FASEB J, 2000 Jul, 14(10), 1380 - 8 A bacteria-induced switch of sympathetic effector mechanisms augments local inhibition of TNF-alpha and IL-6 secretion in the spleen; Straub RH et al.; It is believed that an inflammation-induced activation of the CNS leads to an inhibition of overshooting immune responses to prevent extensive local cytokine secretion . However, immunosuppression by the sympathetic nervous system may be unfavorable when bacteria are present locally and when TNF-alpha is necessary to overcome infection . We now report in a superfusion model, using mouse spleen slices, that although local Pseudomonas aeruginosa increased splenic TNF-alpha and IL-6 secretion severalfold over basal levels, electrically released neurotransmitters attenuated cytokine secretion to similar basal level as under bacteria-free conditions . Bacteria reversed noradrenergic inhibitory effector mechanisms: Under bacteria-free conditions, TNF-alpha secretion was very low and IL-6 secretion was mainly inhibited by alpha2-adrenoreceptor ligation . In the presence of bacteria, TNF-alpha and IL-6 secretion were high and IL-6 secretion was mainly inhibited by beta-adrenoreceptor ligation . The alpha- to beta-adrenoswitch of IL-6 inhibition in the presence of bacteria was mediated by the prior adrenergic regulation of TNF-alpha . In vivo, chemical abrogation of sympathetic inhibition reduced accumulation of bacteria in the spleen, which depended at least in part on TNF-alpha . This suggests that activation of the sympathetic nervous system may be a forerunner for accumulation of bacteria in tissue and consecutively sepsis due to intensified inhibition of TNF-alpha secretion. Pediatr Infect Dis J, 2000 Jun, 19(6), 495 - 8 Liver enzyme abnormalities in gram-negative bacteremia of premature infants; Shamir R et al.; BACKGROUND: Hyperbilirubinemia and liver enzyme abnormalities are commonly observed in sepsis . However, the frequency in premature neonates and the specific relation to gram-negative bacteria are not known . PATIENTS AND METHODS: Charts of all preterm infants who had positive blood cultures for either gram-negative bacteria or coagulase-negative staphylococci were reviewed . Neonates with gram-negative bacteremia (n = 54) were compared with neonates with coagulase-negative staphylococcal bacteremia (n = 31) . In addition infants with gram-negative bacteremia and elevated liver enzymes (n = 25) were compared with infants with gram-negative bacteremia and normal liver enzymes (n = 29) . RESULTS: Liver enzyme abnormalities accompanied 46.3% (25 of 54) of gram-negative bacteremia and 12.9% (4 of 31) of episodes of coagulase-negative staphylococcal bacteremia (P = 0.002) . Serum concentrations of liver enzymes were significantly higher in infants with gram-negative bacteremia than in those with coagulase-negative staphylococcal bacteremia (P < 0.0001), but no difference in alkaline phosphatase serum values was observed . Infants with gram-negative bacteremia and elevated liver enzymes were not fed for a longer period than infants with gram-negative bacteremia and normal liver enzymes (7.3 +/- 6.3 days vs . 4.0 +/- 4.3 days, P = 0.03), and this was accompanied by significant conjugated hyperbilirubinemia (P < 0.0001) . Ventilation, total parenteral nutrition and medications were not responsible for the observed differences . Klebsiella pneumoniae bacteremia was commonly associated with elevated liver enzymes (12 of 18), whereas none of the infants with Pseudomonas aeruginosa bacteremia had elevated liver enzymes . CONCLUSIONS: Gram-negative bacteremia is commonly associated with cholestasis in premature neonates . Liver enzyme abnormalities are more common than elevated conjugated bilirubin, not all gram-negative bacteria have the same effect and the lack of enteral feeding seems to play a more significant role than the administration of parenteral nutrition. Zh Mikrobiol Epidemiol Immunobiol, 2000 Jan-Feb, (1), 31 - 3 {The description of an esculin-positive biovar of Pseudomonas aeruginosa}; Sivolodskii EP; In the study of 280 P . aeruginosa strains isolated in different hospitals of St . Petersburg for the first time 48 strains capable of hydrolyzing esculin have been detected . The hydrolysis of esculin is determined in plates with the use of the microvolume techniques the results were evaluated after 3-hour incubation at 37 degrees C . The data confirming the existence of the exculin-positive biovar of P . aeruginosa have been obtained; these data show the wide spread of esculin-positive strains in hospitals of different specialization (17.1 +/- 5.1% of P . aeruginosa strains), the characteristic combination of the sign of esculin hydrolysis with such signs as the absence of the smell of trimethylamine and the phenomenon of "iridescent lysis" of the colonies, the stability of the sign of esculin hydrolysis in strains, repeatedly isolated from patients, after the storage of the cultures and their treatment with plasmid-eliminating preparation . The name "esculinolytica" has been proposed for this biovar . The typing strain of biovar esculinolytica has been deposited in the culture collection of the Russian Research Institute of Agricultural Microbiology as P . aeruginosa ARRIAM 64-A . This biovar been found to be most widely spread in urological hospitals, where esculin-positive strains are isolated 3 times more frequently (32.2 +/- 5.1% of P . aeruginosa strains) than in surgical hospitals (10.7 +/- 2.2%). Polim Med, 1999, 29(1-2), 27 - 33 Antimicrobial properties of copper-coated electroconductive polyester fibers; Grzybowski J et al.; Three synthetic copper-coated EURO-static fibers (PET--polyester, PA--polyamide, and PAC--polyacrylamide) manufactured by EUROPA Corporation S.C., Poland, were tested as potential antimicrobial agents . The inhibitory properties of the fibers were examined using different microorganisms as follows: i . Staphylococcus aureus ATCC 25293, and Pseudomonas aeruginosa ATCC 27853 reference strains, ii . 8 strains of S . aureus (4 MRSA and 4 MSSA) and 5 strains of P . aeruginosa isolated from infected wounds, and iii . fungal pathogen Scopulariopsis sp . isolated from onychomycosis case . The results of experiments have evidenced that polyester (PET) copper-coated EURO-static fibers inhibit the growth of all the strains used. Virology, 2000 Jul 5, 272(2), 331 - 7 The three-dimensional structure of bacteriophage PP7 from Pseudomonas aeruginosa at 3.7-A resolution; Tars K et al.; The three-dimensional structure of phage PP7 from Pseudomonas aeruginosa has been determined to 3.7-A resolution . A comparison with distantly related small RNA phages showed that the biggest differences were found in the FG loops, forming the contacts around the fivefold and threefold axes . In contrast to the situation in other phages, the FG loops of phage PP7 are very similar in all three subunits . This supports the hypothesis that no switches are needed for the assembly control in these viruses . Some of the most conserved residues lie within the region involved in RNA binding in the related phages MS2 and seem to have the same function . Virology, 2000 Jun 20, 272(1), 85 - 97 Characterization of the lysogenic repressor (c) gene of the Pseudomonas aeruginosa transposable bacteriophage D3112; Salmon KA et al.; Bacteriophage D3112 is a Mu-like temperate transposable phage of Pseudomonas aeruginosa . Genetic mapping and DNA sequence analysis have identified the left end of the phage genome as encoding the transposase enzyme (A) and the lysogenic (c) repressor . The c open reading frame (ORF), located at the leftmost end of the phage genome and transcribed from right to left, has four possible GTG initiation codons . Using site-directed mutagenesis, each of the four GTG codons was modified to GTA, which cannot serve as an initiation codon . Plasmids were constructed expressing either the wild-type repressor ORF or the ORFs containing the mutated GTA codons . When introduced into Pseudomonas aeruginosa, no immunity to superinfection by D3112 was observed when the second GTG had been mutated . Northern blotting analysis demonstrated that the D3112 c repressor is transcribed as a 900-nt mRNA . The promoter region was defined by transcriptional lacZ fusions and primer extension analyses to bp 972-940 from the left end of the phage genome . When the D3112 c repressor was overexpressed and purified as a fusion protein with a C-terminal six-histidine extension (cts15-His6), it showed high affinity for a 261-bp PvuII fragment localized directly upstream of the c repressor ORF . Our results indicate that although D3112 c shows higher amino acid similarity to the lambda family of repressors than it does to those of Mu and D108, it appears that its structure and function more accurately reflect an evolutionary ancestry with those from transposable coliphages Mu and D108 . Biochem Biophys Res Commun, 2000 Jul 5, 273(2), 578 - 83 Characterization of an ECF sigma factor protein from Pseudomonas aeruginosa; Wilson MJ et al.; The PvdS protein is essential for synthesis of the siderophore pyoverdine by Pseudomonas aeruginosa . PvdS has some sequence similarity to a family of alternative sigma factor proteins (the ECF {extracytoplasmic factor} family) that direct bacterial RNA polymerases to transcribe genes encoding extracytoplasmic factors . PvdS was purified as a His-tagged protein (hPvdS) and this was used to test the hypothesis that PvdS is a sigma factor protein . The purified protein caused core RNA polymerase from Escherichia coli to bind specifically to the promoters of pyoverdine synthesis genes and enabled transcription from these promoters in vitro . In addition, PvdS was found to co-purify with RNA polymerase from P . aeruginosa, indicating that PvdS associates with RNA polymerase inside the bacteria . These results show that PvdS is a sigma factor protein . Am J Surg, 2000 Feb 1, 179(2 Suppl 1), 18 - 23 Empiric therapy for pneumonia in the surgical intensive care unit; Fabian TC; Empiri c therapy of ventilator-associated pneumonia (VAP) in surgical patients should be based on intensive care unit (ICU)-specific surveillance data, because microbial flora patterns vary widely between geographic regions as well as within hospitals . Surgical ICUs have higher VAP rates than other units . Data from the National Nosocomial Infection Surveillance (NNIS) System report Pseudomonas aeruginosa and Staphylococcus aureus to be the most frequent isolates (each 17.4%) . Data from the NNIS documents high resistance patterns in ICUs compared with hospitals at large, as well as unit-specific patterns . VAP risk factors for surgical patients include thoracoabdominal surgery, altered level of consciousness, advanced age, diabetes mellitus, malnutrition, chronic obstructive pulmonary disease, and prior antibiotic administration . Promising prevention strategies include restricting ventilator circuit changes, in-line heat moisture exchange filters, semi-recumbant positioning, and continuous subglottic aspiration . Pharmacodynamics should be considered when choosing antibiotic regimens . Postantibiotic effect and time-dependent versus concentration-dependent killing should be studied in clinical trials . Current guidelines for choosing regimens have been well developed by the American Thoracic Society. Lung, 2000, 178(3), 181 - 9 Detection of elastase from Pseudomonas aeruginosa in sputum and its potential role in epithelial cell permeability; Azghani AO et al.; Most clinical strains of Pseudomonas aeruginosa produce elastase, a zinc metalloprotease that is implicated in the pathogenesis of infections related to these organisms . To better understand the physiologic role of this protease in the regulation of airway permeability, we developed a panel of specific monoclonal antibodies (mAb) against purified Pseudomonas elastase (PE) that do not react with either neutrophil elastase or porcine pancreatic elastase . These mAbs were used in a competitive enzyme-linked immunosorbent assay to determine the concentrations of PE in sputum samples from patients with pulmonary infections . Sputum from patients infected with P . aeruginosa showed a varying amount of PE, whereas others indicated no signals . We also found that the mAbs blocked the effect of PE on epithelial barrier function in vitro on the basis of measurement of transmonolayer electrical resistance of polarized epithelial cells as an index of paracellular permeability. J Reconstr Microsurg, 2000 May, 16(4), 297 - 301 Effect of distant septic foci on the patency of microvascular anastomoses; Topalan M et al.; Despite all technical improvements, some free-tissue transfers are still subject to failure in the early postoperative period due to anastomotic occlusion . Local or distant foci of sepsis may be present in most of these cases . Using 40 Sprague-Dawley adult small female rats, distant septic abscesses were formed with Pseudomonas aeruginosa (including 2 x 10(8) bacteria/ml) suspension . Microarterial and microvenous anastomoses were carried out and animals were divided into experimental and control groups . With the use of postoperative exploration, anastomotic patency was evaluated, cultures of wound, blood, and tissue were taken, and the anastomosis line and thrombi were evaluated using different staining techniques on prepared histopathologic sections . Compared to the control groups, thrombus formation in animals with distant septic foci, both with arterial and microvenous anastomosis, was found to be greater, and the difference was statistically significant . Colonies of Pseudomonas were not demonstrated in samples taken from wound infections, hemocultures, cultures of vessel wall and thrombi, and in histopathologic sections . In this experimental model, it appears that distant septic foci have an occluding effect on the microanastomosis. J Formos Med Assoc, 2000 May, 99(5), 435 - 7 Pediatric endogenous endophthalmitis; Tsai YY et al.; Pediatric endogenous endophthalmitis is a rare disease that can cause serious ophthalmic damage . We describe two cases of pediatric endogenous endophthalmitis . The first occurred in an 8-month-old boy and the second in a 7-day-old girl . These two patients had developed pneumonia due to Pseudomonas aeruginosa infection prior to the onset of ocular symptoms . The interval between the onset of pneumonia and ocular symptoms was 1 week, but endophthalmitis was diagnosed 9 days after the onset of ocular symptoms in the first case and 3 days after the onset of ocular symptoms in the second case . The ocular manifestations included eyelid swelling, purulent discharge, redness, corneal edema, hypopyon, and poor red reflex . Despite treatment with aggressive antimicrobial therapy, both patients became totally blind with eyeball atrophy. J Bacteriol, 2000 Jul, 182(14), 4051 - 8 Influence of deletions within domain II of exotoxin A on its extracellular secretion from Pseudomonas aeruginosa; Voulhoux R et al.; Pseudomonas aeruginosa is a gram-negative bacterium that secretes many proteins into the extracellular medium via the Xcp machinery . This pathway, conserved in gram-negative bacteria, is called the type II pathway . The exoproteins contain information in their amino acid sequence to allow targeting to their secretion machinery . This information may be present within a conformational motif . The nature of this signal has been examined for P . aeruginosa exotoxin A (PE) . Previous studies failed to identify a common minimal motif required for Xcp-dependent recognition and secretion of PE . One study identified a motif at the N terminus of the protein, whereas another one found additional information at the C terminus . In this study, we assess the role of the central PE domain II composed of six alpha-helices (A to F) . The secretion behavior of PE derivatives, individually deleted for each helix, was analyzed . Helix E deletion has a drastic effect on secretion of PE, which accumulates within the periplasm . The conformational rearrangement induced in this variant is predicted from the three-dimensional PE structure, and the molecular modification is confirmed by gel filtration experiments . Helix E is in the core of the molecule and creates close contact with other domains (I and III) . Deletion of the surface-exposed helix F has no effect on secretion, indicating that no secretion information is contained in this helix . Finally, we concluded that disruption of a structured domain II yields an extended form of the molecule and prevents formation of the conformational secretion motif. J Bacteriol, 2000 Jul, 182(14), 3934 - 41 The Pseudomonas aeruginosa devB/SOL homolog, pgl, is a member of the hex regulon and encodes 6-phosphogluconolactonase; Hager PW et al.; A cyclic version of the Entner-Doudoroff pathway is used by Pseudomonas aeruginosa to metabolize carbohydrates . Genes encoding the enzymes that catabolize intracellular glucose to pyruvate and glyceraldehyde 3-phosphate are coordinately regulated, clustered at 39 min on the chromosome, and collectively form the hex regulon . Within the hex cluster is an open reading frame (ORF) with homology to the devB/SOL family of unidentified proteins . This ORF encodes a protein of either 243 or 238 amino acids; it overlaps the 5' end of zwf (encodes glucose-6-phosphate dehydrogenase) and is followed immediately by eda (encodes the Entner-Doudoroff aldolase) . The devB/SOL homolog was inactivated in P . aeruginosa PAO1 by recombination with a suicide plasmid containing an interrupted copy of the gene, creating mutant strain PAO8029 . PAO8029 grows at 9% of the wild-type rate using mannitol as the carbon source and at 50% of the wild-type rate using gluconate as the carbon source . Cell extracts of PAO8029 were specifically deficient in 6-phosphogluconolactonase (Pgl) activity . The cloned devB/SOL homolog complemented PAO8029 to restore normal growth on mannitol and gluconate and restored Pgl activity . Hence, we have identified this gene as pgl and propose that the devB/SOL family members encode 6-phosphogluconolactonases . Interestingly, three eukaryotic glucose-6-phosphate dehydrogenase (G6PDH) isozymes, from human, rabbit, and Plasmodium falciparum, contain Pgl domains, suggesting that the sequential reactions of G6PDH and Pgl are incorporated in a single protein . 6-Phosphogluconolactonase activity is induced in P . aeruginosa PAO1 by growth on mannitol and repressed by growth on succinate, and it is expressed constitutively in P . aeruginosa PAO8026 (hexR) . Taken together, these results establish that Pgl is an essential enzyme of the cyclic Entner-Doudoroff pathway encoded by pgl, a structural gene of the hex regulon. Transplantation, 2000 Jun 15, 69(11), 2360 - 6 Significance of blood stream infection after lung transplantation: analysis in 176 consecutive patients; Palmer SM et al.; BACKGROUND: Although infection is a leading cause of death after lung transplantation, very little is known about the incidence, epidemiology, and clinical significance of bloodstream infections in lung transplant recipients . METHODS: All blood cultures were reviewed in 176 consecutive lung transplant recipients over a 6-year period . Data were obtained from a prospectively collected microbiological database . RESULTS: Bloodstream infection (BSI) occurred in 25% (44/176) of all lung transplant recipients over the 6-year study period . Staphylococcus aureus, Pseudomonas aeruginosa, and Candida species were the most common bloodstream isolates after lung transplantation . The epidemiology of posttransplant BSI, however, varied considerably between early and late posttransplant time periods and also differed between cystic fibrosis (CF) and non-CF patients . BSI infection after transplantation was associated with significantly worse survival by Kaplan-Meir analysis (P value log rank test=0.0001) . In a multivariable logistic regression model, posttransplant BSI was a significant predictor of posttransplant death (odds ratio 5.62, CI 2.41-13.11, P=0.001), independent of other pre- and posttransplant factors . CONCLUSIONS: Bloodstream infection represents a serious complication after lung transplantation, occurring more frequently than previously recognized, and independently contributing to posttransplant mortality. Jpn J Antibiot, 2000 Apr, 53(4), 194 - 200 {In vitro interaction of piperacillin and imipenem/cilastatin combined with aminoglycosides against Pseudomonas aeruginosa}; Yamashiro Y et al.; The in vitro interactions of piperacillin (PIPC) and imipenem/cilastatin (IPM/CS) combined with 5 kinds of aminoglycosides (gentamicin, tobramycin, amikacin (AMK), isepamicin and netilmicin) were investigated against IPM/CS-susceptible (MIC of IPM/CS was < or = 3.13 micrograms/ml) and IPM/CS-resistant (MIC of IPM/CS was > or = 12.5 micrograms/ml) Pseudomonas aeruginosa . The following results were obtained . 1 . In the checkerboard dilution studies, the combinations of PIPC with aminoglycosides showed synergistic effect for more than 50% of the each 54 strains of IPM/CS-susceptible and IPM/CS-resistant P . aeruginosa . The synergistic/additive effects of PIPC with aminoglycosides were demonstrated for all tested strains . 2 . In the checkerboard dilution studies, the combinations of IPM/CS with aminoglycosides showed no antagonism against any strains . The synergistic effects of IPM/CS with aminoglycosides were demonstrated for 0 to 14.8%, and these values were smaller than the combinations of PIPC with aminoglycosides . 3 . Corresponding to the results of checkerboard dilution studies, the combination of PIPC with AMK was more effective than the combination of IPM/CS with AMK on the killing curve for IPM-resistant P . aeruginosa . In conclusion, PIPC showed the synergistic effects in combinations with aminoglycosides against IPM/CS-resistant P . aeruginosa . These results suggest that the combination therapies of PIPC with aminoglycosides are useful for the clinical treatment of serious infections due to P . aeruginosa. Harefuah, 2000 Jan 2, 138(1), 14 - 7, 87 {Corneal infection, cause and effect on vision}; Domniz Y et al.; We conducted a retrospective 5-year survey of corneal infections treated in the ophthalmology ward of Hasharon Hospital . The most frequent type of corneal infection was corneal abscess; the most frequent cause was Staphylococcus albus, although this bacterium is not reported as a frequent cause of corneal infections . There was improvement in visual acuity in 69.2% and no change in 15.4% . Corneal infection by Pseudomonas aeruginosa was the most frequent cause of worsening of corneal acuity (23.08%) . The greatest improvement of visual acuity was in those with corneal ulcers . The worst visual acuity was in those with corneal abscesses . Pseudomonas aeruginosa was the main cause of infection in contact-lens wearers . In the world medical literature, Staphylococcus albus is considered of very low virulence . This bacterium was the most frequent cause of corneal infections in our study so it may have greater virulence in Israel. Respiration, 2000, 67(3), 239 - 47 Diagnosis and treatment of cystic fibrosis; Koch C et al.; This review discusses some diagnostic aspects of cystic fibrosis (CF) including direct mutational analysis . Treatment of major disease manifestations is discussed in more detail with an emphasis on lung disease, in particular chronic infection with Pseudomonas aeruginosa which is responsible for the majority of excess morbidity and mortality . Centralised care and aggressive antimicrobial treatment have led to increased life expectancy and this may be even further increased by the demonstration that chronic P . aeruginosa infection may be prevented, or at least postponed for many years in a majority of patients . Adjunct treatment such as the use of local and systemic anti-inflammatory agents and inhalation of human recombinant DNase are also briefly touched upon . It is emphasised that important questions concerning the link(s) between the mutated gene and lung disease are still missing but that current research raises hope of a more causal treatment in the near future . FEMS Immunol Med Microbiol, 2000 Jul, 28(3), 241 - 5 Decreased virulence of a strain of Pseudomonas aeruginosa O12 overexpressing a chromosomal type 1 beta-lactamase could be due to reduced expression of cell-to-cell signaling dependent virulence factors; Ramisse F et al.; Pseudomonas aeruginosa produces a large variety of virulence factors and is characterized by its capacity to rapidly develop resistance when exposed to antibiotics . In order to evaluate a possible correlation between acquired resistance to antibiotics and virulence, we examined the virulence of four isogenic variants of P . aeruginosa O12 that differ in their resistance phenotypes to various beta-lactam antibiotics in a mouse model of acute pneumonia . Strains overproducing a chromosomal type 1 beta-lactamase were less virulent in both immunocompetent and immunosuppressed animals . Whereas the production of the exopolysaccharide alginate was similar between the four strains, extracellular virulence factors (elastase, rhamnolipid) that are controlled by the cell-to-cell signaling system circuit were detected in reduced amounts in the supernatant of the two isolates overproducing type 1 beta-lactamase . These results suggest that strains overexpressing the chromosomal type 1 beta-lactamase could be less virulent because of a reduction of cell-to-cell signaling dependent virulence factor production. Enzyme Microb Technol, 2000 Jul 1, 27(1-2), 3 - 10 Protease produced by Pseudomonas aeruginosa K-187 and its application in the deproteinization of shrimp and crab shell wastes; Oh Y et al.; In addition to chitinase/lysozyme, Pseudomonas aeruginosa K-187 also produced a protease useful for the deproteinization of shrimp and crab shell wastes . The optimal culture conditions for P . aeruginosa K-187 to attain the highest protease activity were investigated and discussed . The highest protease activity was as high as 21.2 U/ml, 10-fold that (2.2 U/ml) obtained prior to optimization . The protease of P . aeruginosa K-187, produced under the optimal culture conditions, was tested for crustacean waste deproteinization . The percent of protein removal for shrimp and crab shell powder (SCSP) after 7-day incubation was 72%, while that of natural shrimp shell (NSS) and acid-treated SCSP was 78% and 45%, respectively . In contrast, with the protease produced under pre-optimization conditions, the percent of protein removal for SCSP, NSS, and acid-treated SCSP was 48%, 55%, and 40%, respectively . For comparison, three other protease-producing microbes were tested for crustacean waste deproteinization . However, they were shown to be less efficient in deproteinization than P . aeruginosa K-187 . The crude protease produced by P . aeruginosa K-187 can be covalently immobilized on a reversibly soluble polymeric support (hydroxypropyl methycellulose acetate succinate) . The immobilized enzyme was soluble above pH 5.5 but insoluble below pH 4.5 . Immobilization efficiency was 82% . The immobilized enzyme was stable between pH 6 and 9 and at temperatures below 60 degrees C . The optimum pH and temperature for the immobilized enzyme was pH 8 and 50 degrees C . The half-life of the immobilized enzyme was 12 days, longer than that of free protease (8 days) . The utilization of the immobilized enzyme for the deproteinization of SCSP has resulted in a 67% protein removal . By contrast, SCSP protein removal by using free enzymes was 72% . The protease was further purified and characterized . The purification steps included ammonium sulfate precipitation, DEAE-Sepharose CL-6B ion-exchange chromatography, and Sephacryl S-200 gel-permeation chromatography . The enzyme had a molecular weight estimated to be 58.8 kDa by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis . The purified enzyme was active from pH 7 to 9 and its optimal pH was 8. Indian J Pathol Microbiol, 1999 Jul, 42(3), 317 - 20 Bacteraemia in a tertiary care urban hospital in south India; Chaudhury A et al.; A total of 1727 blood samples were cultured aerobically over a one year period, of which 201(11.8%) were positive . The ratio of Gram positive to Gram negative bacteraemia was 1:1 . The three antimicrobials having the highest activities against the Gram positive isolates were amikacin, cefotaxime, and ciprofloxacin to which 88.5, 81.7 and 80.7 percent of the strains were susceptible: and the same agents were equally effective against Gram negative organisms with 84.5, 75.3 and 70.1 percent efficacy respectively . Coagulase negative Staphylococcus spp . was the most frequent organism isolated(60; 29.8%), followed by Pseudomonas aeruginosa (40; 19.9%), and Staphylococcus aureus (34; 16.9%). Ann Surg, 2000 Jul, 232(1), 133 - 42 The key role of Pseudomonas aeruginosa PA-I lectin on experimental gut-derived sepsis; Laughlin RS et al.; OBJECTIVE: To examine the effect of Pseudomonas aeruginosa on intestinal barrier function and its lethal potential when introduced into the intestinal tract of mice . SUMMARY BACKGROUND DATA: The mere presence of P . aeruginosa in the intestinal tract of critically ill patients is associated with a threefold increase in death compared with matched cohorts without this pathogen . Whether this effect is a cause or a consequence of the critically ill state has not been previously addressed . METHODS: Transepithelial electrical resistance, a measure of tight junction permeability, was evaluated in Caco-2 intestinal epithelial cells cells apically inoculated with live P . aeruginosa, exotoxin A, or purified PA-I lectin, an adhesin of P . aeruginosa . Lethality studies to P . aeruginosa were carried out in mice undergoing 30% surgical hepatectomy by injecting the bacteria or its various components directly into the cecum . RESULTS: Only cells exposed to P . aeruginosa or its PA-I lectin developed alterations in barrier function . P . aeruginosa or the combination of PA-I and exotoxin A was lethal to mice when injected into the cecum after partial hepatectomy . Alterations in epithelial barrier function and death in mice were prevented when Pseudomonas was pretreated with N-acetyl D-galactosamine (GalNAc), a binder of PA-I . CONCLUSIONS: P . aeruginosa may act as a pathogen in the gastrointestinal tract, resulting in altered epithelial barrier function and death in a susceptible host . The PA-I lectin of P . aeruginosa may play a key role in its pathogenicity to the intestinal epithelium by inducing a permeability defect to its cytotoxic exoproducts such as exotoxin A. Pediatr Pulmonol, 2000 Jul, 30(1), 25 - 31 French multicenter randomized double-blind placebo-controlled trial on nebulized amiloride in cystic fibrosis patients . The Amiloride-AFLM Collaborative Study Group; Pons G et al.; The effect of amiloride, a sodium channel blocker, has been evaluated in a multicenter randomized double-blind placebo-controlled trial in cystic fibrosis patients more than 5-years-old (n = 137) whose forced vital capacity (FVC), forced expiratory volume in 1 sec (FEV(1)), and forced mid-expiratory flow (FEF(25-75)) were not below 50%, 50%, and 30% of reference values, respectively . Patients were randomly allocated to two parallel groups . Sixty-four patients were chronically colonized with Pseudomonas aeruginosa; they received either amiloride or placebo as a nebulized solution three times daily for 6 months . Routine treatments were continued . Patients chronically colonized with Pseudomonas received nebulized colimycine twice a day for a month during the third and sixth months of treatment . Bronchopulmonary exacerbations were treated in the usual way . The effects of the amiloride treatment were assessed at the end of the 6-month treatment period . The effects on FVC and secondarily on FEV(1), FEF(25-75), the number of days on antibiotic therapy, the Shwachman score, a nutritional index (weight/height(2)), the change in sputum bacterial flora, and nocturnal cough were assessed . For the patients not chronically colonized with Pseudomonas, the effect of the treatment was also evaluated by counting chronic colonizations with pathogens appearing during the trial period . The present study failed to demonstrate any significant benefit of amiloride over placebo on FVC, FEV(1), and the other secondary endpoints in the studied population . Neither the chronically colonized, nor the noncolonized patients benefited . The confidence intervals of the differences between treatment groups indicated small differences that were most likely of no clinical significance . Complementary analyses taking into account the gender, the type of mutation, the subpopulations whose FVC and FEV(1) were below 80% of normal values at the beginning of the study, and also patients less than 10 years old, did not show any statistically or clinically significant improvements following amiloride therapy . Phytother Res, 2000 Jun, 14(4), 278 - 80 Studies on antibacterial activity of Ficus racemosa Linn . leaf extract; Mandal SC et al.; Extracts of Ficus racemosa Linn . leaves were tested for antibacterial potential against Escherichia coli ATCC 10536, Basillus pumilis ATCC 14884, Bacillus subtilis ATCC 6633, Pseudomonas aeruginosa ATCC 25619 and Staphylococcus aureus ATCC 29737 . The effects produced by the extracts were significant and were compared with chloramphenicol . The petroleum ether extract was the most effective against the tested organisms . Ugeskr Laeger, 2000 May 15, 162(20), 2893 - 4 {Pseudomonas aeruginosa as the cause of meningitis in a patient with epidural catheter}; Jensen GH et al.; A case of epidural infection following epidural catheterization is presented . The clinical signs initially were backpain and a small swelling at the site of insertion . Treatment with dicloxacillin was begun, presuming a Staphylococcus-infection . The symptoms persisted . Weeks later the patient developed meningitis and Pseudomonas aeruginosa was cultivated . Antibiotic treatment was changed to ceftazidime, netilmycin and ciprofloxacin . Complete recovery followed. Chemotherapy, 2000 Jul-Aug, 46(4), 229 - 34 In vitro activity of ceftazidime, cefepime and imipenem on 1,005 Pseudomonas aeruginosa clinical isolates either susceptible or resistant to beta-lactams; Bonfiglio G et al.; BACKGROUND: Recently, new 'fourth-generation' cephalosporins, such as cefepime and cefpirome, were introduced into antibacterial chemotherapy . METHODS: In order to explore whether these new cephalosporins offer real advantages against Pseudomonas aeruginosa, we matched the in vitro activity of cefepime with that of ceftazidime and imipenem as reference compounds . RESULTS: Among the 1,005 clinical isolates tested, 86.6% were susceptible to ceftazidime, whereas 80.7 and 76.9% were susceptible to imipenem and cefepime, respectively . Furthermore, the activity of the three compounds against a significant number of clinical isolates of P . aeruginosa expressing different resistance mechanisms to beta-lactam antibiotics was investigated . Among these isolates, 62.5% were still susceptible to ceftazidime, and 52.1 and 38.7% were inhibited by imipenem and cefepime, respectively . CONCLUSION: Ceftazidime and imipenem retained their activity against the majority of clinical P . aeruginosa isolates collected in Italy . Cefepime did not offer competitive advantages in terms of in vitro activity . Antimicrob Agents Chemother, 2000 Jul, 44(7), 1983 - 5 Role of putative loops 2 and 3 in imipenem passage through the specific porin OprD of Pseudomonas aeruginosa; Ochs MM et al.; Mutant proteins with eight amino acid deletions in putative surface loops 2 and 3 of the imipenem-specific porin OprD of Pseudomonas aeruginosa failed to reconstitute imipenem susceptibility in an oprD-deficient background . The loop 3 deletion prevented the ability of imipenem to inhibit KCl conductance through the OprD channel, as previously shown for a loop 2 deletion . This suggests that both loops 2 and 3 have a role in imipenem binding to the OprD channel. Infect Immun, 2000 Jul, 68(7), 4331 - 4 Pseudomonas aeruginosa cell-to-cell signaling is required for virulence in a model of acute pulmonary infection; Pearson JP et al.; Cell-to-cell signaling controls many virulence genes in Pseudomonas aeruginosa . We tested the virulence of las and rhl quorum-sensing mutants in neonatal mice . A lasI rhlI double mutant was nearly avirulent, and the respective single mutant strains were reduced in virulence compared with the wild-type strain . Quorum sensing plays a role in P . aeruginosa pneumonia in neonatal mice. Infect Immun, 2000 Jul, 68(7), 4289 - 96 CXC chemokine receptor CXCR2 is essential for protective innate host response in murine Pseudomonas aeruginosa pneumonia; Tsai WC et al.; Pulmonary infection due to Pseudomonas aeruginosa has emerged as a leading cause of mortality . A vigorous host response is required to effectively clear the organisms from the lungs . This host defense is dependent on the recruitment and activation of neutrophils and macrophages . A family of chemotactic cytokines (chemokines) has been shown to participate in this protective response . In this study, we assessed the role of the ELR(+) (glutamic acid-leucine-arginine motif positive) CXC chemokines and their CXC chemokine receptor (CXCR2) in lung antibacterial host defense . The intratracheal administration of Pseudomonas to mice resulted in the time-dependent influx of neutrophils to the lung, peaking at 12 to 24 h after inoculation . The influx of neutrophils was associated with a similar time-dependent expression of the ELR(+) CXC chemokines, KC, macrophage inflammatory protein 2 (MIP-2), and lipopolysaccharide-induced CXC chemokine (LIX) . Selective neutralization of MIP-2 or KC resulted in modest changes in neutrophil influx but no change in bacterial clearance or survival . However, neutralization of CXCR2 resulted in a striking increase in mortality, which was associated with a marked decrease in neutrophil recruitment and bacterial clearance . Conversely, the site-specific transgenic expression of KC resulted in enhanced clearance of bacteria after Pseudomonas challenge . This study indicates that ELR(+) CXC chemokines are critical mediators of neutrophil-mediated host defense in Pseudomonas pneumonia. Infect Immun, 2000 Jul, 68(7), 3998 - 4004 Acquisition of expression of the Pseudomonas aeruginosa ExoU cytotoxin leads to increased bacterial virulence in a murine model of acute pneumonia and systemic spread; Allewelt M et al.; Pseudomonas aeruginosa is the nosocomial bacterial pathogen most commonly isolated from the respiratory tract . Animal models of this infection are extremely valuable for studies of virulence and immunity . We thus evaluated the utility of a simple model of acute pneumonia for analyzing P . aeruginosa virulence by characterizing the course of bacterial infection in BALB/c mice following application of bacteria to the nares of anesthetized animals . Bacterial aspiration into the lungs was rapid, and 67 to 100% of the inoculum could be recovered within minutes from the lungs, with 0.1 to 1% of the inoculum found intracellularly shortly after infection . At later time points up to 10% of the bacteria were intracellular, as revealed by gentamicin exclusion assays on single-cell suspensions of infected lungs . Expression of exoenzyme U (ExoU) by P . aeruginosa is associated with a cytotoxic effect on epithelial cells in vitro and virulence in animal models . Insertional mutations in the exoU gene confer a noncytotoxic phenotype on mutant strains and decrease virulence for animals . We used the model of acute pneumonia to determine whether introduction of the exoU gene into noncytotoxic strains of P . aeruginosa lacking this gene affected virulence . Seven phenotypically noncytotoxic P . aeruginosa strains were transformed with pUCP19exoUspcU which carries the exoU gene and its associated chaperone . Three of these strains became cytotoxic to cultured epithelial cells in vitro . These strains all secreted ExoU, as confirmed by detection of the ExoU protein with specific antisera . The 50% lethal dose of exoU-expressing strains was significantly lower for all three P . aeruginosa isolates carrying plasmid pUCP19exoUspcU than for the isogenic exoU-negative strains . mRNA specific for ExoU was readily detected in the lungs of animals infected with the transformed P . aeruginosa strains . Introduction of the exoU gene confers a cytotoxic phenotype on some, but not all, otherwise-noncytotoxic P . aeruginosa strains and, for recombinant strains that could express ExoU, there was markedly increased virulence in a murine model of acute pneumonia and systemic spread. J Leukoc Biol, 2000 Jun, 67(6), 808 - 16 Exoenzyme S from Pseudomonas aeruginosa induces apoptosis in T lymphocytes; Bruno TF et al.; Exoenzyme S from Pseudomonas aeruginosa is a unique T cell mitogen; it is a powerful immunostimulus that activates a large proportion of T cells, but results in delayed and reduced lymphocyte proliferation . This study was performed to explain the discrepancy between early T cell activation and subsequent proliferation . Studies revealed that exoenzyme S induced rapid and unsustained surface expression of CD69, but could not induce interleukin-2 receptor alpha (IL-2R alpha) up-regulation on T cells . IL-2 was undetectable in supernatants and addition of rIL-2 could not reverse the unresponsiveness, indicating that anergy was not involved . Exoenzyme S induced membrane phosphatidylserine translocation, DNA hypodiploidy, and DNA fragmentation, implicating apoptosis as the mechanism for the unresponsiveness . Exoenzyme S-induced apoptosis shows features of both propriocidal and death by neglect, suggesting shared characteristics of an intermediate pathway . Thus, a Pseudomonas exoproduct induces T cell apoptosis, which may contribute to the pathogenesis of Pseudomonas infections in diseases such as cystic fibrosis. Int J Antimicrob Agents, 2000 Jun, 15(1), 73 - 6 In vitro activity of four fluoroquinolones against clinical isolates of Pseudomonas aeruginosa determined by the E test; Swiatlo E et al.; Ciprofloxacin, levofloxacin, ofloxacin, and trovafloxacin were tested by the E-test against 100 clinical isolates of Pseudomonas aeruginosa . Ciprofloxacin was the most active of the tested agents with 82% of isolates having a MIC </=1 mg/l (range: 0.094->8) . Levofloxacin and trovafloxacin had nearly identical potency: 75% and 76% of the isolates were inhibited by </=2 mg/l of these agents, respectively (range: 0.125->8 for levofloxacin; 0.19->8 for trovafloxacin) . Ofloxacin was the least active of the four quinolones, with 43% of the isolates having a MIC >2 mg/l . All isolates resistant to ciprofloxacin were also resistant to the other agents, i.e . resistance to ciprofloxacin predicted resistance to all the quinolones tested in every case . This data demonstrates that fluoroquinolones are active agents against P . aeruginosa . In vitro susceptibility testing, however, is crucial to assess the resistance pattern in any specific location and for each individual agent. FEMS Microbiol Lett, 2000 Jun 15, 187(2), 161 - 5 Protective effects of alpha-tocopherol and beta-carotene on para-nonylphenol-induced inhibition of cell growth, cellular respiration and glucose-induced proton extrusion of bacteria; Okai Y et al.; para-Nonylphenol (NP) showed a dose-dependent inhibition against the cell growth of Bacillus subtilis, Micrococcus luteus, Pseudomonas aeruginosa and Staphylococcus aureus at 5-100 microM . However, other typical plastic-derived endocrine disruptors such as bisphenol A and di-2-ethylhexyl phthalate (DEHP) did not significantly affect the cell growth of these bacteria at 5-100 microM . The NP-induced cell growth inhibition was restored when concomitantly supplemented with lipophilic antioxidants such as alpha-tocopherol and beta-carotene, but not with hydrophilic antioxidants, ascorbic acid and (-)-epigallocatechin gallate (EGCG) . NP also suppressed in a dose-dependent manner cellular oxygen consumption and glucose-induced proton extrusion of these bacteria at 10-100 microM . Both effects were prevented when added with alpha-tocopherol and beta-carotene, but not with ascorbic acid and EGCG . The significance of these results is discussed from the viewpoint of environmental microbiology and a possible biochemical mechanism of the inhibitory effect of NP is suggested. J Bacteriol, 2000 Jun, 182(12), 3400 - 4 Identification and characterization of two chemotactic transducers for inorganic phosphate in Pseudomonas aeruginosa; Wu H et al.; Two chemotactic transducers for inorganic phosphate (P(i)), designated CtpH and CtpL, have been identified in Pseudomonas aeruginosa . The corresponding genes (ctpH and ctpL) were inactivated by inserting kanamycin and tetracycline resistance gene cassettes into the wild-type genes in the P . aeruginosa PAO1 genome . Computer-assisted capillary assays showed that the ctpH single mutant failed to exhibit P(i) taxis when the concentration of P(i) in the capillary was higher than 5 mM . Conversely, the ctpL single mutant could not respond to P(i) at the concentration of 0.01 mM . The ctpH ctpL double mutant was defective in P(i) taxis at any concentration ranging from 0.01 to 10 mM . To investigate regulation of P(i) taxis, the ctpH and ctpL genes were also disrupted individually in the P . aeruginosa phoU and phoB single mutants . The ctpH phoU and ctpH phoB double mutants were defective in P(i) taxis, regardless of whether the cells were starved for P(i) . The ctpL phoU double mutant was constitutive for P(i) taxis, whereas the ctpL phoB double mutant was induced by P(i) limitation for P(i) taxis . The region upstream of ctpL, but not ctpH, contained a putative pho box sequence . Expression of ctpL::lacZ was induced by P(i) limitation in PAO1, while it was constitutive in the phoU mutant . In contrast, the phoB mutant showed only background levels of ctpL::lacZ expression . These results showed that ctpL is involved in the pho regulon genes in P . aeruginosa . The ctpH phoU mutant, which failed to exhibit P(i) taxis, was constitutive for ctpL::lacZ expression, suggesting that the P(i) detection by CtpL requires PhoU . Like PAO1, the phoB and phoU single mutants were constitutive for expression of ctpH::lacZ . Thus, the evidence that the ctpL phoU mutant, but not the ctpL phoB mutant and PAO1, was constitutive for P(i) taxis raised the possibility that PhoU exerts a negative control on P(i) detection by CtpH at the posttranscriptional level. Zh Mikrobiol Epidemiol Immunobiol, 1999 Jul-Aug, (4), 3 - 7 {The reciprocal effect of the causative agents in a mixed infection in burn injury}; Bel'skii VV et al.; In vitro experiments on the joint cultivation of Pseudomonas aeruginosa, Escherichia coli, staphylococci and in vivo experiments carried out on mice with the experimental mixed infection of a burn injury revealed the pronounced antagonistic action of P . aeruginosa with respect to staphylococci and E . coli . Under the same conditions, in the joint cultivation of P . aeruginosa and fungi of the genus Candida and in the mixed infection of a burn wound caused by the same microorganisms the mutual stimulation of multiplication and a considerable aggravation of the clinical course of a burn wound were observed . The mutual influence of associates in the mixed infection of a burn injury is manifested by an increase in the number of an antibiotic-resistance microorganisms in their populations and, so far as the macroorganism is concerned, in the aggravation of the course of the infectious process and the formation of the pronounced state of immunodeficiency. Neurology, 2000 Jun 13, 54(11), 2153 - 5 Status epilepticus associated with cefepime; Dixit S et al.; ARTICLE ABSTRACT: Cefepime is a fourth-generation cephalosporin widely used for gram-negative sepsis . The authors report two patients in whom nonconvulsive status epilepticus developed while they were on treatment with cefepime for Pseudomonas aeruginosa infection . The status epilepticus resolved completely once the drug was withdrawn . Cefepime therapy can result in status epilepticus, especially if given in higher doses than required. J Mol Biol, 2000 Jun 16, 299(4), 1005 - 17 Crystal structure of Pseudomonas aeruginosa PAK pilin suggests a main-chain-dominated mode of receptor binding; Hazes B et al.; Fibers of pilin monomers (pili) form the dominant adhesin of Pseudomonas aeruginosa, and they play an important role in infections by this opportunistic bacterial pathogen . Blocking adhesion is therefore a target for vaccine development . The receptor-binding site is located in a C-terminal disulphide-bonded loop of each pilin monomer, but functional binding sites are displayed only at the tip of the pilus . A factor complicating vaccination is that different bacterial strains produce distinct, and sometimes highly divergent, pilin variants . It is surprising that all strains still appear to bind a common receptor, asialo-GM1 . Here, we present the 1.63 A crystal structure of pilin from P . aeruginosa strain PAK . The structure shows that the proposed receptor-binding site is formed by two beta-turns that create a surface dominated by main-chain atoms . Receptor specificity could therefore be maintained, whilst allowing side-chain variation, if the main-chain conformation is conserved . The location of the binding site relative to the proposed packing of the pilus fiber raises new issues and suggests that the current fiber model may have to be reconsidered . Finally, the structure of the C-terminal disulphide-bonded loop will provide the template for the structure-based design of a consensus sequence vaccine . J Bacteriol, 2000 Jul, 182(13), 3843 - 5 Positive correlation between virulence of Pseudomonas aeruginosa mutants in mice and insects; Jander G et al.; Strain PA14, a human clinical isolate of Pseudomonas aeruginosa, is pathogenic in mice and insects (Galleria mellonella) . Analysis of 32 different PA14 mutants in these two hosts showed a novel positive correlation in the virulence patterns . Thus, G . mellonella is a good model system for identifying mammalian virulence factors of P . aeruginosa. J Bacteriol, 2000 Jul, 182(13), 3826 - 31 Secretion of nucleoside diphosphate kinase by mucoid Pseudomonas aeruginosa 8821: involvement of a carboxy-terminal motif in secretion; Kamath S et al.; Nucleoside diphosphate kinase (Ndk) is a ubiquitous enzyme which functions in balancing the nucleotide pool of the cell . We have recently reported that in addition to being intracellular in both mucoid and nonmucoid Pseudomonas aeruginosa, Ndk is also secreted into the extracellular environment by mucoid P . aeruginosa cells . This secreted Ndk has biochemical activity similar to the intracellular Ndk and is 16 kDa in size . To demonstrate that Ndk is indeed secreted and to localize the secretion motif, we constructed an ndk knockout mutant, which lacks both intracellular and extracellular forms of Ndk . In this study, we report the construction of deletion derivatives made from the carboxy-terminal region of Ndk . These deletion derivatives were introduced into the ndk::Cm knockout mutant and were examined for the intracellular and extracellular presence of Ndk . It was observed that the carboxy-terminal 8-amino-acid region is required for the secretion of Ndk into the extracellular region . This region has the sequence DXXX, where X is a predominantly hydrophobic residue . Such sequences represent a conserved motif in proteins secreted by the type I secretory pathway in gram-negative microorganisms . To investigate the significance of this motif in the secretion of Ndk, we constructed a fusion protein of Ndk and the blue fluorescent protein (BFP) as well as a fusion protein of mutated Ndk (whose DTEV motif has been changed to AAAA) and the BFP . The presence of extracellular Ndk was detected only in the ndk::Cm knockout mutant harboring the wild-type BFP-Ndk protein fusion . We could not detect the presence of extracellular Ndk in the ndk::Cm knockout mutant containing the mutated BFP-Ndk protein fusion . In addition, we have also used immunofluorescence microscopy to localize the wild-type and mutated BFP-Ndk proteins in the cell . The significance of these observations is discussed. Wien Klin Wochenschr, 2000 Apr 7, 112(7), 329 - 33 Risk of Pseudomonas aeruginosa cross-colonisation in patients with cystic fibrosis within a holiday camp--a molecular-epidemiological study; Hunfeld KP et al.; OBJECTIVE: A study on the molecular epidemiology of Pseudomonas aeruginosa in patients with cystic fibrosis (CF) from Germany (N = 18) and Israel (N = 12) is presented . The aim is to provide an answer to the question as to whether or not social contact outside the hospital environment involves a potential risk for person-to-person spread of this pathogen . METHODS: Sputa from German and Israeli patients were obtained while these were attending a holiday camp in Israel . The sputum samples were analysed with regard to Pseudomonas aeruginosa . Strains dissimilar in macroscopic appearance and/or antibiotic resistance patterns were genotyped using pulsed-field gel electrophoresis after digestion of genomic DNA with restriction endonuclease Spel . The genetic polymorphism of DNA fragment patterns of all strains (N = 146) was studied for their overall relatedness using a fingerprint software system . RESULTS: Most of the German patients (77.7%) were colonised persistently by a unique clonal type during the four-week screening period . Isolates obtained from Israeli patients displayed a very close clonal relationship and a higher antibiotic resistance as a result of preceding epidemic spread of certain clones before the camp . Additionally, isolates showing identical PFGE patterns were demonstrated once in a single male Israeli patient and in one female German patient, suggesting previous cross-colonisation . CONCLUSION: The occurrence of person-to-person spread through social contact in patients with CF is supported by our findings, but remains a rare event outside the hospital environment, provided appropriate hygienic measures are applied. Eur J Clin Invest, 2000 Jun, 30(6), 553 - 9 Metabolic and inflammatory responses to pulmonary exacerbation in adults with cystic fibrosis; Bell SC et al.; BACKGROUND: We hypothesized that increased resting energy expenditure in adults with cystic fibrosis was related to chronic inflammation secondary to pulmonary infection and could be modified by treatment of the underlying infection . METHOD: To determine the relationship between resting energy expenditure and the inflammatory and metabolic responses, we studied 22 adults with cystic fibrosis and chronic Pseudomonas aeruginosa infection before and after treatment of a respiratory exacerbation . Resting energy expenditure was measured by indirect calorimetry . Spirometry and circulating concentrations of C-reactive protein, neutrophil elastase alpha1-antiproteinase complex, catecholamines, non-esterified fatty acids and glycerol were determined . RESULTS: The mean (95% confidence interval)% predicted FEV1 was 28.5% (20.6, 36.4) and mean body weight 50.7 kg (47.4, 54.1) . Following treatment, 1-s forced expiratory volume (FEV1) and weight increased, while C-reactive protein (P<0.0001) and neutrophil elastase alpha1-antiproteinase complex concentrations (P<0.0001) were reduced . Resting energy expenditure decreased from 6.8 (6.3, 7.2) to 6.25 (5.9, 6.6) MJ day-1 by day 15 (P<0.001) . Changes in resting energy expenditure and C-reactive protein were related (r = 0.66, P< 0.0001) . Weight gain was inversely related to resting energy expenditure (r = 0.43, P = 0.02) and unrelated to energy intake (r = 0.02, P = 0.47) . Post-treatment reduction in norepinephrine was related to changes in heart rate (r = 0.57, P<0.01), resting energy expenditure (r = 0.51, P = 0.001) and non-esterified fatty acids (r = 0.42, P< 0.05) . CONCLUSIONS: A parallel reduction in the host inflammatory and catabolic responses followed treatment of a respiratory exacerbation and may have contributed to weight gain. Kaohsiung J Med Sci, 2000 Mar, 16(3), 162 - 5 Otogenic brain abscess--a case report; Chen PT et al.; Brain abscess is one of the life-threatening complications of otitis media . Mortality and morbidity have decreased with the advent of antibiotic therapy . More frequently encountered in cases of acute otitis media in the preantibiotic era, in recent years otogenic brain abscess was noticed almost only in patients of chronic otitis media with cholesteatoma . A case of brain abscess in a 49-year-old female was initially diagnosed as a headache . A high resolution computed tomography (HRCT) scan of the temporal bones later revealed that there were two abscesses over the right side temporal lobe . A modified radical mastoidectomy was performed . Cultures of the middle ear cholesteatoma later grew Pseudomonas aeruginosa and Strenotrophomonas maltophilia . Antibiotic therapy was carried on for three months postoperatively . The patient improved but retained a conductive hearing loss. Microbiology, 2000 Jun, 146 ( Pt 6), 1429 - 35 The Pseudomonas aeruginosa hscA gene encodes Hsc66, a DnaK homologue; Campos-Garcia J et al.; Under heat-stress conditions bacteria induce, among other heat-shock proteins, the Hsp70 molecular chaperone (DnaK), which is involved in protein stabilization . It has been shown in Escherichia coli that an Hsp70 homologue called Hsc66, which is widespread in bacteria, functions as a chaperone in vitro . This paper reports the isolation of a Pseudomonas aeruginosa W51D mutant (W51M22) by insertion of the mini-Tn5-Hg transposon, which was unable to grow on ethanol and other short-chain alcohols as sole source of carbon . The transposon insertion in this mutant was shown to be located in the hscA gene encoding Hsc66 . The inability of mutant W51M22 to use ethanol was complemented by the E . coli hscBA-fdx operon . The authors characterized the transcriptional arrangement of hscA, showing that it forms part of an operon with the upstream hscB gene, and that it is also expressed from its own promoter . These results are compatible with the P . aeruginosa Hsc66 protein being a functional molecular chaperone involved in the stabilization, in the presence of ethanol, of some proteins required for bacterial growth on short-chain alcohols. J Immunol, 2000 Jun 15, 164(12), 6576 - 82 Prolonged elevation of IL-1 in Pseudomonas aeruginosa ocular infection regulates macrophage-inflammatory protein-2 production, polymorphonuclear neutrophil persistence, and corneal perforation; Rudner XL et al.; The kinetics of IL-1 (alpha and beta) production after Pseudomonas aeruginosa corneal infection was examined in susceptible (cornea perforates) C57BL/6J (B6) and resistant (cornea heals) BALB/cByJ (BALB/c) mice . IL-1alpha and -1beta (mRNA and protein) were elevated in both mouse strains, and levels peaked at 1 day postinfection (p.i . ) . Significantly greater amounts of IL-1 protein were detected in B6 vs BALB/c mice at 1 and 3 days p.i . At 5 days p.i., IL-1alpha and -1beta (mRNA and protein) remained elevated in B6, but began to decline in BALB/c mice . To test the significance of elevated IL-1 in B6 mice, a polyclonal neutralizing Ab against IL-1beta was used to treat infected B6 mice . A combination of subconjunctival and i.p . administration of IL-1beta polyclonal Ab significantly reduced corneal disease . The reduction in disease severity in infected B6 mice was accompanied by a reduction in corneal polymorphonuclear neutrophil number, bacterial load, and macrophage inflammatory protein-2 mRNA and protein levels . These data provide evidence that IL-1 is an important contributor to P . aeruginosa corneal infection . At least one mechanism by which prolonged and/or elevated IL-1 expression contributes to irreversible corneal tissue destruction appears to be by increasing macrophage inflammatory protein-2 production, resulting in a prolonged stimulation of polymorphonuclear neutrophil influx into cornea . In contrast, a timely down-regulation of IL-1 appears consistent with an inflammatory response that is sufficient to clear the bacterial infection with less corneal damage. J Nat Prod, 2000 May, 63(5), 605 - 10 Bioactive 12-oleanene triterpene and secotriterpene acids from Maytenus undata; Muhammad I et al.; The aerial parts of Maytenus undata yielded four new 12-oleanene and 3,4-seco-12-oleanene triterpene acids, namely, 3-oxo-11alpha-methoxyolean-12-ene-30-oic acid (1), 3-oxo-11alpha-hydroxyolean-12-ene-30-oic acid (2), 3-oxo-olean-9(11), 12-diene-30-oic acid (3), and 3,4-seco-olean-4(23),12-diene-3, 29-dioic acid (20-epi-koetjapic acid) (5), together with the known 3, 11-dioxoolean-12-ene-30-oic acid (3-oxo-18beta-glycyrrhetinic acid) (4), koetjapic acid (6), and the 12-oleanene artifact 3-oxo-11alpha-ethoxyolean-12-ene-30-oic acid (7) . Koetjapic acid (6) inhibited the growth of Staphylococcus aureus, methicillin-resistant S . aureus, and Pseudomonas aeruginosa, with an MIC range of 3.125-6.25 microg/mL . The new 3,4-secotriterpene acid 20-epi-koetjapic acid (5) potently inhibited rat neonatal brain microglia phorbol ester-stimulated thromboxane B(2) (IC(50) = 0.5 microM) and superoxide anion (IC(50) = 1.9 microM) generation. J Ethnopharmacol, 2000 Jun, 70(3), 343 - 9 Screening of some Palestinian medicinal plants for antibacterial activity; Essawi T et al.; Antibacterial activity of organic and aqueous extracts of 15 Palestinian medicinal plants were carried against eight different species of bacteria: Bacillus subtilis, two Escherichia coli species, Staphylococcus aureus (methicillin resistant), two S . aureus (methicillin sensitive) species, Pseudomonas aeruginosa, and Enterococcus fecalis . Of the 15 plants tested, eight showed antibacterial activity . Each plant species has unique against different bacteria . The most active antibacterial plants against both gram-positive and gram-negative bacteria were Thymus vulgaris and Thymus origanium . The organic and aqueous extract from the same plants showed different activities; the organic extract showed the same or greater activity than the aqueous extract . Finally, the hole-plate diffusion method showed larger activity than the disc diffusion method. J Antimicrob Chemother, 2000 Jun, 45(6), 899 - 901 In vitro activity of meropenem, imipenem, cefepime and ceftazidime against Pseudomonas aeruginosa isolates from cystic fibrosis patients; Christenson JC et al.; We studied 67 Pseudomonas aeruginosa isolates from cystic fibrosis patients, and compared their in vitro susceptibility to two carbapenems (meropenem and imipenem) and two cephalosporins (cefepime and ceftazidime) . The carbapenems were more effective in vitro than the cephalosporins: 92.5% of isolates were susceptible to the former and 77.6% to the latter . Essentially no difference was found between meropenem and imipenem . More discrepancies were seen between cefepime and ceftazidime: four of 67 isolates (6.0%) were more susceptible to cefepime than to ceftazidime, while eight (11 . 9%) were more susceptible to ceftazidime than to cefepime. Am J Respir Cell Mol Biol, 2000 Jun, 22(6), 714 - 21 Mucoid Pseudomonas aeruginosa, TNF-alpha, and IL-1beta, but not IL-6, induce human beta-defensin-2 in respiratory epithelia; Harder J et al.; Cultured lung epithelial cells release antibacterial activity upon contact with Pseudomonas aeruginosa (PA), which is impaired in cystic fibrosis (CF) . In order to identify the factors responsible for killing PA by a biochemical approach, we purified antimicrobial activity from supernatants of the A549 lung epithelial cell line, previously stimulated with PA bacteria, by subsequent high performance liquid chromatography . NH(2)-terminal sequencing of a major bactericidal compound revealed it to be identical with human beta-defensin-2 (hBD-2) . A mucoid phenotype of PA, but not two nonmucoid PA strains, high concentrations (> 10 microg/ml) of PA lipopolysaccharide, tumor necrosis factor alpha, and interleukin (IL)-1beta, but not IL-6, dose-dependently induced hBD-2 messenger RNA in cultured normal bronchial, tracheal, as well as normal and CF-derived nasal epithelial cells . Genomic analysis of hBD-2 revealed a promoter region containing several putative transcription factor binding sites, including nuclear factor (NF) kappaB, activator protein (AP)-1, AP-2, and NF-IL-6, known to be involved in the regulation of inflammatory responses . Thus, hBD-2 represents a major inducible antimicrobial factor released by airway epithelial cells either on contact with mucoid PA or by endogenously produced primary cytokines . Therefore, it might be important in lung infections caused by mucoid PA, including those seen in patients with CF. Eur Respir J, 1999 Jan, 13(1), 100 - 2 Genotype-phenotype correlations in cystic fibrosis: clinical severity of mutation S549R(T-->G); Frossard PM et al.; With a view to assessing genotype-to-phenotype correlations in cystic fibrosis (CF), the clinical presentation of CF children from the United Arab Emirates (UAE) who were homozygous for cystic fibrosis transmembrane conductance regulator (CFTR) mutation S549R(T-->G was investigated . This mutation is localized in intron 11 (nucleotide binding domain 1 of the CFTR protein) and had so far been described as a private mutation only . The associations between the R549/R549 genotype and 20 outcome variables, including age at diagnosis, sweat chloride concentrations, growth percentiles, meconium ileus, pancreatic sufficiency, pulmonary disease, associated complications and micro-organism colonization were examined in a group of 15 CF children (9 females and 6 males) . Mean current age and age at diagnosis were both low (5.4+/-3.5 and 1.0+/-1.1 yrs, respectively) . Although none of the 15 CF patients had presented with meconium ileus at birth, all were pancreatic insufficient and had very severe lung disease, with a high rate of Pseudomonas aeruginosa and Staphylococcus aureus . Two patients died during the course of this investigation (one was 5 months and the other, 6 yrs old) . The clinical presentation associated with S549R(T-->G) homozygosity in the United Arab Emirates is quite homogeneous and shows an extreme degree and course of cystic fibrosis severity. Am J Physiol Lung Cell Mol Physiol, 2000 Jun, 278(6), L1213 - 20 Airway surface liquid composition in mice; Cowley EA et al.; Airway surface liquid (ASL) lines the conducting airways of the respiratory tract . We collected small samples of this liquid from the lower tracheae of anesthetized C57BL/6 mice and determined its ionic composition (in mM: 87.2 Na(+), 4.7 K(+), and 57.0 Cl(-)) . Intravenous methacholine produced significant increases in the concentrations of Na(+), K(+), and Cl(-) within ASL . A limited analysis of liquid from cystic fibrosis transmembrane conductance regulator (CFTR) knockout mice revealed no significant differences compared with littermate controls; however, Pseudomonas aeruginosa infection led to an increase in the salt concentration of ASL in cftr(+/+) mice . Morphometric measurements of tracheal submucosal gland volume revealed significant differences between inbred mouse strains, corresponding to ease of ASL collection . We conclude that although submucosal glands may be responsible for the production of some ASL, the ionic composition of this liquid is actively regulated by the underlying epithelial cells. Protein Eng, 2000 May, 13(5), 329 - 37 Linker insertion mutagenesis based on IS21 transposition: isolation of an AMP-insensitive variant of catabolic ornithine carbamoyltransferase from Pseudomonas aeruginosa; Seitz T et al.; The bacterial insertion sequence IS21 when repeated in tandem efficiently promotes non-replicative cointegrate formation in Escherichia coli . An IS21-IS21 junction region which had been engineered to contain unique SalI and BglII sites close to the IS21 termini was not affected in the ability to form cointegrates with target plasmids . Based on this finding, a novel procedure of random linker insertion mutagenesis was devised . Suicide plasmids containing the engineered junction region (pME5 and pME6) formed cointegrates with target plasmids in an E.coli host strain expressing the IS21 transposition proteins in trans . Cointegrates were resolved in vitro by restriction with SalI or BglII and ligation; thus, insertions of four or 11 codons, respectively, were created in the target DNA, practically at random . The cloned Pseudomonas aeruginosa arcB gene encoding catabolic ornithine carbamoyltransferase was used as a target . Of 20 different four-codon insertions in arcB, 11 inactivated the enzyme . Among the remaining nine insertion mutants which retained enzyme activity, three enzyme variants had reduced affinity for the substrate ornithine and one had lost recognition of the allosteric activator AMP . The linker insertions obtained illustrate the usefulness of the method in the analysis of structure-function relationships of proteins. Jpn J Antibiot, 2000 Mar, 53(3), 171 - 8 {Evaluation of the activity and effects of combinations of various antibacterial agents against methicillin-resistant Staphylococcus aureus in vitro}; Kouda M et al.; MICs of various antibacterial agents against methicillin-resistant Staphylococcus aureus (MRSA) were measured . Furthermore, we evaluated the effects of combinations of antibacterial agents against MRSA in vitro . In 24 cases out of 37, in which MRSA was isolated from inpatients, other microorganisms, such as Candida spp., Entrococcus spp., and Pseudomonas aeruginosa, were simultaneously isolated . From the results of minimum inhibitory concentrations (MICs), obtained from micro broth-dilution method, of various antibacterial agents against MRSA, range of MICs of arbekacin (ABK), vancomycin (VCM) and teicoplanin (TEIC) were < or = 0.25-4.0, 0.5-1.0 and 0.25-4.0 micrograms/ml respectively, and no strains of MRSA showed resistance to ABK, VCM and TEIC, so that we concluded that these three antibacterial agents were effective for MRSA infection . On the in vitro study of combination-effect of antibacterial agents, significant synergistic effects were achieved in the combination of VCM and flomoxef (FMOX) (Synergism rate was 97.3%) or VCM and imipenem (IPM) (Synergism rate was 97.2%) . From the results that the fractional inhibitory concentration index in the combination of VCM with IPM was smaller than that with FMOX and that P . aeruginosa or Enterococcus spp . were simultaneously isolated in high frequency in the MRSA-isolated cases, we thought that the combination of VCM with IPM is more useful, because IPM is effective against P . aeruginosa but FMOX is not. Tidsskr Nor Laegeforen, 2000 Feb 20, 120(5), 568 - 71 {Cystic fibrosis in the Western health region}; Fluge G; 78 patients with cystic fibrosis have been diagnosed in western Norway (Health Region 3) since 1960 . 31 were born before 1971 (40%) . 44 patients were born in the county of Hordaland (62%), including the city of Bergen where the western Cystic Fibrosis Centre is located . 24 (31%) patients were born in Rogaland, and five (7%) in Sogn & Fjordane . 12 patients have died, 9 of them born before 1971 (survival in this cohort of patients is 71%) . During two decades from 1971 to 1990, 3 out of 33 patients have died (survival 91%) . All patients born after 1990 are alive . The median age at death was 18.5 years . Among patients alive in 1999, 32 (41.5%) are more than 20 years and 19 (24.7%) are more than 30 years old . Median age among all patients is 19.5 years (range 3-51) . Chronic lung infection with Pseudomonas aeruginosa has been diagnosed in 20.8% of all patients, which is lower than the average of Norwegian cystic fibrosis patients (35%) . Survival among cystic fibrosis patients in Western Norway is good compared to data from other countries . It seems realistic to expect that our patients should achieve a median survival to more than 30 years of age. Cornea, 2000 May, 19(3), 353 - 4 Antibacterial activity of anesthetic solutions and preservatives: an in vitro comparative study; Dantas PE et al.; PURPOSE: To investigate the antibacterial activity of topical anesthetic solutions and their preservatives individually in vitro, to determine involvement with bacterial growth inhibition . MATERIAL AND METHODS: Proparacaine and tetracaine (in concentrations of 0.125%, 0.25%, and 0.50%), edetate disodium (EDTA), benzalkonium chloride, EDTA + benzalkonium chloride, and sterile saline solution were used . Five microliters of each solution were applied to standard filter paper disks and placed in Mueller-Hinton agar previously inoculated with known strains of Pseudomonas aeruginosa and Staphylococcus aureus . Zones of growth inhibition were measured 24 hours later and analyzed . RESULTS: There were no zones of inhibition in the agar inoculated with P . aeruginosa to all tested solutions . Benzalkonium chloride alone and associated with EDTA inhibited growth of S . aureus . All other solutions did not inhibit S . aureus . CONCLUSION: Preservative-free anesthetic solutions seemed not to interfere with bacterial development in culture media . Benzalkonium chloride alone and associated with EDTA inhibited development of gram positive bacteria, S . aureus, but did not inhibit P . aeruginosa. Biochim Biophys Acta, 2000 May 31, 1485(2-3), 145 - 52 Mass spectrometry monitoring of rhamnolipids from a growing culture of Pseudomonas aeruginosa strain 57RP; Deziel E et al.; Two rapid and simple methods for the characterisation and quantification of rhamnolipids produced by a growing culture of the Pseudomonas aeruginosa strain 57RP were developed . Two rhamnolipids were purified and their response factors determined . The various rhamnolipids produced were then measured using liquid chromatography/mass spectrometry . The culture supernatants were injected directly, without prior purification, in a HPLC equipped with a C(18) reverse-phase column . The complete profile of rhamnolipid congeners produced during a 2 week cultivation period was monitored . In order to shorten the analysis time, another method was developed which did not require chromatographic separation of the rhamnolipids prior to their detection . Quantification of rhamnolipids using the direct infusion method gave results very similar to those obtained with HPLC separation . These two methods were very well correlated with the standard colorimetric orcinol method . The rhamnolipid profiles obtained show that the various rhamnolipid congeners are secreted simultaneously, and that their relative proportion remained unchanged throughout the cultivation period. Biochim Biophys Acta, 2000 May 31, 1485(2-3), 111 - 20 Reverse hydrolysis reaction of a recombinant alkaline ceramidase of Pseudomonas aeruginosa; Kita K et al.; Recently, we purified an alkaline ceramidase (CDase) of Pseudomonas aeruginosa and found that the enzyme catalyzed a reversible reaction in which the N-acyl linkage of ceramide was hydrolyzed or synthesized {J . Biol . Chem . 273 (1998) 14368-14373} . Here, we report the characterization of the reverse hydrolysis reaction of the CDase using a recombinant enzyme . The reverse hydrolysis reaction of the CDase was clearly distinguishable from the reaction of an acyl-coenzyme A (CoA) dependent N-acyltransferase, because the CDase catalyzed the condensation of a free fatty acid to sphingosine (Sph) without cofactors but did not catalyze the transfer of a fatty acid from acyl-CoA to Sph . The reverse hydrolysis reaction proceeded most efficiently in the presence of 0.05% Triton X-100 at neutral pH, while the hydrolysis reaction tended to be favored with an increase in the concentration of the detergent at alkaline pH . The specificity of the reverse reaction for fatty acids is quite broad; saturated and unsaturated fatty acids were efficiently condensed to Sph . In contrast, the stereo-specificity of the reverse reaction for the sphingoid bases is very strict; the D-erythro form of Sph, not the L-erythro or D/L-threo one, was only acceptable for the reverse reaction . Chemical modification of the enzyme protein affected or did not affect both the hydrolysis and reverse reactions to the same extent, suggesting that the two reactions are catalyzed at the same catalytic domain. FEMS Microbiol Lett, 2000 Jun 1, 187(1), 59 - 63 The wbpM gene in Pseudomonas aeruginosa serogroup O17 resides on a cryptic copy of the serogroup O11 O antigen gene locus; Dean CR et al.; The Pseudomonas aeruginosa serogroup O11 strain PA103 O antigen gene locus consists of 11 genes designated wzz, wzx, wbjA, wzy, wbjB-F, wbpL, and wbpM . The distribution of each of these genes amongst the 20 P . aeruginosa international antigenic typing system (IATS) serogroups was analyzed by Southern blot . As shown previously, wbpM was present in all 20 serogroups . The remaining O11 O antigen genes, with the exception of wzy, were present in the serogroup O17 strain IATSO17, despite the structural unrelatedness of the O11 and O17 O antigens . Sequencing revealed the presence of a cryptic serogroup O11 locus in the IATSO17 interrupted by two copies of a 1.1-kb insertion element . Introduction of plasmid pLPS2, containing the complete O11 O antigen locus from strain PA103, into IATSO17 resulted in production of both the O11 and O17 O antigens . The results of insertional inactivation of wbpM in IATSO17 are discussed. Biochem, Eng . J. . 2000 Jul 1, 5(3), 219 - 223 The synthetic rate of dipeptide catalyzed by organic solvent-stable protease from Pseudomonas aeruginosa PST-01 in the presence of water-soluble organic solvents; Ogino H et al.; The initial synthetic rates of peptide Cbz-Arg-Leu-NH(2) from Cbz-Arg and Leu-NH(2) using PST-01 protease in the presence and absence of organic solvents were investigated under various conditions . The synthetic rates of Cbz-Arg-Leu-NH(2) in the presence of 50% (v/v) methanol, 50% (v/v) N,N-dimethylformamide (DMF) and 60% (v/v) dimethyl sulfoxide (DMSO) were 1.6-, 2.4-, and 5.1-times higher than that in the absence of organic solvent, respectively . The PST-01 protease was not only stable in the presence of organic solvents but also exhibited high reaction rates in the presence of methanol, DMF, and DMSO . When the Cbz-Arg concentration was lower than 60mM or the Leu-NH(2) concentration was lower than 400mM, the initial rates increased lineally with increase in their concentrations . However, the rates did not increase when the Leu-NH(2) concentration was more than 500mM . The optimum temperature and pH of the reaction were 40 degrees C and 7.0, respectively. Biochem, Eng . J. . 2000 Jul 1, 5(3), 191 - 200 Cloning and sequencing of a gene of organic solvent-stable protease secreted from Pseudomonas aeruginosa PST-01 and its expression in Escherichia coli; Ogino H et al.; A gene of organic solvent-stable protease (PST-01 protease) secreted by Pseudomonas aeruginosa PST-01 was cloned and its nucleotide was sequenced . The nucleotide sequence analysis revealed that the PST-01 protease was a pseudolysin, which was an elastase produced by P . aeruginosa and was well characterized by the previous investigators . The PST-01 protease produced in recombinant Escherichia coli was not secreted into the extracellular medium, but its proenzyme was released by the lysis of the cells and became a 33.1kDa mature enzyme autoproteolytically . Its characteristics including organic solvent stability were as same as those of the PST-01 protease secreted by P . aeruginosa PST-01. J Wound Care, 1999 Nov, 8(10), 499 - 502 Infection control properties of some wound dressings; Bowler PG et al.; The ability of some wound dressings to sequester and retain micro-organisms associated with wound fluid is perceived to provide beneficial properties regarding infection control . This study used an in vitro model to investigate and compare such properties in a range of fibrous absorbent dressings (alginate, hydrofibre and hydrophobic) . Dressings were challenged with a simulated wound fluid containing common wound pathogens (Staphylococcus aureus or Pseudomonas aeruginosa) . Bacterial sequestering and binding levels were monitored over time . A hydrofibre dressing and two calcium alginate dressings were shown to effectively sequester challenge organisms from a simulated wound fluid . However, the hydrophobic and hydrofibre dressings produced statistically significant results in their ability to adsorb and retain challenge organisms (p < 0.05) . These investigations have demonstrated that a hydrofibre dressing effectively sequesters and retains micro-organisms upon exposure to simulated wound fluid, and may therefore provide a passive mechanism for reducing the microbial load in wounds and in the surrounding environment . Further in vivo studies are required to investigate these dressing properties. Scand J Infect Dis, 2000, 32(2), 197 - 9 Ceftazidime versus aztreonam in the treatment of pseudomonal chronic suppurative otitis media in children; Somekh E et al.; In a prospective, open, randomized trial, ceftazidime was given to 15 children and aztreonam to another 15 children with chronic suppurative otitis media without cholesteatoma (CSOM) . Patients enrolled to the study had a pure culture of Pseudomonas aeruginosa growing in middle ear discharge . The subjects' mean age was 56 months in the ceftazidime group and 48 months in the aztreonam group . Success rate, defined as complete disappearance of discharge, was 84.6% in the ceftazidime treated patients and 67% in the aztreonam group (p value not significant) . The number of days required for complete dryness of the ear was 7.9 and 8.4 respectively . Two patients in each group had recurrence of suppurative discharge within 90 d from discontinuation of antibiotic treatment . These results suggest that aztreonam is an optional alternative systemic treatment for paediatric patients with pseudomonal CSOM, especially in subjects allergic to penicillin. Acta Virol, 1999 Dec, 43(6), 395 - 8 Transduction of antibiotic resistance in Pseudomomas aeruginosa: relationship between lytic and transducing activity of phage isolate AP-423; Blahova J et al.; Isolation and propagation of a wild type phage, isolate AP-423, from an apparently lysogenic strain of Pseudomonas aeruginosa, resistant to a series of anti-pseudomonadal antibiotics, and its use for transduction of resistance determinants is described . The phage isolate AP-423 showed a phenomenon of host restriction, i.e . it was lysogenic only for some of the recipient strains tested . Its transduction capacity, both in sets of genes transduced and frequency of transduction, was different in two recipient strains of P . aeruginosa . This phage showed also some restriction in titers, to which it could be propagated, only in certain recipient strains. J Biomed Mater Res, 2000 Aug, 51(2), 224 - 32 Pooled human immunoglobulins reduce adhesion of Pseudomonas aeruginosa in a parallel plate flow chamber; Poelstra KA et al.; The influence of pooled polyclonal immunoglobulin (IgG) interactions with both bacteria and model substrates in altering Pseudomonas aeruginosa surface adhesion is reported . Opsonization of this pathogen by polyclonal human IgG and preadsorption of IgG to glass surfaces both effectively reduce initial deposition rates and surface growth of P . aeruginosa IFO3455 from dilute nutrient broth in a parallel plate flow chamber . Polyclonal IgG depleted of P . aeruginosa-specific antibodies reduces the initial deposition rate or surface growth to levels intermediate between exposed and nonexposed IgG conditions . Bacterial surface properties are changed in the presence of opsonizing IgG . Plateau contact angle analysis via sessile drop technique shows a drop in P . aeruginosa surface hydrophobicity after IgG exposure consistent with a more hydrophilic IgG surface coat . Zeta potential values for opsonized versus nonopsonized bacteria exhibit little change . X-ray photoelectron spectroscopy measurements provide surface compositional evidence for IgG attachment to bacterial surfaces . Surface elemental ratios attributed to IgG protein signals versus those attributed primarily to bacterial polysaccharide surface or lipid membrane change with IgG opsonization . Direct evidence for antibody-modified P . aeruginosa surface properties correlates both with reduction of bacterial adhesion to glass surfaces under flow in nutrient medium reported and previous reports of IgG efficacy against P . aeruginosa motility in vitro and infection in vivo . Acta Crystallogr D Biol Crystallogr, 2000 Jun, 56 ( Pt 6), 761 - 2 Crystallization and initial crystallographic analysis of phosphomannomutase/phosphoglucomutase from Pseudomonas aeruginosa; Regni CA et al.; The enzyme phosphomannomutase/phosphoglucomutase (PMM/PGM) catalyzes the conversion of mannose 6-phosphate to mannose 1-phosphate in the second step of the alginate biosynthetic pathway of Pseudomonas aeruginosa . PMM/PGM has been crystallized by hanging-drop vapor diffusion in space group P2(1)2(1)2(1) . Crystals diffract to 1.75 A resolution on a synchrotron X-ray source under cryo-cooling conditions . PMM/PGM substituted with selenomethionine has been purified and crystallizes isomorphously with the native enzyme . Structure determination by MAD phasing is under way . Because of its role in alginate biosynthesis, PMM/PGM is a potential target for therapeutic inhibitors to combat P . aeruginosa infections. Science, 2000 May 19, 288(5469), 1251 - 4 High frequency of hypermutable Pseudomonas aeruginosa in cystic fibrosis lung infection; Oliver A et al.; The lungs of cystic fibrosis (CF) patients are chronically infected for years by one or a few lineages of Pseudomonas aeruginosa . These bacterial populations adapt to the highly compartmentalized and anatomically deteriorating lung environment of CF patients, as well as to the challenges of the immune defenses and antibiotic therapy . These selective conditions are precisely those that recent theoretical studies predict for the evolution of mechanisms that augment the rate of variation . Determination of spontaneous mutation rates in 128 P . aeruginosa isolates from 30 CF patients revealed that 36% of the patients were colonized by a hypermutable (mutator) strain that persisted for years in most patients . Mutator strains were not found in 75 non-CF patients acutely infected with P . aeruginosa . This investigation also reveals a link between high mutation rates in vivo and the evolution of antibiotic resistance. Exp Lung Res, 2000 Apr-May, 26(3), 163 - 78 The effect of alginate on the invasion of cystic fibrosis respiratory epithelial cells by clinical isolates of Pseudomonas aeruginosa; Massengale AR et al.; Chronic infection in the cystic fibrosis (CF) lung is characterized by Pseudomonas aeruginosa strains that overproduce the mucoid exopolysaccharide, alginate . Previous experiments have shown that long-term survival of P . aeruginosa in the CF lung may be facilitated by increased adherence and decreased invasion of respiratory epithelial cells . Therefore, mucoid and nonmucoid clinical isolates of P . aeruginosa were assayed for their ability to associate with and invade the CF respiratory epithelial cell line, CF/T43 . Association assays and gentamicin exclusion assays demonstrated that mucoid P . aeruginosa associates with and invades CF/T43 cell monolayers significantly less than nonmucoid P . aeruginosa strains (P = .004, .02) . Fluorescence microscopy invasion assays confirmed these results . The differences in association and invasion by the P . aeruginosa strains were not due to differences in lipopolysaccharide phenotype or cytotoxicity for CF/T43 respiratory epithelial cells . Exogenous bacterial alginate had no effect on the invasion of CF respiratory epithelia by a nonmucoid strain . Invasion assays with the wild-type P . aeruginosa strain PAO1 and isogenic algU and mucA mutant strains failed to show differences in invasion (P = .25) . We conclude that (i) mucoid P . aeruginosa isolates associate with and invade CF/T43 respiratory epithelial cells with less efficiency than nonmucoid P . aeruginosa, (ii) these differences are not due to variations in lipopolysaccharide phenotype between strains, (iii) neither exogenous nor endogenous alginate affects the ability of P . aeruginosa to invade CF/T43 respiratory epithelial cells, and (iv) invasion of CF/T43 respiratory epithelial cells by a laboratory reference strain of P . aeruginosa does not appear to be regulated by AlgU. Biomaterials, 2000 Jun, 21(12), 1235 - 46 Synthesis and characterization of a novel biodegradable antimicrobial polymer; Woo GL et al.; Bacterial infection is a frequent complication associated with the use of medical devices . In an effort to address this problem, antibacterial agents have been incorporated or applied directly onto the surfaces of numerous types of medical devices . This study assessed the feasibility of using a novel biodegradable polymer to release antibiotic drugs in response to inflammatory related enzymes . A model drug polymer was synthesized using 1,6-hexane diisocyanate (HDI), polycaprolactone diol (PCL), and a fluoroquinolone antibiotic, ciprofloxacin . Polymers were characterized by size-exclusion chromatography (SEC), and elemental analysis . Biodegradation studies were carried out by incubating the polymers with solutions of cholesterol esterase (CE) or phosphate buffer (pH 7.0) for 30 days at 37 degrees C . The degradation was assessed by high-performance liquid chromatography (HPLC), mass spectrometry (MS) and 14C radiolabel release . Subsequently, the activity of the released antibiotic was assessed against a clinical isolate of Pseudomonas aeruginosa . HPLC analysis showed the release of multiple degradation products which were identified, by tandem MS, to include ciprofloxacin and derivatives of ciprofloxacin . The microbiological assessment showed that the released ciprofloxacin possessed antimicrobial activity; 1 microg/ml was measured after 10 days . The results of this study suggest that these novel bioresponsive antimicrobial polymers or similar analogs show promise for use in the control of medical device associated infections. Int J Dermatol, 2000 Apr, 39(4), 270 - 3 Pseudomonas aeruginosa folliculitis after shower/bath exposure; Zichichi L et al.; BACKGROUND: Pseudomonas aeruginosa folliculitis (PF) can develop after exposure to contaminated water in heated swimming pools, whirlpools, and hot-tubes, or after diving suit dressing . METHODS: We observed and studied 14 cases of PF after shower/bath exposure, an underestimated pathogenic event . Cutaneous and environmental microbiological evaluations were performed . RESULTS: In our cases, the clinical expression of dermatitis was constant, PF being a clinically well recognizable skin infection, presenting with follicular, macular, and papulopustular lesions located on the lateral aspect of the trunk, axillary folds, hips, buttocks, and suprapubic area . In all cases, Pseudomonas aeruginosa was isolated from lesional skin; seven cases were serotyped revealing, in three cases, serotype 0 : 1, in two cases 0 : 8, in one case 0 : 10, and in one case 0 : 11 . In three families, Pseudomonas aeruginosa was isolated in the well water . In a further three families, Pseudomonas aeruginosa was isolated from bathroom and kitchen components . CONCLUSIONS: Based on our experience, we suggest that shower/bath exposure should be definitively included amongst the possible pathogenic events causing PF . Pseudomonas aeruginosa is responsible for a number of clinical pictures, e.g . otitis externa, conjunctivitis, toe web intertrigo, green nail syndrome, infection of burns and wounds, and folliculitis . Pseudomonas aeruginosa folliculitis (PF) has been reported to develop as a consequence of exposure to contaminated water in heated swimming pools, whirlpools, and hot-tubes, or related to diving suits and leg waxing.1-4 We observed 14 cases of PF after shower/bath exposure . This is probably an underestimated pathogenic event; to our knowledge, only one case has been reported to date.5 In our patients, the clinical expression of dermatitis was constant, PF being a clinically well recognizable skin condition. J Bacteriol, 2000 Jun, 182(11), 3142 - 50 Interplay between efflux pumps may provide either additive or multiplicative effects on drug resistance; Lee A et al.; The effects of simultaneous expression of several efflux pumps on antibiotic resistance were investigated in Escherichia coli and Pseudomonas aeruginosa . Several combinations of efflux pumps have been studied: (i) simultaneous expression of a single-component efflux pump, which exports antibiotics into the periplasm, in combination with a multicomponent efflux pump that accomplishes efflux directly into the external medium; (ii) simultaneous expression of two single-component pumps; and (iii) simultaneous expression of two multicomponent pumps . It was found that when efflux pumps of different structural types were combined in the same cell (the first case), the observed antibiotic resistance was much higher than that conferred by each of the pumps expressed singly . Simultaneous expression of pairs of single-component or multicomponent efflux pumps (the second and third cases) did not produce strong increases in antibiotic resistance. Perit Dial Int, 2000 Mar-Apr, 20(2), 227 - 31 Pharmacokinetics of intraperitoneal piperacillin/tazobactam in patients on peritoneal dialysis with and without pseudomonas peritonitis; Zaidenstein R et al.; OBJECTIVE: The objective of this study was to assess the pharmacokinetics of intraperitoneal (IP) administration of the antibiotic combination piperacillin/tazobactam (PIP/TAZ) to patients on chronic ambulatory peritoneal dialysis (CAPD) with and without pseudomonas peritonitis . DESIGN: Open-labeled study . SETTING: The study was carried out in the CAPD unit of Assaf Harofeh Medical Center, Zerifin, Israel . PATIENTS AND METHODS: Six patients participated in the study, 4 had pseudomonas peritonitis, all were given an IP loading dose of 4 g/0.5 g PIP/TAZ . Twenty-four hours after the initial dose, a maintenance dose of 0.5 g/0.0625 g PIP/TAZ was administered with each dialysate exchange for a period of 1 week . The patients without peritonitis received only the loading dose . High performance liquid chromatography was used to determine the concentrations of PIPITAZ in plasma obtained at 0, 30, 60, 90, 120, 360, 480, 600, 720, and 1440 minutes after administration . Samples of the dialysate fluid for determination of PIP/TAZ concentration were collected at 6,10,14, 24, and 72, 120, and 168 hours . RESULTS: After the loading dose, the highest plasma PIP concentration (Cmax) was 51.6 t 21.25 Lig/mL and appeared at 1.5 = 0.45 hours (t,,a) . During the maintenance period plasma PIP concentration was 5.2 t 4.75 Lg/mL . Tazobactam was detected in the plasma of 1 patient only . The concentration of TAZ in the dialysate fluid during the maintenance period was 2.3 t 0.5 ig/mL . CONCLUSIONS: Piperacillin administered IP at 4 g reached plasma concentrations comparable to intravenous administration and considered therapeutic (above the MIC90 for Pseudomonas aeruginosa) in CAPD patients with or without peritonitis.The maintenance dose, however, should be augmented . Tazobactam could not be detected in the plasma of most patients and the therapeutic implications of IP administration of TAZ cannot be directly correlated to intravenous administration. Perit Dial Int, 2000 Mar-Apr, 20(2), 209 - 14 Exit-site care with ciprofloxacin otologic solution prevents polyurethane catheter infection in peritoneal dialysis patients; Montenegro J et al.; OBJECTIVE: Mupirocin ointment and antiseptics are standard cleansing agents in routine exit-site care of peritoneal dialysis (PD) catheters, but these agents have a deleterious effect on polyurethane devices . We assessed the effectiveness of topical use of ciprofloxacin otologic solution for preventing exit-site infection (ESI) in PD patients with polyurethane catheters . DESIGN: Prospective study . SETTING: Service of Nephrology of an acute-care teaching hospital in Galdacano, Bizkaia, Spain . PATIENTS: A total of 164 patients with polyurethane catheters inserted was studied from start of continuous ambulatory PD to the end of a 24-month period . Patients were divided into two groups according to exit-site treatment protocols . INTERVENTION: Patients in group 1 (n = 86) were instructed on daily exit-site care with soap and water only; whereas patients in group 2 (n = 78) cleansed with soap and water, followed by application of a single-dose vial of 0.5 mL ciprofloxacin (1 mg) for application around the insertion site . MAIN OUTCOME MEASURES: Episodes of ESI and peritonitis . RESULTS: There were 67 episodes of ESI among patients in group 1 versus 9 episodes among patients in group 2 (p < 0.05), resulting in a rate of 0.41 and 0.06 episodes per patient-year of exposure, respectively (p < 0.001) . Staphylococcus aureus ESI rate was 0.34 in group 1 versus 0.06 in group 2 (p = 0.001) . Infections caused by Pseudomonas aeruginosa and other pathogens occurred in 11 patients in group 1 and in no patients in group 2 (p = 0.05) . Peritonitis due to S . aureus ESI was significantly less frequent among patients treated with ciprofloxacin (1 vs 9 cases, p = 0.001) . Removal of the catheter was necessary in 5 patients in group 1 and in no patients in group 2 (p < 0.05) . CONCLUSION: Daily application of ciprofloxacin otologic solution at the exit site of PD patients with polyurethane catheters inserted significantly reduces the rate of ESI caused by S . aureus and other organisms, particularly P . aeruginosa. Rev Panam Salud Publica, 2000 Mar, 7(3), 179 - 84 A bacteriological study of hospital beds before and after disinfection with phenolic disinfectant; de Andrade D et al.; In hospitals, one of the ways to control microbial contamination is by disinfecting the furniture used by patients . This study's main objective was to evaluate the microbiological condition of hospital mattresses before and after such disinfection, in order to identify bacteria that are epidemiologically important in nosocomial infection, such as Staphylococcus aureus and Pseudomonas aeruginosa . RODAC plates with two different culture media were used to collect specimens . Patient beds were selected according to previously established criteria, and surface areas on the mattresses were chosen at random . From the total of 1,040 plate cultures from 52 mattresses, positive results were obtained from 500 of them (48.1%), 263 before disinfection and 237 after disinfection . Considering the selectivity of the culture media, the positivity rate was high . There were high prevalences of S . aureus both before and after mattress disinfection . The study results suggest that the usual disinfection procedures, instead of diminishing the number of microbes, merely displace them from one part of the mattress to another, and the number of microorganisms remains the same. J Tongji Med Univ, 1998, 18(3), 172 - 6 The bacterial inhibitory ability and in vivo drug release pattern of a new drug delivery system: ciprofloxacine/tricalcium phosphate delivery capsule; Zheng Q et al.; The bacterial inhibitory ability of a new drug delivery system (DDS): Ciprofloxacine/tricalcium phosphate delivery capsule (CTDC), its in vivo drug release pattern, and the influence of ultrasonic irradiation on its drug release were investigated . It was found that CTDC had a strong and sustained inhibitory ability to some common pathogens of bone and joint infections, such as staphylococcus aureus, escherichia coli and pseudomonas aeruginosa . In vivo drug-release study in animals demonstrated a high concentration of ciprofloxacine in the bone tissue surrounding CTDC which was placed in the greater trochanter of the rabbit and continued to release ciprofloxacine for at least 5 weeks and the blood level of ciprofloxacine was low . In vivo study also showed ultrasonic irradiation could increase the amount of ciprofloxacine released from CTDC, which may be an economical, effecient and safe new method to achieve the control of drug release from DDS. Curr Eye Res, 2000 Apr, 20(4), 260 - 7 Perlecan in the basement membrane of corneal epithelium serves as a site for P . aeruginosa binding; Chen LD et al.; PURPOSE: To determine whether binding of Pseudomonas aeruginosa (P . aeruginosa) to the scarified mouse cornea depends on interaction with proteoglycans (PGs) . METHODS: Scarified corneas were treated with anti-proteoglycan monoclonal antibodies (MAbs), glycosaminoglycans (GAGs), heparinase III or chondroitin ABC lyase before inoculation with P . aeruginosa strain ATCC 19660 or PAO1 . Scanning electron microscopy (SEM) was used to quantitate adherent bacteria . Frozen sections of unwounded and wounded mouse cornea, the latter treated or not treated with heparinase III were stained to spatially localize perlecan {core protein or heparan sulfate (HS) side chains} . Anti-perlecan MAb against the heparan sulfate proteoglycan core protein and succinyl wheat germ agglutinin (sWGA), a lectin which recognizes N-acetyl-glucosamine in heparan sulfate, respectively were used . RESULTS: Anti-perlecan MAb, as well as heparan sulfate, heparin and heparinase III decreased the binding of both bacterial strains to cornea, and the decrease was concentration-dependent . Fluorescence microscopic analysis of sections of mouse cornea immunostained with anti-perlecan MAb showed that perlecan was localized to the epithelial basement membrane . Scarification of the mouse cornea exposed perlecan in the basement membrane and increased bacterial binding to this site was consistent with this exposure . Lectin staining revealed that heparinase treatment removed heparan sulfate side chains of perlecan from the exposed basement membrane, and this removal was consistent with a decrease in bacterial binding . CONCLUSIONS: These studies provide evidence that perlecan, core protein and its heparan sulfate side chains serve as a binding site for Pseudomonas aeruginosa when the basement membrane of the cornea is exposed. Cytokine, 2000 Apr, 12(4), 417 - 21 Dual effect of IL-4 on resistance to systemic gram-negative infection and production of TNF-alpha; Giampietri A et al.; To determine the effect of interleukin 4 (IL-4) administration in a live sepsis model characterised by high-level production of tumour necrosis factor a (TNF-alpha), mice infected systemically with lethal or sublethal inocula of Pseudomonas aeruginosa were given the recombinant cytokine at different times before infection . Improved survival and decreased TNF-alpha production were observed in lethally infected mice treated with the cytokine 1 day before challenge . In contrast, increased mortality and overproduction of TNF-alpha were observed in sublethally infected mice given IL-4 at the time of infection. Rinsho Byori, 2000 Feb, 48(2), 167 - 73 {Antimicrobial susceptibility testing for Mycobacterium tuberculosis by the bioluminescence assay of mycobacterial ATP using filamentous cell treatment}; Yamazaki T et al.; The antimicrobial susceptibility testing for Mycobacterium tuberculosis by the bioluminescence assay of adenosine triphosphate(ATP) derived from living mycobacteria was improved introducing filamentous cell treatment(FCT) reported for beta-lactam susceptibility test of Pseudomonas aeruginosa by Hattori . Before ATP extraction, bacterial cells were treated with the FCT reagent for 30 minutes at room temperature . Adenosine phosphate deaminase in the FCT reagent simultaneously digested the extracted ATP and released ATP in a liquid culture of M . tuberculosis H37Rv and the RLU level was decreased markedly . Using this improved ATP method, we determined the ATP contents of M . tuberculosis inoculated into Middle-brook 7H9 broth medium with or without drugs . In ethambutol(EB) susceptibility, the ATP method reported previously, showed false-resistance when judged within 7 days . To eliminate false-resistance in EB susceptibility we applied the modified ATP method with FCT treatment to strains determined EB susceptible by reference methods . Using this modified ATP method, we could judge EB susceptibility of 5 ATCC reference strains within 3 days, and these of 15 clinical isolates of M . tuberculosis within 5 days . And all the results obtained were coincident between the ATP method and the reference methods . The reproducibility of this modified ATP method was evaluated with six ATCC reference strains at the concentrations of 0.1 microgram/ml of isoniazid(INH), 2.0 micrograms/ml of rifampicin(RFP), 2.5 micrograms/ml of EB, 2.0 micrograms/ml of streptomycin(SM), and 5.0 micrograms/ml of kanamycin(KM) . The test was repeated six times . Reduction of ATP contents were observed in susceptible strains but not in resistant ones within 3 days of cultivation and susceptibilities to drugs could be determined within 3 days at every time when combined FCT to the ATP method . And highly reproducible results were obtained . It is strongly suggested that this modified method is simple, rapid, highly reproducible and nonradiometric, and could be used for the assessment of drug susceptibility for M . tuberculosis. Rinsho Byori, 2000 Jan, Suppl 111, 100 - 8 {Gene examination methods (detection and genotyping of resistant genes)--multiple-drug-resistant Pseudomonas aeruginosa}; Arakawa Y; Pseudomonas aeruginosa has been regarded as one of the most stubborn pathogens that easily acquire resistance to various antimicrobial agents . Recently, several clinical isolates that have acquired multiple-resistance to carbapenems, fluoroquinolones, and newly developed aminoglycosides such as amikacin have been reported . Thus, the "drug-resistant P . aeruginosa" was designated as a "class 4 pathogen" in the new "Law Concerning the Prevention of Infectious Diseases and Medical Care for Patients with Infections" in Japan . It is generally considered that no medical facility can escape from severe attack by both multiple-drug resistant gram-positive cocci such as MRSA and gram-negative rods including P . aeruginosa in the 21st century . Therefore, emergency countermeasures should be anticipated to prevent further proliferation of these multiple-drug resistant pathogens. Med Dosw Mikrobiol, 1999, 51(3-4), 339 - 45 {Bactericidal activity of human, swine and cattle serum against pseudomonas aeruginosa strains}; Wolska K et al.; The aim of this study was to assess of bactericidal activity of human, swine and cattle serum against 136 of Pseudomonas aeruginosa strains isolated from people, fishes, domestic and fur animals . The mechanism of the bactericidal activity of serum against gram-negative bacteria is complex and involves the participation of complement, antibodies and lysozyme (1, 5, 7, 8, 10, 11, 13, 14, 24, 25, 27, 30) . The susceptibility of gram-negative rods to serum is differentiated . Pseudomonas aeruginosa strains are the most resistant (17, 25, 30) . This opportunistic pathogen produce proteases that destroy complement components and immunoglobulins (3, 18, 19) . The bactericidal activity of serum was determined after 3 hours incubation of bacteria in 50% serum by the method of Jankowski (1981) (5) . The results of this study indicate that 71% of this strains were resistant to swine serum action, 68% of this strains were resistant to bovine serum and 57% of the Pseudomonas aeruginosa strains were sensitive to human serum . The P . aeruginosa strains isolated from fishes were the most sensitive to serum action and the strains isolated from people and cattle were most resistant to the bactericidal activity of serum. Am J Surg, 2000 Feb, 179(2A Suppl), 18S - 23S; discussion 24S-25S Empiric therapy for pneumonia in the surgical intensive care unit; Fabian TC; Empiri c therapy of ventilator-associated pneumonia (VAP) in surgical patients should be based on intensive care unit (ICU)-specific surveillance data, because microbial flora patterns vary widely between geographic regions as well as within hospitals . Surgical ICUs have higher VAP rates than other units . Data from the National Nosocomial Infection Surveillance (NNIS) System report Pseudomonas aeruginosa and Staphylococcus aureus to be the most frequent isolates (each 17.4%) . Data from the NNIS documents high resistance patterns in ICUs compared with hospitals at large, as well as unit-specific patterns . VAP risk factors for surgical patients include thoracoabdominal surgery, altered level of consciousness, advanced age, diabetes mellitus, malnutrition, chronic obstructive pulmonary disease, and prior antibiotic administration . Promising prevention strategies include restricting ventilator circuit changes, in-line heat moisture exchange filters, semi-recumbant positioning, and continuous subglottic aspiration . Pharmacodynamics should be considered when choosing antibiotic regimens . Postantibiotic effect and time-dependent versus concentration-dependent killing should be studied in clinical trials . Current guidelines for choosing regimens have been well developed by the American Thoracic Society. Clin Pharmacol Ther, 2000 Apr, 67(4), 368 - 72 Pharmacokinetics of cefpirome during continuous venovenous hemofiltration: rationale for an 8-hour dosing interval; Banyai M et al.; OBJECTIVE: Cefpirome is a new semisynthetic cephalosporin, primarily eliminated by the kidneys, that requires dosage adjustment in patients with kidney failure . The optimal dosing regimen of cefpirome in patients with continuous veno-venous hemofiltration (CVVH) is unknown . METHODS: Pharmacokinetic properties of cefpirome were investigated in eight anuric patients with acute kidney failure treated by CVVH . All patients received a dosage of 2 g cefpirome every 8 hours after starting the hemofiltration with high-flux polysulfone membranes . Concentrations of cefpirome in plasma and ultrafiltrate were measured by HPLC . RESULTS: Total clearance and hemofiltration clearance of cefpirome were 589.1 +/- 164.5 mL/min and 43.3 +/- 7.8 mL/min, respectively . Serum elimination half-life was 2.36 +/- 0.59 hours . The highest plasma drug concentration was 14.8 +/- 3.2 microg/mL, and it declined to trough levels of 3.1 +/- 0.8 microg/mL at the end of the dosing interval . CONCLUSION: On the basis of previously published pharmacodynamic characteristics of cefpirome and the pharmacokinetic parameters obtained in this study, we calculated a required total daily dose of 2 g every 8 hours to achieve sufficient plasma antibiotic levels to cover the majority of target pathogens . However, this dosage may be insufficient during CVVH for intermediate resistant strains of Pseudomonas aeruginosa. Mem Inst Oswaldo Cruz, 2000 May-Jun, 95(3), 367 - 73 Biological screening of Brazilian medicinal plants; Alves TM et al.; In this study, we screened sixty medicinal plant species from the Brazilian savanna ("cerrado") that could contain useful compounds for the control of tropical diseases . The plant selection was based on existing ethnobotanic information and interviews with local healers . Plant extracts were screened for: (a) molluscicidal activity against Biomphalaria glabrata, (b) toxicity to brine shrimp (Artemia salina L.), (c) antifungal activity in the bioautographic assay with Cladosporium sphaerospermum and (d) antibacterial activity in the agar diffusion assay against Staphylococcus aureus, Escherichia coli, Bacillus cereus and Pseudomonas aeruginosa . Forty-two species afforded extracts that showed some degree of activity in one or more of these bioassays. J Antimicrob Chemother, 2000 May, 45(5), 639 - 43 Time-kill studies of tea tree oils on c