Epidemiol Infect, 1989 Apr, 102(2), 205 - 14 Pseudomonas aeruginosa cross-colonization and persistence in patients with cystic fibrosis . Use of a DNA probe; Wolz C et al.; To investigate cross-colonization with and persistence of Pseudomonas aeruginosa in cystic fibrosis (CF), 181 isolates from 76 CF patients were typed using a P . aeruginosa-specific DNA probe . Whereas sibling pairs predominantly harboured genotypically identical P . aeruginosa strains, all of the other patients harboured different strains . Seventy-nine per cent (22/31) of the infected CF patients harboured the same strains at the beginning and the end of a summer camp . A change of strains was seen in 10% (3/31) of the patients at the end of the camp . Forty-six per cent (6/13) of the patients who were apparently initially uninfected, acquired P . aeruginosa by the end of the period . Genotyping proved that strain change or acquisition was due to cross-colonization in four of nine cases . Very little P . aeruginosa was isolated from the inanimate environment . Persistence of P . aeruginosa after a temporary loss due to antibiotic therapy was seen in 12/16 paired patient strains before and after antibiotic therapy . Thus, suppression followed a flare-up seemed to occur in these patients rather than eradication and a new infection . When 35 patients were followed over a period of 6 months, 7 (20%) changed the strain in their sputum . Only one of 43 patients harboured two different P . aeruginosa strains simultaneously over a long period. J Virol, 1989 Apr, 63(4), 1587 - 94 Characterization of the genome of Pseudomonas aeruginosa bacteriophage phi PLS27 with particular reference to the ends of the DNA; Allan BJ et al.; The DNA of Pseudomonas aeruginosa rough-specific bacteriophage phi PLS27 was studied . The genome size as determined by summing the sizes of restriction fragments was 42.7 kilobase pairs . Of particular interest was the fact that the DNA was insensitive to certain common restriction endonucleases including EcoRI, BamHI, and HindIII . The ends of the phage DNA were cloned and sequenced, revealing direct repeats of 318 nucleotides . The left end of the genome when cloned into the promoter selection vector pKK232-8 exhibited promoter activity in Escherichia coli . Two promoters bearing greater than 70% sequence homology to the plasmid pNM74 TOL operon and PAK pilin promoters were identified. Infect Immun, 1989 Apr, 57(4), 1299 - 304 Whole, submandibular, and parotid saliva-mediated aggregation of Pseudomonas aeruginosa in cystic fibrosis; Komiyama K et al.; The aggregation of mucoid and nonmucoid Pseudomonas aeruginosa by submandibular, parotid, and whole saliva from patients with cystic fibrosis (CF) and non-CF subjects was investigated . There were significant differences (P less than 0.01) in aggregation of mucoid and nonmucoid variants of P . aeruginosa by submandibular and whole saliva from CF patients and non-CF subjects . However, the differences in the parotid secretion were not as pronounced . Patients with CF who were colonized with P . aeruginosa demonstrated a significantly higher (P less than 0.05) percent aggregation of the mucoid variants by the submandibular secretion and of both mucoid and nonmucoid variants by whole saliva, compared with corresponding secretions from patients with CF not colonized with this pathogen . The parotid saliva aggregation activity was not markedly different for the two groups with CF . From patients with CF, whole saliva demonstrated a higher percent P . aeruginosa aggregation than did the submandibular saliva . In non-CF subjects, however, the percent aggregation of P . aeruginosa by submandibular saliva was higher than that by whole saliva . Our results indicate that the sero-mucous products of the submandibular gland have a more significant role in P . aeruginosa aggregation than the serous secreting parotid cells and that the submandibular secretion is possibly responsible for the differences in oral colonization by this pathogen in subjects with and without CF. Infect Immun, 1989 Apr, 57(4), 1050 - 3 Adherence of Pseudomonas aeruginosa to tracheal epithelium; Marcus H et al.; Adherence of mucoid and nonmucoid strains of Pseudomonas aeruginosa to tracheal epithelium was studied with a perfused-trachea model . The species specificity of adherence was studied by infecting tracheas from hamsters, guinea pigs, or mice . Perfused tracheas from hamsters were infected with strains of P . aeruginosa in the presence of various sugars, lectins, cations, or charged polymers . Adherence of mucoid strains of P . aeruginosa was greatest for guinea pigs; that for hamsters and mice was approximately the same . Nonmucoid strains did not adhere well to epithelium from any of the species tested . N-Acetylglucosamine, galactose, and N-acetylneuraminic acid were the best inhibitors of adherence of mucoid strains of P . aeruginosa . Phaseolus vulgaris agglutinin and Arachis hypogaea agglutinin enhanced adherence of mucoid strains . Adherence of mucoid strains was also enhanced by the presence of Ca2+ in the incubation medium . Poly-L-lysine, poly-L-aspartic acid, and polyglycine inhibited adherence of a mucoid strain by 96, 86, and 52%, respectively . In general, the adherence of nonmucoid strains was not affected . The results indicate that carbohydrates are involved in the interaction of mucoid strains of P . aeruginosa with tracheal cells and that divalent cations may enhance this interaction . The lectin data show that lectins can interact with the mucoid organisms and the host and suggest that lectins may play a role in the adhesion process. J Surg Res, 1989 Apr, 46(4), 311 - 6 Hepatocellular membrane function during chronic burn injury; Minei JP et al.; Hepatocellular membrane dysfunction, as indicated by depolarization of the membrane potential, occurs after acute injury and early bacteremia . To determine whether hepatocellular membrane dysfunction occurs in the setting of ongoing thermal injury and infection, Wistar rats were divided into four groups: (1) sham-burned, freely fed controls (FF); (2) rats sustaining approximately 30% total body surface area dorsal full-thickness scald burn (Burn); (3) rats sustaining burns as in group 2 followed by immediate inoculation of 1 x 10(8) CFU Pseudomonas aeruginosa (Burn/Inf); and (4) sham-burned rats pair-fed to the food intake of the Burn/Inf group (PF) . On the third and seventh days postburn, body and liver weights were determined . In vivo hepatocellular transmembrane potentials were measured and hepatic ATP, RNA, DNA, and protein contents were assayed . By Day 7, despite greater weight loss in the Burn/Inf group than due to starvation alone (P less than 0.01 Burn/Inf vs FF and PF), hepatic mass was conserved . This was associated with hyperpolarization of the hepatic transmembrane potential (-46.6 +/- 1.5 vs -32.1 +/- 0.6 mV, Burn/Inf vs FF, P less than 0.01) and increases in RNA (141 +/- 9 vs 91 +/- 4 mg/liver, Burn/Inf vs FF, P less than 0.01) and DNA (37 +/- 5 vs 22 +/- 2 mg/liver, Burn/Inf vs FF, P less than 0.05) contents, with no change in ATP or hepatic protein contents . There was a significant hypercorticosteronemia observed in the Burn/Inf group (43 +/- 9 vs 2.8 +/- 0.7 micrograms/dl, Burn/Inf vs FF, P less than 0.01) . This hepatic membrane hyperpolarization and augmented RNA content were not secondary to burn or starvation alone as the response in these groups was significantly less than that of the Burn/Inf group . It is suggested that this hepatic membrane hyperpolarization is one mechanism by which hepatic function is maintained during ongoing burn infection in the rat. FEBS Lett, 1989 Mar 27, 246(1-2), 211 - 7 Expression of the blue copper protein azurin from Pseudomonas aeruginosa in Escherichia coli; Karlsson BG et al.; The structural gene for the blue copper protein azurin from Pseudomonas aeruginosa has been subcloned in different expression plasmid vectors . The highest yield of expression was obtained when the gene with its native ribosome-binding site was placed downstream of the lac promoter in plasmid pUC18 . The protein is exported to the periplasmic space in Escherichia coli and the amount corresponds to 27% of the total protein content in the periplasmic space . The preprotein is cleaved correctly according to N-terminal sequencing of the purified protein . Azurin has been purified in large amounts and is spectroscopically indistinguishable from the protein purified from P . aeruginosa. FEBS Lett, 1989 Mar 27, 246(1-2), 39 - 43 Nucleotide sequence of the aliphatic amidase regulator gene (amiR) of Pseudomonas aeruginosa; Lowe N et al.; The nucleotide sequence of a 1001 bp ClaI/XhoI DNA fragment encoding the amidase regulator gene (amiR) from Pseudomonas aeruginosa has been determined . The sequence derives from strain PAC433, a constitutive high expressing amidase mutant, and contains two overlapping open reading frames . Analysis of the sequence has identified one of the reading frames as amiR . The gene encodes a 196 amino acid polypeptide which shows a strong bias towards codons with G or C in the third position . The amiR gene shows no sequence homology with other bacterial regulator proteins. Vrach Delo, 1989 Mar, (3), 111 - 3 {Indices of lysozyme activity and of liver metabolic function in staphylococcal and Pseudomonas aeruginosa infection}; Pavlenko VA et al.; Experimental investigations indicate that localized forms of staphylococcal, Pseudomonas aeruginosa and associated (Pseudomonas aeruginosa-staphylococcal) infection inhibit the lysozyme activity, cause disorders in the metabolism of P-450 cytochrome, free-radical and iron-containing liver proteins. Jpn J Med, 1989 Mar-Apr, 28(2), 189 - 95 Secretory IgA(S-IgA) levels in sera from patients with diffuse panbronchiolitis; Noda Y et al.; The serum S-IgA levels of 33 patients with diffuse panbronchiolitis (DPB) were compared with those of 13 patients with chronic bronchitis (CB) and 24 patients with bronchiectasis (BE), to obtain information on differences in the pathologic states in DPB and other chronic bronchial diseases . The S-IgA levelw as elevated in all three bronchial diseases, being significantly higher in DPB than in CB, and intermediate in BE . Persistent bacterial infections developed in most of the patients with DPB and two-thirds of those with BE, but in few of those with CB . Serum S-IgA levels were especially high in patients expectorating Pseudomonas aeruginosa-positive sputum, who constituted two-thirds of the patients with DPB and about one-third of those with BE . The highest levels over (100 micrograms/ml) were observed in far-advanced patients with DPB who expectorated P . aeruginosa-positive sputum . The increase in the serum level of IgA was less than that of S-IgA in all three diseases . These results indicate that the marked elevation of the serum S-IgA level in patients with DPB is due to extensive, chronic infection of the airways of the lungs, especially the peripheral airways, and that serum S-IgA is a useful marker for determining the clinical stage and the pathologic state of patients with diffuse peripheral airway diseases. Eur J Biochem, 1989 Mar 1, 180(1), 1 - 8 The histidine residue of codon 715 is essential for function of elongation factor 2; Omura F et al.; Several mutant cDNAs of elongation factor 2 (EF-2) were constructed by site-directed mutagenesis and their products expressed in mouse cells were investigated . Amino acid substitution for the histidine residue of codon 715, which is modified post-translationally to diphthamide, resulted in non-functional EF-2 and this substitution did not render EF-2 resistant to Pseudomonas aeruginosa exotoxin A, which inactivates EF-2 transferring ADP-ribose to the diphthamide residue . These non-functional EF-2s with replacements of the histidine-715 residue showed various extents of inhibition of protein synthesis by competing with functional EF-2 in vivo . These results suggest that histidine-715 is essential for the translocase activity of EF-2 and that the region around diphthamide functions in recognition of, and/or binding to ribosomes . Substitution of proline for the alanine-713 residue and substitution of glutamine for the glycine-717 residue converted EF-2 to partially toxin-resistant forms . Two-dimensional gel analysis with fragment A of diphtheria toxin of these toxin-resistant EF-2s revealed that their ADP-ribosylations by toxin were much less than that of wild-type EF-2. Circ Shock, 1989 Mar, 27(3), 211 - 7 Thallium 201 uptake in kidneys and heart as an indicator of prognosis in septic shock in the rat; Senda M et al.; The change in distribution of cardiac output in septic shock was examined by radionuclide imaging with thallium 201 thallous chloride (201Tl) which allows noninvasive evaluation of relative blood flow to various organs except for the brain . Pseudomonas aeruginosa (1 X 10(9)-2 X 10(10) organisms) were inoculated into the thigh of rats 18-24 hr before the study . The mean arterial pressure was measured with an intracarotid catheter . Fractional blood flow to the heart, kidneys, and liver was evaluated as organ uptake of 201Tl . Those with zero or less than 5% kidney uptake (n = 8) had a high heart uptake and all died within 3 hr even if their pressure was maintained . In contrast, 20 out of 24 rats with kidney uptake greater than 5% survived for more than 6 hr . Those results suggest that the kidney uptake, representing fractional renal blood flow, is an excellent indicator of short-term prognosis in septic shock. Chem Pharm Bull (Tokyo), 1989 Mar, 37(3), 811 - 2 Protease-catalyzed semisynthesis of human neuropeptide Y; Sakina K et al.; Human neuropeptide Y was semisynthesized by enzymatic condensation of des-Tyr36-NH2 human neuropeptide Y and H-Tyr-NH2 using Pseudomonas aeruginosa elastase, a metalloenzyme possessing a hydrolytic specificity for the imino side of hydrophobic amino acids . The optimum pH for this enzymatic synthesis was judged to be around 7 in a high concentration of an organic solvent. Eur Respir J, 1989 Mar, 2(3), 234 - 7 N-acetylcysteine in cystic fibrosis and Pseudomonas aeruginosa infection: clinical score, spirometry and ciliary motility; Stafanger G et al.; The effect of peroral N-acetylcysteine (NAC) in patients with cystic fibrosis (CF) and chronic pulmonary Pseudomonas aeruginosa infection was studied in 52 patients in a double-blind, placebo-controlled, cross-over trial of two, 3 month durations . Active treatment consisted of NAC, 200 mg x 3 daily (patients weighing less than 30 kg) or 400 mg x 2 daily (greater than 30 kg) . The effect was evaluated by a subjective clinical score, weight, sputum bacteriology, blood leucocyte count, sedimentation rate, titres of specific antimicrobial antibodies, lung function parameters and measurement of nasal ciliary function in vitro . 31 patients completed the study . No significant differences in lung function or subjective clinical scores were seen between NAC and placebo for the study group as a whole . Patients with peak expiratory flow rate (PEFR) below 70% of predicted normal values showed a satisfactory significant increase in PEFR, forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) during NAC treatment . No effect of NAC on ciliary activity was observed. Infect Immun, 1989 Mar, 57(3), 771 - 8 Purification and characterization of an extracellular protease from Pseudomonas cepacia; McKevitt AI et al.; An extracellular proteinase (PSCP) produced by Pseudomonas cepacia was purified from culture supernatants by ammonium sulfate precipitation, anion exchange chromatography on DEAE-Sephacel, and G200 gel filtration chromatography . The protease has an apparent Mr of 34,000 by electrophoresis . Substrates cleaved by the protease include gelatin, hide powder, and collagen but not human immunoglobulin G (IgG), IgM, secretory IgA, or IgA . The enzyme had the characteristics of a metalloprotease, a pH optimum of 6, and a temperature optimum of 45 degrees C . Intratracheal instillation of purified PSCP into rat lungs produced a bronchopneumonia characterized by polymorphonuclear cell infiltration and proteinaceous exudation into large airways . Rats responded immunologically to active immunization with PSCP, but this response was not protective against subsequent lung infection with P . cepacia . PSCP was shown to have antigenic similarity with Pseudomonas aeruginosa elastase by an immunoblotting technique . Sera from 10 cystic fibrosis patients, with and without a previous history of P . cepacia colonization, were shown to possess antibody reactive against PSCP. Bull Chest Dis Res Inst Kyoto Univ, 1989 Mar, 22(1-2), 43 - 9 {Three cases of chronic respiratory failure}; Kurasawa T; Three patients with chronic respiratory failure of various etiology, who have (had) been in the hospital for some years, were presented . The case 1 is male and was 57 years old on admission . He has been suffered from right thoracic emphysema from October '83, which is under chronic infection of Pseudomonas aeruginosa, with bronchial fistula and aortic valve insufficiency . His pulmonary function is severely restrictive and the grade of his dyspnea has been V of Hugh-Jone's criteria and he is now unable to leave from bed . The case 2 is male and was 41 y.o . on admission . He has been ill with diffuse cystic bronchiectasis from 33 y.o . and bronchorrhea (greater than 200 ml/day) with chronic infection of Pseudomonas aeruginosa has been lasting and recurrent attacks of infection have progressively worsened of his pulmonary and cardiac functions . He is now indispensable to assist ventilation by artificial respirator every 2-3 days . The case 3 was male and 27 y.o . on admission . He had admitted because of severe dyspnea due to familial pulmonary fibrosis on August '86 . His disorder had been progressive and resistant against repeated corticosteroid therapy . He died of respiratory failure at 30 years old . The transplantations of lung and heart-lung for critical patients with respiratory failure have been challenged in North America and Europe, but in Japan, many social and medical problems about transplantation have yet been unresolved . The indications for and against lung or heart-lung transplantation to these three patients was discussed with reference to English literatures. Proc Natl Acad Sci U S A, 1989 Mar, 86(6), 1954 - 7 Formation of pilin in Pseudomonas aeruginosa requires the alternative sigma factor (RpoN) of RNA polymerase; Ishimoto KS et al.; The promoter region of the Pseudomonas aeruginosa pilin gene has a high degree of similarity to the nitrogen-regulated promoters of enteric bacteria . These promoters are recognized by the alternative sigma factor of RNA polymerase, termed RpoN (NtrA or GlnF) . This observation suggested that the P . aeruginosa pilin gene may be transcribed by the RpoN-containing RNA polymerase . We, therefore, cloned the RpoN gene from P . aeruginosa into Escherichia coli (where it formed a functional product) and used that cloned gene to construct a mutant of P . aeruginosa that was insertionally inactivated in its RpoN gene . This mutant failed to synthesize pilin, indicating that the RpoN sigma factor is required for transcription of the pilin gene. Nihon Kyobu Shikkan Gakkai Zasshi, 1989 Mar, 27(3), 293 - 8 {Therapy against intractable respiratory infections with Pseudomonas aeruginosa}; Nakata K; Patients with diffuse panbronchiolitis (DPB) are frequently affected by Pseudomonas aeruginosa superinfection . To elucidate the predisposing factors of Pseudomonas aeruginosa superinfection in patients with DPB, we analyzed the age of onset, duration of the disease, chest X-ray findings, blood gas levels, lung function, and bacteriological findings . These data were compared with those of patients who had not developed Pseudomonas aeruginosa superinfection . The administration of antibiotics and corticosteroids did not influence the incidence of Pseudomonas aeruginosa superinfection in DPB patients . The patients with long duration, more severe lung function and more deteriorated roentgenological findings developed Pseudomonas aeruginosa superinfection more easily . These infections in the lower respiratory tract significantly affect the prognosis of DPB patients . Using long-term administrations of a new quinolone antibacterial agent against DPB, acute exacerbations were controlled in some patients and the frequency of their admission to hospital was lessened . A multicomponent vaccine raised antibody titers against OEP, elastase, protease and exotoxin in DPB patients . Further clinical investigations are under way in our hospital to confirm the clinical usefulness of the Pseudomonas aeruginosa multicomponent vaccine in DPB from prophylactic and therapeutic points of view. Mol Microbiol, 1989 Mar, 3(3), 371 - 81 Regulation of exotoxin A synthesis in Pseudomonas aeruginosa: characterization of toxA-lacZ fusions in wild-type and mutant strains; Vasil ML et al.; A mobilizable plasmid which carries the promoter for the exotoxin A (ETA) structural gene fused to lacZ was integrated into the chromosome of wild-type and mutant strains of Pseudomonas aeruginosa at the toxA locus by homologous recombination . beta-galactosidase synthesis in the strains (cointegrates) carrying the toxA-lacZ fusions was regulated like ETA synthesis is in P . aeruginosa . Two multicopy plasmids carrying a positive regulatory gene designated toxR were constructed which are identical except with respect to the orientation of toxR to the lacZ promoter on the plasmid . These plasmids were then introduced into P . aeruginosa cointegrate strains . When toxR was using its own promoter, synthesis of beta-galactosidase in the cointegrate strains was increased but the pattern of iron regulation was not altered . In contrast, when the lacZ promoter was directing synthesis of the toxR product in the cointegrate strains, iron regulation of beta-galactosidase and ETA synthesis were abolished. Mol Gen Genet, 1989 Mar, 216(1), 75 - 80 Cloning and sequencing of the Pseudomonas aeruginosa 1244 pilin structural gene; Castric PA et al.; The pilin structural gene of Pseudomonas aeruginosa 1244 was cloned in both cosmids and lambda . Expression of the cloned gene was detected in P . aeruginosa strains PAO2003, PA103, and 653A by an immunoblot reaction utilizing monoclonal antibodies . Western blot analysis showed that pilin expressed from the cloned gene was slightly larger than native 1244 pilin when produced in strains PAO2003 and 653A, but distinctly smaller in PA103 . Bacteriophages specific for the 1244 pilus did not lyse strain PAO2003 containing the cloned 1244 pilin gene, indicating that functional 1244 pili were not assembled in this recombinant strain . Nucleotide sequencing revealed a coding region which when translated would produce a 15,615 dalton peptide . The amino-terminal region of this peptide is identical with published pilin sequences . While the rest of the peptides are generally dissimilar, common residues are seen within potentially antigenic regions. J Clin Microbiol, 1989 Mar, 27(3), 558 - 60 Clinical evaluation of a direct fluorescent monoclonal antibody test for detection of Pseudomonas aeruginosa in blood cultures; Pfaller MA et al.; A direct fluorescent monoclonal antibody test (DFA; Genetic Systems Corp., Seattle, Wash.) was evaluated for the detection of Pseudomonas aeruginosa in 178 blood culture broths obtained from 128 patients . The DFA identified 44 (98%) of 45 blood cultures positive for P . aeruginosa and was negative in 131 (98%) of 133 blood cultures which grew gram-negative rods other than P . aeruginosa . Upon further investigation, saline suspensions of the organism from the false-negative blood culture were strongly (4+) DFA positive . The false-positive reactions were not due to cross-reactivity, as shown by lack of DFA staining of the non-P . aeruginosa isolates following subculture to agar media . The specificity of the reagent was further demonstrated by directly staining culture isolates including 10 serotypes of P . aeruginosa (all positive) and 57 selected gram-negative bacilli including eight species of Pseudomonas that were not P . aeruginosa (all negative) . DFA staining of blood culture broths was easy to perform and read with minimal background fluorescence . The DFA method can be performed in 50 min and appears promising as a rapid method for the identification of P . aeruginosa bacteremia. J Clin Microbiol, 1989 Mar, 27(3), 490 - 4 Comparative evaluation of mitogenicity and basement-membrane-degrading activity of Pseudomonas aeruginosa slime glycolipoprotein and alginate; Anastassiou ED et al.; Alginate from a heavily mucoid Pseudomonas aeruginosa strain and slime glycolipoprotein obtained from the revertant nonmucoid variant of the mucoid strain were tested for mitogenic activity on human peripheral lymphocytes and for degradation of 3H-labeled basement membranes of the anterior lens capsule of bovine eyes . Slime glycolipoprotein exerted mitogenic activity in concentrations from 50 to 200 micrograms/ml, whereas alginate was not mitogenic as shown by {3H}thymidine uptake . Alginate did not show any basement membrane degradation, whereas slime glycolipoprotein exhibited basement-membrane-degrading activity from 35 to 450 micrograms/ml in a dose-related manner . This activity was inhibited by metal chelators but not thiol protease inhibitors . The results suggest that alginate lacks the mitogenic and biodegrading activities of slime glycolipoprotein; these activities nevertheless need further investigation. Eur J Clin Microbiol Infect Dis, 1989 Mar, 8(3), 233 - 7 Once-daily versus thrice-daily administration of netilmicin in combination therapy of Pseudomonas aeruginosa infection in a man-adapted neutropenic animal model; Gerber AU et al.; A granulocytopenic mouse model was used to elucidate the impact of dose spacing on the activity of netilmicin against Pseudomonas aeruginosa . A thigh infection was produced and then treated with netilmicin combined with azlocillin . Netilmicin was injected subcutaneously at decreasing doses every 20 min to result in plasma-concentration-time curves similar to those observed in patients on intravenous netilmicin treatment . A once-daily regimen was simulated and compared to a simulated conventional schedule of every 8 h . Identical total amounts of drug were used in both groups of comparatively treated mice . Therapeutic efficacy was quantitated by repeated determinations of surviving organisms in thigh homogenates . Combination therapy was significantly more effective than azlocillin treatment alone . In combination regimens the simulated once-daily netilmicin schedule killed the target organisms faster than the simulated thrice-daily regimen and was significantly more efficacious by 24 and 32 h in two out of three strains of Pseudomonas aeruginosa tested . It is concluded that the results of combination therapy of severe Pseudomonas aeruginosa infections in the immunocompromised host might be improved by choosing an aminoglycoside dosage interval of 24 h instead of the conventional 8 h. Pneumologie, 1989 Mar, 43(3), 147 - 58 {Phagocytosis of Staphylococcus aureus and Pseudomonas aeruginosa by pulmonary macrophages and granulocytes}; Hurter T; The elimination (by human alveolar macrophages and pulmonary granulocytes) of one laboratory strain and one wild-type strain each of Staphylococcus aureus and Pseudomonas aeruginosa has been investigated in vitro . Administering 10(6) alveolar macrophages to 5 x 10(6) staphylococci leads to a reduction of viable colony-forming units (CFU) to 5.8 x 10(5) (L} (laboratory strain (L) and 6.9 x 10(5) (wild-type strain (W)) after 60 minutes . The corresponding values of Ps . aeruginosa are 4.3 x 10(6) (L) and 4.4 x 10(6) (W) . The peak chemiluminescence values are 9, 160 (L) and 12,300 (W) counts/10(5) cells for Staph . aureus and 7,670 (L) and 8,950 (W) for Ps . aeruginosa . The addition of heat-inactivated serum increases the number of staphylococci observed within alveolar macrophages from 1.57 to 3.18 (L) and from 1.32 to 2.79 (W) . Fresh serum enhances ingestion to 8.15 (L) and 9.73 (W) staphylococci/macrophage . The number of phagocytized pseudomonads in pure culture medium is 0.51 (L) and 0.46 (W) bacteria/macrophage . An increase by the addition of serum is not possible . While staphylococci adhere to the membrane of the macrophage over a wide area, the pseudomonads adhere to it in a highly circumscribed manner . The following ingestion steps do not differ from one organism to the other . After 60 minutes, cultivation of 10(6) granulocytes with 5 x 10(6) staphylococci leads to a reduction of the viable CFU to 7.5 x 10(5) (L) and 9.4 x 10(5) (W) . In the case of Ps . aeruginosa, the number of surviving CFU is 4.5 x 10(6) (L) and 3.8 x 10(6) (W) . After the addition of Staph . aureus, peak values of 37,800 (L) and 35,500 (W) counts/10(5) cells are measured . The corresponding values for Ps . aeruginosa are 28,900 (L) and 33,100 counts/10(5) cells (W) . The addition of heat-inactivated serum leads, in the case of Staphylococcus aureus, to an increase in the ingestion of the laboratory strain from 1.07 to 1.94 and of the wild-type strain from 0.94 to 2.41 bacteria/granulocyte . Fresh serum brings about a further increase to 5.52 (L) and 4.82 (W) . In the case of Ps . aeruginosa . 0.75 (L) and 0.69 (W) ingested bacteria/granulocyte are observed; the addition of serum fails to bring about any increase . In contrast to the staphylococci, pseudomonads adhere to the granulocyte membrane in a punctate manner only . No differences of ingestion are seen. J Burn Care Rehabil, 1989 Mar-Apr, 10(2), 131 - 7 Combined host and specific anti-Pseudomonas-directed therapy for Pseudomonas aeruginosa infections in burned mice: experimental results and theoretic considerations; Holder IA et al.; Four strains of Pseudomonas aeruginosa showed significant variation in the elaboration of the virulence-associated exoproducts, exotoxin A and proteases . These variations appeared to affect the mortality observed when these strains were used to infect burned mice . Further, these variations seemed to influence the efficacy of individual after sepsis treatments designed to (1) reduce total microbial load in the host (HIg treatment), (2) reduce total circulating protease load in the host (alpha 1-PI treatment), or (3) neutralize the specific effects of exotoxin A (antitoxin treatment) . Combined treatment, using all of these approaches, was significantly better than individual treatment. Isr J Med Sci, 1989 Mar, 25(3), 123 - 6 Bone and joint infections due to Pseudomonas aeruginosa: clinical aspects and treatment; Finkelstein R et al.; The occurrence of nine cases of Pseudomonas septic arthritis and osteomyelitis within a 12-month period may be considered an uncommon experience even in a large, tertiary referral center . These cases are reported here and the various clinical patterns and therapeutic implications associated with Pseudomonas bone infections are discussed . Epidemiologic serotyping of the isolated strains indicated that they do not represent a single nosocomial outbreak. J Med Microbiol, 1989 Mar, 28(3), 183 - 9 Isolation of mucoid strains of Pseudomonas aeruginosa from non-cystic-fibrosis patients and characterisation of the structure of their secreted alginate; McAvoy MJ et al.; When the incubation period of primary isolation plates was extended to 48 h, mucoid strains of Pseudomonas aeruginosa were found in specimens from various infected sites in patients who did not have cystic fibrosis . The 17 mucoid isolates were characterised in terms of mucoid type, pyocin type, and their sensitivity or resistance to seven beta-lactam and two aminoglycoside antibiotics . The carbohydrate, uronic acid (alginate) and protein content of the water-soluble extracellular material of 15 strains was determined . This material was fractionated by ion-exchange chromatography, and the presence of alginate confirmed by the chemical assay of uronic acids and their quantitation by gas-liquid chromatography . Uronic acids were absent from a non-mucoid revertant of one strain . The strains produced alginate with a high content of mannuronic acid and substituted with O-acetyl groups . By proton nuclear magnetic resonance (1H-nmr) analysis the alginate from three strains was shown to lack polyguluronate blocks in its structure . These properties are also found in the alginate of mucoid P . aeruginosa strains from patients with cystic fibrosis. J Bacteriol, 1989 Mar, 171(3), 1698 - 704 Structural gene and complete amino acid sequence of Pseudomonas aeruginosa IFO 3455 elastase; Fukushima J et al.; The DNA encoding the elastase of Pseudomonas aeruginosa IFO 3455 was cloned, and its complete nucleotide sequence was determined . When the cloned gene was ligated to pUC18, the Escherichia coli expression vector, bacteria carrying the gene exhibited high levels of both elastase activity and elastase antigens . The amino acid sequence, deduced from the nucleotide sequence, revealed that the mature elastase consisted of 301 amino acids with a relative molecular mass of 32,926 daltons . The amino acid composition predicted from the DNA sequence was quite similar to the chemically determined composition of purified elastase reported previously . We also observed nucleotide sequence encoding a signal peptide and "pro" sequence consisting of 197 amino acids upstream from the mature elastase protein gene . The amino acid sequence analysis revealed that both the N-terminal sequence of the purified elastase and the N-terminal side sequences of the C-terminal tryptic peptide as well as the internal lysyl peptide fragment were completely identical to the deduced amino acid sequences . The pattern of identity of amino acid sequences was quite evident in the regions that include structurally and functionally important residues of Bacillus subtilis thermolysin. J Bacteriol, 1989 Mar, 171(3), 1278 - 83 The algR gene, which regulates mucoidy in Pseudomonas aeruginosa, belongs to a class of environmentally responsive genes; Deretic V et al.; The Pseudomonas aeruginosa capsule, composed of polysaccharide alginate, is an important Pseudomonas virulence factor encountered primarily in cystic fibrosis . The regulatory algR gene positively controls transcription of a key alginate biosynthetic gene, algD . The algR gene was subcloned and sequenced by creating a set of nested deletions in M13 bacteriophage . DNA sequence analysis of algR revealed the homology of its gene product with a recently recognized class of environmentally responsive bacterial regulatory genes, including ompR, phoB, sfrA, ntrC, spoOA, dctD, and virG; these transcriptional activators control cellular reactions to osmotic pressure, phosphate limitations, or specific chemical compounds present in the medium or released from wounded host tissue . These findings indicate that novel conditions in lungs affected by cystic fibrosis may be participating in the control of mucoidy. Arch Intern Med, 1989 Mar, 149(3), 630 - 4 Analysis of amikacin-resistant Pseudomonas aeruginosa developing in patients receiving amikacin; Maloney J et al.; During a 36-month period, 28 patients treated for infections due to amikacin-susceptible Pseudomonas aeruginosa subsequently developed infections or colonization with amikacin-resistant P aeruginosa at the same site . Eleven amikacin-susceptible/-resistant pairs of isolates were analyzed for aminoglycoside-inactivating enzymes, plasmid profiles, cellular proteins, outer membrane proteins (OMPs), lipopolysaccharide (LPS) profiles, and amikacin uptake . While clearly distinct from isolates of other patients, sensitive and resistant isolates from the same patients were indistinguishable in plasmid profile, LPS profiles, and OMPs . These results suggest that the resistant P aeruginosa isolates were derived from the sensitive isolates . None of the resistant isolates produced enzymes known to inactivate amikacin . In nine of 11 resistant isolates tested, transport of amikacin into P aeruginosa was reduced . A major mechanism of in vivo development of amikacin resistance in P aeruginosa is alteration in permeability to amikacin, but the aquisition of plasmids or changes in OMPs or LPS profile may not account for this phenomenon. Plast Reconstr Surg, 1989 Mar, 83(3), 494 - 9 Suction lipoplasty: biohazardous aerosols and exhaust mist--the clouded issue; Cukier J et al.; With the increased popularity of suction lipoplasty procedures, attention has been focused on their safety . One significant concern involves the rotary vane aspirators used to provide the suction required for the procedure . A series of experiments was carried out to determine whether aerosols are produced during the use of a rotary vane aspirator, since aerosols are known to be hazardous under appropriate conditions . Using a viable strain of Pseudomonas aeruginosa, we challenged the system through the suction port, and the exhaust from the aspirator was then cultured in a particle sampler . Results indicate that viable pathogens are released from the exhaust in physiologically significant particles capable of penetrating to the level of the alveolus in the normal human lung . These infectious particles were produced for 3 hours after the initial challenge . When an appropriate filtration device was attached to the aspirator outflow, the aspirator pump and environment were protected . In the absence of an appropriate filtration device, the aerosolized particles may constitute a hazard to patients or medical workers in the vicinity of the aspirator. Infect Immun, 1989 Mar, 57(3), 996 - 8 Exoenzyme S of Pseudomonas aeruginosa ADP-ribosylates the intermediate filament protein vimentin; Coburn J et al.; Exoenzyme S, which had been thought to be unselective, catalyzes the ADP-ribosylation of only a subset of cellular proteins . The intermediate filament protein vimentin is one of the more abundant substrates . Disassembled vimentin, and proteolytic fragments of vimentin that cannot form filaments, is more readily ADP-ribosylated than is filamentous vimentin. Infect Immun, 1989 Mar, 57(3), 882 - 6 Pseudomonas aeruginosa cytotoxin: periplasmic localization and inhibition of macrophages; Kluftinger JL et al.; Pseudomonas aeruginosa cytotoxin has been isolated previously from cell autolysates . Both purified cytotoxin and periplasmic contents (osmotic shock fluid) cross-reacted on Western immunoblots with antibodies specific for cytotoxin . In addition, both preparations caused a significant reduction in antibody-mediated phagocytosis of P . aeruginosa M2 by mouse macrophage cell line P388D1 . Phagocytosis was restored in each case on preincubation of cytotoxin or periplasmic contents with anti-cytotoxin serum . Both cytotoxin and periplasmic contents caused depolarization of the P388D1 cell membrane, as demonstrated with a polarization-sensitive fluorescent probe . Similar correlations were not observed for other P . aeruginosa cell fractions or for osmotic shock fluid from Escherichia coli C600 . These data indicate that P . aeruginosa cytotoxin is localized in the periplasm and has the potential to inhibit macrophage-mediated phagocytosis, possibly by perturbing ion gradients across the macrophage plasma membrane. Infect Immun, 1989 Mar, 57(3), 817 - 22 Stimulation by fibronectin of macrophage-mediated phagocytosis of Pseudomonas aeruginosa; Kluftinger JL et al.; In a previous investigation it was determined that Pseudomonas aeruginosa cells taken directly from a mouse in vivo growth system were significantly more susceptible to nonopsonic phagocytosis by macrophages than were similar cells after being washed in buffer (N . M . Kelly, J . L . Battershill, S . Kuo, J . P . Arbuthnott, and R . E . W . Hancock, Infect . Immun . 55:2841-2843, 1987) . It was demonstrated that a phagocytosis-promoting factor was found in the supernatant obtained from chambers incubated in the peritoneal cavities of laboratory mice or rats . The phagocytosis-promoting factor was effective with both strains of P . aeruginosa tested, using both unelicited mouse peritoneal macrophages and the P388D1 mouse macrophage cell line as the phagocytic cells . Phagocytosis enhancement was observed with in vivo-grown bacteria and with bacteria grown in vitro on agar plates, but not with bacteria grown in vitro with rapid agitation . Supernatants from mice and rats were fractionated using a fast pressure liquid chromatography gel exclusion column . The phagocytosis-promoting factor copurified with fibronectin . Furthermore, antifibronectin sera negated the phagocytosis-promoting activities of in vivo chamber supernatant, while commercial bovine fibronectin was itself capable of promoting phagocytosis . The concentrations of fibronectin increased in both rat and mouse peritoneal chambers with time, coincident with the ability of chamber supernatants to promote phagocytosis . It was concluded that fibronectin was the phagocytosis-promoting factor of chamber supernatants . Bacterial presence in the peritoneal chambers was not required to elicit fibronectin uptake into the chambers. FEMS Microbiol Immunol, 1989 Mar, 1(4), 245 - 51 Saccharides of seven Pseudomonas aeruginosa immunotypes; Stanislavsky ES et al.; The structures of O-specific polysaccharides obtained by mild acid degradation of lipopolysaccharides (LPS) from seven Pseudomonas aeruginosa Fisher's immunotypes have been studied . The polysaccharides consist mainly of monoamino and diamino sugars, frequently also carrying acidic functions . Some of the sugars were detected in nature for the first time in these organisms . The structures of the O-specific polysaccharides of the immunotypes 2, 3, 4, 5 and 6 are identical to those of the polysaccharides of the 011; 0(2a)2c; 01; 010a, 10b and 07a, 7d Lanyi-Bergan serological subgroups respectively, whereas no analogues have been found for the immunotypes 1 and 7 . Some cross-reactions between the LPS of different immunotypes were observed in passive haemagglutination tests; the results of inhibition of passive haemagglutination and agar gel immunoprecipitation point, however, to a specificity of the LPS . Many of the LPS of the seven Pseudomonas aeruginosa immunotypes manifest rather a high cross-protective activity in active immunization tests in mice . The nature of the cross-protective activity of the LPS is discussed. Mol Microbiol, 1989 Mar, 3(3), 421 - 8 Cloning and analysis of the gene for the major outer membrane lipoprotein from Pseudomonas aeruginosa; Cornelis P et al.; The gene for the Pseudomonas aeruginosa outer membrane lipoprotein I was isolated from a genomic library in the phage lambda EMBL3 vector and subsequently subcloned in the low copy-number, wide host-range plasmid vector, pKT240 . The cloned gene was highly expressed, resulting in the production of a low molecular-weight protein (8 kD) that was found to be associated with the outer membrane . Sequence analysis showed an open reading frame of 83 amino acids with a putative N-terminal hydrophobic signal peptide of 19 residues immediately followed by the lipoprotein consensus sequence, GLY-CYS-SER-SER (residues 19-22) . The predicted amino acid composition of the mature polypeptide and that of the purified lipoprotein I of P . aeruginosa (Mizuno and Kageyama, 1979) were identical . In contrast with other Gram-negative outer membrane lipoproteins, conformation predictions suggested that the mature protein was a single alpha helix. Eur J Biochem, 1989 Feb 15, 179(3), 667 - 75 Permeability to cefsulodin of the outer membrane of Pseudomonas aeruginosa and discrimination between beta-lactamase-mediated trapping and hydrolysis as mechanisms of resistance; Hewinson RG et al.; A pair of strains of Pseudomonas aeruginosa (3-Pre: cefsulodin-sensitive, inducible beta-lactamase; and 3-Post: cefsulodin-resistant, elevated beta-lactamase, derived from 3-Pre by subculture in the presence of cefsulodin) were taken as representative of the class of bacteria resistant to third-generation cephalosporins due to elevated synthesis of the normally inducible, chromosomally encoded beta-lactamase . These two strains were used to differentiate between 'trapping' and 'hydrolytic' mechanisms of cefsulodin resistance by (a) measuring the outer-membrane permeabilities to cefsulodin, (b) measuring the kinetics of cefsulodin hydrolysis and the stoichiometry of cefsulodin trapping by the periplasmic beta-lactamase, and (c) comparing the predictions of the trapping and hydrolysis hypotheses with the minimum inhibitory concentrations (MIC) of cefsulodin . The MIC of cefsulodin for strains 3-Pre and 3-Post were 2.35 microM (1.25 micrograms ml-1) and 37.6 microM (20.0 micrograms ml-1) respectively . The permeability parameter for cefsulodin of the outer membrane of the resistant strain was 0.0034 cm3 min-1 mg dry mass-1, so the flux of cefsulodin across its outer membrane at the MIC was calculated to be 0.120 nmol min-1 mg dry mass-1 . Hydrolysis of cefsulodin by the beta-lactamase in the periplasm occurred at a rate of 0.118 nmol min-1 mg dry mass-1 which can thus account for resistance by matching the above rate of inflow . Trapping by the beta-lactamase, even with a 1:1 stoichiometry, would require the enzyme to be synthesized at 5.0 micrograms protein min-1 mg dry mass-1 or about 40% of the dry mass/generation . We conclude that hydrolysis, but not trapping, adequately explains the resistance to cefsulodin in P . aeruginosa 3-Post . A similar calculation for latamoxef resistance, using data taken from the literature, led to the same conclusion. J Clin Pharm Ther, 1989 Feb, 14(1), 29 - 34 Microbial contamination of cosmetics and personal care items in Egypt--shaving creams and shampoos; Abdelaziz AA et al.; We examined a total of 192 samples, including eight different brands of shaving cream and eight brands of shampoo, for their total aerobic bacterial, coliforms and fungal counts . Shaving creams were more heavily contaminated with bacteria than shampoos . Viable bacterial were not recovered from 57% and 10% of shampoos and shaving creams, respectively . Only 3% of shaving creams were heavily contaminated with more than 10(4) c.f.u./g, while none of the shampoos contained such a high number of bacteria . With regard to the medium range contamination levels, 52% of shaving creams showed bacterial counts ranging from 10(2) to 10(3) c.f.u./g or ml, compared to 15% of shampoos which were contaminated to the same level . Fourteen per cent of shaving creams were contaminated with greater than 10(3)-10(4) c.f.u./g or ml, compared to 1% of the shampoos . No coliforms were recovered from either the shaving creams or the shampoos; however, Staphylococcus spp . were detected in six samples of both shampoos and shaving creams . Some of these Staphylococci, were aureus type . One isolate of Pseudomonas aeruginosa was also detected in a sample of shampoo . The incidence of fungal contamination was much less than the bacterial contamination . No viable fungi were recovered from 88% and 76% of the shaving creams and shampoos, respectively . The majority of the remaining samples, for both products, were contaminated with less than 100 fungal cell/g or ml . The pH of all the tested samples was alkaline (pH 7.2-9), which is well known to inhibit fungal contamination. J Pediatr, 1989 Feb, 114(2), 309 - 14 Intravenous immune globulin treatment of pulmonary exacerbations in cystic fibrosis; Winnie GB et al.; The effect of intravenously administered immune globulin (IVIG) on patients with cystic fibrosis with an acute exacerbation of pulmonary infection was evaluated in a double-blind study . Patients at least 12 years of age, with chronic respiratory tract colonization with Pseudomonas aeruginosa and hospitalized with a reduction in pulmonary function, were randomly assigned to receive 20% dextrose (control subjects: n = 8) or 100 mg/kg IVIG (Gamimune) (experimental subjects: n = 8) on days 1, 2, and 3; all patients received intravenous antibiotics and chest physiotherapy . There were no differences between groups on admission; patients had moderate to severe disease as measured by Shwachman-Kulczycki scores and pulmonary function tests . Both groups improved clinically . The IVIG treatment was associated with significant increases in forced vital capacity and forced expiratory volume in 1 second (p less than 0.01) and with greater percent improvement in forced expiratory volume and forced expiratory flow (25% to 75%) (p less than 0.05) . There was no effect on length of hospitalization (18.3 +/- 11.9 days control vs 17.6 +/- 6.5 experimental) . The C3 level was decreased at discharge in IVIG-treated patients; circulating immune complex levels were unchanged . One patient in each group experienced side effects . There were no differences on follow-up at 6 weeks . We conclude that IVIG infusion early in treatment for pulmonary exacerbations in cystic fibrosis patients with moderate to severe disease may be associated with greater improvement in pulmonary function than standard treatment alone. J Infect Dis, 1989 Feb, 159(2), 232 - 8 Piroxicam treatment protects mice from lethal pulmonary challenge with Pseudomonas aeruginosa; Sordelli DO et al.; The effect of treatment with the nonsteroidal anti-inflammatory agent piroxicam on leukocyte migration to the lungs was investigated after aerosol administration of sublethal doses of Pseudomonas aeruginosa to mice . Piroxicam decreased, in a dose-related fashion, the polymorphonuclear leukocyte recruitment to, and the degree of perivascular and peribronchial infiltration in, the lungs . Piroxicam treatment also protected the animals in a dose-dependent manner from challenge with lethal doses of P . aeruginosa . The effect of piroxicam was not related to direct action of the drug on the microorganisms . Piroxicam treatment maintained the animal's pulmonary defenses against infection while diminishing inflammatory responses against P . aeruginosa, an occurrence decreasing the potential for tissue damage due to phagocytes migrating from circulation. Kinderarztl Prax, 1989 Feb, 57(2), 81 - 7 {The incidence of lung infection in patients with mucoviscidosis and the disease course with various pathogen spectra}; Dietzsch HJ et al.; A retrospective review on the frequency of lung infections by the most important organisms Staphylococcus aureus and Pseudomonas aeruginosa in patients with cystic fibrosis in the GDR during the period from 1981 to 1985 revealed an average infectious rate of 58.6 per cent by Staphylococcus and of 26.9 per cent by Pseudomonas respectively . A distinct increase of the infections by Pseudomonas could only be documented from 1981 to 1982, later on the infectious rate remained nearly equal and showed only a peak value of 35.6 per cent in 1983 . Thus the frequency of infections by Pseudomonas is significantly lower in the GDR than in most other countries . The comparison of the course of the disease in patients with permanent lung infection during 4 or 5 years established a mortality rate of a double amount in the group of patients with Pseudomonas colonisation versus the group with Staphylococcus colonisation . Otherwise no significant difference could be stated in the mean age of the beginning of lung infection (Staphylococcus = 9.7 years of age - Pseudomonas = 10.1 years of age) . Analysing the pulmonary x-ray findings we found a significantly more rapid deterioriation during the follow-up period in patients with permanent Pseudomonas infection than in patients with permanent Staphylococcus infection, whereas the evolution of body height and weight did not take different course. Rinsho Ketsueki, 1989 Feb, 30(2), 158 - 63 {Sepsis in patients with hematological diseases}; Kanamori H et al.; Sepsis is one of the important complications on the treatment of severe hematological diseases . In this report, we analyzed sepsis in 309 patients with hematological diseases who were admitted to the First Department of Internal Medicine of Yokohama City University Hospital from 1979 to 1986 . Positive blood culture were found in 17.8% (55/309 cases) and total positive cases were 73 including recurrent patients . Positive rate by underlying diseases was 30.3% in acute leukemia, 20.8% in chronic myelocytic leukemia, 17.2% in aplastic anemia, 8.0% in multiple myeloma, 6.0% in malignant lymphoma and 6.5% in others . The organisms causing sepsis were as follows; gram negative bacilli 56.4%, gram positive organisms 34.6%, fungus 6.4% and anaerobic bacteria 2.6% . Pseudomonas aeruginosa was found in 19.2% . The mortality rate of patients with sepsis was 34.2% (25/73 cases) . The significant prognostic factors in patients with sepsis were the degree of neutropenia, duration of neutropenia (500 less than microliters), the species of organisms, simultaneous complication with shock and the site of other infections. Br J Exp Pathol, 1989 Feb, 70(1), 1 - 8 In-vivo blockage of neutrophil migration by LPS is mimicked by a factor released from LPS-stimulated macrophages; Cunha FQ et al.; The present study was performed to determine the effect of an intravenous injection of the macrophage-derived neutrophil chemotactic factor (MNCF) (Cunha & Ferreira 1986) on neutrophil migration to rat peritoneal cavities, which were challenged with chemotactic stimuli . Macrophage monolayers stimulated by LPS release a factor (MW greater than 10,000 D) which, when injected intravenously, blocked neutrophil migration in carrageenin-induced peritonitis . This inhibition was dependent on dose and lasted more than 2 h . It was not due to neutropaenia, hypotension or LPS contamination . Neutrophil migration induced by LPS, MNCF, the Gram-negative bacterium Pseudomonas aeruginosa was also blocked by intravenous administration of the factor . Intravenous injection of recombinant interleukin 1 beta or tumour necrosis factor-alpha, present in the samples of the factor, failed to reproduce the described inhibitory effect on neutrophil migration . The release of this factor by LPS-stimulated macrophage monolayers was inhibited by dexamethasone but not by indomethacin . It is suggested that the failure of neutrophils to migrate during septicaemia may be the result of a continuous release of chemotactic factors in the circulation, particularly of the macrophage-derived neutrophil chemotactic factor(s). J Med Microbiol, 1989 Feb, 28(2), 101 - 8 Antibody-independent protection against Pseudomonas aeruginosa infection in mice after treatment with a homologous strain vaccine; Fujimura T et al.; Formalin-killed cells of Pseudomonas aeruginosa strain M-24 elicited an antibody-independent protective effect against P . aeruginosa infection in mice . The effect was observed as early as 6 h after administration and 100% protection was obtained by 48 h . The protective effect could not be attributed to the production of specific antibody . In M-24-treated mice, the bacteria in the peritoneal cavity, blood and liver were eliminated 12 h after P . aeruginosa infection . This suggested that the protective effect was due to enhanced bacterial elimination . The percentage of macrophages in the peritoneal cavity was increased after M-24 administration . Furthermore, the enhanced bacterial elimination was abrogated by treatment of mice with 60Co-irradiation or carrageenan . These findings suggest the involvement of macrophages in the enhanced bacterial elimination observed . The chemiluminescence of peritoneal exudate cells from M-24-treated mice was markedly increased when compared with that of cells from untreated mice . The ability to kill P . aeruginosa in vitro was also greater in macrophages from mice treated with killed M-24 than in cells from proteose-peptone-treated mice . The M-24-treated mice showed enhanced nonspecific protection against infection with lethal doses of P . aeruginosa, Escherichia coli or Listeria monocytogenes . However, susceptibility to LPS in mice was not increased by M-24 treatment . These results suggest that macrophage activation without increasing LPS susceptibility was responsible for the antibody-independent protection induced by killed M-24. J Hosp Infect, 1989 Feb, 13(2), 109 - 15 Bacterial contamination of dialysate in dialysis-associated endotoxaemia; Watzke H et al.; Bacteriological investigations and endotoxin (ET) determinations were performed during a routine haemodialysis session for six patients . The glucose free dialysate was prepared with untreated tap water . All patients were dialysed for 5 h . Pseudomonas aeruginosa was regularly isolated in numbers up to 10(7) cfu ml-1 from samples of the dialysate inflow, the dialysate site and the dialysate outflow . ET levels in the plasma of the patients increased continuously during haemodialysis and were always higher in the blood outflow line of the dialyzer than in the blood inflow . Despite the high bacterial counts in the dialysate and the increasing ET levels in the patients plasma neither bacteraemia nor fever was observed . The former is due to the impermeability of the dialyzer membrane for bacteria, the latter is explained by low pyrogenicity of P . aeruginosa endotoxin . Inspection of the dialyzer machines revealed that air-traps and heater-unit for the incoming (untreated) tap water before mixing with the dialysate concentrate were the only sites where high bacterial release was feasible, as this part of the machine escaped disinfection due to the construction of these devices . We recommend the regular disinfection of all parts of a dialyzer machine, including heating units, air traps and valves. Infect Immun, 1989 Feb, 57(2), 520 - 6 Immunological studies of the disulfide bridge region of Pseudomonas aeruginosa PAK and PAO pilins, using anti-PAK pilus and antipeptide antibodies; Lee KK et al.; Pseudomonas aeruginosa is an opportunistic pathogen that attaches to host cells via their pili . The pilus of P . aeruginosa PAK consists of a polymer of a single subunit, pilin, which is a 144-residue polypeptide . The C-terminal end of this protein is semiconserved in a number of strains and contains a disulfide bridge . We have synthesized the C-terminal peptide PAK (128-144)-OH in both its reduced and oxidized forms and the analog PAK(A-129) (128-144)-OH, in which cysteine-129 was substituted by alanine . These three peptides were used to immunize rabbits and prepare antipeptide antisera . It was found that antipeptide antisera to reduced peptide (17-R) and to oxidized peptide (17-O) bound to native PAK pili and cross-reacted with strain PAO pili in direct enzyme-linked immunosorbent assay (ELISA) and immunoblot experiments . However, the antiserum to the peptide immunogen PAK(A-129)(128-144)-OH, which does not have the ability to form the disulfide bridge, did not bind to either PAK or PAO pili . Competitive ELISA experiments with reduced and oxidized peptides of Ac-PAK(128-144)-OH showed that there was no difference in binding between the two peptides for 17-R or 17-O immunoglobulin G . When immunoglobulin G from native PAK antipilus antiserum was used in competitive or direct ELISA experiments, there was also no preference in binding to reduced or oxidized Ac-PAK(128-144)-OH or to PAK(A-129)(128-144)-OH . This result showed that the disulfide bridge in Pseudomonas pili is not critical to the immunogenicity of this region . However, the disulfide bridge is important in the immunogenicity of the C-terminal peptide when preparing antipeptide antisera that are cross-reactive with pili from different strains, since only the disulfide bridge peptide antisera cross-reacted well with the PAO pili as shown by competitive ELISA, suggesting that this region could be an important candidate for development of a synthetic vaccine. Singapore Med J, 1989 Feb, 30(1), 63 - 5 In vitro activity of some newer quinolone compounds; Lim VK; The quinolones are a group of antimicrobial agents that act by inhibiting bacterial DNA gyrases, enzymes essential in DNA replication . Several newer quinolone agents have been introduced recently . These are broad spectrum agents which may be administered orally . In-vitro susceptibility testing of five quinolone agents namely norfloxacin, pefloxacin, enoxacin, ofloxacin and ciprofloxacin against recent clinical bacterial isolates at the General Hospital Kuala Lumpur was performed . The results confirm the broad spectrum and high activity of these agents against these isolates which included Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus . The quinolones would provide valuable alternatives in the treatment of infections caused by organisms resistant to the more commonly used antibiotics. J Bacteriol, 1989 Feb, 171(2), 983 - 90 Role of protein F in maintaining structural integrity of the Pseudomonas aeruginosa outer membrane; Gotoh N et al.; To investigate the functional role of protein F of the outer membrane of Pseudomonas aeruginosa, we isolated mutants devoid of protein F, and the defective gene was transferred to a wild-type strain by plasmid FP5-mediated conjugation . Chemical analyses of the protein F-deficient outer membrane revealed that the amount of outer membrane protein was reduced to 72 to 74% of that of the protein F-sufficient strain and that lipopolysaccharides and phospholipids increased to 117 to 123% and 135 to 136%, respectively . The mutants and the transconjugant showed the following characteristics: (i) growth rates of protein F-deficient strains in low-osmolarity medium (e.g., L broth containing 0.1% NaCl) were less than 1/10 the rate of the protein F-sufficient strain; (ii) protein F-deficient cells were rounded, and the outer membrane formed large protruded blebs; and (iii) the outer membrane became physically fragile, since a significant amount of periplasmic proteins leaked out and the cells became highly sensitive to osmotic shock . The results suggested that protein F plays an important role in morphogenesis and in maintaining the integrity of the outer membrane . Determination of the diffusion rates of saccharides and beta-lactam antibiotics showed that the protein F-deficient outer membrane had no detectable transport defect compared with the protein F-sufficient outer membrane . The MICs of antibiotics for the protein F-deficient strains were nearly identical to those for the protein F-sufficient strain. Res Microbiol, 1989 Feb, 140(2), 125 - 37 Purification and peptidase activity of a bacteriolytic extracellular enzyme from Pseudomonas aeruginosa; Brito N et al.; A bacteriolytic enzyme excreted by Pseudomonas aeruginosa Paks I was purified: samples were found to be homogeneous by gel filtration chromatography, ion exchange chromatography using CM-cellulose, immunoelectrophoresis, PAGE and SDS-PAGE . The molecular weight of the lytic enzyme was estimated to be 15,000-19,000 . The enzyme was active on Gram-positive bacteria with glycine-containing interpeptide bridges in their murein layers . In addition, this lytic enzyme showed peptidase activity catalysing the hydrolysis of pentaglycine peptides into tri- and diglycine peptides. Mol Microbiol, 1989 Feb, 3(2), 261 - 5 Cloning of xcp genes located at the 55 min region of the chromosome and involved in protein secretion in Pseudomonas aeruginosa; Filloux A et al.; Pleiotropic mutations (xcp) affecting secretion of proteins in Pseudomonas aeruginosa have been previously characterized and mapped at 0 min, 55 min and 65 min . Genomic libraries of this organism have been constructed and the genes xcp-5 and xcp-54, located at the 55 min region, were cloned using the adjacent met allele as a marker, and complementation of xcp strains . From our linkage and cloning analysis, the most probable gene order in this region appears to be pyrD.. . xcp-5/xcp-54/met-9011/oru-314/trpF/leu-10 . Restriction mapping and transposon (Tn1725) insertion mutagenesis demonstrated that: (i) the overall size of DNA necessary for xcp expression was 9kb, (ii) the two loci are not adjacent on the chromosome, and (iii) the two loci are expressed independently . The xcp-5 gene has been subcloned on a 4kb EcoRI fragment. Bioorg Khim, 1989 Feb, 15(2), 231 - 48 {Synthesis of the polysaccharide common antigen of Pseudomonas aeruginosa and its L-analog in the form of 6-aminohexyl glycosides}; Tsvetkov IuE et al.; A synthesis of trisaccharide monomers built of D- or L-rhamnose residues is described . Their polycondensation in the presence of a 6-aminohexyl rhamnoside derivative afforded 6-aminohexyl glycosides of the natural common polysaccharide antigen of Pseudomonas aeruginosa and its L-analogue, respectively. Zh Mikrobiol Epidemiol Immunobiol, 1989 Feb, (2), 32 - 6 {Erythrocyte diagnostic kits for detection of Pseudomonas aeruginosa exotoxin A}; Deriabin PN et al.; The use of formulated chick red blood cells loaded with IgG preparations and affinity-purified antibodies, in comparison with initial immune serum to P . aeruginosa exotoxin A (ETA), has been shown to increase the sensitivity of antibody erythrocyte diagnosticum (AbED) 17-fold and to ensure the detection of ETA at a concentration of 1.2 mg of protein per ml . The passive hemagglutination (PHA) test with AbED has proved to be a more sensitive method for the detection of ETA than the antibody neutralization test with the use of antigenic erythrocyte diagnosticum, the latex agglutination test, the coagglutination test and the enzyme immunoassay . The PHA test has permitted the detection of ETA in the culture fluid of 80% of P . aeruginosa cultures under study. FEMS Microbiol Lett, 1989 Feb, 48(3), 335 - 8 Choline and betaine as inducer agents of Pseudomonas aeruginosa phospholipase C activity in high phosphate medium; Lucchesi GI et al.; In Pseudomonas aeruginosa, choline or betaine employed as the sole carbon and nitrogen source in a high phosphate medium induced a phospholipase C and an acid phosphatase activity but not an alkaline phosphatase activity . The P . aeruginosa strain utilized in this work does not possess a constitutive phospholipase C, since under culture conditions identical to those utilized by other authors (J . Bacteriol . 93, 670-674 (1967) and J . Bacteriol . 150, 730-738 (1982), our phospholipase C proved to be an inorganic phosphate-repressible enzyme . These findings enable us to conclude that although the phosphate control for the synthesis of phospholipase C may exist, it is expressed only under certain favorable culture conditions. Eur J Clin Microbiol Infect Dis, 1989 Feb, 8(2), 144 - 6 Detection of soluble Pseudomonas aeruginosa antigens in bronchial secretions by a coagglutination test; Sofianou D et al.; The coagglutination test was used for the detection of soluble Pseudomonas aeruginosa antigens in 165 bronchial secretions collected from critically ill patients with pulmonary infection . Of 57 cultures positive for Pseudomonas aeruginosa, 41 were positive by the coagglutination test . The test had a specificity of 97%, a sensitivity of 72%, a positive predictive value of 93% and a negative predictive value of 87% when compared with agar culturing . Quantitation of bacterial growth indicated that some Pseudomonas aeruginosa isolates were probably strains colonizing the bronchial tree of intubated patients rather than the etiologic agent of infection . When these specimens were excluded, the sensitivity and negative predictive value were 87% and 94.6%, respectively . The coagglutination test cannot replace isolation methods, but it is a rapid and useful procedure to screen patients suspect for pulmonary infection caused by Pseudomonas aeruginosa, providing a presumptive diagnosis when the result is positive. Eur J Clin Microbiol Infect Dis, 1989 Feb, 8(2), 136 - 41 Postantibiotic and bactericidal effect of imipenem against Pseudomonas aeruginosa; Odenholt I et al.; The postantibiotic effect of imipenem on Pseudomonas aeruginosa was studied at different inocula using one ATCC strain and four clinical isolates . The postantibiotic effect was measured using two different methods: viable counts and bioluminescence assay of intracellular bacterial ATP . The postantibiotic effect could be demonstrated with both methods (viable counts 1-2 h, ATP assay 3-5 h) for all strains at an inoculum of 10(6) CFU/ml . When the inoculum was raised to 10(8) CFU/ml, no postantibiotic effect could be observed with either method using routine growth conditions . This disappearance of the postantibiotic effect coincided with a loss of bactericidal effect of imipenem when high inocula were used . Improved oxygenation of the cultures restored the bactericidal and postantibiotic effects of imipenem at high inocula. Appl Environ Microbiol, 1989 Feb, 55(2), 511 - 3 Pseudomonas pellicle in disinfectant testing: electron microscopy, pellicle removal, and effect on test results; Cole EC et al.; Pseudomonas aeruginosa ATCC 15442 is a required organism in the Association of Official Analytical Chemists use-dilution method for disinfectant efficacy testing . When grown in a liquid medium, P . aeruginosa produces a dense mat or pellicle at the broth/air interface . The purpose of this investigation was to examine the pellicle by scanning electron microscopy, to evaluate three pellicle removal methods, and to determine the effect of pellicle fragments on disinfectant efficacy test results . The efficacies of three methods of pellicle removal (decanting, vacuum suction, and filtration) were assessed by quantifying cell numbers on penicylinders . The Association of Official Analytical Chemists use-dilution method was used to determine whether pellicle fragments in the tubes used to inoculate penicylinders affected test results . Scanning electron micrographs showed the pellicle to be a dense mass of intact, interlacing cells at least 10 microns thick . No significant differences in pellicle removal methods were observed, and the presence of pellicle fragments usually increased the number of positive tubes in the use-dilution method significantly. Microb Pathog, 1989 Feb, 6(2), 103 - 12 Purification and characterization of cytotoxin from the crude extract of Pseudomonas aeruginosa; Hayashi T et al.; Pseudomonas aeruginosa cytotoxin is a protein toxin exhibiting cytotoxic effects on various eukaryotic cells . The toxin was purified from a crude extract of Pseudomonas aeruginosa 158 by trypsin treatment and serial chromatography . The purified cytotoxin was electrophoresed as a single protein band on polyacrylamide gel electrophoresis (PAGE) in the presence or absence of sodium dodecyl sulfate (SDS) . The molecular weight of the purified toxin was determined to be 29,000 by SDS-PAGE and gel filtration chromatography in the presence of 6 M guanidine hydrochloride; the isoelectoric point was pH 6.0 . The N-terminal amino acid sequence of the purified toxin was determined . The purified toxin showed the strongest cytotoxic effect on rabbit polymorphonuclear leukocytes and diverse degrees of cytotoxic effects on various eukaryotic cells including red blood cells and cultured cells. Curr Eye Res, 1989 Feb, 8(2), 195 - 202 The contribution of bacterial surface hydrophobicity to the process of adherence of Pseudomonas aeruginosa to hydrophilic contact lenses; Klotz SA et al.; Ten isolates of Pseudomonas aeruginosa obtained from the corneas of patients with Pseudomonas keratitis adhered to soft contact lenses in significantly greater numbers than did six isolates from other body sites (P less than .05) . However, there was no predominant serotype among the 10 corneal isolates tested . Isolates grown statically in broth at 37 degrees C formed a pellicle and adhered two times as much to contact lenses as did isolates grown in broth while shaking which did not form a pellicle (P less than .01) . The more adherent isolates (grown at 37 degrees C) were shown to be more hydrophobic than the less adherent bacteria (grown at 26 degrees C) by their propensity to accumulate at the interface between hexadecane and saline and their movement into polyethylene glycol from dextran . These corneal isolates agglutinated erythrocytes, a process that was inhibited by dilute solutions (as low as 0.01%) of three commonly used surfactants . These same surfactants inhibited the adherence of Pseudomonas aeruginosa to soft contact lens surfaces by as much as 52% . It is concluded that hydrophobic interactions may significantly contribute to the ability of Pseudomonas aeruginosa to adhere to contact lenses. Eur J Immunol, 1989 Feb, 19(2), 413 - 6 Comparison of the effects of recombinant interleukin 6 and recombinant interleukin 1 on nonspecific resistance to infection; Van der Meer JW et al.; Interleukin 1 (IL 1) is a potent enhancer of nonspecific resistance to infection in mice . Since IL 1 also induces interleukin 6 (IL 6), we tested the hypothesis that IL 6 mediates the effect of IL 1 on nonspecific resistance . In a lethal Pseudomonas aeruginosa infection in granulocytopenic mice, in which 80 ng of recombinant human IL 1 alpha protects against death, IL 6 appeared to be much less effective . Dosages of 8 ng, 80 ng and 320 ng IL 6 did not differ from the control, whereas 800 ng had a marginal protective effect (0.05 less than p less than 0.1) . IL 1 and IL 6 did not potentiate each other in animals treated with suboptimal dosages of both cytokines . Numbers of bacteria cultured from the blood, thigh muscle, liver, spleen, and kidney were similar in animals treated with 800 ng IL 6 and in control animals, arguing against activation of microbicidal mechanisms . The serum concentration profile of IL 6 after an i.p . injection of 80 ng IL 1 was similar to that after 80 ng IL 6 i.p . Only minute amounts of IL 1 were detected in serum after an i.p . injection of IL 6 . Taken these data together, it appears that increased resistance to infection induced by IL 1 is not mediated by IL 6. Zentralbl Bakteriol Mikrobiol Hyg {B}, 1989 Feb, 187(3), 266 - 8 The use of arginine brilliantgreen glucose peptone broth (ABGP medium) as a primary culture medium for Pseudomonas aeruginosa; Schubert R; A new medium is described for the isolation of P . aeruginosa organisms which have been damaged by chlorination . This medium utilises the fact that with arginine . P . aeruginosa produces a strongly alkaline reaction resulting in an easily identified colour change from gray-green to blue-violet . The indicators used for this purpose are bromothymol blue and cresol red . The medium may be used with either the membrane filter or the liquid enrichment technique . Incubation is at 37 degrees C for 24 or 48 h. J Med Microbiol, 1989 Feb, 28(2), 109 - 12 Age-dependent changes in protease and elastase production in cultures of Pseudomonas aeruginosa; Umeki S; The intracellular and extracellular protease and elastase contents of Pseudomonas aeruginosa were studied in relation to the age of the culture . The intracellular protease and elastase content of the organisms decreased as the culture grew older, whereas extracellular protease and elastase greatly increased 24 h after incubation commenced, suggesting that host tissue injury by P . aeruginosa infection may increase in proportion to the duration of tissue colonisation. Infect Immun, 1989 Feb, 57(2), 426 - 31 Isolation, structure, and immunogenicity of Pseudomonas aeruginosa immunotype 4 high-molecular-weight polysaccharide; Pier GB et al.; A high-molecular-weight, immunogenic form of the lipopolysaccharide O side chain of Pseudomonas aeruginosa Fisher immunotype 4 (type 4, International Antigenic Typing System 1, Lanyi O:6) was isolated and characterized . Analysis by nuclear magnetic resonance spectroscopy confirmed the structural similarity of this high-molecular-weight polysaccharide and the type 4 O side chain . The polysaccharide was immunogenic in rabbits and mice, eliciting opsonophagocytic killing antibodies . Immunization with the polysaccharide produced significant protection against homologous challenge in both burned and granulocytopenic mice . Naturally acquired opsonic killing antibodies to type 4 polysaccharide were present in sera from unimmunized normal adults at levels comparable to postimmunization levels achieved after immunization with other type-specific polysaccharides . The specificity of the naturally occurring antibodies for the O side chain was documented by immunoblot analysis and inhibition studies . Naturally occurring polysaccharide-specific antibodies were comparable in their protective activity against live challenge in neutropenic animals to immunization-induced murine antibodies with similar specificity . These data suggest that naturally occurring serum antibody to P . aeruginosa type 4 lipopolysaccharide O side chains in most adults is not distinguishable in quantity or quality from immunization-induced antibodies in mice; evaluation of type 4-specific vaccines in humans may be complicated by this finding. Infect Immun, 1989 Feb, 57(2), 344 - 50 Surface characteristics of Pseudomonas aeruginosa grown in a chamber implant model in mice and rats; Kelly NM et al.; Pseudomonas aeruginosa PAO1 was grown in vivo in chambers implanted into the peritoneums of mice and rats . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of extracts of bacterial cells taken from the chambers and washed to remove loosely bound host proteins revealed the presence of the major outer membrane proteins D2, E, F, G, and H2 . Western immunoblotting with specific antisera confirmed the presence of porin protein F and lipoprotein H2 . However, there was no apparent induction of the phosphate starvation-inducible porin P or the divalent cation starvation-inducible protein H1 . Small amounts of proteins with molecular weights similar to those of the iron-regulated outer membrane proteins were found in cells grown in vivo; however, their presence could not be confirmed immunologically . The presence of pili and flagella on the cells grown in vivo was demonstrated by electron microscopy and Western immunoblotting . A consistent alteration in the lipopolysaccharide banding pattern was observed after growth in vivo . Compared with cells of strain PAO1 grown in vitro, cells grown in vivo appeared to lack a series of high-molecular-weight O-antigen-containing lipopolysaccharide bands and gained a new series of lower-molecular-weight lipopolysaccharide bands . This alteration in the lipopolysaccharide after growth in vivo did not affect the O-antigen serotype or the resistance of the bacteria to serum. Kansenshogaku Zasshi, 1989 Feb, 63(2), 145 - 55 {Study of the prophylactic and therapeutic effect of human granulocyte-colony stimulating factor (G-CSF) on experimental pyelonephritis induced by pseudomonas aeruginosa in neutropenic mice}; Tanaka N et al.; We investigated the prophylactic and therapeutic effect of human granulocyte-colony stimulating factor (G-CSF) on mice with ascending pyelonephritis induced by Pseudomonas aeruginosa (G-group) . This experimental model was established by a two course administration of cyclophosphamide, so that it kept the mice in a neutropenic status (around 2000 white blood cells/mm3) from the time of infection to the time of sacrifice . The cyclophosphamide-treated group increased their susceptibility more than the control group . In the cyclophosphamide-treated group, the prophylactic administration of G-CSF (2 micrograms/day/mouse) yielded a lower incidence of infection and of infection-induced mortality than that of saline alone . However, the therapeutic administration of G-CSF did not produce significant decreases of these rates, suggesting that this type of administration had no effect on infection . At the time of sacrifice, the prophylactic administration of G-CSF increased the number of neutrophils, while at the time of induced infection, no increase of neutrophils was found . G-CSF therapeutic administration was not able to increase neutrophils during the experiment . An investigation of the bacterial capacity of peritoneal exudate neutrophils revealed that G-CSF prophylactic administration accelerated its capacity, although cyclophosphamide alone did not . These results suggest that G-CSF has a prophylactic effect on bacterial infection in neutropenic mice, and that this effect, in part, depends upon both the increase of neutrophils and the acceleration of bactericidal capacity produced by G-CSF. Biochim Biophys Acta, 1989 Jan 30, 978(2), 267 - 75 Electric pulse induced membrane permeabilization . Spatial orientation and kinetics of solute efflux in freely suspended and dielectrophoretically aligned plant mesophyll protoplasts; Mehrle W et al.; Asymmetric breakdown (occurring in only one hemisphere of the cell) was induced in freely suspended and dielectrophoretically aligned vacuole-containing or evacuolated plant protoplasts as well as in isolated vacuoles . In suspended cells breakdown was restricted to the hemisphere facing the anode and in isolated vacuoles to the opposite hemisphere . This difference in the orientation of the asymmetric breakdown can be explained by the opposite direction of the intrinsic membrane potentials of isolated vacuoles and of cells on which the generated potential difference is superimposed . The ensuing permeabilization of the membrane was microscopically monitored by dye uptake and by release of chloroplasts and of cytoplasmic and/or vacuolar solutes . The asymmetric release of intracellular substances (organic acids and/or amino acids) was detected by accumulation of chemotactic bacteria (Pseudomonas aeruginosa) close to the permeabilised membrane area of the cells or vacuoles . Maximum bacteria accumulation required about 5 min and subsequently disappeared after a further 20 min presumably because of the restoration of the original membrane impermeability . With vacuoles retention of the accumulated bacteria was shorter indicating that the resealing process of the tonoplast membrane was faster than that of the plasmalemma . From the kinetics of bacteria accumulation and retention it is therefore possible to deduce information about the life-span and the resealing properties of electropermeabilized membrane areas on the single-cell level . Symmetric breakdown in both hemispheres of the cells could be achieved by electric field-mediated cell rotation of about 180 degrees between two pulses of the same polarity or by application of two pulses of alternating polarity . In dielectrophoretically aligned protoplasts of comparable diameter, breakdown occurred in both hemispheres, even though the breakdown was still asymmetric . It could be demonstrated by the uptake of the vital dye neutral red that the size of the membrane area which was permeabilized was much larger in that hemisphere oriented to the anode than in the other one . The relevance of these observations for further improvement of electroinjection of macromolecules and of electrofusion is discussed . In particular, it is pointed out that positioning of differently sized cells in electric field-mediated hybridisation and the polarity of the breakdown pulse is of great importance with respect to hybrid yield. Mol Cell Biochem, 1989 Jan 23, 85(1), 81 - 9 Choline transport in Pseudomonas aeruginosa; Salvano MA et al.; Choline used as the sole carbon or carbon and nitrogen source induces in Pseudomonas aeruginosa an active transport system . The induction of the choline uptake is repressed by succinate independently of the presence of ammonium ion in the culture medium . The repression mediated by succinate was insensitive to cyclic AMP . Substitution for dibutyryl-cyclic AMP was without effect . Choline metabolites that also support the growth of Pseudomonas aeruginosa were poor inducer agents of the choline transport . Kinetic evidence and the employment of choline metabolites as effectors indicated that the choline uptake system of this bacterium is formed by at least two components: one of high affinity (Km = 3 microM) and another of low affinity (Km = 400 microM) . Contrary to what occurs in the synaptosome system, the high affinity form for the choline uptake was not dependent on Na+ ions and is not inhibited by hemicholinium-3 . Since Pseudomonas aeruginosa can utilize choline as the sole carbon and nitrogen source, the induction of the choline transport with two components in this bacterium may be related to its own strategy to survive and grow in an adverse environment. J Mol Biol, 1989 Jan 20, 205(2), 343 - 53 Regulation of transcription in Escherichia coli from the mer and merR promoters in the transposon Tn501; Lund PA et al.; We report studies on deletion mutants of the regulatory region of the mercuric ion resistance (mer) genes of transposon Tn501, isolated from Pseudomonas aeruginosa . Transcription of the mer genes in Escherichia coli from the promoter Pmer is regulated both positively (in the presence of mercuric salts) and negatively (in their absence) by the product of the merR gene . The merR gene is transcribed divergently with respect to the other mer genes, and negatively regulates its own synthesis . The experiments described here suggest that both positive and negative regulation by MerR, as well as its autoregulation, are largely mediated by MerR binding to a single site on DNA . This site contains a hyphenated dyad symmetrical sequence centred 24 base-pairs before the start of the mer transcript . Additional sites may be involved in full repression of the mer and merR promoters . Studies on deletions of the Pmer promoter show that the -35 sequence is not required for constitutive activity . An alternative -10 sequence may be used in the absence of the -35 and normal -10 sequences, but the properties of a point mutation indicate that, in the presence of the -35 sequence, the normal -10 sequence is required for promoter activity . A model for the regulation of expression of the mercury resistance genes by mercuric ions and the MerR protein is discussed. Z Hautkr, 1989 Jan 15, 64(1), 17 - 20 {Infections of the skin caused by gram-negative pathogens . Foot infections--wound infections--folliculitis}; Wassilew SW; Gram-negative bacilli are ubiquitous . They are found in 10-15% of the intertriginous bacterial flora and the most important one is Pseudomonas aeruginosa . Heat, moisture, mazeration, and reduction of the normal Gram-positive flora favor a rapid establishment of Gram-negative bacilli and the ensuing development of clinical infections . The diagnosis depends of the characteristic clinical features and localization, the patient's history, as well as the result of bacteriological investigation . The significance of the isolation of Gram-negative bacilli from non-specific lesions must be carefully evaluated . Cutaneous lesions respond well to therapy if they can be dried . Systemic antibiotics acting against Gram-negative bacilli have been found helpful . In patients with acne and Gram-negative folliculitis, isotretinoin has a good effect. Eur J Biochem, 1989 Jan 15, 179(1), 53 - 60 Sequence analysis and expression of the arginine-deiminase and carbamate-kinase genes of Pseudomonas aeruginosa; Baur H et al.; The arcABC operon of Pseudomonas aeruginosa encodes arginine deiminase, catabolic ornithine carbamoyltransferase and carbamate kinase, respectively . We have determined the nucleotide sequences of the arcA and arcC genes . The arcA open reading frame specifies a polypeptide of 46.3 kDa . The same molecular mass was obtained for the subunit of purified arginine deiminase after electrophoresis under denaturing conditions . The N-terminal amino acid sequence of arginine deiminase was in agreement with the corresponding nucleotide sequence . The native arginine deiminase had an estimated molecular mass of 175-180 kDa, suggesting a tetrametric structure . The enzyme was activated by Mg2+ or Mn2+ and strongly inhibited by Zn2+ . The apparent Km for L-arginine was 0.04 mM in the presence of Mg2+ and 0.47 mM without Mg2+ . The arcC open reading frame codes for a 33-kDa protein, confirming the molecular mass previously reported for the subunit of carbamate kinase . The translation-initiation site of arcC was determined by deletion mapping . Two regions of dyad symmetry found between arcA and arcC might stabilize the putative arcABC transcript in the upstream (arcA) region; this might contribute to the high level of arcA expression as compared to the moderate level of arcC expression . Carbamate kinase had 37% sequence similarity (and 13.5% identity) with the C-terminal part of carbamoyl-phosphate synthetase (large subunit) from Escherichia coli . Arginine deiminase had no apparent similarity with argininosuccinate lyase . Thus, the arcA and arcC genes do not appear to be closely related to arginine biosynthetic genes, whereas it had previously been shown that the arcB gene has a high degree of identity with the arginine biosynthetic argF genes of P . aeruginosa and E . coli. Eur J Biochem, 1989 Jan 15, 179(1), 195 - 200 The azurin gene from Pseudomonas aeruginosa . Cloning and characterization; Arvidsson RH et al.; We have cloned and sequenced the Pseudomonas aeruginosa azurin structural gene and its flanking regions . The DNA sequence predicts a pre-protein with a signal peptide of 19 amino acids followed by the 128-amino-acid mature azurin protein . Nuclease-S1 mapping and primer elongation experiments indicated two 5' termini of the azurin transcript . The major transcript of the azurin gene is initiated around 35 base pairs upstream from the translational start . The minor transcript, with a promoter region sharing homology with a consensus nif promoter of Klebsiella pneumoniae and also with other Pseudomonas genes, is initiated 145 base pairs upstreams of the azurin initiation codon . Downstream from the azurin structural gene a sequence similar to a transcriptional terminator is found . Northern blot analysis indicated two sizes of the azurin mRNA (0.54 kb and 0.65 kb) confirming the S1 mapping and the predictions from the nucleotide sequence. FEMS Microbiol Lett, 1989 Jan 15, 48(2), 227 - 30 Does aminoglycoside-acetyltransferase in rapidly growing mycobacteria have a metabolic function in addition to aminoglycoside inactivation? Udou T, Mizuguchi Y, Wallace RJ Jr. All the rapidly growing mycobacteria tested, Mycobacterium fortuitum complex, M . smegmatis, M . phlei, and M . vaccae, contained one of two characteristics, but were different from previously recognized aminoglycoside-acetyltransferases . The acetylation reaction of both the enzymes from M . fortuitum and Pseudomonas aeruginosa (3-N-acetyltransferase-III) with radiolabeled acetyl coenzyme A was inhibited severely by oxalacetate . It was suggested that the inhibitory effect of oxalacetate is due to the condensation reaction between oxalacetate and acetyl coenzyme A resulting in the generation of citrate. Hosp Pharm, 1989 Feb, 24(2), 110 - 4 Retrospective evaluation of piperacillin use in a university hospital; Volger BW et al.; A retrospective review was conducted to evaluate the appropriateness of empiric and definitive piperacillin use and to determine if less expensive, more appropriate antibiotics could have been used . The criteria were approved by the chief of infectious diseases and the drug utilization review committee at this institution . One hundred courses of piperacillin use in adult patients were reviewed . Therapy was categorized as appropriate in 78 of the 100 courses: 21 were appropriate empiric, 32 were appropriate empiric changed to definitive due to culture and sensitivity reports, and 25 were appropriate definitive . Reasons for this high percentage of appropriate use include: 1) 40% of the infections involved Pseudomonas aeruginosa; 2) 18% of the organisms that were sensitive to piperacillin were resistant to ticarcillin and mezlocillin; and 3) 64% of the courses of therapy involved critically ill patients with diagnoses such as neutropenia secondary to cancer chemotherapy, burns, sepsis, and hospital-acquired pneumonia . Although only 19.7% of the 11,845 grams of piperacillin used were categorized as inappropriate, the cost is relatively high (annualized to $19,648) and cost savings could be realized if piperacillin use were monitored more closely. J Antibiot (Tokyo), 1989 Jan, 42(1), 54 - 62 Synthesis and structure-activity relationships of 3-(2-imidazolyl)thiomethyl cephalosporins; Nakanishi E et al.; The synthesis and structure-activity relationships of a series of 3-(2-imidazolyl)thiomethyl cephalosporins are described . Among the compounds, 7 beta-{2-(2-amino-1,3-thiazol-4-yl)-(Z)-2-methoxyiminoacetamido}-3- {(4,5-dicarboxyimidazol-2-yl)thiomethyl}-3-cephem-4-carboxylic acid (1) exhibited potent activity against both Gram-positive and Gram-negative bacteria including Pseudomonas aeruginosa . We also estimated lipophilicity of the compounds from the chromatographic log k' value of reversed-phase HPLC . The relationship between lipophilicity and biological activity showed that compound 1 had the most suitable lipophilicity. Pediatr Res, 1989 Jan, 25(1), 49 - 54 Serum mucin-associated antigen levels of cystic fibrosis patients are related to their ages and clinical statuses; Frates RC Jr et al.; Mucin levels are generally elevated in sera from many cystic fibrosis (CF) patients as measured by radioimmunoassay using monoclonal antibody 19-9, which is directed against the mucin-associated sialyl Lea antigen . Antibody 19-9 can only be used to measure mucin-associated antigen levels in those patients who are genetically able to make detectable levels of mucin-associated sialyl Lea epitope . Serial studies of 20 patients followed over 3-5 y showed that their serum mucin-associated antigen levels varied directly with respect to the severity of their disease and inversely with their Shwachman-Kulczycki clinical scores (p less than 0.001) and Brasfield chest roentgenographic scores (p less than 0.02) . Serum mucin-associated antigen levels in samples from 89 CF patients were generally higher in the older patients (p less than 0.025) . Serum mucin-associated antigen levels of CF patients who were colonized with Pseudomonas aeruginosa did not significantly differ from those of uninfected CF patients . The mean serum mucin-associated antigen level of CF patients colonized with Pseudomonas was higher than the mean mucin level of six non-CF bronchiectatic patients whose lungs were colonized with Pseudomonas (p = 0.053) . Serum mucin-associated antigen levels are thus related to CF patients' ages and clinical statuses. Lab Anim Sci, 1989 Jan, 39(1), 37 - 43 Cardiopulmonary responses to Pseudomonas septicemia in swine: an improved model of the adult respiratory distress syndrome; Mustard RA et al.; Bacteremia with resultant damage to multiple organ systems remains a serious problem in intensive care of human patients . We have developed a clinically relevant swine model of sepsis-induced adult respiratory distress syndrome (ARDS) . Twenty-three animals were given various doses of Pseudomonas aeruginosa intravenously . Low cardiac output septic shock was prevented with massive fluid infusion . It was found that a dose of 1.0 X 10(7) colony forming units per 20 kg/min for 2 hours reliably produced respiratory failure in a setting of hyperdynamic sepsis which meets the diagnostic criteria of human ARDS. In Vitro Cell Dev Biol, 1989 Jan, 25(1), 44 - 8 A continuous alveolar macrophage cell line: comparisons with freshly derived alveolar macrophages; Helmke RJ et al.; Responses of a recently developed rat alveolar macrophage cell (NR8383.1) line were compared to those of freshly derived alveolar macrophages in vitro . Marked inter- and intraspecies heterogeneity in levels of phagocytosis of unopsonized Pseudomonas aeruginosa or zymosan was noted among freshly derived alveolar macrophages from rats, rabbits, and baboons . In contrast, phagocytic responses of alveolar macrophage cell line were predictable and highly reproducible . Similar results were obtained in measuring oxidative burst, as indicated by the production of H2O2 and luminol-enhanced chemiluminescence . Responses were again highly variable in freshly derived alveolar macrophages stimulated with zymosan or phorbol myristic acetate; moreover, freshly derived alveolar macrophages exhibited a wide range of chemiluminescence activity in unstimulated cultures . Results strongly suggest that data derived from the continuous alveolar macrophage culture NR8383.1 can be extrapolated to freshly derived alveolar macrophages of various species, and in many experiments will be useful in avoiding the significant animal-to-animal variance observed among freshly derived cell preparations. Sov Med, 1989, (6), 23 - 6 {Phagotherapy of postoperative suppurative-inflammatory complications in patients with neoplasms}; Kochetkova VA et al.; The authors assess the efficacy of phage therapy ofsuppurative and inflammatory complications in oncological patients . A clinical and laboratory analysis has involved 131 patients whose etiotropic therapy consisted of bacteriophages (65 patients) and antibiotics (66) . Medicinal phages, manufactured by the Tbilisi Research Institute for Vaccines and Sera, have been administered according to 3 schemes: (1) parallel with antibiotics, (2) after long ineffective antibiotic therapy, (3) phages alone starting from the onset of the purulent complication . The preparations have been prescribed with due consideration for the isolated microflora sensitivity . Incorporation of phages in combined therapy of infectious complications has yielded positive results in 81.5% of cases, whereas antibiotics have proved effective in but 60.6% . The efficacy of phage therapy depends on the type of pyoinflammatory complications (the results are the best in the management of wound infections), the microflora pattern of the purulent foci (phages are the most effective with a corresponding monoinfection), characteristics of the therapeutic phages proper (Pseudomonas aeruginosa phage is characterized by the highest therapeutic activity, as compared to staphylococcal and other phages). Eur J Pediatr, 1989 Jan, 148(4), 330 - 2 Cystic fibrosis in Saudi Arabia; Nazer H et al.; Cystic fibrosis (CF) is generally believed to be rare or nonexistent in Saudi Arabia . The aim of this report is to document the occurrence of CF in Saudi Arabia . Thirteen Saudi children were diagnosed as having CF, evidenced by typical clinical features and elevated sweat chloride concentrations (greater than 60 mmol/l) . Duration of symptoms prior to diagnosis varied from 1 month-5 years (mean 23 months) . The main clinical manifestations of the children were abdominal distention, failure to thrive, steatorrhoea, hepatomegaly, rectal prolapse and recurrent respiratory infections, often with Pseudomonas aeruginosa . In addition, eight patients with symptoms and a family history highly suggestive of CF, but without confirmatory sweat test results are presented . We hope that this report will increase the awareness of CF and ensure an earlier diagnosis of the disease in Saudi Arabia. J Comp Pathol, 1989 Jan, 100(1), 37 - 46 The frog palate mucosa as a model for studying bacterial adhesion to mucus-coated respiratory epithelium; Plotkowski MC et al.; Most of the methods proposed to quantify bacterial adherence to respiratory mucosa differ mainly from in vivo conditions in the absence of the mucus blanket and in the exposure of the sub-mucosal connective tissue (SMCT) to the micro-organisms . We propose the frog palate as a model to study bacterial adhesion to the respiratory mucosa, with a system which allows the mucus to be preserved and the bacterial adhesion to be quantified in a standardized mucosal area, where mucociliary transport is still active . In order to evaluate the role of respiratory mucus in bacteria-mucosa interaction, we compared the adhesion of radiolabelled pneumococci to 12 mucus-coated and 10 non-mucus-coated frog palate mucosae . The presence or absence of mucus was controlled by scanning electron microscopy (SEM) . After a 10 min incubation period, the bacterial adhesion to mucus-coated palate mucosa was five times greater (P less than 0.01) than that to uncoated mucosa . By SEM, bacteria were never seen attached to ciliated cells but could be detected on small areas where mucus was not totally eliminated . Even after a 120 min contact of bacteria to uncoated mucosa, bacterial adhesion remained only half that to mucus-coated epithelium . In order to ascertain whether the exposure of the SMCT represented a means of attraction to bacteria, we incubated the frog palate mucosa face-down with radiolabelled Pseudomonas aeruginosa . As much as 44 per cent of added bacteria adhered to exposed SMCT and, by SEM, numerous micro-organisms were seen attached to connective tissue . In contrast, only a few bacteria were observed adhering to the mucosa, mainly to granules of mucus. Int J Immunopharmacol, 1989, 11(4), 349 - 58 Enhancement of nonspecific resistance to bacterial infections and tumor regressions by treatment with synthetic lipid A-subunit analogs . Critical role of N- and 3-O-linked acyl groups in 4-O-phosphono-D-glucosamine derivatives; Nakatsuka M et al.; Enhancement of nonspecific resistance against Pseudomonas aeruginosa infection and regression of growth of Meth A fibrosarcoma by chemically synthesized lipid A-subunit analogs, 4-O-phosphono-D-glucosamine derivatives carrying 3-O- and N-linked acyl groups, were investigated . Compounds carrying an (R)-3-hydroxytetradecanoyl (C14-OH) group at the 2-N-position with (R)-3-tetradecanoyloxytetradecanoyl {C14-O-(C14)} or (R)-3-dodecanoyloxytetradecanoyl {C14-O-(C12)} groups at the 3-O-position, termed GLA-60 or GLA-63, respectively, showed strong activity about one-tenth that of natural lipid A . The protective activity of compounds carrying an (R)-3-hexadecanoyloxytetradecanyl group instead of a C14-O-(C14) or C14-O-(C12) group was very weak . GLA-59 carrying the same acyl components as those of GLA-60 but with reversed binding sites showed significant but not so strong protective activity . The activity of compounds possessing a tetradecanoyl group instead of a C14-OH group in GLA-60 or GLA-63 was weaker than that of GLA-60 or GLA-63 . Intravenous or intratumoral administration of GLA-59, GLA-60 and GLA-63 induced significant regression of Meth A fibrosarcoma in terms of tumor size, tumor weight and number of cured mice . The activity of GLA-59 was almost equivalent to that of GLA-60 . None of the tested compounds exhibited significant pyrogenicity at a dose of 10 micrograms/kg in rabbits. Chemotherapy, 1989, 35(4), 237 - 41 Pharmacokinetics of cefsulodin in rat cerebrospinal fluid during experimental Pseudomonas aeruginosa meningitis; Meulemans A et al.; Experimental meningitis was induced in rats with Pseudomonas aeruginosa . Bacteria were inoculated in the second ventricle . Twenty hours later cefsulodin penetration was studied in CSF by on-line cannula system which permitted sampling of CSF in the third ventricle . Comparison with healthy animals indicated breakdown of the blood-CSF barrier and high concentrations of cefsulodin were found in CSF. Scand J Infect Dis Suppl, 1989, 60, 84 - 8 Comparison of efficacy and tolerance of intravenously and orally administered ciprofloxacin in cystic fibrosis patients with acute exacerbations of lung infection; Strandvik B et al.; Twenty patients (17-27 yr) with cystic fibrosis were given ciprofloxacin at 30 pulmonary infectious exacerbations . All patients were chronically colonized with Pseudomonas aeruginosa . Twenty-five courses were completed, 13 orally (15 mg/kg b.i.d.) and 12 intravenously (4-6 mg/kg b.i.d.) . Clinical efficacy was excellent or good in 85-90% of the courses and growth of P . aeruginosa was markedly reduced in 33-46% . Body weight and clinical score improved significantly . White blood cell count decreased and pulmonary function was improved . Reversible adverse effects, mainly rash and urticaria, appeared at seven occasions, five severe enough to cause interruption of treatment . Clinical efficacy and tolerance were better with oral than intravenous administration at the dosages used in this study . Excellent bioavailability provides additional basis for oral treatment with ciprofloxacin in cystic fibrosis patients. Microb Pathog, 1989 Jan, 6(1), 75 - 80 Octavalent Pseudomonas aeruginosa O-polysaccharide-toxin A conjugate vaccine; Cryz SJ Jr et al.; An octavalent Pseudomonas aeruginosa conjugate vaccine was synthesized by covalently coupling the O-polysaccharide (O-PS) moiety derived from lipopolysaccharides of Habs serotypes 1, 2, 3, 4, 5, 6, 11 and 12 to toxin A . Adipic acid dihydrazide was used as a spacer molecule to facilitate conjugation . The vaccine was composed of 37% (w/w) O-PS and 63% toxin A, devoid of enzymatic activity characteristic of toxin A, non-toxic for mice and guinea pigs, and non-pyrogenic . The vaccine elicited a significant rise in immunoglobulin G antibody levels to all serotypes of lipopolysaccharide contained in the vaccine and to toxin A . Serotypes 6, 10 and 11 were most immunogenic in mice whereas serotypes 1 and 5 engendered the lowest antibody response . Antitoxin A antibody was able to neutralize the cytotoxicity of toxin A . Immunization of mice with the vaccine conferred significant protection against subsequent challenge with all P . aeruginosa serotype strains contained in the vaccine. An Otorrinolaringol Ibero Am, 1989, 16(1), 5 - 11 {Malignant otitis externa}; Perez Garrigues H et al.; This is an aggressive infectious disease caused by Pseudomonas aeruginosa, especially seen in diabetics and elderly, sometimes coming to and end in those cases in which the diagnosis has been protracted or the treatment misleading . But it must be said that the diagnosis is not easy, either because of the insidiousness of the onset or the hidden difficulties when dealing with malignant growths of the meatus . The AA report a case of the kind emphasizing the importance of the real diagnosis as well as the right treatment . This should be prudently prolonged beyond the apparent healing until the cultures show a total sterilization. Allerg Immunol (Leipz), 1989, 35(1), 59 - 64 {Stimulation of nonspecific immune defense and antibody synthesis with zymosan}; von Baehr R et al.; A local sterile inflammation was induced in mice of different inbred strains by means of an i.p . injection of zymosan, a water-insoluble polysaccharide of the cell wall of Saccharomyces cerevisiae . This resulted in a strong enhancement of the unspecific resistance against infections with a LD 90 of gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae and Bordetella bronchiseptica) . Furthermore, the treatment with zymosan before or after an immunization with sheep erythrocytes (SE) resulted in a significantly increased specific IgM and IgG primary response in the spleen . However, it was particularly striking that the number of plaque-forming cells against SE was very drastically enhanced in non-immunized mice, in particular cases even up to more than 70 times over the background . Consequently, zymosan possesses immunomodulating effects both on the unspecific resistance and on immune reactions . These results are discussed in relation to the Lyt+-B-cells and an acute phase reaction of B-cells. Z Exp Chir Transplant Kunstliche Organe, 1989, 22(1), 27 - 37 {Experimental animal studies of the healing of burn wounds with the use of local antimicrobials and hyperbaric O2 therapy}; Kaiser W; Standardized burns without experimental infection and such with infection by a constant number of bacteria of a fixed Pseudomonas-aeruginosa-strain were treated differently . Silver sulfadiazine- and Cefsulodin-cream, Polyvidon-iodine-ointment (PVP-Jod), and hyperbaric oxygen (OHP) were used to them . The courses of healing were checked by determination of the wound area daily . The best results showed the experimental uninfected untreated wounds . Wounds infected by Pseudomonas were healing most quickly by immediate and continuous application of hyperbaric oxygen . By silver sulfadiazine- and Cefsulodin-cream treated burns showed statistical significant better results than the control group . The courses of healing were significantly poorer in delayed application of OHP (only from the 8th day after the burn) or in case of therapy with PVP-iodine alone. Dev Comp Immunol, 1989 Summer, 13(3), 205 - 16 Effect of parasitism by Cotesia congregata on the susceptibility of Manduca sexta larvae to bacterial infection; Ross DR et al.; Eggs and larvae of the braconid Cotesia congregata are not encapsulated within the hemocoel of their habitual host, Manduca sexta . Experiments were performed to evaluate the status of antibacterial defensive responses in M . sexta larvae parasitized by this gregarious endoparasitoid . Previous investigations have shown that immunologically naive, nonparasitized M . sexta larvae are resistant to infection by Pseudomonas aeruginosa . Studies using naive, parasitized larvae demonstrated that inoculation with 10 colony forming units of P . aeruginosa 9027, one hour after oviposition, resulted in greater than 90% host mortality . Examination of the fate of P . aeruginosa and Escherichia coli injected into parasitized larvae suggested that the cellular antibacterial defenses of the host, nodule formation, and phagocytosis, were impaired. Arch Microbiol, 1989, 152(3), 302 - 8 Isolation and characterization of an alginate lyase from Klebsiella aerogenes; Lange B et al.; The bacterium Klebsiella aerogenes (type 25) produced an inducible alginate lyase, whose major activity was located intracellularly during all growth phases . The enzyme was purified from the soluble fraction of sonicated cells by ammonium sulfate precipitation, anion- and cation-exchange chromatography and gel filtration . The apparent molecular weight of purified alginate lyase of 28,000 determined by gel filtration and of 31,600 determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the active enzyme was composed of a single polypeptide . The alginate lyase displayed a pH optimum around 7.0 and a temperature optimum around 37 degrees C . The purified enzyme depolymerized alginate by a lyase reaction in an endo manner releasing products which reacted in the thiobarbituric acid assay and absorbed strongly in the ultraviolet region at 235 nm . The alginate lyase was specific for guluronic acid-rich alginate preparations . Propylene glycol esters of alginate and O-acetylated bacterial alginates were poorly degraded by the lyase compared with unmodified polysaccharide . The guluronate-specific lyase activity was applied in an enzymatic method to detect mannuronan C-5 epimerase in three different mucoid (alginate-synthesizing) strains of Pseudomonas aeruginosa . This enzyme which converts polymannuronate to alginate could not be demonstrated either extracellularly or intrac