Chem Pharm Bull (Tokyo), 1992 Sep, 40(9), 2555 - 6 Biologically active complexes of nickel(II), copper(II) and zinc(II) with Schiff-base ligand derived from the reaction of 2-aminopyridine and pyrrol-2-carboxaldehyde--their synthesis and characterisation; Chohan ZH et al.; A new Schiff-base ligand N-(2'-pyrrylmethylidene)2-aminopyrimidine derived from the reaction of 2-amino pyrimidine and pyrrol-2-carboxaldehyde and its nickel(II), copper(II) and zinc(II) complexes have been synthesised and characterised on the basis of elemental analysis, molar conductance, infrared, electronic and proton nuclear magnetic resonance (1H-NMR) and magnetic susceptibility data . The ligand and its complexes when screened for antibacterial activity against bacterial species such as, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae . In all cases, the activity substantially increased on complexation with metals. Appl Environ Microbiol, 1992 Sep, 58(9), 2886 - 93 Metal regulation of siderophore synthesis in Pseudomonas aeruginosa and functional effects of siderophore-metal complexes; Visca P et al.; Pseudomonas aeruginosa synthesizes two siderophores, pyochelin and pyoverdin, characterized by widely different structures, physicochemical properties, and affinities for Fe(III) . Titration experiments showed that pyochelin, which is endowed with a relatively low affinity for Fe(III), binds other transition metals, such as Cu(II), Co(II), Mo(VI), and Ni(II), with appreciable affinity . In line with these observations, Fe(III) and Co(II) at 10 microM or Mo(VI), Ni(II), and Cu(II) at 100 microM repressed pyochelin synthesis and reduced expression of iron-regulated outer membrane proteins of 75, 68, and 14 kDa . In contrast, pyoverdin synthesis and expression of the 80-kDa receptor protein were affected only by Fe(III) . All of the metals tested, except Mo(VI), significantly promoted P . aeruginosa growth in metal-poor medium; Mo(VI), Ni(II), and Co(II) were more efficient as pyochelin complexes than the free metal ions and the siderophore . The observed correlation between the affinity of pyochelin for Fe(III), Co(II), and Mo(VI) and the functional effects of these metals indicates that pyochelin may play a role in their delivery to P . aeruginosa. Pediatr Pulmonol, 1992 Sep, 14(1), 44 - 51 IgG subclass antibody responses to alginate from Pseudomonas aeruginosa in patients with cystic fibrosis and chronic P . aeruginosa infection; Pressler T et al.; Chronic bronchopulmonary infection with alginate-producing, mucoid Pseudomonas aeruginosa is characteristically associated with cystic fibrosis (CF) . A significant correlation between the antibody response to alginate and poor lung function has been reported . Enzyme-linked immunosorbent assays were developed for the quantitation of human IgG1, IgG2, IgG3, and IgG4 antibodies to P . aeruginosa alginate . We investigated the pattern of IgG subclass antibodies against P . aeruginosa alginate in serum of patients with CF, others with chronic P . aeruginosa infection, and healthy controls . Healthy controls and patients with CF, before they acquired P . aeruginosa infection, had no or very low titers of antibodies against P . aeruginosa alginate . The latter with chronic infection had significantly higher antibody levels than all others groups, including patients with chronic P . aeruginosa infection but no CF . CF with chronic P . aeruginosa infection led to an inverse correlation between lung function parameters and levels of IgG3 and IgG4 . Fifty-seven patients with CF have been followed for an average of 12 years with multiple antibody assays covering the preinfection, early, and late stage of chronic infection . All of them developed IgG1 and IgG3 antibodies to alginate at the start of infection . IgG2 antibodies developed later and showed only a slow increase during the chronic infection . Patients who died had significantly higher IgG2 anti-alginate antibody levels than other investigated groups . Elevated levels of IgG2 and IgG3 antibodies to P . aeruginosa alginate are a sign of poor prognosis in CF. J Antibiot (Tokyo), 1992 Sep, 45(9), 1526 - 32 Role of the aminothiadiazolyl group in the antipseudomonal activity of cefclidin; Watanabe N et al.; Cefclidin (E1040), which has an aminothiadiazolyl group in the 7 beta-side chain, showed about four-fold higher activity against Pseudomonas aeruginosa than its aminothiazolyl counterpart . Cefclidin had lower affinity and a higher Vmax value for the chromosomal type I beta-lactamase (cephalosporinase) from P . aeruginosa than its aminothiazolyl counterpart . No differences between the affinities of both compounds for the most of the sensitive essential PBPs were observed . Hydrophilicity of cefclidin was higher than that of its counterpart . The antipseudomonal activity of cefclidin, which was increased by the introduction of the aminothiadiazolyl group, was suggested to have resulted mainly from higher resistance to cephalosporinase hydrolysis at pharmacologically relevant low concentrations due to its low affinity for cephalosporinase, and secondarily from good penetration of cefclidin through the outer membrane due to increased hydrophilicity. Cornea, 1992 Sep, 11(5), 393 - 7 Fibrin-enmeshed tobramycin liposomes: single application topical therapy of Pseudomonas keratitis; Frucht-Perry J et al.; Treatment of bacterial keratitis requires frequent application of topical antibiotics . We studied the efficacy of a single topical administration of tobramycin incorporated in large multivesicular liposomes and enmeshed in a fibrin sealant on rabbit corneas infected with Pseudomonas aeruginosa . One cornea each of 25 New Zealand albino rabbits was infected with P . aeruginosa . Twenty-four hours later, the animals were randomly divided into five groups of five . Group A received single hourly drops (50 microliters) of fortified tobramycin (14.5 mg/ml, total of 17.4 mg) . Group B received a single topical application of 3.5 mg tobramycin, in 0.1 ml multivesicular liposomes, enmeshed in a fibrin sealant with an overlaying bandage contact lens . Group C was treated in the same manner as group B without the addition of fibrin sealant . Groups D and E served as nondrug-treated controls, with group D receiving topical fibrin-enmeshed liposomes devoid of tobramycin and group E receiving hourly topical balanced salt solution (BSS) drops . All animals were killed 24 h after initiation of therapy . Significantly fewer colonies of Pseudomonas were present in corneas of all three treated groups, as compared with the two nondrug-treated control groups (p less than 0.02) . There were significantly fewer colonies of Pseudomonas in groups A and B as compared with group C (p less than 0.02) . No significant difference was noted between a single administration of topical fibrinen-meshed tobramycin-encapsulated liposomes (group B) and 24 doses of hourly fortified topical tobramycin (group A, p greater than 0.05) . Tobramycin-encapsulated megaliposomes may serve as a useful adjunct in treatment of Pseudomonas keratitis. Acta Paediatr, 1992 Sep, 81(9), 720 - 2 Noma neonatorum: an unusual case of noma involving a full-term neonate; Lin JY et al.; Noma neonatorum is a gangrenous process that occurs in the oral, nasal or anal area and occasionally the eyelids and scrotum of the newborn . The disease is caused by Pseudomonas aeruginosa and usually affects premature ill babies during the first few weeks of life . A full-term neonate with nasal and scrotal noma is uncommon and is therefore reported. Arch Dis Child, 1992 Sep, 67(9), 1086 - 8 The value of serum IgG titres against Pseudomonas aeruginosa in the management of early pseudomonal infection in cystic fibrosis; Brett MM et al.; We report the results of a clinical trial . Patients enrolled had serum IgG titres against Pseudomonas aeruginosa above the control range . Assignment to the observation or treatment group was by minimisation . Significant signs or symptoms in any patient prompted antipseudomonal treatment . In addition, the treatment group received antipseudomonal treatment at intervals of four months until the serum IgG titre returned to the control range . P aeruginosa was isolated intermittently from patients in the main trial . Nineteen patients were enrolled (12 observation, seven treatment) . After one year in the trial changes in parameters studied, including forced expiratory volume in one second, IgG titre, serum IgG concentrations, and frequency of P aeruginosa isolation had improved in the treated group and worsened in the observation group. Antimicrob Agents Chemother, 1992 Sep, 36(9), 1951 - 7 Adaptive resistance following single doses of gentamicin in a dynamic in vitro model; Barclay ML et al.; Adaptive resistance is a phenomenon recently described for Pseudomonas aeruginosa and other gram-negative bacilli following exposure to aminoglycoside antibiotics . It is a reversible form of resistance which develops within 1 to 2 h of initial exposure to an aminoglycoside and disappears several hours after removal of the antibiotic . We investigated adaptive resistance in P . aeruginosa ATCC 27853 following single doses of gentamicin by using a dynamic in vitro model which mimics in vivo pharmacokinetics . The initial peak gentamicin concentrations were 2.5, 8, and 25 mg/liter, and these were followed by an exponential decay in the concentration, with a half-life of 2.5 h . The degree of adaptive resistance was greater and the duration was longer with higher initial gentamicin concentrations . Maximal adaptive resistance occurred between 2 and 10 h following 8 mg/liter and between 2 and 16 h following 25 mg/liter . Full recovery of susceptibility occurred at approximately 36, 39, and 43 h following 2.5, 8, and 25 mg/liter, respectively, at which times the gentamicin concentrations were extremely low . Longer dosing intervals for aminoglycosides may improve efficacy by allowing time for adaptive resistance to resolve. Antimicrob Agents Chemother, 1992 Sep, 36(9), 1922 - 7 Potential effects of erythromycin on host defense systems and virulence of Pseudomonas aeruginosa; Hirakata Y et al.; We evaluated several potential effects of erythromycin (EM) on host defense systems and the virulence of Pseudomonas aeruginosa . Peritoneal macrophages obtained from mice given 250 mg of EM per kg of body weight for 7 days by the intraperitoneal, intravenous, subcutaneous, or oral route produced significantly greater amounts of thymocyte-activating factors . These data suggest that EM enhances the in vivo production of cytokines, such as interleukins 1 and 6 . Treatment of P . aeruginosa D4 with subinhibitory concentrations of EM enhanced the association of bacteria with murine Kupffer cells in vitro and increased bacterial clearance from the blood in mice . EM suppressed the in vitro production of exotoxin A, total protease, elastase, and phospholipase C by P . aeruginosa D4; exotoxin A production by P . aeruginosa PA-103; and total protease production by P . aeruginosa B16 and PAO1 in a generally dose-dependent manner . These data demonstrate that EM produces various effects in addition to its direct antimicrobial activity, suggesting that it has potential as an immunomodulator or bacterial virulence-suppressing agent against P . aeruginosa and other infections. Antimicrob Agents Chemother, 1992 Sep, 36(9), 1902 - 8 Diffusion of meropenem and imipenem through the outer membrane of Escherichia coli K-12 and correlation with their antibacterial activities; Cornaglia G et al.; The outer membrane permeability to meropenem and imipenem in Escherichia coli K-12 was investigated, and its porin-deficient mutants were transformed with a constructed vector carrying the carbapenem-hydrolyzing CphA metallo-beta-lactamase gene . By using the method of Zimmermann and Rosselet, meropenem was shown to penetrate through the outer membrane of E . coli K-12 five times faster than cephaloridine but twice as slowly as imipenem . Lack of one or both porins significantly reduced the penetration of both carbapenems . No evidence of specific porin pathways of the type described in Pseudomonas aeruginosa was found . Despite its slower penetration, meropenem was two to eight times more active than imipenem against both parent and porin-defective mutants, whether harbouring CphA beta-lactamase or not . Meropenem was also more active than imipenem against E . coli DC2, a strain with a breakdown in the outer membrane permeability which made periplasmic concentrations of beta-lactams similar to the external concentrations . In this strain, meropenem caused a more than 50% reduction in cell number increase at a concentration very close to the 50% inhibitory concentration for penicillin-binding protein type 2 (PBP 2), whereas imipenem, at the same concentration, did not significantly inhibit cell growth . This result was explained by the higher affinity of meropenem for PBP 3 compared with imipenem and supports the conclusion that synergistic inhibition of both PBPs was the main mechanism in the better antibacterial activity of meropenem. Antimicrob Agents Chemother, 1992 Sep, 36(9), 1847 - 51 Cross-resistance to meropenem, cephems, and quinolones in Pseudomonas aeruginosa; Masuda N et al.; Multiple-drug-resistant mutants were isolated from Pseudomonas aeruginosa PAO1 on agar plates containing ofloxacin and cefsulodin . These mutants were four to eight times more resistant to meropenem, cephems, carbenicillin, quinolones, tetracycline, and chloramphenicol than the parent strain was . In contrast, these mutants showed no significant changes in their susceptibilities to all carbapenems except meropenem . In these mutants, the amounts of an outer membrane protein with an apparent molecular weight of 49,000 (designated OprM) were increased compared with the amount in PAO1 . Multiple-drug-resistant mutants of this type were also isolated from PAO1 on agar plates containing meropenem . Approximately 5% of clinical isolates showed cross-resistance to meropenem, cephems, and quinolones, concomitant with overproduction of OprM . Moreover, these two phenotypes, i.e., multiple-drug resistance and overproduction of OprM, were cotransferable by transduction . These data suggest that overproduction of OprM is associated with cross-resistance to meropenem, cephems, and quinolones in P . aeruginosa . The ofloxacin-cefsulodin-resistant mutant required higher concentrations of meropenem to induce beta-lactamase than PAO1 did, indicating the possibility that this mutation involves decreased outer membrane permeability to meropenem. Ann Pediatr (Paris), 1992 Sep, 39(7), 443 - 6 {Gangrenous ecthyma of the diaper area in infants}; Taieb A et al.; Ecthyma gangrenosum due to Pseudomonas aeruginosa is a skin infection in which necrotic ulcerations surrounded by a red areola develop . The diaper area is the region most often involved in infants . Typically, ecthyma gangrenosum occurs in patients with septicemia and risk factors (chemotherapy, neutropenia) . However, transient bacteremia or an infection confined to the skin may be the cause in some patients, with maceration in the diaper area and previous antibiotic therapy as risk factors. Mol Gen Genet, 1992 Sep, 234(3), 475 - 80 Molecular cloning, characterization and purification of ornithine carbamoyltransferase from Mycobacterium bovis BCG; Timm J et al.; A genomic library of Mycobacterium bovis BCG has been constructed by cloning DNA partially digested with Sau3A into the Escherichia coli expression vector pAS1 . The gene coding for ornithine carbamoyl-transferase (EC.2.1.3.3; OTCase), hereafter referred to as argF, was isolated from the library by complementation of a double argF-argI mutant of E . coli and its sequence was determined . The translation initiation codon used, GTG, was identified by comparing the amino acid sequence deduced from the gene with the N-terminal sequence of the corresponding purified protein . On this basis, the M . bovis BCG OTCase monomer consists of 307 amino acid residues and displays about 44% identity with other OTCases, the most closely related homologue being the anabolic enzyme of Pseudomonas aeruginosa . The native enzyme has an estimated molecular mass of 110 kDa, suggesting a trimeric structure as is the case for most of the anabolic OTCases known from various organisms. Leber Magen Darm, 1992 Sep, 22(5), 173 - 6 {Antibiotic prevention and therapy of infectious complications in ERCP}; Sauter G et al.; Endoscopic retrograde cholangiopancreatography (ERCP) may be complicated by bacteremia, cholangitis, or biliary sepsis . Bacteremia during ERCP implies a potential risk of endocarditis in patients with valvular prostheses or a previous history of infectious endocarditis . For these patients antibiotic prophylaxis prior to ERCP is recommended . Cholangitis or biliary sepsis may develop after ERCP in patients with obstructed bile ducts . In these patients antibiotics should be administered until adequate drainage of biliary obstructions is achieved . Antibiotic prophylaxis and antibiotic therapy must consider the spectrum of micro-organisms which is normally found in each of these situations . Regarding bacteremias associated with ERCP gram-positive cocci predominate, whereas cholangitis and biliary sepsis are caused mainly by gram-negative rods like Escherichia coli, Pseudomonas aeruginosa, or Klebsiella spp. J Am Acad Dermatol, 1992 Sep, 27(3), 415 - 8 Perineal ecthyma gangrenosum in infancy and early childhood: septicemic and nonsepticemic forms; Boisseau AM et al.; BACKGROUND: Ecthyma gangrenosum is characterized by necrotic ulcerations surrounded by an erythematous halo . It is secondary to Pseudomonas aeruginosa infection . Most lesions are located in the anogenital and axillary areas, but the route of infection is generally difficult to establish . OBJECTIVE: We report six children with perineal ecthyma gangrenosum and discuss predisposing factors, origin, and route of infection . METHODS: This was a retrospective clinical study . RESULTS: Three children had blood cultures positive for P . aeruginosa, and one died . Predisposing factors were present in all cases; two had received chemotherapy (neuroblastoma, acute lymphoblastic leukemia), and two had idiopathic granulocytopenia . The last two patients previously had received treatment with systemic antibiotics and had abnormal granulocyte killing several months later . CONCLUSION: Septicemic ecthyma gangrenosum can be rapidly fatal in young children and requires aggressive antibiotic therapy . Benign ecthyma gangrenosum in healthy infants may result from a modification of bowel microflora after antibiotic therapy in conjunction with maceration in the diaper area . However, careful evaluation and long-term follow-up must be done to detect neutropenia, functional abnormalities of granulocytes, or a possible immune deficiency. Eur J Pediatr, 1992 Sep, 151(9), 684 - 7 Influence of the development of diabetes mellitus on clinical status in patients with cystic fibrosis; Lanng S et al.; The impact of pre-diabetes on clinical status was retrospectively studied in 38 cystic fibrosis (CF) patients with diabetes mellitus (DM) and 38 non-diabetic CF patients (control patients), matched in pairs for age, sex, and chronic Pseudomonas aeruginosa lung infection . Quarterly parameters of CF clinical status were collected for 6 years prior to the diagnosis of DM in the index case . Compared to the control patients, decreases in body weight, body mass index (BMI), forced expiratory volume in 1s (FEV1), and forced vital capacity (FVC) and an increase in the daily intake of pancreatic enzyme capsules were found in the pre-diabetic patients . Statistically significant differences in body weight, BMI, FEV1, FVC, and intake of pancreatic enzyme capsules between pre-diabetic and control patients emerged 4, 4, 1.25, 3 and 4.5 years prior to the diagnosis of DM, respectively . The number of lung infections did not differ between the two groups of patients . Thus, when DM develops in CF patients, an insidious decline in overall clinical status is observed for years prior to its diagnosis . Whether clinical deterioration in CF leads to DM, or pre-diabetes results in declining CF clinical status is presently unknown . Accumulating evidence suggests that the latter may be the case since insulin therapy seems to improve lung function in CF. FEMS Microbiol Rev, 1992 Sep, 9(1), 73 - 90 Protein secretion in Pseudomonas aeruginosa; Tommassen J et al.; The Gram-negative bacterium Pseudomonas aeruginosa secretes many proteins into the extracellular medium . At least two distinct secretion pathways can be discerned . The majority of the exoproteins are secreted via a two-step mechanism . These proteins are first translocated across the inner membrane in a signal sequence-dependent fashion . The subsequent translocation across the outer membrane requires the products of at least 12 distinct xcp genes . The exact role of one of these proteins, the XcpA protein, has been resolved . It is a peptidase that is required for the processing of the precursors of four other Xcp proteins, thus allowing their assembly into the secretion apparatus . This peptidase is also required for the processing of the precursors of type IV pili subunits . Two other Xcp proteins, XcpR and XcpS, display extensive homology to proteins involved in pili biogenesis, which suggests that the assembly of the secretion apparatus and the biogenesis of type IV pili are related processes . The secretion of alkaline protease does not require the xcp gene products . This enzyme, which is encoded by the aprA gene, is not synthesized in a precursor form with an N-terminal signal sequence . Secretion across the two membranes probably takes place in one step at adhesion zones that may be constituted by three accessory proteins, designated AprD, AprE and AprF . The two secretion pathways found in P . aeruginosa appear to have disseminated widely among Gram-negative bacteria. J Formos Med Assoc, 1992 Sep, 91(9), 879 - 85 Management of infected tibial intramedullary nailing using an organized treatment protocol; Ueng WN et al.; Twenty cases of osteomyelitis, following intramedullary nailing of tibial shaft fractures, were managed with a prospective treatment protocol, comprising intramedullary reaming debridement, antibiotic-bead depot, external skeletal fixation, microvascular muscle flap and early cancellous bone grafting . The follow-up period ranged from 25 to 48 months (average, 34.3 months) . Pseudomonas aeruginosa (37.5%) and Staphylococcus aureus (20.8%) were the organisms most commonly involved . There were eight united and 12 ununited fractures after reaming debridement surgery . Nineteen infections were initially arrested by one debridement . One infection was arrested by two Sequential debridements . All 12 ununited fractures were stabilized by Hoffmann unilateral external fixation until the fracture healed . The time spent in external fixation ranged from three to seven months (average, 5.2 months) . Early cancellous bone grafting was successfully accomplished for nine ununited fractures with major debridement bone loss . The average union time of the nine fractures with bone grafting was 6.5 months (range, from six to eight months) . We believe that this treatment protocol gives a predictable and rapid recovery . The complications were infection recurrence in two cases at the old tibial shaft fracture sites, minor Hoffmann pin tract infection in two cases and stiffness in two ankles and one knee. Antimicrob Agents Chemother, 1992 Sep, 36(9), 1915 - 21 Tn21-specific structures in gram-negative bacteria from clinical isolates; Zuhlsdorf MT et al.; A total of 807 gram-negative clinical isolates were treated with five different probes: intragenic segments for the transposase gene tnpA; the resolvase gene tnpR; the modulator of the resolvase, tnpM; the integraselike factor gene tnpI; and a 20-mer oligonucleotide for the recombinational site of action for the integrase . A total of 8% of the isolates hybridized with all five Tn21-related probes, and another 11% represented transposons in which one or more of the tested genes were missing . This 11% included groups whose descriptions have been published as well as groups that have not yet been described . The not-yet-described groups include various deletion products and some precursor structures, as is predicted for the evolution of Tn21-like transposons . The integration system appears to be coupled with Tn21-like structures and yet independent from these structures, implying an independent evolution of this system from Tn21-like transposons . The structures were found with similar incidence levels in all species tested except Pseudomonas aeruginosa, for which a novel separate family of class II transposons has been described before. Kansenshogaku Zasshi, 1992 Sep, 66(9), 1236 - 42 {Effect of azithromycin on human serum sensitivity of Pseudomonas aeruginosa}; Tateda K et al.; We examined the effect of azithromycin (AZM), a 15-membered azalide newly synthesized from erythromycin (EM), on serum sensitivity of 6 strains of Pseudomonas aeruginosa . Incubation for 48 h on agar with EM 12 micrograms/ml or AZM 1.6 micrograms/ml induced increased serum sensitivity in 2 of 6 strains (S-6, PA-103), but there were no changes in any strains with josamycin (JM) 12 micrograms/ml . Although EM 12 micrograms/ml induced increased serum sensitivity of S-6 after more than 36 h incubation, AZM 1.6 micrograms/ml induced increased serum sensitivity of this strain at 12 h incubation . AZM 0.8 microgram/ml (1/62.5 MIC) showed more potent activity to enhance serum sensitivity of S-6 than that of EM 12 micrograms/ml (1/8 MIC) after 48 h incubation . P . aeruginosa S-6 incubated with EM 12 micrograms/ml or AZM 1.6 micrograms/ml for 48 h was less hydrophobic than that of control bacteria, but there was little change in the hydrophobicity of the strain incubated with JM 12 micrograms/ml . These results show that AZM has more potent activity to enhance serum sensitivity of P . aeruginosa than that of EM . Since decrease of cell surface hydrophobicity of P . aeruginosa S-6 was correlated with increased serum sensitivity, EM and AZM may induce enhanced serum sensitivity by changing cell surface structure of P . aeruginosa. Antimicrob Agents Chemother, 1992 Sep, 36(9), 2046 - 8 Interplay of impermeability and chromosomal beta-lactamase activity in imipenem-resistant Pseudomonas aeruginosa; Livermore DM; Mutational loss of the D2 porin causes imipenem resistance in Pseudomonas aeruginosa . It was found that this mechanism could function only when the chromosomal beta-lactamase was expressed . Mutants lacking both the beta-lactamase and the D2 porin were almost as susceptible as those that lacked the beta-lactamase but retained the porin . Thus, imipenem resistance reflected an interplay of the enzyme and impermeability, not either factor alone . These findings suggest that the activity of a carbapenem more beta-lactamase stable than imipenem should be less affected by the porin loss . Meropenem approached this behavior. Biochem J, 1992 Sep 1, 286 ( Pt 2), 361 - 7 E.p.r . and magnetic circular dichroism spectroscopic characterization of bacterioferritin from Pseudomonas aeruginosa and Azotobacter vinelandii; Cheesman MR et al.; The e.p.r . and magnetic circular dichroism (m.c.d.) spectra of bacterioferritin (BFR) extracted from Pseudomonas aeruginosa and Azotobacter vinelandii have been studied over a wide temperature range down to liquid-helium temperature . The e.p.r . spectra show the presence of low-spin Fe3+ haem with g values of 2.86, 2.32, 1.48 (P . aeruginosa) and 2.88, 2.31, 1.46 (A . vinelandii), in both the presence and absence of the BFR core . Together with evidence from the porphyrin-to-Fe3+ charge-transfer band at 2240 and 2270 nm the axial haem ligands are identified as two methionines . The low-temperature m.c.d . spectra in the region 300-1000 nm of P . aeruginosa and A . vinelandii BFR are identical with one another and unaffected by removal of the iron core . Hence it can be concluded that the presence of the iron core has no detectable effect on the electronic states and on the stereochemistry of the haem group . This was unexpected, in view of the observations by Watt, Frankel, Papaefthymiou, Spartalian & Stiefel {(1986) Biochemistry 25, 4330-4336} that the redox potential of the haem group in A . vinelandii BFR shifts from -475 mV to -225 mV on removal of the core . The e.p.r . spectra of holoBFR show a broad symmetrical derivative-shaped band centred at g = 2.0 which decreases in bandwidth as the temperature is raised . This signal is assigned to the uncompensated electron spins of the iron core. J Infect Dis, 1992 Sep, 166(3), 568 - 73 Enhanced release of elastase and oxidative inactivation of alpha-1-protease inhibitor by stimulated human neutrophils exposed to Pseudomonas aeruginosa pigment 1-hydroxyphenazine; Ras GJ et al.; The in vitro effects of the Pseudomonas aeruginosa-derived phenazine pigments pyocyanin and 1-hydroxyphenazine (1-hp) on neutrophil elastase release and myeloperoxidase-induced inactivation of alpha-1-protease inhibitor (alpha 1-PI) were investigated . 1-hp (6-25 microM), but not pyocyanin, caused a dose-dependent enhancement of elastase release by FMLP:cytochalasin B (CB)-activated human neutrophils . 1-hp (0.78-6.25 microM) also increased the oxidative inactivation of the elastase inhibitory capacity of alpha 1-PI exposed to FMLP:CB-activated neutrophils . Methionine, a scavenger of hypochlorous acid, completely protected alpha 1-PI from inactivation by stimulated neutrophils in the presence or absence of 1-hp . Similar protective effects were observed with sodium azide, an inhibitor of myeloperoxidase . P . aeruginosa-derived 1-hp may promote an elastase-antielastase imbalance in vivo by increasing the release of neutrophil elastase and by enhancing the oxidative inactivation of alpha 1-PI, thereby contributing to the development of tissue destruction in P . aeruginosa-infected patients. Infect Immun, 1992 Sep, 60(9), 3771 - 9 Genetic analysis of Pseudomonas aeruginosa adherence: distinct genetic loci control attachment to epithelial cells and mucins; Simpson DA et al.; Infection of mucosal tissues by the opportunistic pathogen Pseudomonas aeruginosa is initiated by attachment of the bacterium to host tissues . To gain a better understanding of this interaction, we used two methods to isolate mutants of P . aeruginosa with altered adherence to cultured A549 cells and to mucins . First, from a population of nonpiliated mutants of P . aeruginosa mutagenized with transposon Tn5G, we have isolated variants that are defective in binding to both A549 cells and respiratory mucins . Using a cloned transposon plus flanking DNA from one such mutant as a DNA probe, we have isolated plasmids from a cosmid bank, which, upon reintroduction to the original mutants, restored adhesion to both A549 cells and mucin . The second strategy to identify genes involved in adhesion used mutagenesis of P . aeruginosa N1G, an rpoN mutant which is unable to bind to either A549 cells or mucin, with transposon Tn5 containing an outward-directed promoter . From this bank of mutagenized P . aeruginosa N1G, two classes of adhesion variants were isolated; one class attached to A549 cells and to mucin, and the other class restored binding of the rpoN mutant to mucin but not to A549 cells . These findings suggest that P . aeruginosa can express at least two adhesins distinct from pili, one recognizing receptors shared by epithelial cells and mucins and the other recognizing mucins alone. Zhonghua Zheng Xing Shao Shang Wai Ke Za Zhi, 1992 Sep, 8(3), 177 - 8, 246 {The pH value of granulating wound and skin graft in burn patients}; Chai JK; The relationship between pH of granulation of burn wound and take rate of skin graft was presented . The bacteria in the granulation tissue were also quantified . The results showed: (1) The optimal pH of granulation wound for the take of skin graft is 7.2-7.5; (2) pH of granulation of burn wound is related to quantity and species of bacteria in the granulation tissue . The wound pH is 6.7 or lower when the number of Escherichia coli or Staphylococcus aureus is over 10(7)/gm of granulation tissue . The wound pH is 8.0 when the number of Pseudomonas aeruginosa is 10(8)/gm of granulation tissue . Because the measurement of wound pH is rapid, simple and noninvasive, it might be useful in predicting the take rate of skin graft. Eur J Clin Microbiol Infect Dis, 1992 Sep, 11(9), 817 - 22 Genome fingerprinting by pulsed-field gel electrophoresis and ribotyping to differentiate Pseudomonas aeruginosa serotype O11 strains; Poh CL et al.; The performance of ribotyping and pulsed-field gel electrophoresis was compared in the differentiation of a collection of 44 Pseudomonas aeruginosa serotype O11 strains isolated in seven hospitals in Singapore . Digestion of genomic DNA by EcoRI and SacI followed by Southern hybridization with the Pseudomonas aeruginosa 16S and 23S rRNA gene revealed seven distinct ribotypes . Ribotyping using a combination of both enzymes revealed 11 ribotypes . In contrast, electrophoretic analysis differentiated 41 different strain types among the 44 clinical isolates using either SpeI or DraI . Pulsed-field gel electrophoresis demonstrated greater sensitivity than ribotyping in the differentiation of Pseudomonas aeruginosa strains of the same ribotype and could thus be used alone in epidemiological investigations of hospital outbreaks of Pseudomonas aeruginosa serotype O11 infection. Eur J Clin Microbiol Infect Dis, 1992 Sep, 11(9), 810 - 6 Multilocus enzyme electrophoresis of major O-antigen reference strains of Pseudomonas aeruginosa; Charnock C et al.; Multilocus enzyme electrophoresis was used to characterize 27 Pseudomonas aeruginosa serogroup reference strains used in the major O-antigen schemes according to which Pseudomonas aeruginosa has been typed . Sixteen enzyme loci were assayed, ten of which showed electrophoretic variation . Genetic diversity was expressed for each enzyme locus, and as the mean allelic diversity of loci . Ten electrophoretic types were identified among the strains . The genetic distance between pairs of electrophoretic types was expressed as the proportion of loci at which similar alleles occurred . More than 80% similarity was observed between any pair of electrophoretic types, reflecting the homogeneity of multilocus genotypes within this species . Similarity between electrophoretic types was represented in the form of a dendrogram and by multi-dimensional scaling . Three distinct clusters of electrophoretic types were revealed; within each the serogroups appeared to be randomly distributed. J Mol Biol, 1992 Aug 20, 226(4), 943 - 57 RNA processing modulates the expression of the arcDABC operon in Pseudomonas aeruginosa; Gamper M et al.; Anaerobic growth of Pseudomonas aeruginosa on arginine depends on the arcDABC operon encoding the enzymes of the arginine deiminase pathway . The co-ordinate, anaerobic induction of these enzymes requires the FNR-like regulatory protein ANR, which activates the arc promoter lying upstream from arcD . By Northern hybridization experiments, three abundant arcA, arcAB and arcABC transcripts and three minor arcDA, arcDAB and arcDABC transcripts could be detected . The 5' ends of the arcA, arcAB and arcABC mRNAs were determined by S1 and primer extension mapping . These 5' ends appear to be generated by endonucleolytic cleavage (processing) in arcD mRNA rather than by a second promoter; this was concluded from the effects of insertion and deletion mutations in arcD . Intergenic inverted repeats between arcA and arcB as well as between arcB and arcC were shown to be involved in the formation of 3' ends of arc transcripts . Deletion of either intergenic region in the P . aeruginosa chromosome led to the loss of the arcA or arcAB transcript, respectively . Dot blot experiments revealed that arc mRNAs extracted from the wild-type strain had similar chemical half-lives in the arcA, arcB and arcC regions, ranging from 16 to 13 minutes . The half-life of arcD mRNA, by contrast, was significantly shorter, suggesting that this mRNA segment may be destabilized by the processing cuts within arcD . Deletion of the putative intergenic stem-loop structures did not result in a dramatic loss of arc mRNA stability . Thus, the intergenic hairpin structures do not contribute importantly to the overall mRNA stability; they might act primarily as partial transcription terminators and locally protect the 3' ends from exonuclease action . The expression levels of the four Arc proteins correlated approximately with the relative abundance of the corresponding mRNA segments . In conclusion, mRNA processing and, presumably, partial termination of transcription contribute to differential gene expression within the arc operon. J Inorg Biochem, 1992 Aug 15-Sep, 47(3-4), 175 - 81 Haem binding to ferritin and possible mechanisms of physiological iron uptake and release by ferritin; Moore GR et al.; Haem binding to horse spleen ferritin and Pseudomonas aeruginosa bacterioferritin has been studied by spectroscopic methods . A maximum of 16 haems per ferritin molecule, and 24 haems per bacterioferritin molecule, has been shown to bind . The influence of the bound haem on the rate of reductive iron release has been investigated . With a range of reductants and in the absence of haem the rate of release varied with the reductant, but in the presence of haem the rate was both independent of the reductant and faster than with any of the reductants alone . This indicates the rate-limiting step for iron release in the absence of haem was electron-transfer across the protein shell . Based on the results obtained with the in vitro assay system and from a consideration of data currently in the literature, plausible schemes for ferritin and bacterioferritin iron uptake and release are described. J Steroid Biochem Mol Biol, 1992 Aug, 42(7), 721 - 7 Identification of an estrogen-binding protein in Pseudomonas aeruginosa; Rowland SS et al.; A constitutive estrogen-binding protein (EBP) has been identified in the cytosol of Pseudomonas aeruginosa, a Gram-negative bacterium . All 14 strains tested contained the EBP . Estradiol binding was rapid and maximal binding occurred by 90 min at 0 degrees C . Dissociation of estradiol from the binding protein occurred at a rate of 4.6 fmol/min with a t1/2 of 42 min . EBP binding was destroyed by protease treatment and at high temperature . Sodium molybdate had no effect on binding . The Kd determined by Scatchard analysis was 3.9 nM and the Bmax was 323 fmol/mg protein . The EBP sedimented at 8.9 S on sucrose density gradients . The presence of 0.4 M KCl increased estradiol binding 6-fold but did not cause a shift in the sedimentation value . Gel filtration of the native protein gave an estimated molecular weight of 215,000 and a Stokes radius of 50.2 A . Steroid binding specificity, in order of decreasing affinity, was estradiol, estrone, dihydrotestosterone, estriol, testosterone, progesterone and promegestone . Other steroid hormones tested did not compete for estradiol binding . Identification of an EBP in a bacterium allows a comparative analysis of other steroid-binding proteins in unicellular microorganisms. Biochemistry, 1992 Aug 11, 31(31), 7152 - 4 Inhibition of human skin fibroblast collagenase, thermolysin, and Pseudomonas aeruginosa elastase by peptide hydroxamic acids; Grobelny D et al.; The hydroxamic acid HONHCOCH2CH(i-Bu)CO-L-Trp-NHMe, isomer 6A (GM 6001), inhibits human skin fibroblast collagenase with Ki of 0.4 nM using the synthetic thiol ester substrate Ac-Pro-Leu-Gly-SCH(i-Bu)CO-Leu-Gly-OEt at pH 6.5 . The other isomer, 6B, which has the opposite configuration at the CH2CH(i-Bu)CO alpha-carbon atom, has a Ki of 200 nM for this enzyme . GM 6001 is one of the most potent inhibitors of human skin fibroblast collagenase yet reported . GM 6001 has a Ki of 20 nM against thermolysin and Pseudomonas aeruginosa elastase . Isomer 6B has a Ki of 7 nM against thermolysin and 2 nM against the elastase . 6A and 6B are the most potent hydroxamate inhibitors reported for these bacterial enzymes . The pattern of inhibition for all three enzymes suggests that isomer 6A is the (R,S) compound, stereochemically analogous to the L,L-dipeptide, and isomer 6B is the (S,S) compound, analogous to the DL-dipeptide . The tolerance of the D configuration by thermolysin and the elastase allows these inhibitors to discriminate between the human and bacterial enzymes simply by inversion of configuration at the CH2CH(i-Bu)CO alpha-carbon atom . Substitution of the potential metal liganding groups carboxylate and hydrazide for the hydroxamate group yields much weaker inhibitors for all three enzymes. J Mol Biol, 1992 Aug 5, 226(3), 651 - 60 Functional structures of the recA protein found by chimera analysis; Ogawa T et al.; We developed a novel genetic method for finding functional regions of a protein by the analysis of chimeras formed between homologous proteins . Sets of chimeric genes were made by intramolecular homologous recombination in a linearized plasmid DNA carrying both recA genes of Escherichia coli and Pseudomonas aeruginosa . A recBCsbcA strain of E . coli was used for isolation of plasmids carrying recombinants between these genes . Examination of properties of E . coli strains deleting the recA gene and carrying a plasmid with a chimeric gene shows that chimera formation at certain positions inactivates a RecA function . Frequently, all chimeras with a junction in a certain region of the protein inactivate a function . Rather than a direct effect of the presence of the junction at a particular position, mismatching of the regions both sides of the junction that are derived from the different species is responsible for the inactivation . For a chimeric protein to be functional, certain pairs of sequences in different regions of the protein must derive from the same parent . Four pairs of such sequences were found: two are involved in activities for genetic recombination and for resistance to ultraviolet light irradiation and the others in formation of active oligomers . Regions defined by these sequences are located in the looped regions of the protein . A pair of regions may co-operate to form a functional folded structure. Infect Immun, 1992 Aug, 60(8), 3332 - 8 Characterization of antibody-mediated inhibition of Pseudomonas aeruginosa adhesion to epithelial cells; Sexton M et al.; An enzyme-linked immunosorbent assay system was developed and used to study adhesion of Pseudomonas aeruginosa to human epithelial cells and the abilities of specific antibodies to inhibit this process . Human buccal epithelial cells coated onto microtiter plates were incubated with P . aeruginosa suspensions, and adherent bacteria were detected by using anti-P . aeruginosa serum and a horseradish peroxidase-conjugated secondary antiserum . Adhesion, quantitated as an increase in A405, varied linearly with increasing numbers of bacterial CFU added per well in the range of 10(5) to 10(8) CFU per well . Adhesion of P . aeruginosa increased following trypsinization of buccal epithelial cells . Preincubation of bacteria with monoclonal antibodies directed against P . aeruginosa outer membrane protein H2 inhibited adhesion with all eight of the isolates tested . Preincubation of P . aeruginosa with sera from infected cystic fibrosis patients also resulted in inhibition of adhesion in the enzyme-linked immunosorbent assay system . This inhibitory activity was shown to be due to two factors: P . aeruginosa-specific immunoglobulin G and a non-immunoglobulin G serum component . These data support the hypothesis that bacterial components other than pili are involved in adhesion and suggest that anti-P . aeruginosa antibodies may be of use in preventing adhesion and subsequent colonization with P . aeruginosa. Infect Immun, 1992 Aug, 60(8), 3162 - 8 Induction of tumor necrosis factor (TNF) and interleukin-1 (IL-1) by Pseudomonas aeruginosa and exotoxin A-induced suppression of lymphoproliferation and TNF, lymphotoxin, gamma interferon, and IL-1 production in human leukocytes; Staugas RE et al.; Pseudomonas aeruginosa is a dominant pathogen in infection in cystic fibrosis . This bacterium is thought to play a major role in the chronic bronchial infection-induced pathophysiology . Our data showed that whole formalin-fixed heat-killed P . aeruginosa was mitogenic for human lymphocytes and induced production of substantial amounts of tumor necrosis factor alpha (TNF) in peripheral blood mononuclear leukocytes in cultures . Significant amounts of TNF were produced at 10(3) bacteria per 2 x 10(5) mononuclear leukocytes . Treatment of P . aeruginosa with polymixin B did not affect its ability to stimulate TNF production, suggesting that bacterial lipopolysaccharide is not involved . P . aeruginosa, however, did not stimulate production of the T-cell lymphokine lymphotoxin (TNF beta) . Exotoxin A, considered to be an important virulence factor produced by P . aeruginosa, did not stimulate either lymphoproliferation or production of TNF . In fact, this toxin, at nontoxic concentrations, was found to depress lymphoproliferation induced by phytohemagglutinin and Staphylococcus aureus and decreased production of TNF, lymphotoxin, and gamma interferon in either lymphocytes or macrophages . This toxin similarly inhibited the production of interleukin-1 beta (IL-1 beta) and IL-1 alpha, but for the inhibition of the latter, 25-fold-less toxin was required than for inhibition of the former . Inhibition of production of TNF was as sensitive as the IL-1 alpha to exotoxin A . The effects of exotoxin A on lymphoproliferation and cytokine production could be neutralized by the addition of anti-exotoxin A antibodies . These results suggest that two mechanisms by which P . aeruginosa could contribute to the chronic bronchial infection-induced pathophysiology are the nonspecific stimulation of TNF and IL-1 and the release of exotoxin A, a toxin which depresses immune responses. Infect Immun, 1992 Aug, 60(8), 3150 - 5 Induction of inflammatory mediator release (serotonin and 12-hydroxyeicosatetraenoic acid) from human platelets by Pseudomonas aeruginosa glycolipid; Konig B et al.; Purified glycolipid from Pseudomonas aeruginosa induced the generation of significant amounts of 12-hydroxyeicosatetraenoic acid (12-HETE) and serotonin release from human platelets . The release of serotonin was first observed 2 min after addition of the glycolipid and increased with time . Significant serotonin release was obtained at glycolipid concentrations above 5 micrograms/ml and increased dose-dependently up to 100% at glycolipid concentrations above 40 micrograms/ml . Glycolipid induced 12-HETE in a time- and dose-dependent manner . 12-HETE formation was first measured after 10 min of incubation and increased with time . Optimal 12-HETE formation was obtained at a glycolipid concentration of 50 micrograms/ml; higher concentrations of glycolipid led to a decrease in 12-HETE formation, indicating a cytotoxic effect . Stimulation of platelets with glycolipid (12-HETE formation and serotonin release) was accompanied by calcium influx, translocation of protein kinase C, activation of guanylylimidodiphosphate binding, and increased GTPase activity in platelet membranes within the same concentration range. Arch Biochem Biophys, 1992 Aug 1, 296(2), 505 - 13 Purification, molecular cloning, and expression of lipase from Pseudomonas aeruginosa; Chihara-Siomi M et al.; An extracellular lipase secreted by Pseudomonas aeruginosa TE3285 was purified . A genomic library of this strain was constructed in lambda EMBL3, and a DNA fragment 2.7 kb long containing the lipase gene, lipA, was isolated with an oligonucleotide probe synthesized on the basis of the partial amino acid sequence of a purified preparation of the enzyme . Nucleotide sequence analysis showed an open reading frame of 933 bases, and the deduced amino acid sequence agreed well with the molecular mass and partial amino acid sequences of mature lipase . The results of alignment of the amino acid sequences of five lipases from Pseudomonas species considered together with the published crystal structure studied with human pancreatic lipase showed that Ser82, His251, and Asp209 were catalytic residues and that a surface loop from residues 172 to 204 was responsible for the substrate specificity . About 50 bases downstream of lipA, there was another gene, lipB . The sequence of lipB was highly homologous to that of putative modulators of the production of active lipases in other Pseudomonas species . Expression plasmids encoding lipA followed by the complete or incomplete lipB gene downstream of the lac promoter of pUC18 were constructed . lipA was expressed in Escherichia coli 1100 only in the presence of the complete lipB gene. J Bacteriol, 1992 Aug, 174(15), 5156 - 60 Activation defects caused by mutations in Escherichia coli rpoA are promoter specific; Gussin GN et al.; Escherichia coli RNA polymerases containing mutated alpha subunits were tested for their ability to respond to three different positive regulators (activators) in vitro . The two alpha (rpoA) mutants, alpha-256 and alpha-235, have deletions of the C-terminal 73 and 94 amino acids, respectively . In runoff transcription assays catalyzed by reconstituted holoenzyme, the effects of the mutations on each of three promoters tested were different: activation of the lambda pRM promoter by cI protein (repressor) was nearly normal, activation of the lambda pRE promoter by cII protein was reduced approximately fivefold, and direct activation of the trpPB promoter of Pseudomonas aeruginosa was completely inhibited . We also found that the reconstituted mutant enzyme was defective in recognition of trpPI in the absence of activator . The differential responses of the three promoters to their activators in the presence of the mutant enzymes indicate that the location of an activator-binding site does not by itself determine the region of RNA polymerase with which the activator interacts. J Bacteriol, 1992 Aug, 174(15), 5149 - 51 Phosphate taxis in Pseudomonas aeruginosa; Kato J et al.; Pseudomonas aeruginosa was shown to be attracted to phosphate . The chemotactic response was induced by phosphate starvation . The specificity of chemoreceptors for phosphate was high so that no other tested phosphorus compounds elicited a chemotactic response as strong as that elicited by phosphate . Competition experiments showed that the chemoreceptors for phosphate appeared to be different from those for the common amino acids . Mutants constitutive for alkaline phosphatase showed the chemotactic response to phosphate regardless of whether the cells were starved for phosphate. J Gen Microbiol, 1992 Aug, 138 ( Pt 8), 1665 - 70 Heterologous expression of an alginate lyase gene in mucoid and non-mucoid strains of Pseudomonas aeruginosa; Gacesa P et al.; A 1.95 kb DNA fragment containing the aly gene from Klebsiella pneumoniae, which encodes an alginate lyase, has been ligated into the broad-host-range vector pLAFR3 . Transfer of the resultant recombinant plasmid, pALY8, into mucoid and non-mucoid strains of Pseudomonas aeruginosa resulted in expression of the alginate lyase . The heterologously expressed alginate lyase, which had the same isoelectric point and substrate specificity as the native enzyme, altered the morphology of mucoid strains . Analysis of the extracellular material from mucoid strains revealed that lyase expression reduced the M(r) and overall yield of alginate produced . The mature form of the recombinant enzyme was the same as that produced extracellularly by Klebsiella pneumoniae; however, most of the alginate lyase was retained intracellularly by P . aeruginosa. J Oral Pathol Med, 1992 Aug, 21(7), 299 - 304 The role of saliva in aggregation and adherence of Pseudomonas aeruginosa in patients with cystic fibrosis; Tumber-Saini SK et al.; The aggregation and adherence activity of P . aeruginosa, mediated by whole saliva from cystic fibrosis (CF) patients and non-CF subjects, was investigated . CF saliva-mediated aggregation of P . aeruginosa was stronger than the activity of non-CF saliva . Likewise, P . aeruginosa adherence to buccal epithelial cells (BEC) of CF patients was stronger than to BEC of non-CF subjects . Adherence of non-mucoid P . aeruginosa to BEC of CF patients was increased by saliva, whereas the mucoid variant was not . CF patients colonized with P . aeruginosa showed higher adherence of the non-mucoid variant than non-colonized CF patients . CF patients with high saliva-mediated adherence of non-mucoid P . aeruginosa also had high salivary aggregation activity . Increased CF saliva-mediated aggregation activity may be linked to the increased non-mucoid P . aeruginosa adherence to BEC of CF patients. Jpn J Antibiot, 1992 Aug, 45(8), 958 - 64 {Combined effects of arbekacin with other antibiotics against methicillin-resistant Staphylococcus aureus . II . The combined effect of arbekacin with imipenem or cefminox}; Deguchi K et al.; Combined antibacterial effects of imipenem (IPM)+arbekacin (ABK) and cefminox (CMNX)+ABK against methicillin-resistant Staphylococcus aureus (MRSA) were examined and the obtained results are summarized below . 1 . Either combination, IPM+ABK or CMNX+ABK, showed a strong antibacterial effect against MRSA when blood concentration of ABK were sustained at MIC as could be expected in clinical situations . While at sub MICs of ABK the antibacterial effect of these combination was slightly less than those of the previously reported combinations of ABK and other antibiotics . 2 . Antibacterial effects of the combinations against MRSA were strongly dependent on the concentration of ABK and less dependent on the concentration of IPM or CMNX . As were observed in the previously tested combinations of ABK with other antibiotics, the antibacterial effect of the combination appeared to be highly dependent on the antibacterial activity and the concentration of ABK . 3 . As IPM has potent antibacterial activities against Gram-negative bacteria (GNB) including Pseudomonas aeruginosa while CMNX has potent antibacterial activities against GNB except P . aeruginosa, it is likely that the combinations of IPM+ABK or CMNX+ABK are useful for treatment of infections with MRSA together with GNB. Jpn J Antibiot, 1992 Aug, 45(8), 949 - 57 {Combined effects of arbekacin with other antibiotics against methicillin-resistant Staphylococcus aureus . I . The combined effect of arbekacin with fosfomycin or clavulanic acid/ticarcillin}; Deguchi K et al.; As arbekacin (ABK) has a highly potent antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA), its combined effects with fosfomycin (FOM) and clavulanic acid/ticarcillin (CVA/TIPC) against MRSA were examined . The obtained results are summarized as follows . 1 . Against MRSA either combination, FOM+ABK or CVA/TIPC+ABK showed a strong antibacterial effect at the MIC or the sub MIC of ABK in the blood expected from clinical observations . The MIC of ABK by the combination use seemed to be equivalent to the MBC value . 2 . Effective concentrations of antibiotics in these combinations appeared to be strongly dependent on the effective concentration of ABK and less dependent on that of FOM or CVA/TIPC . Therefore, the antibacterial activity of a combination seems to mostly depend on the antibacterial activity and the concentration of ABK . 3 . As FOM and CVA/TIPC have antibacterial activities against Pseudomonas aeruginosa, combinations of ABK with these antibiotics are likely to be effective against double infection with P . aeruginosa in MRSA infected patients. Eur J Clin Microbiol Infect Dis, 1992 Aug, 11(8), 715 - 21 Quantitative assay for testing susceptibility of HIV isolates to zidovudine and sCD4 (178)-PE40; Verhoef J et al.; A quantitative coculture assay is described for determining susceptibility of HIV-1 isolates to zidovudine and sCD4 (178)-PE40 (a 60 kDa hybrid protein consisting of the gp120 binding region of CD4 linked to the translocation and ADP-ribosylation regions of Pseudomonas aeruginosa exotoxin) . The assay was relatively simple to perform and gave highly reproducible results often within three to five days . IC50 values of zidovudine for HIV-1 strains isolated from two AIDS patients and an asymptomatic seropositive individual were in the range 0.001-0.002 mumol . Isolates obtained after six months of zidovudine treatment had zidovudine IC50 values of 0.01-0.5 mumol . All isolates were equally sensitive to sCD4 (178)-PE40 (IC50 1-5 micrograms/ml) . HIV-1 activation in the chronically infected cell line U-1 was inhibited by sCD4 (178)-PE40 but not by zidovudine. Curr Eye Res, 1992 Aug, 11(8), 727 - 38 Systemic and topical protection studies using Pseudomonas aeruginosa flagella in an ocular model of infection; Rudner XL et al.; Purified flagella from P . aeruginosa ATCC 19660 were used for active, passive, or topical immunization prior to corneal challenge with strain 19660 . At 30 days post-infection, a significant number of mice actively or passively immunized with flagella and infected with the homologous bacterial strain were protected from ocular disease when compared to control animals . In topical immunization studies, premixing of 19660 flagella with the bacterial inoculum prior to ocular challenge with strain 19660, provided results similar to those of the active or passive immunization studies . A reduced lipopolysaccharide (LPS:1 E.U./mg) flagella preparation was also produced and used similarly . Again, significant protection was achieved in mice immunized by flagella regardless of the immunization route . An in vitro adherence assay also was performed to examine quantitatively the effect of exogenously applied flagella, or an antiflagella monoclonal antibody (MAb) on bacterial adhesion . Premixing of the bacterial inoculum with flagella or the MAb prior to applying it topically to corneas in organ culture all significantly inhibited bacterial binding . These results strongly suggest that significant ocular protection is achieved with either active or passive immunization, or premixing of the bacterial inoculum with flagella from strain 19660 prior to ocular challenge with the homologous bacterial strain . They also indicate that topical application of flagella or antiflagella MAb provide protection against ocular disease by decreasing bacterial adhesion to cornea. Antimicrob Agents Chemother, 1992 Aug, 36(8), 1601 - 5 Quinolone accumulation in Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus; McCaffrey C et al.; The accumulation of quinolones by Escherichia coli JF568, Pseudomonas aeruginosa PAO1, and Staphylococcus aureus ATCC 29213 was measured by a modified fluorometric assay (J . S . Chapman and N . H . Georgopapadakou, Antimicrob . Agents Chemother . 33:27-29, 1989) . The quinolones examined were fleroxacin, pefloxacin, norfloxacin, difloxacin, A56620, ciprofloxacin, ofloxacin, and Ro 09-1168 . In all three organisms, uptake was complete in less than 5 min and was proportional to extracellular quinolone concentrations between 2 and 50 micrograms/ml, which is consistent with simple diffusion . Washing cells with quinolone-free buffer decreased accumulation by up to 70% in E . coli and P . aeruginosa but not in S . aureus . Similarly, incubation with the uncouplers 2,4-dinitrophenol and carbonyl cyanide m-chlorophenylhydrazone increased accumulation up to fourfold in E . coli and P . aeruginosa, though not in S . aureus, suggesting endogenous, energy-dependent efflux . High quinolone hydrophobicity was generally associated with decreased accumulation in E . coli and P . aeruginosa (except in the case of pefloxacin) but was associated with increased accumulation in S . aureus (except in the case of difloxacin) . Ciprofloxacin had the highest accumulation in E . coli and P . aeruginosa, while pefloxacin had the highest accumulation in S . aureus. Mol Microbiol, 1992 Aug, 6(16), 2319 - 26 Polyphosphate-selective porin OprO of Pseudomonas aeruginosa: expression, purification and sequence; Siehnel RJ et al.; The oprO gene of Pseudomonas aeruginosa codes for a polyphosphate-specific porin and terminates 458 bp upstream of the start codon for the phosphate-specific porin OprP . OprO was found to be expressed only under phosphate-starvation conditions in both wild-type and oprP::Tn501 mutant P . aeruginosa strains . However, unlike the rest of the genes of the Pho regulon, including oprP, expression of oprO required cells to be in the stationary growth phase in addition to phosphate starvation . Wild-type P . aeruginosa cells were grown in fermentor culture under these conditions and fractionated by selective solubilization in octylpolyoxyethylene detergent solution . Solubilized OprO was separated from OprP by application to a Mono Q FPLC column and elution with a salt gradient and shown to be functionally identical to cloned OprO produced in Escherichia coli . DNA sequencing of oprO showed the gene product to be highly homologous to OprP, with 76% identity and 16% conserved substitutions . Most genes of the Pho regulon possess a modified -35 region called the Pho box . Two such elements, separated by 4 bp were found in oprO . DNA sequencing also revealed a second Pho box in the oprP gene with the same spacing. Rinsho Ketsueki, 1992 Aug, 33(8), 1041 - 5 {Malignant lymphoma developing from the wall of chronic empyema following artificial pneumothorax}; Tachibana J et al.; A 60-year-old man who had had chronic empyema following an artificial pneumothorax for pulmonary tuberculosis when he was 26 years old developed malignant lymphoma of the chest wall . The patient was admitted because of right pyothorax as a result of pseudomonas aeruginosa infection and underwent right thoracotomy . During lavage of the right thoracic cavity a tumor was found arising from the empyematic wall . Pathologic examination revealed that it was malignant lymphoma (diffuse large, immunoblastic, B cell type) . Treatment with VEAP-Bleomycin elicited a good response . Seven months after chemotherapy, the patient underwent thoracoplasty in addition to packing the cavity with the latissimus dorsi and the greater omentum . Following this, the patient received chemotherapy once a month for one and a half years, after which he was kept under close observation without treatment . Complete remission has now lasted for 49 months since the initial treatment . This is the first reported lymphoma case with closure of the empyematic wall and is remarkable since this patient has remained in complete remission for the last two years without any treatment. J Chemother, 1992 Aug, 4(4), 225 - 7 Subcutaneous nodules caused by Pseudomonas aeruginosa: healing without incision and drainage; Korten V et al.; In a patient with multiple myeloma, numerous indurated, subcutaneous nodules and pyomyositis due to Pseudomonas aeruginosa were noted . These lesions resolved with ciprofloxacin plus ceftazidime therapy without surgical incision and drainage . Despite another course of cancer chemotherapy after total disappearance, there were no recurrences at the end of 3 months . Quinolones initially combined with other antipseudomonal beta-lactam agents may be the drugs of choice in the management of patients with subcutaneous nodules caused by P . aeruginosa. Kansenshogaku Zasshi, 1992 Aug, 66(8), 1097 - 104 {Long-term chemotherapy using erythromycin (EM) for chronic lower airway infection: effectiveness of clarithromycin in EM ineffective cases--the fourth report}; Mikasa K et al.; The effectiveness of long-term chemotherapy using Erythromycin (EM) for chronic lower airway infection has been practically proved . However, there still exist some ineffective cases, or cases in which the clinical effect was scarcely seen . In the present study was administered Clarithromycin (CAM) to such cases and found that CAM was effective in alleviating symptoms in some of them . The results were presented along with clinical findings and other basic studies . The subjects were 4 cases in which EM was either ineffective or low in its clinical effect . The subjects consisted of 1 case of DPB and 3 cases of bronchiectasis . EM was clinically ineffective in 2 cases and slightly effective in the two others . The pathogen was Pseudomonas aeruginosa in all cases . The dosage of EM was 200-1200 mg/day . The period of administration ranged from 2 years to 6 years 9 months . CAM was given orally after meals at a dose of either 200 or 400 mg/day . Chemotherapy had continued for 3-8 months at the time of final observation (Feb . 1992) . Clinical effectiveness was evaluated on the basis of sputum volume and PaO2 examination, as well as evaluation by the patients themselves . As a result, in all 4 cases, reduction in sputum volume and improvement of PaO2 were observed . All subjects evaluated CAM therapy as being more effective than EM therapy . Moreover, it was found that CAM inhibited both the elastase and leucocidin produced in one of the ineffective cases by P . aeruginosa, whereas EM didnt.(ABSTRACT TRUNCATED AT 250 WORDS) Chest, 1992 Aug, 102(2), 647 - 9 Systemic hypersensitivity vasculitis associated with bronchiectasis; Tanaka E et al.; Systemic hypersensitivity vasculitis developed in a 53-year-old man during acute exacerbation of bronchiectasis infected with Pseudomonas aeruginosa . High grade fever, mononeuropathy multiplex, cutaneous vasculitis, and biopsy specimen-proved mesangioproliferative glomerulonephritis with crescent formation and leukocytoclastic vasculitis associated with circulating immune complex occurred . Corticosteroid and cyclophosphamide therapy was effective for vasculitis and bronchiectasis. Am J Clin Pathol, 1992 Aug, 98(2), 222 - 6 Systemic polyclonal B-immunoblastic proliferation with marked peripheral blood and bone marrow plasmacytosis; Poje EJ et al.; The clinical and pathologic features of a case of acute systemic polyclonal B-immunoblastic proliferation characterized by pronounced peripheral blood and bone marrow plasmacytosis and infiltration of the hepatic portal areas by immunoblasts, plasma cells, and lymphocytes are reported . Clinical and laboratory findings during the acute phase and long-term follow-up support the diagnosis of a benign process, possibly related to Pseudomonas aeruginosa septicemia . The patient experienced a dramatic clinical recovery on administration of high-dose intravenous corticosteroids . Pathologists should be aware of this entity so as not to confuse it with non-Hodgkin's lymphoma or a form of plasma cell dyscrasia. Pediatr Res, 1992 Aug, 32(2), 175 - 8 Genotype/phenotype association in cystic fibrosis: analyses of the delta F508, R553X, and 3905insT mutations; Liechti-Gallati S et al.; A striking clinical phenomenon of cystic fibrosis is the heterogeneous disease expression . It must therefore be assumed that the nature of the mutations associated with cystic fibrosis might partly determine the phenotypic manifestations . The relation between the cystic fibrosis mutations delta F508, R553X, and 3905insT and clinical parameters such as sweat test electrolytes, age at chronic Pseudomonas aeruginosa colonization, Chrispin-Norman x-ray scores, and relative underweight have been investigated in 45 patients homozygous for delta F508 (delta F2), in 12 compound heterozygotes for delta F508/R553X (delta F1/RX1), in three R553X homozygotes (RX2), and in 13 patients compound heterozygous for delta F508/3905insT (delta F16) . We have found significant differences between the genetically defined subgroups concerning the mean age at onset and the cumulative incidence of chronic P . aeruginosa colonization and Chrispin-Norman x-ray scores . The significant results as well as some trends regarding the relative underweight demonstrate a milder clinical course in R553X heterozygotes and more severe disease in the delta F16 group compared to delta F508 homozygotes . The three patients homozygous for R553X presented with a two-stage course showing mild progression before P . aeruginosa infection and as severe a course as the delta F16 patients after P . aeruginosa colonization at the age of 12 y . The findings presented here indicate that specific mutations can influence the severity and progression of the disease, implicating the importance of mutation and haplotype analyses . However, wide variations within the genetically homogeneous subgroups illustrate that other determinants of the clinical status do exist. Crit Care Med, 1992 Aug, 20(8), 1127 - 33 Effects of granulocyte colony-stimulating factor in modifying mortality from Pseudomonas aeruginosa pneumonia after hemorrhage; Abraham E et al.; BACKGROUND AND METHODS: Alterations in immune function occurring after hemorrhage and trauma may contribute to the high occurrence rates of nosocomial pneumonia, multiorgan system failure, morbidity, and mortality in this setting . Therapy with granulocyte colony-stimulating factor (G-CSF) can increase neutrophil numbers and function, and enhance resistance to infection in experimental and clinical settings associated with abnormal immune function . To investigate whether treatment with G-CSF could increase resistance to pneumonia after hemorrhage, we bled mice 30% of the blood volume and treated them with various doses of G-CSF, starting either immediately or 2 days after hemorrhage . Pseudomonas aeruginosa pneumonia was induced by the intratracheal instillation of 2 x 10(7) colony-forming units of P . aeruginosa 4 days after blood loss, and mortality was assessed over the next 7 days . RESULTS: Treatment of mice with 100 or 500 micrograms/kg/day G-CSF, but not with 50 micrograms/kg/day, resulted in significant increases in the numbers of circulating polymorphonuclear cells . Platelet counts significantly decreased in mice given 500 micrograms/kg/day G-CSF . Mice given 100 micrograms/kg/day G-CSF starting 2 days after blood loss had improved outcome compared with vehicle-treated controls (38% survival rate in the G-CSF treated group vs . 8% in controls, p less than .05) . There also was a trend toward an improved survival rate in mice treated with 50 micrograms/kg/day G-CSF for 4 days after hemorrhage (46% survival rate in G-CSF treated vs . 17% in controls) . CONCLUSIONS: G-CSF prophylactically administered after hemorrhage can improve survival from pneumonia due to P . aeruginosa . However, the protection afforded by G-CSF was highly dependent on the dosing schedule used. J Bacteriol, 1992 Aug, 174(15), 4977 - 85 Analysis of the Pseudomonas aeruginosa major outer membrane protein OprF by use of truncated OprF derivatives and monoclonal antibodies; Finnen RL et al.; TnphoA mutagenesis of the cloned oprF gene was utilized to generate 16 classes of fusions encoding differing lengths of the amino terminus of OprF fused to either alkaline phosphatase or to peptide tags of 1 to 20 amino acids, depending on the orientation and reading frame into which TnphoA was inserted . Representatives of each of the 16 classes were sequenced to determine the precise fusion joint . Four of these 16 representatives which produced in-frame fusions to alkaline phosphatase and another 8 with fusion joints in the amino-terminal half of OprF failed to react with a panel of 10 specific monoclonal antibodies . In contrast, OprF derivatives with predicted fusion joints at amino acids 180, 204, 289, and 299 reacted with one to five of the monoclonal antibodies . Four other immunoreactive OprF derivatives were created by subcloning and encoded amino acids 1 to 187, 188 to 326, 1 to 273 and 1 to 170 plus 301 to 326 . On the basis of reactivity with the TnphoA-truncated derivatives and subclones of oprF, the epitopes for all 10 monoclonal antibodies were localized, in part, to specific regions of OprF . Nnie of the 10 monoclonal antibodies, 8 of which recognize surface-exposed epitopes, mapped within the carboxy-terminal region of OprF that is homologous to the Escherichia coli outer membrane protein OmpA . Thus, we concluded that parts of the carboxy terminus of OprF are exposed on the external face of the outer membrane . In addition, a clone containing only the first two cysteine residues of OprF demonstrated reactivity with monoclonal antibodies MA4-4 and MA7-8 that was destroyed by 2-mercaptoethanol treatment, as was reactivity with intact OprF . Thus, we conclude that this first pair of cysteines at residues 176 and 185 of mature OprF form a disulfide bond. Biotechnology (N Y), 1992 Aug, 10(8), 910 - 2 Antibody production in silkworm cells and silkworm larvae infected with a dual recombinant Bombyx mori nuclear polyhedrosis virus; Reis U et al.; We have examined the efficiency of coexpression of two heterologous genes from a recombinant Bombyx mori nuclear polyhedrosis virus for the production of antibodies in silkworm larvae . The cDNAs encoding the light and the heavy chains of a murine immunoglobulin, directed against lipoprotein I of Pseudomonas aeruginosa, were brought under the control of two separate copies of the viral polyhedrin promotor . Infection of silkworm cells with the recombinant baculovirus yielded a maximum of 6.4 micrograms/ml IgG2A in the culture supernatant 72 hours post infection, while 800 micrograms/ml IgG2A was found in the hemolymph of infected fifth instar silkworm larvae seven days after infection with the same construct . The recombinant antibody exhibited a similar antigen specificity and avidity to that of the monoclonal antibody derived from ascites fluid. Antimicrob Agents Chemother, 1992 Aug, 36(8), 1791 - 3 Nucleotide sequence of the protein D2 gene of Pseudomonas aeruginosa; Yoneyama H et al.; Protein D2 of the outer membrane of Pseudomonas aeruginosa was shown to form the imipenem-permeable pore . We cloned and sequenced the protein D2 gene . The protein D2 gene encodes a polypeptide with 443 amino acids consisting of 23 and 420 amino acid residues for the signal peptide and mature polypeptide (M(r), 46,010), respectively . Protein D2 contains the highest molar ratio of glycine and no cysteine . The polar amino acids are scattered throughout the sequence. J Biochem (Tokyo), 1992 Aug, 112(2), 290 - 8 A novel terminal oxidase, cytochrome baa3 purified from aerobically grown Pseudomonas aeruginosa: it shows a clear difference between resting state and pulsed state; Fujiwara T et al.; A novel type of cytochrome c oxidase was purified to homogeneity from Pseudomonas aeruginosa which was grown aerobically . The purified oxidase contained two molecules of heme a, two atoms of copper, and one molecule of protoheme per molecule . One of the two heme a molecules in the oxidase reacted with carbon monoxide, so that the enzyme was of baa3-type . The oxidase molecule was composed of three subunits with molecular weights of 38,000, 57,000, and 82,000 . Although the oxidase oxidized ferrocytochrome c-550 obtained from the bacterial cells grown aerobically, the oxidizing activity was not high . The "resting form" and the "pulsed form" of the oxidase were observed clearly with this enzyme, and the transition from the resting form to the pulsed form was accompanied by a distinct change of the enzymatic activity . The difference in the kinetics of the catalytic reactions between the two forms is discussed. J Bacteriol, 1992 Aug, 174(16), 5196 - 203 Reevaluation, using intact cells, of the exclusion limit and role of porin OprF in Pseudomonas aeruginosa outer membrane permeability; Bellido F et al.; Earlier studies that used model membrane reconstitution methods have come to different conclusions regarding the exclusion limit of the outer membrane of Pseudomonas aeruginosa and whether OprF is the major channel-forming protein in the outer membrane . In this study, a 6.2-kbp SalI fragment, encoding only two cytoplasmic enzymes, alpha-galactosidase and sucrose hydrolase, and the inner membrane raffinose permease, was cloned behind the m-toluate-inducible tol promoter of vector pNM185 to create plasmid pFB71 . P . aeruginosa strains harboring pFB71, when grown with inducer, produced both enzymes encoded by the insert and had acquired the ability to grow on the disaccharide melibiose and the trisaccharide raffinose . The rate of growth was dependent on the concentration and size of the saccharide and was decreased three- to fivefold by the absence of OprF, as examined by measuring the growth on melibiose and raffinose of an isogenic OprF-deficient omega insertion derivative, H636(pFB71) . At high concentrations, di-, tri-, and tetrasaccharides could pass across the outer membrane to plasmolyze P . aeruginosa, as measured by light scattering and confirmed by electron microscopy . The initial rate kinetics of light-scattering changes were dependent on the size of the saccharide being used . Furthermore, the rates of change in light scattering due to raffinose and stachyose uptake across the outer membrane for strain H636 were fivefold or more lower than for its OprF-sufficient parent H103 . These data are consistent with model membrane studies showing that OprF is the most predominant porin for compounds larger than disaccharides in P . aeruginosa and suggest that the exclusion limit for this porin and the outer membrane is greater than the size of a tetrasaccharide . In addition, these data confirmed the existence of other porins with a predominant function in monosaccharide uptake and a more minor function in the uptake of larger saccharides. Biochem Biophys Res Commun, 1992 Jul 31, 186(2), 645 - 51 Porin of Pseudomonas aeruginosa forms low conductance ion channel in planar lipid bilayers; Obara M et al.; Protein E1, a porin of the outer membrane of Pseudomonas aeruginosa, was reconstituted into planar lipid bilayers . Single channel conductance of the protein appeared to be 230 pS (pico siemens) in 1 M KCl-10 mM Hepes, pH7.2 . This value is approximately 5 times lower than the conductance of the OmpF channel of Escherichia coli . Conductance increased linearly as the membrane potential was raised from -200 mV to +200 mV, and was nearly proportional to the KCl concentration . These results show that protein E1 is probably a genuine porin in the P . aeruginosa outer membrane supporting the earlier conclusion that protein E1 forms a small channel. FEBS Lett, 1992 Jul 20, 306(2-3), 119 - 24 Crystal structure of Pseudomonas aeruginosa apo-azurin at 1.85 A resolution; Nar H et al.; The 3D structure of apo-azurin from Pseudomonas aeruginosa has been determined at 1.85 A resolution . The crystal structure is composed of two different molecular forms of apo-azurin arranged as hetero-dimers in the tetramer of the asymmetric unit . Form 1 closely resembles the holo-protein lacking copper . Form 2 shows differences in the metal binding site region induced by the incorporation of a solvent molecule into this site . The positions of the copper ligands His46 and His117 are shifted by 0.6 A and 1.6 A . The His117 side chain adopts a position at the surface of the protein, thereby facilitating access to the copper site . The presence of two different molecular forms of apo-azurin in the crystal lattice may reflect an equilibrium between the two forms in solution . 1H-NMR spectra of apo-azurin recorded as a function of pH show that at high pH the line broadening of His35, His46 and His117 resonances is consistent with an interconversion between forms 1 and 2 . At low pH, no broadening is observed . This may indicate that here the interconversion is fast on the NMR timescale. FEBS Lett, 1992 Jul 13, 306(1), 5 - 8 Separation of gate- and channel-forming domains in the pore-forming protein of the outer membrane of Pseudomonas aeruginosa; Yoshihara E et al.; The domains of the pore-forming protein responsible for the gate and channel formations were separated and identified in the outer membrane of Pseudomonas aeruginosa . The proteolytic cleavage of the 46K channel protein, protein D2, yielded two major domains with apparent M(r) of 27K and 19K . We identified the 27K polypeptide to be the channel-forming domain by an in vitro permeability assay . The channel size of purified 27K domain was indistinguishable from that of native protein D2 . Degradation of the 19K domain into small subfragments increases the channel activity about ten times suggesting that the 19K polypeptide forms the gate or cap. FEBS Lett, 1992 Jul 13, 306(1), 41 - 5 Properties of Pseudomonas aeruginosa exotoxin A ionic channel incorporated in planar lipid bilayers; Gambale F et al.; Acidic conditions induce the incorporation of Pseudomonas aeruginosa exotoxin A into phospholipid planar bilayers and the formation of pores permeable to electrolytes . Channel openings occur as single events, although they may occasionally cluster in bursts . In 100 mM KCl, the elementary single channel current amplitude is 3.1 pA (at a transmembrane voltage of 100 mV), the mean open time is 1.3 ms, while bursts may last for several seconds . Noise analysis gave results identical to single channel analysis . Voltage pulse protocols and continuous cycling voltage ramps showed that the toxin channel is voltage dependent, having a higher probability of being open at positive voltages. Appl Microbiol Biotechnol, 1992 Jul, 37(4), 446 - 50 Scale-up and optimization of culture conditions of a human heterohybridoma producing serotype-specific antibodies to Pseudomonas aeruginosa; Schurch U et al.; Three different stirred bioreactors of 0.5 to 121 volume were used to scale up the production of a human monoclonal antibody . Inoculation density and stirrer speed were evaluated in batch cultures, whereas dilution rate and pH were optimized in chemostat cultures with respect to high specific antibody production rate and high antibody yield per time and reactor volume . The cell line used for the experiments was a heterohybridoma, producing immunoglobulin M (IgM) against lipopolysaccharide of Pseudomonas aeruginosa . Cells were cultured in spinner flasks of 500 ml liquid volume for adaptation to stirred culture conditions . Subsequently cells were transferred to the 1.5-l KLF 2000 bioreactor and to the 12-l NLF 22 bioreactor for pilot-scale cultures . Chemostat experiments were done in the 1.5-l KLF bioreactor . Cell density, viability, glucose and lactate and antibody concentration were measured during culture experiments . In batch cultures in all three stirred bioreactors, comparable maximal cell densities and specific growth rates were achieved . Chemostat experiments showed that at a pH of 6.9 and a dilution rate of 0.57 per day the specific antibody production rate was threefold higher than similar experiments done at pH 7.2 with a dilution rate of 0.36 per day . By optimizing pH and dilution rate in chemostat cultures the daily yield of human IgM increased nearly threefold from 6 to 16 mg/day and per litre of reactor volume . The yield per litre of medium increased twofold. J Med Chem, 1992 Jul 10, 35(14), 2631 - 42 Synthesis and structure-activity relationships of cephalosporins with C-3' catechol-containing residues; Arnould JC et al.; Cephalosporins with new catechol substituents at C-3' have been synthesized, including novel compounds with C-3' carbon-carbon bonds . Many of these compounds have high potency against Gram-negative bacteria, in particular against resistant strains like Pseudomonas aeruginosa . Structure-activity relationships are discussed in terms of their dependence on the pKa of the C-3' catechol and also in terms of steric and conformational factors of the C-3' substituent . The best overall properties were found in compounds with a bulky and/or conformationally restricted acidic C-3' catechol. J Immunol Methods, 1992 Jul 6, 151(1-2), 157 - 64 Analyses of affinity distributions within polyclonal populations of antigen-specific antibodies . Evaluation of their accuracy in population detection using monoclonal antibodies; Bruderer U et al.; The potential of an ELISA based detection of affinity distributions within polyclonal populations of antigen-specific serum antibodies was assessed by analyzing defined probes composed of monoclonal antibodies (MAb) . In a competitive binding ELISA in which the concentration of antigen in the liquid phase and the solid phase was varied, we analyzed mixtures containing defined percentage compositions of MAb exhibiting apparent affinity constants (aK) between 3 x 10(6) and 2 x 10(9) M-1 for Pseudomonas aeruginosa exotoxin A . Our results indicate that the detectability of antibody populations depends on the antigen concentrations in the solid phase and on the affinity distribution of the probe to be analyzed . In wells coated with high antigen concentrations, antibody titers reflected antibody concentrations, whereas at low antigen concentrations antibody titers primarily reflect antibody affinities . Independent of their affinities, subpopulations less than 10% could not be detected . Low affinity antibodies were preferentially underestimated . The degree of distortion depended on the composition of the probe to be analyzed . In general, the higher the absolute and the relative affinity of a population, the stronger was its capacity to interfere with the detection of other populations . As a consequence, the heterogeneity of affinity distributions in polyclonal samples may be substantially underestimated . The experiments reported provide guidelines for an optimal design and an adequate interpretation of ELISA based qualitative analyses of polyclonal antibody samples. J Biol Chem, 1992 Jul 5, 267(19), 13585 - 92 Characterization of the mmsAB operon of Pseudomonas aeruginosa PAO encoding methylmalonate-semialdehyde dehydrogenase and 3-hydroxyisobutyrate dehydrogenase; Steele MI et al.; A 5417-base pair (bp) region of Pseudomonas aeruginosa PAO chromosomal DNA containing the mmsAB operon and an upstream regulatory gene (mmsR) has been cloned and characterized . The operon contains two structural genes involved in valine metabolism: mmsA, which encodes methylmalonate-semialdehyde dehydrogenase; and mmsB, which encodes 3-hydroxyisobutyrate dehydrogenase . mmsA and mmsB share the same orientation and are separated by a 16-bp noncoding region . The transcriptional start site for the operon has been pinpointed to a cytidine residue located 77 bp from the translational start site of the operon . mmsR is located on the opposite strand and begins 134 bp from the translational start site of mmsA . MmsR has been identified as a member of the XylS/AraC family of transcriptional regulators and appears to act as a positive regulator of the mmsAB operon . Sequence comparison of MmsA to other proteins in the data bases revealed that MmsA belongs to the aldehyde dehydrogenase (NAD+) superfamily . MmsB shares a 44% amino acid identity with 3-hydroxyisobutyrate dehydrogenase from rat liver . Mutants with insertionally inactivated mmsR, mmsA, and mmsB grow slowly on valine/isoleucine medium and exhibit reduced enzyme activity in cell-free extracts compared to P . aeruginosa PAO. Diagn Microbiol Infect Dis, 1992 Jul, 15(5), 435 - 9 Oral ofloxacin for infections caused by bacteria resistant to oral antimicrobial agents; Eron LJ et al.; We conducted an open trial of oral ofloxacin, 400 mg q 12 hr, in the treatment of infections caused by Pseudomonas aeruginosa and other bacteria resistant to traditional oral antibiotics . There were 53 evaluable subjects, 30 having infection of the skin/skin structure, 13 of the respiratory tract, six of bone, and four complicated infections of the genitourinary tract . Most subjects (72%) had previously failed a course of antibiotic therapy for their infections . Ofloxacin therapy was administered an average of 45 days per patient, and was well tolerated . Clinical success was observed in 40 (74%) subjects, with failure in 13 (26%) . Responses ranged from 50% success for six lower respiratory tract infections to 100% success for six cases of osteomyelitis . There were 28 P . aeruginosa with 22 Gram-positive and 24 other Gram-negative infections . Of 28 cases in which P . aeruginosa was implicated as a pathogen, 19 (68%) were successfully treated with ofloxacin therapy . Overall, 63 (85%) of 74 pathogens were eradicated . Of the 11 persistent organisms, five were P . aeruginosa in which resistance to ofloxacin emerged during therapy . Our data imply ofloxacin to be effective for infections due to ofloxacin-susceptible organisms other than P . aeruginosa; for this pathogen, ofloxacin therapy is associated with a high degree of failure . If ofloxacin is used for infection due to P . aeruginosa, microbiologic surveillance is necessary for cases that either fail to respond or relapse clinically. Infect Immun, 1992 Jul, 60(7), 2808 - 14 Comparison of adherence of Pseudomonas aeruginosa to respiratory epithelial cells from cystic fibrosis patients and healthy subjects; Saiman L et al.; The adherence of Pseudomonas aeruginosa PAO1 to primary cultures of cystic fibrosis nasal polyp (CFNP), normal human nasal polyp (NHNP), and immortalized CF and normal cell lines was studied . PAO1 bound significantly more to primary CFNP cells than to NHNP cells as the mean adherence +/- standard deviation of 5 x 10(7) CFU of 35S-labeled bacteria per ml per well was 15.09 x 10(6) +/- 4.25 x 10(6) CFU/ml per well and 7.62 x 10(6) +/- 2.11 x 10(6) CFU/ml per well, respectively (Mann-Whitney U test, P less than 0.0001) . There was no significant difference in PAO1 adherence to the immortalized CF and normal cell lines . The primary CFNP cells had more receptors (115 per cell) than did NHNP cells (34 per cell) . P . aeruginosa binding to CFNP was blocked by GlcNAc, NeuAc, L-Fuc, and D-Gal, while binding to NHNP was blocked only by GlcNAc, suggesting that receptors on the two cell types were qualitatively different . Pseudomonas supernatants containing protease, phospholipase C, and neuraminidase activity increased adherence to CFNP and NHNP cells . The Pseudomonas exoproducts modified epithelial cell glycoconjugates, as characterized by binding of fluorescein isothiocyanate-labeled lectins and the release of sialic acid . There was minimal release of fibronectin by the bacterial supernatants . The affinity of P . aeruginosa for CF epithelial cells appeared to be due to an increased number of receptors and modification of the epithelial cell surface by P . aeruginosa exoproducts that exposed asialoganglioside binding sites. Pediatr Infect Dis J, 1992 Jul, 11(7), 542 - 6 Outpatient management of chronic suppurative otitis media without cholesteatoma in children; Dagan R et al.; Prolonged antipseudomonal parenteral antibiotic therapy combined with daily aural toilet has been effective in resolving long standing ear discharge in children with chronic suppurative otitis media . However, such treatment suffered from the disadvantages of prolonged hospitalization . We conducted a prospective study to investigate the feasibility and efficacy of exclusive outpatient treatment of children with chronic suppurative otitis media without cholesteatoma who had failed ototopical/oral antimicrobial therapy . The treatment consisted of daily aural toilet (suction and debridement) and twice daily parenteral ceftazidime (50 mg/kg/dose) . Thirty-seven children were included . The duration of discharge from the ear before treatment was 6 to 121 months (median, 30 months) . Aerobic cultures yielded Pseudomonas aeruginosa in 97%, often with other organisms . The management and follow-up were performed jointly by otolaryngology and infectious diseases physicians using the hospital ambulatory services . The route of ceftazidime administration (intravenous or intramuscular) was chosen according to the parents' and patients' convenience . Discharge stopped within 3 to 20 days (median, 8 days) in all children but one . Seventy-six percent of the 29 children available for follow-up 12 months after treatment were still free of discharge . Our results demonstrate that a regiment combining daily aural toilet and twice daily parenteral ceftazidime is highly efficacious in resolving ear discharge in children with chronic suppurative otitis media without cholesteatoma and that such a regimen does not require hospitalization. Mikrobiyol Bul, 1992 Jul, 26(3), 271 - 80 {The effectiveness of various disinfection methods on the surface of gloved hands}; Goktas P et al.; The gloved hands were contaminated by using E.coli and Pseudomonas aeruginosa Brain Heart Infusion broth cultures and the efficacy of tap water, unmedicated bar soap, benzalkonium chloride 1% (Zefiran), Na-hypochloride 1% and 5%, alcohol 70%, hexachlorophene 3%, hexachlorophene 3% liquid soap (Solu-heks), chlorhexidine 1.5% liquid soap (Savlon), chlorhexidine 4% liquid soap (Hibiscrub), povidone-iodine 10% solution (Betadine and polyod), povidone-iodine 7.5% liquid soap (Betadine and Polyod) were compared onto the gloved hands . Disinfectants were applied for 30, 45 and 60 seconds . It was found that washing time of 30 seconds with chlorhexidine 4% (Hibiscrub) liquid soap or povidone-iodine 7.5% liquid soap (Betadine and Polyod) was required to eradicate all the organisms inoculated from both glove surfaces . Povidone-iodine and chlorhexidine were more effective washing agents than the other disinfectants . In underdeveloped or developing countries and areas where gloves can not easily supplied, it has been suggested that gloved hands could be washed between patient treatments and gloves re-used in dentistry, gynecology and like the other areas of medicine . Chlorhexidine 4% and povidone-iodine 7.5% liquid soaps are recommended as a hand-washing agents in these areas. Jpn J Antibiot, 1992 Jul, 45(7), 866 - 79 {Clinical evaluation of a new carbapenem, meropenem, in infants and children}; Meguro H et al.; A new carbapenem antibiotic, meropenem (MEPM), was evaluated for its safety and efficacy in 33 infants and children . MEPM was effective in all the 32 evaluable cases including 4 cases of bacterial meningitis and 5 cases of Pseudomonas aeruginosa infections . The mean half life of plasma concentrations of MEPM was 0.84 +/- 0.09 hours after 30 minutes intravenous drip infusion . Mild diarrhea (2 cases), transient elevation of transaminases (8 cases), and transient eosinophilia (2 cases) were associated with the MEPM therapy, but none of them was problematic . These data suggest that MEPM is safe in infants and children and could be one of the therapeutic agents for severe infections or infections in compromised hosts. Jpn J Antibiot, 1992 Jul, 45(7), 799 - 808 {Clinical evaluation of combined therapy of ceftazidime and tobramycin for intractable pulmonary infection mainly caused by Pseudomonas aeruginosa}; Kikuchi N et al.; In an open, multicenter trial, we investigated the clinical efficacy of a combination therapy of ceftazidime (CAZ) and tobramycin (TOB) for intractable pulmonary infections mainly caused by Pseudomonas aeruginosa . Evaluated for the utility of the combination therapy were 33 cases with pneumonia (Group I: pneumonia caused by P . aeruginosa 15, Group II: pneumonia caused by other Gram-negative bacilli 4 and pneumonia which causative organism was not determined 14) and 23 cases with chronic respiratory tract infection caused by P . aeruginosa . The results obtained are summarized as follows . 1 . In Group I pneumonia, included 11 severe cases and 4 moderate cases, with a mean age of 69.3 years . Significant underlying diseases were present in 14 out of the 15 (93.3%): they included 10 cases of pulmonary diseases and 4 cerebrovascular diseases . The overall efficacy rate in these cases was 60.0%: but the efficacy rate in moderate cases was 100% and that in severe cases was 45.5% . 2 . In Group II pneumonia included 16 severe cases and 2 moderate cases with a mean age of 68.2 years . Significant underlying diseases were present in 15 out of 18 (83.3%, all of the underlying diseases were pulmonary diseases) and the overall efficacy rate was 72.2% with 100% efficacy rate among moderate cases and 68.8% among severe cases . 3 . In the cases with chronic respiratory tract infections caused by P . aeruginosa, the efficacy rate was 82.6% and the eradication rate was 65.2% . We consider the combination therapy of CAZ and TOB is useful for intractable pulmonary infections caused by P . aeruginosa. Vet Microbiol, 1992 Jul, 32(1), 63 - 74 Relationship between the immune response of sheep and the population dynamics of bacteria isolated from fleecerot lesions; Chin JC et al.; In sheep wetted by rain, proliferation of bacteria in the skin-fleece microenvironment invariably discolours the fleece and causes a dermatitic condition known as fleecerot . The changes in population dynamics of fleece bacteria were analysed by carrying out skin washings at randomly selected sites on the back of sheep before, and at 48 h and 96 h after exposure to rain . Gram-positive rods belonging to Bacillus species (10(2)-10(4) cfu/cm2) predominated in dry fleece . Gram-positive cocci (e.g . Micrococcus and Staphylococcus species) as well as Gram-negative rods (pseudomonads) were also present but in lower abundance (less than 10(2) cfu/cm2) . Fleece bacterial populations generally increased in numbers during the first 24-48 h of wetting . By 96 h however, skin washings showed a preponderance of Pseudomonas aeruginosa (10(4)-10(6) cfu/cm2) and to a lesser extent, pigmented Micrococcus species . Growth of fleece bacteria was associated with a characteristic green or yellow/orange staining of fleece . Fewer species of bacteria were isolated from sheep showing green staining while those animals with yellow/orange discolourations appeared to have a more mixed microflora composition . The predominance of P . aeruginosa in the wet fleece of sheep displaying either green or yellow/orange bacterial stain, was accompanied by a significant serological response against this species . Since skin bacteria have never been observed to penetrate cutaneously in skin sections biopsied from fleecerot sites, it must be concluded that the sheep skin is sensitized by continuous exposure to antigens that are associated with or released by P . aeruginosa. Kansenshogaku Zasshi, 1992 Jul, 66(7), 909 - 13 {Growth inhibitory activity of clinical isolates of Pseudomonas aeruginosa against Staphylococcus aureus (MRSA and MSSA)}; Ogino J et al.; Anti staphylococcal activity by clinical isolates of Pseudomonas aeruginosa was tested by the reversed agar plate and the filter paper stamp methods . Almost 40% of Pseudomonas aeruginosa inhibited the growth of both Methicillin resistant Staphylococcus aureus (MRSA) and Methicillin sensitive Staphylococcus aureus (MSSA) . Green pigment (Pyocyanin) produced strains showed a strong inhibitory effect against MRSA and MSSA respectively . But some other pigment (Yellow, Red) strains also showed anti staphylococcal activity . These data suggest the colonization of Pseudomonas aeruginosa with anti staphylococcal activity may not be eradicated by the anti pseudomonic antibiotics. Int J Pediatr Otorhinolaryngol, 1992 Jul, 24(1), 25 - 33 Medical treatment of chronic suppurative otitis media without cholesteatoma in children--a two-year follow-up; Leiberman A et al.; A prospective long-term study was carried out in 48 infants and children with chronic suppurative otitis media without cholesteatoma treated initially with wide spectrum intravenous antibiotics and suction and debridement . Patients were followed for a period of two years . All children were cured after completion of therapy . At 3 and 6 months follow-up 75% of the children were still free of discharge and at 12, 18 and 24 months the proportion of dry ears dropped to 71%, 66% and 52%, respectively . Eighty percent of all recurrences developed already during the first 6 months of follow-up . Pseudomonas aeruginosa was the most common pathogen isolated, both in the initial and recurrent bouts of the disease, and was commonly associated with other pathogens . Children with early reappearance of ear discharge were less likely to benefit from further antimicrobial or surgical treatment . The recurrence rate was not affected by the antibiotic regimen, age, duration of drainage before treatment or the presence of granulation tissue . No intracranial or intratemporal complications were observed during the follow-up period. East Afr Med J, 1992 Jul, 69(7), 394 - 7 The bacteriology of chronic suppurative otitis media; Rotimi VO et al.; Specimens obtained directly from the middle ear under direct vision from 40 patients with chronic suppurative otitis media (CSOM) were investigated quantitatively and qualitatively for aerobes and anaerobes . Twenty one of the 40 specimens yielded aerobes only, 17 yielded a mixed flora of anaerobes and aerobes while only one yielded anaerobes only and the remaining one was sterile . Bacteroides fragilis was the commonest anaerobe and the second single most common bacteria generally present in CSOM . Pseudomonas aeruginosa and Staphylococcus aureus were the dominant aerobic flora appearing in 17 and 14 out of 40 specimens respectively . All pathogens isolated were present in very high counts averaging 10(8) ml of the exudate . The use of systemic anti-anaerobic drug combined with an anti-aerobic drug is worthy of a clinical trial. FEMS Microbiol Immunol, 1992 Jul, 4(5), 267 - 72 Identification of a small epitope in domain Ib of Pseudomonas aeruginosa exotoxin A that elicits enzyme-neutralizing antibodies; Rutault K et al.; A peptide corresponding to amino acids 392-404 of the amino acid sequence of Pseudomonas aeruginosa exotoxin A (the last 13 amino acids of domain Ib) was synthesized and coupled to thyroglobulin . The conjugate induced an antiserum in rabbits with high antibody titer against native toxin as measured by ELISA, and this antiserum was highly efficient in inhibiting the ADP-ribosyltransferase activity of exotoxin A . These data corroborate the potential importance of amino acids 400-404 in the enzymatic mechanism of exotoxin A. J Clin Microbiol, 1992 Jul, 30(7), 1848 - 55 Antibody responses to lipid A, core, and O sugars of the Pseudomonas aeruginosa lipopolysaccharide in chronically infected cystic fibrosis patients; Kronborg G et al.; Enzyme-linked immunosorbent assays were developed separately for the three main parts of the Pseudomonas aeruginosa lipopolysaccharide (LPS) molecule, namely, lipid A, core, and O polysaccharide . Anti-lipid A, anticore, and anti-O polysaccharide antibodies were measured in serum samples from 12 patients with cystic fibrosis (CF) in a longitudinal study covering the period before P . aeruginosa infection was established through at least 5 years of chronic infection . The serum antibody response to all parts of the P . aeruginosa LPS molecule increased during the course of chronic infection . The increase in anti-lipid A antibodies was specific for P . aeruginosa lipid A, since no increase in anti-Escherichia coli lipid A antibodies was seen . Immunoglobulin G, A, and M (IgG, IgA and IgM) antibodies were all involved in the specific systemic response to P . aeruginosa lipid A, core, and the O polysaccharides . IgG and IgA levels in particular increased during the course of infection and were significantly higher than the antibody increase seen with age in a healthy control group . The local immune response in the lungs was investigated by measuring IgG, IgA, and IgM antibodies to the separate parts of the P . aeruginosa LPS molecule in sputum samples from 18 CF patients with at least a 5-year history of chronic P . aeruginosa infection . Antibodies detected in sputum were mainly anti-lipid A and anti-O polysaccharide antibodies of the IgG and IgA isotypes . Very high IgA anti-lipid A titers were detected in sputum samples from some CF patients. Antimicrob Agents Chemother, 1992 Jul, 36(7), 1352 - 7 Effects of the combination of lipopolysaccharide-specific monoclonal antibodies and sparfloxacin against Pseudomonas aeruginosa pneumonia in neutropenic mice; Oishi K et al.; The effects of the combination of a murine monoclonal antibody (MAb) specific for the O side chain of Pseudomonas aeruginosa Fisher immunotype 1 lipopolysaccharide and sparfloxacin in a neutropenic mouse model of P . aeruginosa pneumonia were examined . Under the condition that neither MAb at a dose of 500 micrograms per mouse administered intravenously nor a suboptimal dose of oral sparfloxacin (5 mg/kg of body weight) protected mice from challenge with a fatal dose, the combination therapy with MAb and sparfloxacin caused a significant increase in the survival rate (P less than 0.001 compared with either treatment alone) . The effect of the combination was closely correlated to bacterial killing in plasma and lung tissue of infected mice . In vitro, a significant MAb-dependent, complement-mediated killing of P . aeruginosa was documented in the presence of sparfloxacin at one-half the MIC, while the killing was not observed in the absence of sparfloxacin . These in vivo and in vitro data suggest the usefulness of combination therapy with a lipopolysaccharide-reactive immunoglobulin G MAb and sparfloxacin in neutropenic patients with P . aeruginosa pneumonia. Prostaglandins Leukot Essent Fatty Acids, 1992 Jul, 46(3), 203 - 10 Reduction of eicosanoid production by essential fatty acid depletion does not attenuate the inflammatory response induced by Pseudomonas aeruginosa pneumonia in rat lung; O'Sullivan BP et al.; Sipid mediators of inflammation have been implicated in the pathogenesis of Pseudomonas aeruginosa (PA) related pulmonary damage in patients with cystic fibrosis . We studied the role of these mediators in a rat model of PA endobronchitis using essential fatty acid deficient (EFAD) animals . Whole blood from EFAD animals produced significantly less leukotriene B4 (LTB4) and hydroxyheptadecatrienoic acid when stimulated ex vivo than did whole blood from control animals (p less than 0.005) . Similarly, lung lavage fluid from EFAD animals infected with PA contained less LTB4 and thromboxane B2 (TXB2) than that from control animals . Despite these differences, cellular infiltration of airways in response to PA infection was virtually identical in animals from the regular diet and the EFAD groups . Both EFAD and control animals had a significant increase in white blood cells (WBC) in lung lavage fluid at 1, 3 and 6 days following infection with PA when compared to animals receiving sterile beads . Localized areas of consolidation and nodularity were grossly evident in the lungs of all PA infected animals irrespective of their ability to generate the lipid inflammatory mediators . Microscopic examination of lung sections demonstrated similar changes in all infected animals . We conclude that LTB4 and TXB2 production occurs early in the course of PA pulmonary infection in rats . This early rise in lipid mediators is temporally associated with an influx of WBC into the airways . However, attenuation of eicosanoid production by use of an EFAD diet does not lead to a reduction in the inflammatory response to PA infection.(ABSTRACT TRUNCATED AT 250 WORDS) Am Rev Respir Dis, 1992 Jul, 146(1), 224 - 31 Hepatic versus pulmonary uptake of particles injected into the portal circulation in sheep . Endotoxin escapes hepatic clearance causing pulmonary inflammation; DeCamp MM et al.; Removal of circulating particulates (bacteria, cell debris, endotoxin) is accomplished in most species by macrophages resident in the liver and spleen . We have shown that sheep and other species have phagocytic macrophages resident in their pulmonary capillaries . Moreover, these pulmonary intravascular macrophages accomplish the bulk of uptake of injected tracer particles, bacteria, or endotoxin (LPS) . Because bacteria or LPS of intestinal origin enter the portal circulation, they would first encounter hepatic mononuclear phagocytes . We sought to determine the extent to which particulates injected into the portal circulation of sheep would be taken up by liver or by lung macrophages . Sheep (four per group) were injected via a mesenteric vein with radiolabeled gold colloid, magnetic iron oxide particles, live Pseudomonas aeruginosa, or 125I E . coli endotoxin . For each, the uptake pattern was determined 1 h after injection . Lung and liver were also fixed to determine the cells responsible for uptake and subsequent inflammatory changes . We found that for circulating gold colloid, iron oxide particles, or bacteria, hepatic uptake predominated, and Kupffer cells were responsible . After hepatic uptake of bacteria, inflammatory changes were confined to the liver . In contrast, nearly 50% of endotoxin escaped hepatic clearance and was subsequently removed by the lungs . We then saw inflammatory changes in both lungs and liver . Thus, hepatic macrophages are active in species with pulmonary intravascular macrophages, partially sparing the lungs from uptake and acute inflammation . Endotoxin, however, may elude hepatic uptake, be sequestered in the lungs, and initiate inflammation there. J Bacteriol, 1992 Jul, 174(14), 4847 - 9 The pyocin Sa receptor of Pseudomonas aeruginosa is associated with ferripyoverdin uptake; Smith AW et al.; We have used Tn5 mutagenesis to obtain a mutant resistant to pyocin Sa . When grown in iron-deficient succinate medium this mutant lacked an 85-kDa iron-regulated outer membrane protein (IROMP), and expression of a 75-kDa IROMP was increased compared with that in the parent strain . The mutant was deficient in pyoverdin biosynthesis and showed a 95% decrease in transport of ferripyoverdin purified from the parent strain, suggesting that the 85-kDa IROMP is the specific receptor for ferripyoverdin and pyocin Sa . The mutant compensated for the deficiency in pyoverdin biosynthesis and transport by exhibiting a fourfold increase in ferripyochelin transport . The low-level transport of ferripyoverdin in the Sa-resistant mutant, which extended to heterologous pyoverdins from other strains, suggests that Pseudomonas aeruginosa has a second ferripyoverdin uptake system of lower affinity and broader specificity. J Bacteriol, 1992 Jul, 174(13), 4401 - 9 Cloning of the outer membrane high-affinity Fe(III)-pyochelin receptor of Pseudomonas aeruginosa; Ankenbauer RG; Pseudomonas aeruginosa produces the phenolic siderophore pyochelin under iron-limiting conditions . In this study, an Fe(III)-pyochelin transport-negative (Fpt-) strain, IA613, was isolated and characterized . 55Fe(III)-pyochelin transport assays determined that no Fe(III)-pyochelin associated with the Fpt- IA613 cells while a significant amount associated with KCN-poisoned Fpt+ cells . A P . aeruginosa genomic library was constructed in the IncP cosmid pLAFR1 . The genomic library was mobilized into IA613, and a recombinant cosmid, pCC41, which complemented the Fpt- phenotype of IA613, was isolated . pCC41 contained a 28-kb insert of P . aeruginosa DNA, and the Fpt(-)-complementing region was localized to a 3.6-kb BamHI-EcoRI fragment by deletion and subcloning of the insert . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of IA613 revealed that it lacked a 75-kDa outer membrane protein present in Fpt+ strains . IA613 strains bearing plasmid pRML303, which carries the 3.6-kb BamHI-EcoRI fragment of pCC41, expressed the 75-kDa outer membrane protein and demonstrated a 55Fe(III)-pyoch