinin-induced EDRF/NO production, mediated by BK2 receptors, was not altered in Pseudomonas sepsis (97-98% of total relaxation control and 91-95% septic cohort) . Response to acetylcholine was significantly decreased after sepsis (89-95% of total relaxation control and 51-61% of septic cohort relaxation) . Decreased response to acetylcholine could not be attributed to decreased smooth muscle sensitivity to nitric oxide because the response to bradykinin plus sodium nitroprusside was not altered following sepsis . Vessel reactivity was not altered by increasing pressure settings reflective of changing pulmonary pressure in vivo . These results strongly suggest a sepsis-induced alteration in pulmonary artery endothelial cell receptor sensitivity to acetylcholine, independent of changing pulmonary arterial pressures . This is the first time this decrease has been shown in pseudomonas sepsis. Microbiology, 1996 Jan, 142 ( Pt 1), 79 - 86 Physical mapping of 32 genetic markers on the Pseudomonas aeruginosa PAO1 chromosome; Liao X et al.; The Pseudomonas aeruginosa chromosome was fractionated with the enzymes SpeI and DpnI, and genomic fragments were separated by PFGE and used for mapping a collection of 40 genes . This permitted the localization of 8 genes previously mapped and of 32 genes which had not been mapped . We showed that a careful search of databases and identification of sequences that were homologous to known genes could be used to design and synthesize DNA probes for the mapping of P . aeruginosa homologues by Southern hybridization with genomic fragments, resulting in definition of the locations of the aro-2, dapB, envA, mexA, groEL, oprH, oprM, oprP, ponA, rpoB and rpoH genetic markers . In addition, a combination of distinct DNA sources were utilized as radioactively labelled probes, including specific restriction fragments of the cloned genes (glpD, opdE, oprH, oprO, oprP, phoS), DNA fragments prepared by PCR, and single-stranded DNA prepared from phagemid libraries that had been randomly sequenced . We used a PCR approach to clone fragments of the putative yhhF, sucC, sucD, cypH, pbpB, murE, pbpC, soxR, ftsA, ftsZ and envA genes . Random sequencing of P . aeruginosa DNA from phagemid libraries and database searching permitted the cloning of sequences from the acoA, catR, hemD, pheS, proS, oprD, pyo and rpsB gene homologues . The described genomic methods permit the rapid mapping of the P . aeruginosa genome without linkage analysis. Infect Immun, 1996 Jan, 64(1), 37 - 43 Contribution of specific Pseudomonas aeruginosa virulence factors to pathogenesis of pneumonia in a neonatal mouse model of infection; Tang HB et al.; We sought to identify which Pseudomonas aeruginosa products are involved initiating respiratory tract infection . Defined mutants derived from strain PAO i.e., PAOR1 (lasR),PAO-pmm (algC) (an LPS mutant), and AK1152 (which is Fla- and lacks functional pili), were significantly less virulent than PAO1 in a BALBc/ByJ neonatal mouse model of infection as measured by their abilities to cause acute pneumonia, bacteremia, and death . All three mutants were also less adherent to epithelial cells in an in vitro binding assay . PAOR1 and AK1152 were less able to elicit epithelial production of interleukin-8 than PAO1 . LasR was found to be required for the optimal expression of neuraminidase under conditions of increased osmolarity, as might be present in certain pathological conditions . PAO-exsA::omega,, which lacks exoenzyme S expression, was fully virulent, causing at least as much pathology as PAO1 . The expression of several P . aeruginosa virulence factors appears to be required to establish pulmonary infection in the neonatal mouse. Infect Immun, 1996 Jan, 64(1), 182 - 8 Protease cleavage of iron-transferrin augments pyocyanin-mediated endothelial cell injury via promotion of hydroxyl radical formation; Miller RA et al.; Although a number of bacterium- and host-derived factors have been suggested to contribute to the pathogenesis of Pseudomonas aeruginosa-associated tissue injury, the mechanism remains unclear . We have previously shown that protease modification of iron (Fe)-transferrin generates new iron chelates capable of catalyzing hydroxyl radical (.OH) formation from superoxide and hydrogen peroxide . The latter two oxidants are generated during redox cycling of another P . aeruginosa secretory product, pyocyanin . The lung is a major site of P . aeruginosa infection, with damage to local endothelial cells contributing to the pathogenesis of such infections . Endothelial cells are highly susceptible to oxidant-mediated injury . Therefore, we examined whether pseudomonas elastase-cleaved Fe-transferrin and pyocyanin synergistically enhance pulmonary artery endothelial cell injury via .OH formation . By measuring 51Cr release from cultured endothelial cell monolayers, pseudomonas elastase-cleaved Fe-transferrin significantly augmented cell injury resulting from cellular exposure to sublethal concentrations of pyocyanin . This enhancement in injury was not protease specific, as similar results were obtained with pyocyanin in combination with trypsin- or porcine pancreatic elastase-cleaved Fe-transferrin . The association of iron with the transferrin appeared to be necessary in this process . Supporting the involvement of .OH generation via the Haber-Weiss reaction in augmenting cell injury, catalase, dimethyl thiourea, superoxide dismutase, deferoxamine, and dimethyl sulfoxide significantly inhibited cell injury resulting from exposure to pyocyanin and protease-cleaved Fe-transferrin . Furthermore, spin trapping demonstrated the production of .OH in this cellular system . We conclude that .OH formation resulting from the interaction of protease-cleaved Fe-transferrin and endothelial cell redox cycling of pyocyanin may contribute to P . aeruginosa-associated tissue injury via endothelial cell injury. J Bacteriol, 1996 Jan, 178(2), 511 - 23 Two distinct loci affecting conversion to mucoidy in Pseudomonas aeruginosa in cystic fibrosis encode homologs of the serine protease HtrA; Boucher JC et al.; Conversion to a mucoid, exopolysaccharide alginate-overproducing phenotype in Pseudomonas aeruginosa is associated with chronic respiratory infections in cystic fibrosis . Mucoidy is caused by muc mutations that derepress the alternative sigma factor AlgU, which in turn activates alginate biosynthetic and ancillary regulatory genes . Here we report the molecular characterization of two newly identified genes, algW and mucD, that affect expression of mucoidy . The algW gene, mapping at 69 min, was isolated on the basis of its ability to suppress mucoidy and reduce transcription of the alginate biosynthetic gene algD . The predicted primary structure of AlgW displayed similarity to HtrA (DegP), a serine protease involved in proteolysis of abnormal proteins and required for resistance to oxidative and heat stress in enteric bacteria . Inactivation of algW on the chromosome of the wild-type nonmucoid strain PAO1 caused increased sensitivity to heat, H2O2, and paraquat, a redox cycling compound inducing intracellular levels of superoxide . This mutation also permitted significant induction of alginate production in the presence of subinhibitory concentrations of paraquat . Two new genes, mucC and mucD, were identified immediately downstream of the previously characterized portion (algU mucA mucB) of the gene cluster at 67.5 min encoding the alternative sigma factor AlgU and its regulators . Interestingly, the predicted gene product of mucD also showed similarities to HtrA . Inactivation of mucD on the PAO1 chromosome resulted in conversion to the mucoid phenotype . The mutation in mucD also caused increased sensitivity to H2O2 and heat killing . However, in contrast to algW mutants, no increase in susceptibility to paraquat was observed in mucD mutants . These findings indicate that algW and mucD play partially overlapping but distinct roles in P . aeruginosa resistance to reactive oxygen intermediates and heat . In addition, since mutations in mucD and algW cause conversion to mucoidy or lower the threshold for its induction by reactive oxygen intermediates, these factors may repress alginate synthesis either directly by acting on AlgU or its regulators or indirectly by removing physiological signals that may activate this stress response system. J Bacteriol, 1996 Jan, 178(2), 490 - 5 Cloning and characterization of a gene (msdA) encoding methylmalonic acid semialdehyde dehydrogenase from Streptomyces coelicolor; Zhang YX et al.; A homolog of the mmsA gene of Pseudomonas aeruginosa, which encodes methylmalonic acid semialdehyde dehydrogenase (MSDH) and is involved in valine catabolism in pseudomonads and mammals, was cloned and sequenced from Streptomyces coelicolor . Of the two open reading frames (ORFs) found, which are convergently transcribed and separated by a 62-nucleotide noncoding region, the deduced amino acid sequence of the msdA ORF (homologous to mmsA) is similar to a variety of prokaryotic and eukaryotic aldehyde dehydrogenases that utilize NAD+, particularly to the MmsA protein from P . aeruginosa . No significant similarity was found between the deduced product of ORF1 and known proteins in the databases . An S . coelicolor msdA mutant, constructed by insertion of a hygromycin resistance gene (hyg) into the msdA coding region, lost the MSDH activity and the ability to grow in a minimal medium with valine or isobutyrate as the sole carbon source but grew on propionate . The msdA::hyg mutation was complemented by introduction of the msdA gene on a plasmid . When the S . coelicolor msdA gene was overexpressed in Escherichia coli under the control of the T7 promoter, a protein of 51-kDa, corresponding to the approximate mass of the predicted S . coelicolor msdA product (52.6 kDa), and specific MSDH activity were detected . These results strongly suggest that msdA indeed encodes the MSDH that is involved in valine catabolism in S . coelicolor. J Bacteriol, 1996 Jan, 178(2), 410 - 7 Characterization of genes required for pilus expression in Pseudomonas syringae pathovar phaseolicola; Roine E et al.; Nonpiliated, phage phi 6-resistant mutants of Pseudomonas syringae pv . phaseolicola were generated by Tn5 transposon mutagenesis . A P . syringae pv . phaseolicola LR700 cosmid library was screened with Tn5-containing EcoRI fragments cloned from nonpiliated mutants . The cosmid clone pVK253 complemented the nonpiliated mutant strain HB2.5 . A 3.8-kb sequenced region spanning the Tn5 insertion site contained four open reading frames . The transposon-inactivated gene, designated pilP, is 525 bp long, potentially encoding a 19.1-kDa protein precursor that contains a typical membrane lipoprotein leader sequence . Generation of single mutations in each of the three remaining complete open reading frames by marker exchange also resulted in a nonpiliated phenotype . Expression of this gene region by the T7 expression system in Escherichia coli resulted in four polypeptides of approximately 39, 26, 23, and 18 kDa, in agreement with the sizes of the open reading frames . The three genes upstream of pilP were designated pilM (39 kDa), pilN (23 kDa), and pilO (26 kDa) . The processing of the PilP precursor into its mature form was shown to be inhibited by globomycin, a specific inhibitor of signal peptidase II . The gene region identified shows a high degree of homology to a gene region reported to be required for Pseudomonas aeruginosa type IV pilus production. J Bacteriol, 1996 Jan, 178(1), 85 - 93 Comparative genome mapping of Pseudomonas aeruginosa PAO with P . aeruginosa C, which belongs to a major clone in cystic fibrosis patients and aquatic habitats; Schmidt KD et al.; A physical and genetic map was constructed for Pseudomonas aeruginosa C . Mainly, two-dimensional methods were used to place 47 SpeI, 8 PacI, 5 SwaI, and 4 I-CeuI sites onto the 6.5-Mb circular chromosome . A total of 21 genes, including the rrn operons and the origin of replication, were located on the physical map . Comparison of the physical and genetic map of strain C with that of the almost 600-kb-smaller genome of P . aeruginosa reference strain PAO revealed conservation of gene order between the two strains . A large-scale mosaic structure which was due to insertions of blocks of new genetic elements which had sizes of 23 to 155 kb and contained new SpeI sites was detected in the strain C chromosome . Most of these insertions were concentrated in three locations: two are congruent with the ends of the region rich in biosynthetic genes, and the third is located in the proposed region of the replication terminus . In addition, three insertions were scattered in the region rich in biosynthetic genes . The arrangement of the rrn operons around the origin of replication was conserved in C, PAO, and nine other examined independent strains. J Bacteriol, 1996 Jan, 178(1), 46 - 53 Identification of a novel gene, pilZ, essential for type 4 fimbrial biogenesis in Pseudomonas aeruginosa; Alm RA 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 . We have used transposon mutagenesis to identify loci required for fimbrial assembly or function by screening for mutants that lack the spreading colony morphology characteristic of twitching motility . A subset of these mutants is resistant to fimbria-specific phage . One of these mutants (R270) was found to contain a transposon insertion in a new gene, termed pilZ, which is located on chromosomal SpeI fragment I at about 40 min on the P . aeruginosa map, a position remote from other loci involved in fimbrial biogenesis . pilZ appears to be linked to and possibly forms an operon with a gene, holB*, which is homologous to the gene encoding the delta' subunit of Escherichia coli DNA polymerase III . The product of the pilZ gene is a protein of 118 amino acids (predicted molecular weight, 12,895) which probably has a cytoplasmic location . PilZ appears to be a new class of protein which has not hitherto been represented in the sequence databases, and its function is unknown . Complementation studies indicate that pilZ is able to restore the expression of fimbriae on the surface of P . aeruginosa, as well as twitching motility and sensitivity to fimbria-specific phage when provided in trans to the R270 mutant. J Bacteriol, 1996 Jan, 178(1), 103 - 10 Construction and characterization of pyocin-colicin chimeric proteins; Kageyama M et al.; Chimeric proteins were constructed from pyocin S1 or S2 and colicin E3 or E2, and their characteristics were investigated with special reference to the domain structure . The nuclease domains were interchangeable between two bacteriocins so that a new kind of pyocin, with RNase activity, was created . A bacteriocin which can kill both Pseudomonas aeruginosa and Escherichia coli was also constructed . Investigations with various chimeric proteins indicate that the translocation domain as well as the receptor-binding domain is species specific . Inhibition of lipid synthesis, which is characteristic of pyocins, was also observed with chimeric pyocins carrying the DNase domain of colicin E2 but not with those carrying the RNase domain of E3 . Thus, the DNase domain is responsible for the inhibition of lipid synthesis. Invest Ophthalmol Vis Sci, 1996 Jan, 37(1), 20 - 8 Inhibition of pseudomonal ulceration in rabbit corneas by a synthetic matrix metalloproteinase inhibitor; Barletta JP et al.; PURPOSE . To evaluate the effect of the synthetic matrix metalloproteinase inhibitor, Galardin, on proteases produced by Pseudomonas aeruginosa (PA) and on a rabbit model of Pseudomonas keratitis . METHODS . Protease activities of culture broths from Pseudomonas strains PA-28 and W-186 were characterized in vitro by gelatin zymography and by digestion of Azocasein in the presence and absence of Galardin and the serine protease inhibitor, aprotinin . In a noninfectious in vivo experiment, sterile PA culture broth from W-186 was injected intrastromally into rabbit corneas that were treated topically with Galardin or vehicle, then evaluated clinically and histologically . In an infectious in vivo experiment, rabbit corneas were injected with washed PA-28, then treated topically with Galardin or vehicle and clinically scored . RESULTS . Gelatin zymography of culture broth from W-186 and PA-28 detected two proteases that were both inhibited by Galardin . Galardin reduced the digestion of Azocasein by both PA culture broths by 99%, whereas aprotinin did not significantly reduce the protease activity of PA-28 conditioned broth . Intrastromal injection of sterile W-186 culture broth caused rapid corneal destruction that was prevented by topical treatment with Galardin . Intrastromal injection of washed PA-28 bacteria resulted in progressive corneal melting that was significantly (P < 0.005) delayed, but ultimately not prevented, by topical treatment with Galardin . CONCLUSIONS . Pseudomonal protease activity in culture broth consisted predominantly of metalloproteinases and were effectively inhibited by Galardin in vitro . Topical treatment with Galardin prevented destruction of rabbit corneas by bacterial products present in culture broth, and it delayed corneal destruction after injection of PA bacteria . Galardin may be a useful adjuvant when corneal destruction proceeds despite prompt antibiotic treatment. Am J Respir Crit Care Med, 1996 Jan, 153(1), 325 - 30 Effect of sodium nitroprusside and diethylcarbamazine on hypoxic pulmonary vasoconstriction and regional distribution of pulmonary blood flow in experimental pneumonia; Light RB; The interaction between the effects of indomethacin and sodium nitroprusside or diethylcarbamazine infusion on the efficacy of hypoxic pulmonary vasoconstriction (HPV) and regional distribution of lung blood flow was studied in 15 pentobarbital-anesthetized dogs with acute pneumonia caused by Pseudomonas aeruginosa . After induction of pneumonia central hemodynamics, gas exchange, and regional distribution of lung blood flow (radionuclide-labeled microsphere method) were measured during ventilation of both lungs with oxygen and again with one lung ventilated with nitrogen . The dogs were then randomly assigned to one of three treatment groups: Group I (n = 5) received indomethacin alone (2 mg/kg); Group I-D received indomethacin and diethylcarbamazine (50 mg/kg over 20 min followed by 1 mg/kg/min for the rest of the experiment); Group I-N (n = 5) received indomethacin with sodium nitroprusside to achieve a 20- to 30-mm Hg reduction in mean blood pressure . All measurements were then repeated during both oxygen ventilation and one-lung nitrogen ventilation . In all three groups there was no effect of nitrogen inhalation on distribution of lung blood flow prior to drug treatment, indicating absence of HPV . After treatment, in Group I, perfusion of the pneumonic lung fell from 0.27 +/- 0.08 to 0.10 +/- 0.03 (p < 0.05) of total lung blood flow, and nitrogen ventilation of the left lung reduced perfusion to that region from 0.23 +/- 0.02 to 0.13 +/- 0.02 (p < 0.05), indicating restoration of HPV . In Groups I-D and I-N, HPV was persistently absent or markedly attenuated after treatment, but the percentage of the cardiac output perfusing the pneumonia region fell by an amount similar to that in Group I (0.26 +/- 0.07 to 0.11 +/- 0.04 in Group I-D and 0.35 +/- 0.03 to 0.21 +/- 0.06 in Group I-N, both p < 0.05) . Because these two chemically unrelated pulmonary vasodilators effectively blocked HPV restoration but had no effect on vasoconstriction in the pneumonia region after indomethacin, it is concluded that regional lung blood flow redistribution in pneumonia is mediated by a mechanism other than HPV. Gene, 1995 Dec 29, 167(1-2), 87 - 91 Sequencing and characterization of the downstream region of the genes encoding nitrite reductase and cytochrome c-551 (nirSM) from Pseudomonas aeruginosa: identification of the gene necessary for biosynthesis of heme d1; Kawasaki S et al.; The nirC and nirF genes were identified downstream from nirSM, the structural genes encoding nitrite reductase (NIR) and cytochrome c-551 from Pseudomonas aeruginosa (Pa) . The nirC gene encodes a probable c-type cytochrome with a signal sequence for membrane translocation . The nirF gene codes for a protein of 392 amino acids . A nirF mutant of Pa, constructed by marker exchange mutagenesis, synthesized an inactive NIR protein whose activity was restored by adding purified heme d1 . The mutant strain produced an active NIR, when it was transformed by a broad-host-range plasmid carrying nirF . These results showed that the product of nirF was essential for the biosynthesis of heme d1 in Pa. Gene, 1995 Dec 29, 167(1-2), 81 - 6 Cloning and characterization of the gene (rfc) encoding O-antigen polymerase of Pseudomonas aeruginosa PAO1; Coyne MJ Jr et al.; The lipopolysaccharide (LPS) O-antigen polymerase is the product of the rfc gene . Loss of O-antigen polymerase activity due to mutation in rfc gives rise to a characteristic LPS phenotype known as core-plus-one or semi-rough, wherein the LPS core is capped with a single oligosaccharide unit . Pseudomonas aeruginosa (Pa) AK1401, a derivative of strain PAO1 (serogroup O5), expresses a semi-rough LPS; this mutant phenotype was complemented by a 2.2-kb NsiI-SacI fragment of Pa PAO1 DNA . Sequence analysis of this fragment revealed a 1317-bp open reading frame (ORF) potentially encoding a 438-amino-acid (aa) protein of 48,849 Da . This DNA sequence and the inferred aa sequence contain many of the features of other O-antigen polymerases, including an aberrantly low G + C content (particularly apparent in the high-G + C background of Pa), an unusual codon usage pattern, and a hydrophobicity profile indicative of a membrane protein . A 345-bp fragment internal to the ORF hybridized to genomic DNA from two of ten Pa serogroup strains examined by Southern blot; these two strains express O antigens structurally related to that of strain PAO1. Biochim Biophys Acta, 1995 Dec 27, 1264(3), 249 - 53 Characterization of the Pseudomonas aeruginosa transposable bacteriophage D3112 A and B genes; Ulycznyj PI et al.; The left end DNA of Mu-like transposable bacteriophage D3112 was sequenced from bp 2521 to bp 5483 . Two large open reading frames were identified: ORF A (bp 2539-4611) and ORF B (bp 4626-5378) . ORF A can encode a 690 amino acid, 78 kDa protein which is 44.4% similar to Mu transposase and ORF B can encode a 250 amino acid, 27 kDa protein, which is 46.4% similar to, though 62 amino acids shorter than, the Mu B protein . The cloned D3112 A gene exhibited activity on a mini-D3112-containing plasmid in Pseudomonas aeruginosa. Biochemistry, 1995 Dec 19, 34(50), 16255 - 68 Comparison of NMR solution structures of the receptor binding domains of Pseudomonas aeruginosa pili strains PAO, KB7, and PAK: implications for receptor binding and synthetic vaccine design; Campbell AP et al.; The solution structures of peptide antigens from the receptor binding domains of Pseudomonas aeruginosa strains PAO and KB7 have been determined using two-dimensional 1H NMR techniques . Ensembles of solution conformations for the trans forms of these 17-residue disulfide-bridged peptides have been generated using a simulated annealing procedure in conjunction with distance and torsion angle restraints derived from NMR data . Comparison of the NMR-derived solution structures of the PAO and KB7 peptides, with that previously determined (McInnes et al., 1993) and herein refined for the PAK peptide reveals a common structural motif . All three peptide structures contain a type I beta-turn in the conserved sequence Asp134-X-X-Phe137 and a type II beta-turn in the conserved sequence Pro139-X-Gly-Cys142 . However, the overall folds of the three peptides differ as well as the disposition of the side chains comprising the hydrophobic pockets . The similarities and differences between the structures of the three strains which bind to a common cell surface receptor are discussed in light of their contributions to synthetic vaccine design. FEBS Lett, 1995 Dec 18, 377(2), 145 - 9 A reinvestigation of the covalent structure of Pseudomonas aeruginosa cytochrome c peroxidase; Samyn B et al.; The amino acid sequence of cytochrome c peroxidase from Pseudomonas aeruginosa has been determined using classical chemical degradation techniques combined with accurate mass analysis of all the generated peptides . The sequence obtained is composed of 346 amino acids and confirms the recently published cDNA-derived sequence except at one position {Ridout et al . (1995) FEBS Lett . 365, 152-154} . Based on this sequence, we propose a new model for the binding of the peroxide and the cytochrome electron donor to CCP which is in essence the reverse of the one proposed by Ellfolk et al. Presse Med, 1995 Dec 16-23, 24(39), 1882 - 7 {Cystic fibrosis in adults}; Durieu I et al.; OBJECTIVES: At least half cystic fibrosis patients now reach adulthood . METHODS: We report a population of 61 patients above 18 years of age with the clinical pictures at time of diagnosis and the present clinical status . RESULTS: Thirty-five males and 26 females are aged from 18 to 47 years . Mean age at time of diagnosis was 5 years and 5 months, under 10 years in 80% of patients and above 15 years in 9 patients . Diagnosis was suspected because of pulmonary (2/3) or digestive (1/3) symptoms, insufficient height and weight (1/3) or past family history of cystic fibrosis (1/3) . 37% of patients are homozygotes for delta F508 mutation . Adult patients had a normal height but half of them a body weight under 90% of expected weight . Recurrent pulmonary infections were observed in 95% of patients and 62% have chronically infected sputum with Pseudomonas aeruginosa . These patients had lower weight and a poorer radiological score than patients without pseudomonas . 25% of all patients had chronic respiratory insufficiency . 75% had pancreatic insufficiency and 6 patients diabetes mellitus . Thirteen patients had biological cholestasis and three a liver cirrhosis with portal hypertension . Four women underwent 6 normal pregnancies; semen analysis in five men revealed aspermia . Seven patients died during the last two years because of respiratory insufficiency (4), in the three months after pulmonary transplantation (2), and because of digestive haemorrhage (1) . CONCLUSION: Treatment included daily bronchial drainage, adapted antibiotic treatment and pancreatic enzyme substitution. FEMS Microbiol Lett, 1995 Dec 15, 134(2-3), 137 - 41 Isolation and structure of a new integron that includes a streptomycin resistance gene from the R plasmid of Pseudomonas aeruginosa; Kazama H et al.; A new integron, located on the R plasmid of Pseudomonas aeruginosa, was isolated in Japan . This integron was made up of two conserved segments (5'-and 3'-conserved segments) and a single streptomycin resistance gene as a gene cassette . The structure of this integron resembles that of integron InC, the existence of which was postulated by Bissonnette and Roy (J . Bacteriol . 174, 1248-1257, 1992). Mol Gen Genet, 1995 Dec 15, 249(5), 515 - 25 Toxin A secretion in Pseudomonas aeruginosa: the role of the first 30 amino acids of the mature toxin; McVay CS et al.; Toxin A, one of several virulence factors secreted by the gram-negative bacterium Pseudomonas aeruginosa, is synthesized as a 71 kDa precursor with a typical prokaryotic leader peptide (LP), and is secreted as a 68 kDa mature protein . Evidence from a previous study suggested that a signal required for toxin A secretion in P . aeruginosa may reside within the region defined by the toxin A LP and the first 30 amino acids (aa) of mature toxin A . In the present study, we have used exonuclease Bal31 deletion analysis to examine the specific role of the first 30 aa in toxin A secretion . Four toxA subclones, which encode products containing the toxin A LP and different segments of the 30-residue region fused to a toxin A carboxy-terminal region, were identified . In addition, a gene fusion encoding a hybrid protein consisting of the LP of P . aeruginosa elastase and the final 305 residues of toxin A, was generated . The cellular location of the toxA subclone products in P . aeruginosa was determined by immunoblotting analysis . Toxin A CRMs (cross-reacting material) encoded by different subclones were detected in different fractions of P . aeruginosa including the periplasm and the supernatant . Results from these studies suggest that (1) mature toxin A contains two separate secretion signals one within the N-terminal region and one within the C-terminal region; and (2) the first 30 residues of the mature toxin A form part of the N-terminal secretion signal. Biochim Biophys Acta, 1995 Dec 12, 1272(3), 185 - 9 Scavenging of neutrophil-derived superoxide anion by 1-hydroxyphenazine, a phenazine derivative associated with chronic Pseudomonas aeruginosa infection: relevance to cystic fibrosis; Muller M; The airways of cystic fibrosis patients colonised by Pseudomonas aeruginosa contain the redox active phenazine derivative, 1-hydroxyphenazine (OHP) . As the presence of reactive oxygen species is of importance to tissue damage in cystic fibrosis, OHP was investigated for its ability to reduce molecular oxygen to superoxide . In the presence of NADPH, OHP reduced cytochrome c in a dose-dependent manner . This effect was not inhibited by superoxide dismutase and demonstrates an electron transport role for OHP . The OHP/NADPH system was unable to reduce molecular oxygen to superoxide as judged by an inability to oxidase epinephrine to adrenochrome . However, using lucigenin-enhanced chemiluminescence to detect superoxide, it was found that pathophysiologically relevant concentrations of OHP (5-25 microM) effectively scavenged superoxide from a xanthine/xanthine oxidase system . Similarly, in the presence of OHP, superoxide availability from contact-activated neutrophils was substantially reduced . It is concluded that OHP is an efficient scavenger of superoxide and that electron transfer from superoxide to OHP represents a major mechanism for reduction of OHP in vivo . Reduced OHP has the potential to alter cellular function by participating in the reduction of iron-containing proteins and in this manner contribute to the pathogenesis of P . aeruginosa infection in cystic fibrosis. J Hosp Infect, 1995 Dec, 31(4), 261 - 74 Comparative hygienic surveillance of contamination with pseudomonads in a cystic fibrosis ward over a 4-year period; Bosshammer J et al.; In order to study the long-term distribution and population dynamics of Pseudomonas aeruginosa strains in a highly contaminated hospital environment, two 4-week epidemiological studies, with an interval of 4 years, were carried out in the cystic fibrosis (CF) ward of the Paediatric Clinic of the Medical School of Hannover . Out of the 1948 specimens taken, P . aeruginosa was mainly identified in those from moist, inanimate sources (200 isolates) and hospitalized CF patients (168 isolates) . A correlation was established between the frequency with which P . aeruginosa-positive patients came into contact with hospital facilities and the rate of contamination of these facilities . Rooms reserved for colonized patients were more frequently contaminated with P . aeruginosa in contrast to function rooms in the same ward and the outpatient clinic . However, no direct exchange between patients' strains and the inanimate hospital environment was detected . Out of the 11 genotypes of P . aeruginosa found in 1989 and the 13 genotypes found in 1993, four genotypes were present on both occasions . The most predominant clone was found in tap-water, sinks, wash-basins and creams with an incidence of 34 and 68% in the environmental isolates . The strains seemed to have spread into the adjacent control ward during the 4-year interval . Thus, the separation of colonized and non-colonized patients was undermined through the transfer of strains from a highly contaminated environment without additional hygiene precautions. Mol Microbiol, 1995 Dec, 18(5), 877 - 89 The elastase propeptide functions as an intramolecular chaperone required for elastase activity and secretion in Pseudomonas aeruginosa; McIver KS et al.; Several proteases are secreted by Pseudomonas aeruginosa including elastase, an abundantly secreted neutral zinc-metalloprotease . Elastase (encoded by lasB) is first synthesized with a relatively large propeptide (18 kDa) domain . Here, we present evidence that this propeptide functions as an intramolecular chaperone (IMC) essential for proper maturation of elastase into a hydrolytically active enzyme . An altered elastase allele (lasB6) that encoded an elastase precursor with a precise propeptide deletion was expressed in Escherichia coli, and disrupted cells contained only inactive elastase . However, co-expression of an allele (lasB7) expressing the propeptide as an independent, non-covalently linked protein rescued about one-third of the hydrolytic activity when compared with that obtained with wild-type lasB . Thus, the propeptide was essential for elastase activity and so defined elastase as an IMC-containing protease . We examined the possibility that the propeptide of elastase also plays a role in the localization of the mature protein past the outer bacterial membrane . Expression of lasB6 in P . aeruginosa (lasB delta) in the absence of the propeptide resulted in production of inactive elastase that accumulated within the cell and was not secreted to the culture medium . When lasB7 co-expressed the non-covalently linked propeptide in the same cell with lasB6, efficient secretion was restored and active elastase was then found in the supernatant . Thus, the propeptide was needed for secretion of the mature protein as well as enzymatic activity . This chaperone-like activity of the propeptide appears to involve a direct interaction between the mature and propeptide sequences, and evidence for this was obtained by demonstrating that the non-covalently attached 18 kDa propeptide was co-precipitated with elastase using elastase antibodies . These results are consistent with a hypothesis that the propeptide domain acts as an IMC to control both enzymatic activity and competence for secretion. J Antimicrob Chemother, 1995 Dec, 36(6), 1037 - 41 Resistance to imipenem in Pseudomonas aeruginosa; King A et al.; There was a slow increase in imipenem resistance in Pseudomonas aeruginosa at St . Thomas' Hospital in the 1990s, reaching 3% in 1994 . Of the 94 imipenem-resistant strains, 69% were susceptible to all the other agents tested; 15% were also resistant to one other antibiotic, 5.3% to two, 6.4% to three, 4.3% to four or more compounds . 16% of strains were resistant to at least one other beta-lactam in addition to imipenem . Resistance to other antibiotics was more common among the strains for which imipenem MICs were 4 mg/L than among either imipenem-susceptible or imipenem-resistant (MICs > or = 8 mg/L) strains. Am J Infect Control, 1995 Dec, 23(6), 396 - 8 Pseudomonas aeruginosa outbreak in a neonatal intensive care unit: a possible link to contaminated hand lotion; Becks VE et al.; This article describes a prolonged outbreak of Pseudomonas aeruginosa . The attack rate of this outbreak was 8.5%, with no associated mortality . Hand lotion contaminated with P . aeruginosa was implicated in the transmission of organisms; removal of this hand lotion ended the outbreak . Contaminated hand lotion applied to clean hands of health care workers may have led to direct inoculation of infants at high risk for infection. Zhonghua Jie He He Hu Xi Za Zhi, 1995 Dec, 18(6), 357 - 9, 383 {The clinical significance of Pseudomonas aeruginosa typing and R-plasmid and DNA molecule hybridization in patients with cor pulmonale}; Miao J et al.; 600 strains of Pa were typed from the sputa of patients with cor pulmonale in 8 hospitals . P6 type stoods first, accounting for 25.6%, and the types of PA and the difference in distribution of Pseudomonas from different districts and hospitals, might be used as an important parameter in the epidemiological study . 68 of 150 strains from patients with cor pulmonale contained plasmids . 60 isolates harboring plasmids colonies were hybridized with 6 beta-lactamase and 6 aminoglycoside modifying enzyme resistance gene probes . The experiments demonstrated the 3 strains isolated from patients with nosocomial infection came from one same strain and got a new resistance gene during spreading of nosocomial infection, and these isolates contained homologous sequence with OXA-2 and AAC (6')-1b probes. Intensive Care Med, 1995 Dec, 21(12), 996 - 1002 Use of pulsed-field gel electrophoresis as an epidemiologic tool during an outbreak of Pseudomonas aeruginosa lung infections in an intensive care unit; Talon D et al.; OBJECTIVE: A retrospective study was performed to evaluate the use of DNA polymorphism analysis by pulsed-field gel electrophoresis (PFGE) in assessing the rate of exogenous contamination during an outbreak of Pseudomonas aeruginosa lung infections in an intensive care unit ICU . Another goal was to determine the risk factors, involved in the outbreak.DESIGN: Rectal swabs and tracheal secretions were cultured from all patients upon admission and thereafter once a week throughout their stay in the ICU . Resistance patterns were determined in all P . aeruginosa isolates . We determined the serotypes, pyocin types, plasmid profiles and total DNA macrorestriction patterns for isolates . The restriction fragment length polymorphism (RFLP) of Dra I total DNA digest was studied by PFGE . A retrospective case-control study was performed to determine the risk factors for P . aeruginosa bronchopulmonary colonization . SETTING: The study was carried out in the medical ICU of Besancon University Hospital (France) . RESULTS: The typability, stability and reproducibility of phenotypic markers were not completely satisfactory . Only the RFLP profile satisfied all the criteria for a good typing technique . In four of the 17 patients, P . aeruginosa strains with the same DNA pattern were found . Among the previously reported risk factors for hospital-acquired bronchopulmonary infections, only invasive procedures were determined by multivariate analysis to be significant in our study group . The oropharynx and the bronchial tract are the most likely endogenous sources . CONCLUSION: PFGE-RFLP is a valuable tool for the epidemiologic study of P . aeruginosa . This typing method revealed that exogenous contamination is not always the major source of P . aeruginosa lung infections in mechanically ventilated patients in ICUs. FEMS Immunol Med Microbiol, 1995 Dec, 12(3-4), 187 - 94 Modulation of Pseudomonas aeruginosa adherence to collagen type I and type II by carbohydrates; Stepinska M et al.; This study was undertaken to examine if receptor recognizing saccharides may be involved in the adherence of Pseudomonas aeruginosa to collagen type I and type II . We performed an adherence inhibition assay: cells of individual P . aeruginosa isolates attached to immobilized collagen type I or type II in the presence of monosaccharides, which could serve as blockers of bacterial receptors . Bacterial binding to collagen type I molecules was inhibited to the highest degree by sugar composition D-galactose/D-mannose/N-acetylneuraminic acid (5:5:1), whereas attachment of P . aeruginosa to collagen type II was inhibited by composition d-glucose/D-galactose (1:1) . The same strains which were sensitive to inhibition of binding to collagen type II by both collagen types, were also sensitive to blocking by composition D-glucose/D-galactose . It suggests that saccharides play a role in adherence of P . aeruginosa to collagen type I and type II, and a common receptor for both types of collagen may be available on the surface of P . aeruginosa cells. J Cell Sci, 1995 Dec, 108 ( Pt 12), 3695 - 702 Lung surfactant protein A (SP-A) activates a phosphoinositide/calcium signaling pathway in alveolar macrophages; Ohmer-Schrock D et al.; Lung surfactant protein A (SP-A), the main protein component of lung surfactant which lines the alveoli, strongly enhances serum-independent phagocytosis of bacteria by rat alveolar macrophages . We tested if the effect of SP-A is due to interaction with the macrophages or to opsonization of the bacteria . In phagocytosis assays with fluorescein isothiocyanate labeled bacteria, SP-A had no opsonic effect on Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, but enhanced phagocytosis by acting only on the macrophages . We characterized this activation mechanism . With single cell measurements of fura-2 loaded cells we demonstrate that SP-A raises the intracellular free calcium ion concentration 6 to 8 seconds after addition . This calcium mobilization is dose-dependent in that increased SP-A concentrations lead to a higher percentage of responding cells . Additionally, SP-A leads to a dose-dependent and transient generation of inositol 1,4.5-trisphosphate . Release of intracellular stored calcium by SP-A is a prerequisite for its stimulatory effect on phagocytosis, since SP-A-induced enhancement of phagocytosis can be impaired by prior addition of thapsigargin, a Ca(2+)-ATPase inhibitor that leads to depletion of intracellular calcium stores . We conclude that SP-A activates a phosphoinositide/calcium signaling pathway in alveolar macrophages leading to enhanced serum-independent phagocytosis of bacteria. Zhonghua Hu Li Za Zhi, 1995 Dec, 30(12), 707 - 8 {Experimental test of the bacteriostatic and bactericidal effect of six disinfectants for Pseudomonas aeruginosa}; Zhang XL et al.; Minimal inhibitory concentration (MIC) and minimal bacteriocidal concentration (MBC) of six chosen disinfectants against pseudomonas aeruginosa were tested by the tubular liquid double dilution method . Results showed that 0.06% Chang Kou Jing and 1.5% Supertime are fairly good disinfectants against pseudomonas aeruginosa in infected wound surfaces, postoperative wound dressing, disinfection of surgeons' arms and hands before operation, sterilization of instruments and scrub medical workers' hands after contact with infected patients . The MIC and MBC of the former were 7.5 micrograms/ml and 30.0 micrograms/ml separately, while those of the latter were 11.7 micrograms/ml and 93 micrograms/ml respectively. Vaccine, 1995 Dec, 13(18), 1750 - 3 Ability of synthetic peptides representing epitopes of outer membrane protein F of Pseudomonas aeruginosa to afford protection against P . aeruginosa infection in a murine acute pneumonia model; Hughes EE et al.; Three synthetic peptides (Nos 9, 10 and 18) representing surface-exposed, linear B-cell epitopes of outer membrane protein F of Pseudomonas aeruginosa were each conjugated to the carriers keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA), with the conjugates being used to immunize mice intranasally . Mice were also immunized intranasally with a KLH/BSA carrier control or with a peptide No . 8 conjugate as a negative control . An immunoglobulin G response reactive with P . aeruginosa whole cells was demonstrated by enzyme-linked immunosorbent assay (ELISA) of sera from mice immunized with peptide 9, 10 or 18, whereas no whole-cell reactivity by ELISA was detected in sera from mice immunized with peptide 8 . Upon pulmonary challenge of immunized mice with a Fisher-Devlin immunotype 4 strain of P . aeruginosa, only those mice immunized with peptide 9 or peptide 10 had a significantly greater survival rate compared to control mice immunized with the carriers alone . Peptides 9 (TDAYNQKLSERRAN) and 10 (NATAEGRAINRRVE) have potential for further development as a protective vaccine against P . aeruginosa infections. East Afr Med J, 1995 Dec, 72(12), 766 - 9 Empyema thoracis in adults in Saudi Arabia; Grillo IA et al.; Empyema thoracis in adults is an uncommon disease in the Asir region of Saudi Arabia . In a period of seven years (1988 to 1994), 24 patients were treated for empyema thoracis with a hospital incidence of about 23 patients in 100,000 admissions . The community acquired empyemas are more common and less aggressive in non-Saudi patients (six males and one female) as compared to Saudi patients (11 males and 6 females) whose empyemas are mostly nosocomial with an aggressive course . The peak age in both Saudi and non-Saudi patients is 45 years and 25 years respectively, and the right pleura is more commonly affected than the left pleura in both groups . Risk factors include diabetes mellitus, pulmonary tuberculosis, post-pneumonectomy infections, trauma and pneumonia . The commonest organisms grown are Pseudomonas aeruginosa, Klebsiella species and Staphylococcus aureus, although in almost 40% of the patients the empyemas were sterile . The commonest method of treatment was closed thoracostomy tube drainage. Ann Trop Paediatr, 1995 Dec, 15(4), 269 - 72 Cystic fibrosis in Saudi Arabia: common and rare presentations; Al-Mobaireek KF et al.; The clinical presentations of 12 children with cystic fibrosis seen in King Khalid University Hospital are presented . Ten were of Saudi origin and the other two were African . The mean age of onset of symptoms was 2.3 months, and the mean age at diagnosis was 14.3 months (range 3-48 months) . Seven children were boys and five were girls . All children presented with growth failure, recurrent chest infection and chronic diarrhoea . The parents of 83% of our cases were first-degree relatives . Pseudo-Bartter syndrome was seen in eight children . Sixty-seven per cent of our cases were colonized with Pseudomonas aeruginosa by the time of diagnosis, despite their young age (mean 7 months) . Peripheral neuropathy secondary to vitamin E deficiency, meconium ileus, nasal polyps and gall-stones were present, each in one case . On follow-up, one child died and the other 11 are still alive . We concluded that cystic fibrosis is not rare in Saudi Arabia and that increased awareness of the disease is needed to avoid delay in diagnosis . Efforts should be made to prevent early colonization by Pseudomonas aeruginosa. Eur J Clin Microbiol Infect Dis, 1995 Dec, 14(12), 1057 - 62 Fluorescence-based DNA fingerprinting elucidates nosocomial transmission of phenotypically variable Pseudomonas aeruginosa in intensive care units; Grundmann H et al.; DNA fingerprinting based on automated laser fluorescence analysis of randomly amplified polymorphic DNA (RAPD-ALFA) is a rapid and convenient technique for detecting clonal relatedness of bacterial isolates of nosocomial concern . During an outbreak of Pseudomonas aeruginosa among five patients in a medical intensive care unit, transmission was not suspected because of the phenotypic variability of the initial isolates . However, DNA fingerprinting by RAPD-ALFA and macrorestriction analysis identified a single genotype (strain A) for isolates from three patients and another genotype (strain B) for isolates from the remaining two patients . Strain A isolates displayed three phenotypes defined by different antibiotypes and distinct colony appearance . Retrospective analysis of DNA fingerprints demonstrated that strain A had been transmitted to the index patient one year previously in a different intensive care unit . The study demonstrates that genetic typing approaches are warranted should epidemiological relatedness be identified between phenotypically variant pathogens . Automated laser fluorescence analysis of PCR fingerprints may facilitate routine screening of bacterial isolates for in-house epidemiological surveillance . Antibiograms are an unsuitable approach for the typing of Pseudomonas aeruginosa. Biosci Biotechnol Biochem, 1995 Dec, 59(12), 2358 - 9 A toluene-tolerant mutant of Pseudomonas aeruginosa lacking the outer membrane protein F; Li L et al.; The outer membrane protein profiles of a toluene-tolerant mutant, Pseudomonas aeruginosa, strain PAK103, were compared with those of its parent strain PAO1161 . Protein F (OprF), the most abundant outer membrane protein in the parental strain PAO1161, was missing in the toluene-tolerant strain PAK103 . The absence of OprF may lead to the loss of toluene diffusion across in the outer membrane of the mutant cells. Biokhimiia, 1995 Dec, 60(12), 1964 - 76 {Structure and properties of the common polysaccharide antigen of Pseudomonas aeruginosa bacteria (review)}; Knirel' IuA et al.; The review is devoted to a surface carbohydrate antigen of the bacterium Pseudomonas aeruginosa which is common for the majority of the species and has a lipopolysaccharide nature . The occurrence, detection, isolation, structure, expression on the cell surface, interaction with antibodies, an antibiotic and a bacteriophage as well as the immunotherapeutic potential of the common polysaccharide antigen are discussed. Antimicrob Agents Chemother, 1995 Dec, 39(12), 2822 - 6 In vitro antibacterial activity of DU-6859a, a new fluoroquinolone; Nakane T et al.; The in vitro antibacterial activity of DU-6859a, a new fluoroquinolone, against a wide variety of clinical isolates was evaluated and compared with those of tosufloxacin, ofloxacin, ciprofloxacin, and sparfloxacin . DU-6859a showed potent broad-spectrum activity against gram-positive, gram-negative, and anaerobic bacteria, and its activity was greater than those of the control quinolones . By comparison of MICs at which 90% of strains are inhibited, DU-6859a had potent activity against bacteria resistant to the control quinolones . The time-killing curves of quinolones showed that the number of viable cells decreased rapidly during 2 to 4 of incubation, and regrowth was not seen even after 8 h incubation . At a concentration of four times the MIC, the frequencies of appearance of spontaneous mutants of Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa resistant to DU-6859a were < or = 4.0 x 10(-9) to 1.9 x 10(-8) . The 50% inhibitory concentrations of DU-6859a were 0.86 and 1.05 micrograms/ml for the supercoiling activities of DNA gyrases isolated from E . coli and P . aeruginosa, respectively . The rank order of the 50% inhibitory concentrations observed for both DNA gyrases roughly paralleled the MICs. Antimicrob Agents Chemother, 1995 Dec, 39(12), 2813 - 5 Activity of lipopolysaccharide-binding protein-bactericidal/permeability-increasing protein fusion peptide in an experimental model of Pseudomonas sepsis; Opal SM et al.; A chimeric protein consisting of the N-terminal domain of lipopolysaccharide-binding protein and the C-terminal domain of bactericidal/permeability-increasing protein demonstrated a dose-dependent survival benefit (P = 0.001) and reduced endotoxin levels (P < 0.01) in neutropenic rats with Pseudomonas aeruginosa sepsis . This lipopolysaccharide-binding protein-bactericidal/ permeability-increasing peptide has favorable pharmacokinetics and antiendotoxin properties which may be of value for human sepsis. Antimicrob Agents Chemother, 1995 Dec, 39(12), 2702 - 7 Critical influence of timing of administration of granulocyte colony-stimulating factor on antibacterial effect in experimental endocarditis due to Pseudomonas aeruginosa; Vignes S et al.; The effect of human recombinant granulocyte colony-stimulating factor (hrG-CSF) in rabbits with aortic endocarditis due to Pseudomonas aeruginosa was investigated . hrG-CSF significantly increased the number of polymorphonuclear neutrophils in blood and in cardiac vegetations and the expression of the adhesin molecule CD11b on the surface of polymorphonuclear neutrophils compared with those of animals that had not received hrG-CSF . When treatment was started 72 h after bacterial challenge, hrG-CSF alone had no antibacterial effect and did not enhance the efficacy of ciprofloxacin when used in combination, even with the higher dosing regimen used (50 micrograms/kg of body weight subcutaneously every 12 h for 4 days), in terms of number of positive blood cultures, bacterial counts in vegetations, and survival . In contrast, when treatment was started 30 min prior to bacterial challenge, hrG-CSF (50 micrograms/kg injected every 12 h) decreased bacterial titers in vegetations 72 h later (6.5 +/- 0.9 versus 7.9 +/- 0.9 log10 CFU/g of vegetation for hrG-CSF and controls, respectively; P = prophylactic administration of hrG-CSF did not increase the antibacterial effect of ciprofloxacin . We concluded that the antibacterial effect of hrG-CSF in experimental endocarditis was related to the timing of its administration since hrG-CSF demonstrated a significant but transient antimicrobial effect only when treatment was initiated before bacterial challenge. Antimicrob Agents Chemother, 1995 Dec, 39(12), 2645 - 9 Effects of free and liposome-encapsulated antibiotics on adherence of Pseudomonas aeruginosa to collagen type I; Trafny EA et al.; The adherence of 27 clinical Pseudomonas aeruginosa strains to collagen type I was investigated by using a solid-phase assay . The influence of free antibiotics (amikacin, gentamicin, piperacillin, bacitracin, and polymyxin B) and liposome-entrapped antibiotics (amikacin and polymyxin B) on bacterial attachment to collagen type I was examined . The greatest inhibitory effect was shown for free and liposomal amikacin, which decreased the attachment of 74 and 100% of tested strains, respectively . The mean percent attachment (+/- standard deviation) in the presence of free amikacin was 65.7% (+/- 12.0%) as measured by solid-phase assay . In the presence of liposomal amikacin, the attachment ranged from 17.3% (+/- 6.0%) to 42.1% (+/- 9.4%), depending on the antibiotic solvent . In contrast, polymyxin B, even at a subinhibitory concentration, enhanced attachment of all P . aeruginosa isolates to collagen . Liposomal polymyxin B displayed a protective effect only when the encapsulated drug was of a low concentration . Application of liposome-encapsulated amikacin may be advantageous in injured tissues in which extracellular matrix structures become exposed. Jpn J Antibiot, 1995 Dec, 48(12), 1935 - 8 {Active transport of fosfomycin into cells of Escherichia coli, multidrug-resistant Pseudomonas aeruginosa and Staphylococcus aureus}; Tsuruoka T et al.; We examined effect of fosfomycin (FOM) on growth of Escherichia coli (E . coli) NIHJ JC-2, multidrug-resistant Pseudomonas aeruginosa (P . aeruginosa) PFS80 and Staphylococcus aureus (S . aureus) Smith . FOM inhibited the growth of these organisms at 0.1 approximately 0.5, 10 approximately 50 or 5 approximately 50 micrograms ml, respectively . In E . coli, FOM clearly lyzed the cells at 0.5 microgram/ml . These 3 bacteria incorporated radiolabeled FOM proportionately to incubation time . Intracellular concentration of FOM was estimated by considering the water content of E . coli cells . The ratio of intracellular to extracellular concentrations of FOM was larger than 1 after 5 approximately 10 min . incubation with FOM and reached about 7 after 40 min . FOM was incorporated actively into P . aeruginosa over extracellular concentration after 40-min . incubation on the basis of the above-mentioned water content . It was suggested that FOM was incorporated actively into various bacterial cells, and inhibited efficiently cell wall peptidoglycan synthesis, thus inhibited bacterial growth or lyzed the bacterial cells. J Clin Microbiol, 1995 Dec, 33(12), 3191 - 3 Evaluation of E-Test for determination of antimicrobial MICs for Pseudomonas aeruginosa isolates from cystic fibrosis patients; Marley EF et al.; We determined the E-Test and National Committee for Clinical Laboratory Standards standardized agar dilution MICs of ceftazidime, ciprofloxacin, piperacillin, and tobramycin for Pseudomonas aeruginosa during tests of 100 rough and mucoid P . aeruginosa isolates from cystic fibrosis patients . The levels of agreement (+/- 1 log2 dilution) between quantitative E-Test and agar dilution MIC results were 80, 97, 73, and 89% for ceftazidime, ciprofloxacin, piperacillin, and tobramycin, respectively . Comparison of the results after converting the MIC data to qualitative categories (susceptible, intermediate, and resistant) yielded levels of agreement of 84, 96, 88, and 93% for the same agents, respectively . Of the 39 qualitative discrepancies, 36 were minor and 3 were very major . We conclude that use of the E-Test is easier and more practical than use of the agar dilution method for most laboratories and that the E-Test furnishes results which are at least as accurate as those obtained by the agar dilution method . However, the higher cost of the E-Test method would likely discourage most laboratories from selecting it over disk diffusion for routine antimicrobial susceptibility testing of P . aeruginosa isolates from cystic fibrosis patients. Microbiology, 1995 Dec, 141 ( Pt 12), 3193 - 205 A Bordetella pertussis fepA homologue required for utilization of exogenous ferric enterobactin; Beall B et al.; The bfeA (Bordetella ferric enterobactin) receptor gene was cloned from a Bordetella pertussis chromosomal library by using a screen in Escherichia coli to detect iron-repressed genes encoding exported proteins translationally fused to the E . coli phoA gene . The bfeA gene encoded a protein with a molecular mass of approximately 80 kDa and about 50% amino acid sequence identity to both the fepA- and pfeA-encoded enterobactin receptors of E . coli and Pseudomonas aeruginosa, respectively . Enterobactin prepared from iron-starved E . coli cultures supported growth of B . pertussis and Bordetella bronchiseptica in the presence of the iron chelator ethylenediamine-di-(o-hydroxyphenylacetic acid) (EDDA) . Expression of the bfeA gene was induced by low iron availability, and iron-regulated expression appeared to be dependent upon the presence of the sequence contained within 370 bp upstream of the bfeA structural gene . An internal fragment of the bfeA structural gene and flanking regions were shown by Southern analysis to be highly conserved among Bordetella species . Insertional inactivation of bfeA in both B . pertussis and B . bronchiseptica greatly impaired their ability to grow in the presence of enterobactin and EDDA . These findings suggest that enterobactin produced by other respiratory flora could aid in the colonization of the respiratory tract by Bordetella species. J Am Optom Assoc, 1995 Dec, 66(12), 775 - 9 The effects of short-term contact lens wear on adherence of Pseudomonas aeruginosa to human corneal cells; Harding AS et al.; BACKGROUND: Ulcerative keratitis is the most serious complication of contact lens wear, with the majority of infections being attributed to Pseudomonas aeruginosa . The degree to which bacteria adhere to the cornea may indicate its susceptibility to infection . It has been found that long-term contact lens wear leads to increased adherence of Pseudomonas aeruginosa to corneal epithelial cells . This study examines the effect of short-term contact lens wear . MEANS: Cells were collected using a non-contact corneal irrigation system from a total of 16 subjects fitted with tight and loosely-fitted soft contact lenses . The control group consisted of 10 subjects with no recent contact-lens-wearing experience, eight of whom were later fitted with lenses to form a neophyte group . Six contact lens wearers formed the experienced group . After wearing lenses for a 30-hour period, including overnight, the collected corneal cells were incubated with Pseudomonas aeruginosa, differentially stained with Acridine orange and examined under fluorescence microscopy . RESULTS: The number of bacteria adhering to the cells was determined for the control group, neophyte and experienced groups . The frequency distribution profiles were similar for all groups and did not vary significantly between lens-wearing and non-wearing conditions, between neophyte and experienced wearers, or with lens fit . CONCLUSIONS: Using the technique reported, short-term extended contact lens wear produced minimal detectable changes in the potential of corneal epithelium cells to bind bacteria. Epidemiol Mikrobiol Imunol, 1995 Dec, 44(4), 169 - 70 {The effect of subinhibitory concentrations of quinolone and macrolide antibiotics on production of thermolabile hemolysins in Pseudomonas aeruginosa}; Hybenova D et al.; The authors investigated the effect of subinhibitory concentrations (sub-MICs) of selected quinolones (enoxacine, nalidixic acid, norfloxacine) and macrolid antibiotics (erythromycin, roxitromycin) on the production of thermolabile haemolysin (phospholipase C) of P . aeruginosa . The activity of phospholipase C was markedly reduced by norfloxacine (more than 80%) and enoxacine (75 - 80%) in all sub-MICs . Nalidixic acid inhibited phospholipase C in 1/4 of MIC (75%) . As to macrolid antibiotics the greatest inhibition was caused by erthromycin in 1/4 and 1/8 MIC (more than 90%) . 1/16 MIC of roxitromycin did not influence the activity of the investigated enzyme and other tested sub-MICs caused its reduction to cca one half. Epidemiol Mikrobiol Imunol, 1995 Dec, 44(4), 161 - 4 {Mobilization of genetic determinants of antibiotic resistance in strains of Pseudomonas aeruginosa}; Babalova M et al.; The authors describe a phenomenon of mobilisation of antibiotic resistance from non-transferring strains of P . aeruginosa by cultivation with strains of P . aeruginosa capable to transfer determinants of antibiotic resistance to a susceptible recipient strain, by triparental cross . In this report three strains of P . aeruginosa (No . 282, 283 from Bata's Hospital in Zlin and 76 from Frankfurt University Clinics) are described capable to mobilise for transfer the resistance determinants in four strains of P . aeruginosa (No . 76, 229, 47 and 125 from Frankfurt University Clinics) with multiple antibiotic resistance which itself was not transferable to recipient strains . By an indirect selection method it was assessed, that all six antibiotics (cephalotin-cefazoline, carbenicillin, kanamycin, cefotaxime, ceftazidime and aztreonam), present in resistance spectrum of intermediary recipient strains, were mobilised for transfer . Imipenem and ofloxacin were not mobilised for transfer . In the second and third cycles of transfer the authors confirmed the stability and transferability of the block of six antibiotic resistance determinants, which were not previously transferable. Thorax, 1995 Dec, 50(12), 1301 - 4 Genetic and clinical features of patients with cystic fibrosis diagnosed after the age of 16 years; Gan KH et al.; BACKGROUND--Cystic fibrosis is usually diagnosed in childhood, but a number of patients are not diagnosed until adulthood . The aim of this study was to investigate whether patients diagnosed at an older age had a different genetic constitution, manifestations of disease, and prognosis from those diagnosed at an early age . METHODS--Clinical data and results of lung function tests and DNA analysis of 143 adult patients with cystic fibrosis were entered into a computerised database . Patients diagnosed before their 16th birthday (early diagnosis, ED) were compared with those diagnosed at 16 years of age or older (late diagnosis, LD) . RESULTS--Mean age of diagnosis of the ED group was 4.6 years compared with 27.7 years for the LD group . Mean (SD) percentage predicted pulmonary function was better for the LD group than for the ED group: forced expiratory volume in one second (FEV1) 72.5 (31.1)% and 52.0 (24.8)%, and forced vital capacity (FVC) 89.8 (25.7)% and 71.9 (23.0)%, respectively . Colonisation with Pseudomonas aeruginosa was present in 70% of the ED group and 24% of the LD group . In the ED group 81% had pancreatic insufficiency compared with only 12% of the LD group . None of the LD group was homozygous for delta F508 compared with 58% of the ED group . In the LD group 72% were compound AF508 heterozygotes and 28% had two non-delta F508 mutations . CONCLUSIONS--Among this group of 143 adult patients with cystic fibrosis late diagnosis is caused mainly by delayed expression and mild progression of clinical symptoms . Late diagnosis is associated with milder pulmonary disease, less pancreatic insufficiency, and different cystic fibrosis mutations . Since mortality in cystic fibrosis depends on the progression of pulmonary disease, patients with a late diagnosis have a better prognosis than those diagnosed early. Thorax, 1995 Dec, 50(12), 1246 - 52 Long term effect of erythromycin therapy in patients with chronic Pseudomonas aeruginosa infection; Fujii T et al.; BACKGROUND--Diffuse panbronchiolitis is a chronic infection of the lower respiratory tract common among the Japanese people, with a persistent Pseudomonas aeruginosa infection in the late stage and sustained neutrophil retention in the airways . The long term effect of erythromycin was examined retrospectively in a group of patients with diffuse panbronchiolitis, with and without P aeruginosa infection, and the relationship between drug-induced bacterial clearance and clinical improvement was investigated . METHODS--The history, daily volume of sputum, type of organisms in sputum cultures, pulmonary function tests, arterial blood gas tensions, and chest radiographs were compared in 16 patients with diffuse panbronchiolitis with P aeruginosa infection and 12 without . The total and differential cell counts in the bronchoalveolar lavage (BAL) fluid were compared in 14 of the 28 patients (five of whom were infected with P aeruginosa) before and after 1-12 months of treatment with erythromycin (600 mg/day) . The outcome of treatment in patients showing clearance of organisms on repeated sputum cultures was compared with that in those demonstrating persistence of bacteria in the sputum and patients with normal flora . RESULTS--Erythromycin improved respiratory function and arterial blood gas tensions irrespective of the presence or absence of P aeruginosa in the sputum . Treatment also resulted in a reduction in the BAL fluid total cell count and the percentage of neutrophils in both groups of patients . There were no differences between patients in whom the bacteria cleared and those with persistent bacteria or patients with a normal flora with regard to the degree of improvement of respiratory function, arterial blood gas tensions, and BAL fluid cell composition . CONCLUSION--The results suggest that the efficacy of erythromycin in diffuse pan-bronchiolitis may be due to anti-inflammatory effect, independent of P aeruginosa infection or bacterial clearance. Mol Mar Biol Biotechnol, 1995 Dec, 4(4), 331 - 7 Rhamnolipid biosurfactant enhancement of hexadecane biodegradation by Pseudomonas aeruginosa; Shreve GS et al.; Mutants of Pseudomonas aeruginosa that produce and do not produce rhamnolipid biosurfactant are used to investigate the influence of cell-associated biosurfactant on cellular association with the hydrocarbon-water interface and on hydrocarbon uptake . Rhamnolipid-nonproducing mutant 65E12 of P . aeruginosa is unable to grow in minimal media containing hexadecane as a carbon source in the absence of exogenously added surfactant . Mutant PG201::rhlR grows very slowly in the absence of exogenously added surfactants . Both mutants are deficient in the positive regulatory gene controlling the activation of rhamnolipid synthesis . 65E12 is a double mutant that is also deficient in lipopolysaccharide synthesis . However, growth on hexadecane may be restored to varying degrees when small amounts of purified rhamnolipids or the synthetic anionic surfactant alkyl benzene sulfonate (ABS) is added to the cultures . Rhamnolipid biosurfactant is shown to be approximately 9 times more effective than the structurally similar synthetic anionic surfactant ABS in solubilizing hydrocarbon into the aqueous phase . Physical characteristics of the rhamnolipid and ABS micelles as determined by laser light scattering are described to explain the greater effectiveness of the rhamnolipid in solubilizing hexadecane . The cellular attachment to hydrocarbon-water interfaces and cellular aggregation of the wild-type and mutant strains are examined in the presence and absence of rhamnolipid or synthetic ABS surfactants . Differences in observed hexadecane degradation rates are explained on the basis of emulsified hexadecane concentration, cell surface hydrophobicity, and cellular localization in the culture. J Formos Med Assoc, 1995 Dec, 94(12), 760 - 4 Clinical evaluation of ciprofloxacin ophthalmic solution in the treatment of refractory bacterial keratitis; Tsai AC et al.; Ciprofloxacin is a fluoroquinolone antimicrobial agent inhibiting bacterial DNA gyrase, with good in vitro and in vivo activity against many Gram-positive and Gram-negative ocular pathogens . It has low toxicity, low resistance rate and low minimum inhibitory concentration . The purpose of this study was to evaluate the efficacy of ciprofloxacin in treating bacterial keratitis refractory to conventional therapy . Thirty patients with smear-proven bacterial ulcers were treated by conventional therapy . Of these, cultures were positive in 28 (93.3%) patients . Pseudomonas aeruginosa was isolated in 13 (46.4%) patients, nontuberculous mycobacteria in nine (32.1%) and other bacteria in six (21.4%) . Fifteen patients (50%) were cured with conventional therapy . Four patients (13.3%) underwent surgery due to impending corneal perforation . Eleven patients were shifted to ciprofloxacin therapy because of poor results with conventional treatment . Of these, eight (72.7%) patients were treated successfully . No adverse events were encountered except a white crystalline precipitate in two cases which resolved spontaneously after discontinuation of therapy . In view of its effectiveness and low toxicity, ciprofloxacin should be considered in treating bacterial keratitis which is refractory to conventional therapy. J Bacteriol, 1995 Dec, 177(24), 7194 - 201 Role of the ferric uptake regulator of Pseudomonas aeruginosa in the regulation of siderophores and exotoxin A expression: purification and activity on iron-regulated promoters; Ochsner UA et al.; The cloned Pseudomonas aeruginosa fur (ferric uptake regulator) gene was overexpressed in P . aeruginosa by using a T7 expression system, and the Fur protein (PA-Fur) was purified by using a combination of ion-exchange chromatography and metal affinity chromatography . The DNA binding activity of the PA-Fur protein was confirmed by gel mobility shift assays and DNase I footprints of the synthetic DNA fragment GATAAT GATAATCATTATC, representing a perfect "Fur box" . In addition, it was shown that PA-Fur is capable of binding to promoter and operator determinants of the tightly iron-regulated Escherichia coli fepA-fes enterobactin gene system . The activity of PA-Fur on the promoters of iron-regulated genes involved in the production of two siderophores, pyochelin and pyoverdin, and in the expression of exotoxin A was investigated . Data indicating that the promoters of the pchR gene, encoding a transcriptional activator for pyochelin synthesis, and of the pvdS gene, encoding a positive regulator for pyoverdin production, are specifically recognized by Fur-Fe(II) are presented, suggesting that PA-Fur represses expression of pchR and pvdS during growth in an iron-replete environment . However, neither the promoter region of the gene encoding exotoxin A (toxA) nor the promoters of the regAB operon, required for toxA expression, interacted with high concentrations of purified PA-Fur . These data indicate that iron regulation of exotoxin A production involves additional factors which may ultimately be under the control of PA-Fur. J Bacteriol, 1995 Dec, 177(24), 7019 - 25 Molecular cloning and characterization of a chemotactic transducer gene in Pseudomonas aeruginosa; Kuroda A et al.; A Pseudomonas aeruginosa mutant, defective in taxis toward L-serine but responsive to peptone, was selected by the swarm plate method after N-methyl-N'-nitrosoguanidine mutagenesis . The mutant, designated PCT1, was fully motile but failed to show chemotactic responses to glycine, L-serine, L-threonine, and L-valine . PCT1 also showed weaker responses to some other commonly occurring L-amino acids than did the wild-type strain PAO1 . A chemotactic transducer gene, denoted pctA (Pseudomonas chemotactic transducer A), was cloned by phenotypic complementation of PCT1 . Nucleotide sequence analysis showed that the pctA gene encodes a putative polypeptide of 629 amino acids with a calculated mass of 68,042 . A hydropathy plot of the predicted polypeptide suggested that PctA may be an integral membrane protein with two potential membrane-spanning regions . The C-terminal domain of PctA showed high homology with the enteric methyl-accepting chemotaxis proteins (MCPs) . The most significant amino acid sequence similarity was found in the region of MCPs referred to as the highly conserved domain . The pctA gene was inactivated by insertion of a kanamycin resistance gene cassette into the wild-type gene, resulting in the same observed deficiency in taxis toward L-amino acids as PCT1 . In vivo methyl labeling experiments with L-{methyl-3H}methionine showed that this knockout mutant lacked an MCP with a molecular weight of approximately 68,000. Am J Respir Crit Care Med, 1995 Dec, 152(6 Pt 1), 2150 - 6 Neutrophil-induced lung protection and injury are dependent on the amount of Pseudomonas aeruginosa administered via airways in guinea pigs; Terashima T et al.; We investigated the roles of neutrophils in mediating both the protective effect against bacterial infection and the harmful effect of lung injury induced after the intratracheal instillation of live bacteria . We examined the mortality rate, lung injury, and bacterial clearance following the intratracheal instillation of Pseudomonas aeruginosa in low (10(4) colony-forming units {CFU}) and high doses (10(8) CFU) in normal (control) guinea pigs, others made neutropenic with cyclophosphamide (CPA), and guinea pigs made neutrophilic with recombinant granulocyte colony-stimulating factor (rG-CSF) . Lung injury was assessed by the ratio of the concentration of 125I-labeled albumin in lung tissue to that in plasma (T/P) and the animals' lung weight-to-body weight (LW/BW) ratio . With 10(4) CFU, the CPA group showed an increased T/P ratio of 0.22 +/- 0.03 versus 0.14 +/- 0.01 in the control and 0.11 +/- 0.01 (mean +/- SEM) in the rG-CSF groups (p < 0.01) . Viable bacteria were recovered from bronchoalveolar lavage fluid (BALF) in the CPA group . Neutrophil recruitment was observed in the lungs of animals in the control and rG-CSF groups . With 10(8) CFU, the mortality rate was increased in the rG-CSF group (7 of 10) as compared with the control (0 of 9) and CPA groups (1 of 9) (p < 0.05), which reflected an increased LW/BW (g/kg) ratio (16 +/- 2 versus 12 +/- 1) in the CPA group (p < 0.05) . We conclude that neutrophils protect against lung injury during low-level bacterial challenge, but enhance lung injury and contribute to mortality during high-level bacterial challenge. Am J Respir Crit Care Med, 1995 Dec, 152(6 Pt 1), 2111 - 8 Inflammatory cytokines in cystic fibrosis lungs; Bonfield TL et al.; Chronic pulmonary infection with Pseudomonas aeruginosa continues to be the major cause of morbidity and mortality in cystic fibrosis (CF) . Several characteristics of CF, including the excessive influx of neutrophils into the airways, cachexia, and hyperglobulinemia, could reflect the effects of cytokines, such as interleukin-1 (IL-1), IL-6, IL-8, and tumor necrosis factor (TNF-alpha) . We hypothesized that these pro-inflammatory cytokines, produced by alveolar macrophages in response to pseudomonas and/or other microorganisms, promote the destructive inflammatory process in the lung . We evaluated bronchoalveolar lavage (BAL) fluid and BAL macrophages from 22 CF patients and 13 healthy control (HC) subjects, measuring soluble TNF-alpha, IL-1 beta, IL-6, and IL-8 and the regulatory molecules TNF soluble receptor (TNF-sR), IL-1 receptor antagonist (IL-1Ra), and IL-10 (cytokine synthesis inhibitory factor) . Levels of the proinflammatory cytokines were higher in CF versus HC BAL (p < or = 0.05 for IL-1, TNF, and IL-8; p = 0.06 for IL-6) . In contrast, HC BAL contained significantly more IL-10 than CF BAL (p < 0.05), but TNF-sR and IL-1Ra were similar . Immunocytochemistry demonstrated a higher percentage of CF than control BAL macrophages expressing intracellular cytokines (p < 0.05) . Thus, enhanced macrophage production of proinflammatory cytokines and decreased production of the regulatory molecule IL-10 may have important roles in the pathogenesis of CF lung disease. Am J Respir Crit Care Med, 1995 Dec, 152(6 Pt 1), 1910 - 6 Neutrophil-derived long-lived oxidants in cystic fibrosis sputum; Witko-Sarsat V et al.; We evaluated long-lived oxidant potential in the sputum of patients with cystic fibrosis (CF) by quantitating the methionine-inhibitable, long-lived oxidant fraction of sputum, referred to as the chloramines . Taurine, the preferred amino acid substrate for chloramine formation, and myeloperoxidase (MPO), the chlorinated oxidant-generating enzyme, were also quantitated . As compared with the sputum of asthmatic subjects, the sputum of CF patients contained high concentrations of chloramines along with high levels of taurine and active MPO . A negative correlation between chloramine and taurine was found in the sputum of CF patients . No correlation was found between the density of Pseudomonas aeruginosa and the level of chloramines, taurine, or MPO . In contrast, respiratory parameters (%FEV or %FVC) and a nutritional index correlated positively with chloramine levels, whereas negative correlations were observed with taurine and MPO . In addition, the effect of antibiotic therapy, which significantly increased chloramine and decreased taurine levels, supported a beneficial effect of chloramines on overall clinical status . Our findings support a dual role of long-lived oxidants at the site of airway inflammation in CF, one component of which is their ability to mediate oxidative stress and the other a beneficial effect that may be partly explained by their inhibitory effect on antiprotease defense systems. J Infect Dis, 1995 Dec, 172(6), 1519 - 27 Differences in therapeutic efficacy among cell wall-active antibiotics in a mouse model of gram-negative sepsis; Bucklin SE et al.; The in vivo efficacy of three cell wall-active antibiotics, imipenem, meropenem, and ceftazidime, was compared in mice rendered hypersusceptible to the pathophysiologic effects of lipopolysaccharide by treatment with D-galactosamine . When CF-1 mice were administered Escherichia coli, D-galactosamine, and saline intraperitoneally, an LD50 was achieved at an inoculum of approximately 2 x 10(4) cfu . Administration of antibiotic at 20 mg/kg resulted in significant but widely variable protective efficacy from E . coli lethality among the three antibiotics . At this dose, an approximately 3-fold increase in LD50 was observed with either meropenem or ceftazidime, whereas administration of imipenem resulted in an approximately 8-fold increase in LD50 (P = .0053) . When the dose of antibiotic was decreased to 2 mg/kg, neither meropenem nor ceftazidime could provide measurable protection, whereas imipenem was almost fully protective (P < .002) . These differences in protective efficacy were also noted with experimental Pseudomonas aeruginosa but not Staphylococcus aureus infection. J Bacteriol, 1995 Dec, 177(23), 6718 - 26 Structural elucidation of the lipopolysaccharide core region of the O-chain-deficient mutant strain A28 from Pseudomonas aeruginosa serotype 06 (International Antigenic Typing Scheme); Masoud H et al.; The lipopolysaccharide (LPS) of the Pseudomonas aeruginosa serotype 06 rough-type mutant A28 was isolated by a modified phenol-chloroform-petroleum ether extraction method . Deoxycholate-polyacrylamide gel electrophoresis indicated a single band with mobility similar to that of the complete core region of the wild-type parent serotype 06 (International Antigenic Typing Scheme) strain . Compositional analysis of the LPS indicated that the core oligosaccharide was composed of D-glucose (three units), L-rhamnose (one unit), 2-amino-2-deoxy-D-galactose (one unit), L-glycero-D-manno-heptose (two units), 3-deoxy-D-manno-octulosonic acid (two units), L-alanine (one unit), and phosphate (two units) . Under the mild conditions of hydrolysis with methanolic hydrogen chloride, a 7-O-carbamoyl substituent was observed on the second heptose residue . The glycan structure of the LPS was determined by employing one- and two-dimensional nuclear magnetic resonance spectroscopy and mass spectrometry-based methods with a backbone oligosaccharide that was obtained from the LPS by deacylation, dephosphorylation, and reduction of the terminal glucosamine . On the basis of the results of the present study and our earlier work with the P . aeruginosa 06-derived core-defective mutant R5 (H . Masoud, E . Altman, J . C . Richards, and J . S . Lam, Biochemistry, 33:10568-10578, 1994), a structural model for the complete core oligosaccharide is proposed. Infect Immun, 1995 Dec, 63(12), 4921 - 3 Pyocyanin from Pseudomonas aeruginosa inhibits prostacyclin release from endothelial cells; Kamath JM et al.; Pseudomonas aeruginosa pneumonia causes a vasculitis of small pulmonary arteries . While the fully developed lesion demonstrates vessel wall necrosis, the early lesion is remarkable for preservation of viable endothelium despite vessel wall invasion by bacteria . Pyocyanin, an exoproduct of P . aeruginosa, markedly inhibited prostacyclin production by pulmonary artery endothelial cells without causing cell lysis . Pyocyanin might after vascular homeostasis in the absence of cytolysis. Infect Immun, 1995 Dec, 63(12), 4868 - 76 Cloning and phenotypic characterization of fleS and fleR, new response regulators of Pseudomonas aeruginosa which regulate motility and adhesion to mucin; Ritchings BW et al.; This work has identified two genes (designated fleS and fleR) in Pseudomonas aeruginosa which are highly homologous to members of the subclass of two-component systems involved in transcriptional regulation of a diverse array of genes from sigma 54 promoters . The genes are located upstream from fliE, a flagellar gene of P . aeruginosa, and they are arranged in a putative fleSR operon . FleS has a predicted molecular mass of 43.87 kDa and shows strong homology to histidine kinases which in other two-component systems have been shown to be sensor proteins . FleR has a predicted molecular mass of 51.26 kDa and is homologous to other regulatory proteins that bind to specific upstream activating elements to enhance transcription of genes with sigma 54 promoters . The fleSR system is believed to control both flagellar synthesis and adhesion to mucin . Several lines of evidence are presented . (i) A nonpiliated mutant of P . aeruginosa PAK containing a gentamicin cassette in fleR is nonmotile and nonadhesive . (ii) The fleR mutant regained motility and adhesion when complemented with a wild-type copy of fleR . (iii) A Western blot (immunoblot) of the fleR mutant showed no synthesis of flagellin, and electron microscopy of the fleR mutant confirmed the lack of flagella . Previous work has shown that flagellar mutants with mutations in fliA (sigma 28) or fliC (the structural gene for flagellin) retain adhesion; therefore, these new observations suggest that FleSR regulates both the expression of flagella and the nonpilus adhesin(s) for mucin or that one of the flagellar proteins (other than flagellin) may be responsible for adhesion to mucins. J Trauma, 1995 Dec, 39(6), 1141 - 6; discussion 1146-7 Recombinant human granulocyte colony-stimulating factor treatment improves macrophage suppression of granulocyte and macrophage growth after burn and burn wound infection; Gamelli RL et al.; Granulocyte and macrophage production after burn injury or burn wound infection is significantly reduced and further compromised by endotoxin (ET) . Moreover, the macrophage seems to be the major source of this bone marrow suppression . We sought to determine if recombinant human granulocyte colony-stimulating factor (rhG-CSF), a hematopoietic growth factor that is capable of improving survival after experimental burn wound sepsis, altered postburn macrophage-mediated marrow suppression . Groups of male BDF1 mice (n = 6 to 10) receiving a 15% total body surface area burn +/- infection (B or B + I) with Pseudomonas aeruginosa were injected with 100 ng rhG-CSF twice daily . On day 3, peritoneal-elicited macrophages (5 x 10(6) cells/mL) from either rhG-CSF-treated or control (5% dextrose in water) mice were incubated +/- ET (300 ng/mL) . The resultant macrophage supernatant was added to cultures of target marrow granulocyte-macrophage progenitor cells (GM-CFC) at a volume of 1:10 . The GM-CFC growth as a percentage of cultures not containing macrophage supernatant were compared and reductions in the number of GM-CFC taken as an index of marrow suppression . Macrophages obtained from B and B + I animals reduced target GM-CFC growth, compared with macrophages from normal animals (B vs . normal animals p < 0.05) . In addition, ET-stimulated macrophages induced further bone marrow suppression for all three groups (p < 0.01) . Macrophages from granulocyte colony-stimulating factor-treated animals caused significantly less bone marrow suppression, compared with untreated animals for all groups (p < 0.05 to 0.01).(ABSTRACT TRUNCATED AT 250 WORDS) J Biol Chem, 1995 Nov 24, 270(47), 28246 - 50 Characterization of a phosphoprotein phosphatase for the phosphorylated form of nucleoside-diphosphate kinase from Pseudomonas aeruginosa; Shankar S et al.; Nucleoside-diphosphate kinase (ATP:nucleoside-diphosphate phosphotransferase, EC 2.7.4.6; NDP kinase) is an important enzyme for the maintenance of the correct cellular levels of nucleoside triphosphates (NTPs) and their deoxy derivatives (dNTPs) and is involved in the regulation of cellular development . The enzyme is under the dual control of algR2 and algH in Pseudomonas aeruginosa . We report here the purification and characterization of a protein that dephosphorylates the phosphorylated intermediate form of the P . aeruginosa NDP kinase (Ndk) . Dephosphorylation of Ndk phosphate leads to loss of its enzymatic activity . The 10.1-kDa polypeptide shows 77% homology at the N terminus with the Spo0E phosphatase, identified as a negative regulator of sporulation in Bacillus subtilis and 66% with the human Bax protein, identified as an effector of programmed cell death . The phosphatase termed Npp showed varied specificity toward phosphorylated Ndks from different sources including human erythrocyte Ndk phosphate . Its activity toward other histidine phosphates such as CheA or the alpha-subunit of succinyl-CoA synthetase or phosphoesters such as p-nitrophenyl phosphate was quite limited . Npp was stable at room temperature up to 2 h and required Mg2+ for activity . The presence of a phosphatase capable of dephosphorylating the phosphorylated form of P . aeruginosa Ndk represents an interesting and efficient mode of post-translational modification of an enzyme crucial to cellular development. Biochem Biophys Res Commun, 1995 Nov 22, 216(3), 874 - 80 Biochemical characterization and posttranslational modification of AlgU, a regulator of stress response in Pseudomonas aeruginosa; Schurr MJ et al.; AlgU is homologous to the extreme heat shock sigma factor sigma E from enteric bacteria . In this work, AlgU was overproduced and purified and its function investigated at the biochemical level . AlgU was shown to associate with RNA polymera