Am J Infect Control, 1997 Dec, 25(6), 458 - 62 Nosocomial infections in an oncology intensive care unit; Velasco E et al.; INTRODUCTION: Treatment of cancer has contributed to a growing number of immunocompromised patients with life-threatening nosocomial infections (NI) . High mortality with considerable cost is observed when they are admitted to the intensive care unit (ICU) . Few studies on infection control and surveillance have been undertaken in this population group . METHODS: All patients treated at a six-bed medical-surgical oncology ICU for > 48 hours were prospectively observed for the development of an NI and the influence of device utilization on infection rates . The analysis used the standard definitions of the National Nosocomial Infection Surveillance System Intensive Care Unit surveillance component . RESULTS: From September 1993 through November 1995, 370 infections occurred in 623 patients during 4034 patient-days, for an overall rate of 50.0 per 100 patients or 91.7 per 1000 patient-days . Pneumonia (28.9%), urinary tract infections (25.6%), and bloodstream infections (24.1%) were the main types of infection . The most common microorganisms isolated were Enterobacteriaceae (29.7%), fungi (22.2%), and Pseudomonas aeruginosa (13.2%) . The median device utilization ratios were 0.63, 0.83, and 0.86 for ventilator, indwelling urinary catheter, and central venous catheter, respectively . The highest median device-specific associated infection rate was 41.7 for ventilator . The median for the average length of stay was 8.8 days, and the average severity of illness score was 4.0 . There was a strong positive correlation between the overall NI patient rate and device utilization (r = 0.56, p < 0.01), average severity of illness score (r = 0.54, p < 0.01), and average length of stay (r = 0.67, p < 0.01) . No correlations were statistically significant when patient-days were used in the denominator . Among the devices only the number of central venous catheter days was significantly correlated with infections (r = 0.51, p = 0.01) . The NI patient-day rates were progressively higher the longer the patients stayed in the ICU . CONCLUSIONS: The high rates reported in this study may reflect a combination of several factors related to the underlying illness, neutrophil count, and exposure to invasive procedures . The adjusted infection rates described here provide specific surveillance data for further interhospital comparisons and also to assess the influence of invasive medical interventions, allowing the implementation of preventable measures to control infections. Semin Respir Infect, 1997 Dec, 12(4), 294 - 9 Nonantibiotic measures in the prevention of ventilator-associated pneumonia; Stoutenbeek CP et al.; Aspiration of oropharyngeal and/or gastrointestinal (GI) contents is the main cause of ventilator-associated pneumonia . A number of nonantibiotic measures have been proposed to prevent aspiration eg, drainage of subglottic secretions or the semirecumbent position or to prevent gastric microbial overgrowth by stress-ulcer prophylaxis with sucralfate or early enteral feeding . Critical review of the studies shows that subglottic drainage does not prevent colonization or infection of the respiratory tract with intensive care unit-acquired Enterobacteriaceae or Pseudomonas aeruginosa . The effect of subglottic drainage on primary endogenous infections caused by Staphalococcus aureus and Streptococcus spp in patients not receiving antibiotics is only found in a post-hoc subgroup analysis and might reflect differences in carriage of community-acquired potentially pathogenic microorganisms (PPM) caused by previous antibiotic treatment, rather than a true treatment effect . The semirecumbent position may reduce the incidence of aspiration, particularly in patients without a nasogastric tube, but the aspiration rate remains high even in the short observation periods of the studies . There is no evidence that it reduces the ventilator-associated pneumonia rate . Sucralfate may reduce the increased pneumonia rate caused by H2-antagonists and/or antacids, but it remains to be proven whether it is superior to placebo . Sucralfate has no effect on the oropulmonary route of infection and has therefore no effect on early-onset (primary endogenous) pneumonia, which is characteristically caused by PPM carried in the oropharynx . Early enteral feeding is preferable to total parenteral feeding . However, there is limited evidence that it prevents ventilator-associated pneumonia . The studies showing a benefit of early enteral feeding were relatively small studies, partly in nonventilated patients, and used poorly defined criteria for pneumonia . The oropulmonary route is the most important route in the pathogenesis of pneumonia . Preventive strategies (both antibiotic and nonantibiotic strategies) have to block both the oropulmonary route and the gastropulmonary route to be fully effective . Because microaspiration cannot be fully prevented in critically ill patients, preventive strategies should attempt to eliminate PPM from the oropharynx and GI-tract. Scand J Infect Dis Suppl, 1997, 105, 13 - 23 Antimicrobial susceptibility testing in Sweden . III . Methodology for susceptibility testing; Olsson-Liljequist B et al.; A subcommittee of the Swedish Reference Group for Antibiotics, SRGA-M, has worked with standardization of methodology for susceptibility testing . In vitro data obtained with the disk diffusion procedure were collected from 5 clinical laboratories, compiled and presented as histograms of inhibition zones, and compared with data {minimum inhibitory concentrations (MICs) and inhibition zones} obtained from the reference laboratory at the Swedish Institute for Infectious Disease Control on a collection of clinically relevant bacterial species . Results from the reference collection of strains were presented as MIC histograms, and their corresponding inhibition zones were inserted in the compiled zone histograms as identifiable bars . These distributions formed the basis for decisions of breakpoints . Special tests were recommended for the detection of certain resistance mechanisms . A beta-lactamase test should be used for Haemophilus influenzae, Moraxella catarrhalis, Neisseria gonorrhoeae and enterococci . Screening for beta-lactam resistance caused by altered penicillin binding proteins should be done by using oxacillin 1 microgram for Streptococcus pneumoniae and Staphylococcus aureus (MRSA), and by phenoxymethylpenicillin 10 micrograms for H, influenzae . The standardized disk diffusion procedure was helpful in detecting enterobacteria carrying beta-lactamases with extended spectra . Registration of inhibition zones will provide a powerful tool for the epidemiological surveillance of antibiotic resistance. Plasmid, 1997, 38(3), 210 - 9 Isolation and characterization of a ColE1-like plasmid from Enterobacter agglomerans with a novel variant of rom gene; Mikiewicz D et al.; Complete nucleotide sequence of a plasmid isolated from Enterobacter agglomerans has been determined . The plasmid, called pPIGDM1, consists of 2495 base pairs . The analysis of its nucleotide sequence suggested that pPIGDM1 may be a ColE1-like replicon . We confirmed this hypothesis by constructing a pPIGDM1-derived plasmid harboring the cat gene (pBW4), which could be introduced into Escherichia coli cells, and demonstrating that pBW4 cannot replicate in the absence of the polA function and that its copy number is significantly decreased in the pcnB mutant . Like some other ColE1-type replicons (e.g., pBR322), pPIGDM1-derived plasmids can be amplified both by chloramphenicol method and in isoleucine-starved relA mutants but not in relA+ bacteria . Inactivation of the putative rom gene by insertion of an amplicillin-resistance gene resulted in significant increase in pPIGDM1-derived plasmid copy number in E . coli-despite the fact that amino acid sequence of the putative RNA 1 modulator (Rom) protein is only 55.7% identical to the ColE1 analog . The pPIGDM1-derived rom-like coding sequence is also homologous to the rom-like gene present in the Proteus vulgaris plasmid pPvul . We suggest to group all these gene products into a new family called ROMS (RNA one modulators) . Since a pPIGDM1-derived plasmid is compatible with other ColE1-like replicons (pMB1-, p15A, RSF1030-, and CloDF13-derived) in E . coli, one may consider pPIGDM1 as a progenitor of new cloning vehicles compatible with most (if not all) of currently used plasmid vectors . Moreover, this plasmid may serve as a source of the new rom-like gene coding for a protein useful in investigation of RNA-protein interactions . A role for the pPIGDM1 plasmid in the host strain is not known. Crit Care Med, 1998 Jan, 26(1), 31 - 9 Aerosolized antibiotics in mechanically ventilated patients: delivery and response; Palmer LB et al.; OBJECTIVES: To determine whether aerosolized antibiotics can be delivered efficiently to the lower respiratory tract in mechanically ventilated patients and to define possible clinical responses to these agents . DESIGN: Prospective serial study with cases as their own control . SETTING: A 10-bed respiratory care unit for patients with chronic respiratory failure in a tertiary university hospital . PATIENTS: Ventilator dependent patients who are otherwise medically stable . All subjects had a tracheostomy in place, were colonized with gram-negative organisms, and produced purulent secretions which could be sampled daily . INTERVENTIONS: Six patients received nine courses of nebulized therapy, which consisted of treatments every 8 hrs of gentamicin (80 mg) or amikacin (400 mg) for 14 to 21 days . MEASUREMENTS AND MAIN RESULTS: Doses to the lung were measured using radiolabeled aerosols and antibiotic concentrations in sputum . The response was assessed by a) changes in the volume of respiratory secretions; b) effect on bacterial cultures; and c) changes in the inflammatory cells and mediators of inflammation of the respiratory secretions (interleukin-1beta {IL-1beta}, tumor necrosis factor-alpha {TNF-alpha}, soluble intercellular adhesion molecule-1 {sICAM-1}, and human leukocyte elastase) . On average, patients inhaled 35.4 +/- 5.08% (SD) of the initial drug placed in the nebulizer (neb-charge) . Of this neb-charge, 9.50 +/- 2.78% was found on the respirator tubing and tracheostomy tube and 21.9 +/- 7.15% was actually deposited in the lungs . The remainder of the neb-charge was sequestered in the nebulizer or exhaled . Trough sputum concentrations averaged 4.3 +/- 3.2 microg/mL/mg neb-charge (range 234 to 520 microg/mL) and increased to 16.6 +/- 8.1 microg/mL/mg neb-charge (range 1005 to 5839 microg/mL) immediately after therapy (p = .011) . Serum concentrations were undetectable in most determinations except for a single patient who was in renal failure (8.7 microg/mL amikacin) . Treatment caused a significant reduction in the volume of secretions (p = .002) . Weekly cultures revealed eradication of Pseudomonas species, Serratia marcescens, and Enterobacter aerogenes in most of the trials . Before antibiotic treatment, concentrations of IL-1beta were higher than those reported in acute respiratory distress syndrome . Throughout the duration of the study, IL-1beta correlated significantly with the absolute number of macrophages, neutrophils, and lymphocytes, respectively (r2 = .55, p = .002; r2 = .50, p < .0004, r2 = .36, p = .005) . TNF-alpha concentrations correlated with lymphocytes and neutrophils, respectively (r2 =.27, p = .013, r2 = .21, p = .033) . sICAM-1 concentrations increased two-fold (p < .001) during treatment and then returned to baseline . The volume of secretions was related to neutrophil and IL-1beta concentrations, respectively (r2 = .25, p = .008, r2= .35, p = .006) . CONCLUSIONS: Nebulizer delivery of aerosolized aminoglycosides is efficient and predictable . In our clinical model, aerosolized antibiotics can make a significant impact on respiratory secretions . Their efficacy in treatment of critically ill patients remains to be determined. Mol Microbiol, 1997 Dec, 26(5), 1005 - 11 rpoB sequence analysis as a novel basis for bacterial identification; Mollet C et al.; Comparison of the sequences of conserved genes, most commonly those encoding 16S rRNA, is used for bacterial genotypic identification . Among some taxa, such as the Enterobacteriaceae, variation within this gene does not allow confident species identification . We investigated the usefulness of RNA polymerase beta-subunit encoding gene (rpoB) sequences as an alternative tool for universal bacterial genotypic identification . We generated a database of partial rpoB for 14 Enterobacteriaceae species and then assessed the intra- and interspecies divergence between the rpoB and the 16S rRNA genes by pairwise comparisons . We found that levels of divergence between the rpoB sequences of different strains were markedly higher than those between their 16S rRNA genes . This higher discriminatory power was further confirmed by assigning 20 blindly selected clinical isolates to the correct enteric species on the basis of rpoB sequence comparison . Comparison of rpoB sequences from Enterobacteriaceae was also used as the basis for their phylogenetic analysis and demonstrated the genus Klebsiella to be polyphyletic . The trees obtained with rpoB were more compatible with the currently accepted classification of Enterobacteriaceae than those obtained with 16S rRNA . These data indicate that rpoB is a powerful identification tool, which may be useful for universal bacterial identification. Optom Vis Sci, 1997 Dec, 74(12), 1030 - 8 Potential sources of bacteria that are isolated from contact lenses during wear; Willcox MD et al.; PURPOSE . The aim of this paper was to determine the possible contamination sources of contact lenses during wear . METHODS . Potential sources of the microbiota that colonized hydrogel contact lenses during wear were examined . The microorganisms that colonize contact lenses were grown, identified, and compared to those microorganisms that colonized the lower lid margins, upper bulbar conjunctiva, hands, and contact lens cases of contact lens wearers . In addition, the incidence of contamination of the domestic water supply in the Sydney area was obtained, and this was compared to the incidence of colonization of contact lenses by microorganisms in general and gram-negative bacteria in particular . RESULTS . There was a wide diversity of bacteria that were isolated from each site sampled . Coagulase-negative staphylococci and Propionibacterium spp . were the most common isolates from all ocular sites examined, and constituted the normal ocular microbiota . Other bacteria, including members of the families Enterobacteriaceae and Pseudomonadaceae, were isolated infrequently from all sites, but most frequently from contact lens cases . Statistical analysis revealed that there was a correlation between the isolation of bacteria from the contact lens and the lower lid margin (p < 0.001) . Analysis of this correlation revealed that this was true for the normal microbiota . A correlation was also noted between the colonization of contact lenses by gram-negative bacteria and contamination of the domestic water supply . DISCUSSION . This study has demonstrated that the likely route for the normal ocular microbiota colonizing contact lenses is via the lid margins, whereas colonization by gram-negative bacteria, including potential agents of microbial keratitis, is likely to be from the domestic water supply. Antimicrob Agents Chemother, 1997 Dec, 41(12), 2773 - 5 Canadian Multicenter Susceptibility Study, with a focus on cephalosporins, from 15 Canadian medical centers . The Canadian Multicenter Study Group; Blondeau JM et al.; We have previously reported on the in vitro susceptibilities of 4,482 microorganisms to 10 antimicrobial agents tested as part of a Canadian multicenter study . We now report on the remaining 10 agents tested in that study . Of the cephalosporins reported here, ceftriaxone had the greatest activity (82 to 100% susceptible isolates) against Enterobacteriaceae, compared to ceftizoxime (78 to 100%) and cefoperazone (78 to 100%) . Cefoperazone activity against Pseudomonas aeruginosa was 87%, compared to 92% for ticarcillin-clavulanate . All agents had 97% or greater activity against Staphylococcus aureus. Antimicrob Agents Chemother, 1997 Dec, 41(12), 2742 - 8 In vitro and in vivo antibacterial activities of GV129606, a new broad-spectrum trinem; Di Modugno E et al.; GV129606 is a new parenteral trinem antibiotic belonging to the beta-lactam class . It combines broad-spectrum activity (against gram-negative and -positive bacteria, aerobes and anaerobes), with high potency and resistance to beta-lactamases . Comparative in vitro and in vivo antibacterial activities were determined for GV129606 against more than 400 recent clinical isolates (aerobes, including beta-lactamase producers, and anaerobes), using representative antibacterial agents (meropenem, piperacillin, ceftazidime, cefpirome, ciprofloxacin, and gentamicin for aerobes and metronidazole, cefoxitin, piperacillin, and clindamycin for anaerobes) . Against methicillin-susceptible staphylococci and streptococci, GV129606 and meropenem were the most active of the drugs tested . GV129606 showed an MIC for 90% of strains tested (MIC90) ranging from < or =0.015 to 0.06 microg/ml against methicillin-susceptible staphylococci and Streptococcus sanguis, Streptococcus pyogenes, and Streptococcus agalactiae . Against penicillin-susceptible and -resistant Streptococcus pneumoniae isolates, GV129606, meropenem, and cefpirome showed MIC90s of < or =0.015 and 1 microg/ml, respectively . Meropenem was the most active compound against members of the family Enterobacteriaceae with MIC90s of < or =0.5 microg/ml . Against these species, GV129606 possessed activity superior to those of piperacillin, ceftazidime, cefpirome, and gentamicin, with MIC90s of < or =8 microg/ml, but its activity was two- to sixfold less than that of ciprofloxacin (with the exception of Proteus rettgeri and Providencia stuartii) . Haemophilus spp., Moraxella catarrhalis, Neisseria gonorrhoeae, and Pseudomonas aeruginosa were also included in the spectrum of GV129606 . GV129606 showed good antianaerobe activity, similar to metronidazole . It was stable against all clinically relevant beta-lactamases (similar to meropenem) . The in vitro activity was confirmed in vivo against septicemia infections induced in mice by selected gram-positive and -negative bacteria with 50% effective doses (ED50s) of < or =0.05 and < or =0.5 mg/kg of body weight/dose, respectively . GV129606 was as effective as meropenem against septicemia in mice caused by ceftazidime-resistant Pseudomonas aeruginosa, exhibiting an ED50 of 0.33 mg/kg/dose. Harefuah, 1997 Oct 2, 133(7-8), 275 - 81, 335 {Gram-negative enteric bacteremia in children in the Negev (1989-1994)}; Maimon-Greenwald M et al.; During 1989-1994, there were 322 episodes of Gram-negative enteric bacteremia in 308 children . The incidence increased from 31/100,000 in children younger than 15 years of age during 1989-1991, to 50/100,000 during 1992-1994 . The most common pathogens were Klebsiella, E . Coli, Salmonella and Enterobacter . 39% of episodes were nosocomial and a significant increase was recorded for each species during the last 3 years of the study . Klebsiella represented the most common pathogen causing nosocomial bacteremia, while E . coli and Salmonella were the main pathogens causing community-acquired bacteremia . In this study in southern Israel, the incidence of Gram-negative enteric bacteremia was significantly higher in Bedouin children, with the exception of bacteremia due to Salmonella, which occurred mainly in Jewish children. Lett Appl Microbiol, 1997 Nov, 25(5), 309 - 12 Evaluation of three decarboxylating agar media to detect histamine and tyramine-producing bacteria in ripened sausages; Roig-Sagues AX et al.; Histidine- and tyrosine-decarboxylase activity of 175 strains of bacteria isolated from eight retail samples of Spanish ripened sausages was tested in three decarboxylating agars (Niven medium, Joosten and Northolt medium and modified decarboxylating agar of Maijala) and confirmed by an enzymic method (histamine) and thin-layer chromatography (tyramine) . Enterobacteria and pseudomonads showed the highest percentage of positive responses to histamine and tyramine in the three decarboxylating agars, but only enterobacteria were subsequently confirmed as histamine-producing . Confirmed tyramine-producing strains were all identified as enterococci or lactic acid bacteria . The medium described by Joosten and Northolt was more sensitive and faster at detecting tyramine-producing microorganisms . However, all three media failed to detect one histamine-positive strain of lactic acid bacteria used as a control. J Appl Microbiol, 1997 Nov, 83(5), 613 - 8 Spoilage microflora in fresh chicken breast stored at 4 degrees C: influence of packaging methods; Jimenez SM et al.; Chicken breasts with skin were packaged either in air, under vacuum or in modified atmospheres of (i) 30% CO2/70% N2 and (ii) 70% CO2/30% N2 . After 3, 7, 14 and 21 days of storage at 4 degrees C, the samples were evaluated for spoilage microbial growth, odour and overall aspect . As expected, pseudomonads grew well in air or under vacuum, but growth was suppressed in both types of modified atmosphere packaging (MAP) . However, growth of lactobacilli, Enterobacteriaceae and Brochothrix thermosphacta was not inhibited in MAPs . Modified atmosphere packaging (ii) extended shelf-life up to 21 days compared to 5 days for air-packed samples. Acta Oncol, 1997, 36(6), 643 - 9 Resistance pattern of 2816 isolates isolated from 17631 blood cultures and etiology of bacteremia and fungemia in a single cancer institution; Trupl J et al.; The resistance pattern of 2816 isolates from 17631 blood cultures and the etiology of isolates causing bacteremia and fungemia among 14591 admissions were investigated in an 80-bed single cancer institute during seven years (1990-1996) under the same empiric therapeutic antibiotic policy but with different prophylactic strategies . No change was found in the proportion of Gram-positive versus Gram-negative bacteria isolated from bacteremias (70% vs . 30%) during the past seven years . Furthermore, the proportion of coagulase-negative staphylococci and enterococci was about the same before and after the introduction of ofloxacin in prophylaxis . However, the proportion of Pseudomonas aeruginosa and Stenotrophomonas maltophilia causing bacteremia increased . There was no increase in Candida krusei and Candida glabrata after the introduction of fluconazole into our prophylactic regimen in 1992 . Penicillin-resistance in viridans streptococci increased after penicillin was introduced into prophylaxis in acute leukemia in 1993 . Until 1995 no quinolone-resistant Enterobacteriaceae were observed . Susceptibility to quinolones did not significantly change within the past seven years in Enterobacteriaceae after their introduction to prophylaxis in 1991, but Pseudomonas aeruginosa decreased from 90 to 58.2% . Glycopeptide resistance in enterococci and staphylococci was minimal in the observed period (0.9-4.3%). J Dent Res, 1997 Nov, 76(11), 1770 - 5 Isolation of Enterobacteriaceae from the mouth and potential association with malodor; Goldberg S et al.; Bad breath is a common phenomenon, usually the result of bacterial metabolism in the oral cavity . It is generally accepted that Gram-negative bacteria are responsible for this problem, largely through degradation of proteinaceous substances . In initial experiments, screening of malodorous isolates following outgrowth of samples obtained from saliva, periodontal pockets, and the tongue dorsum yielded enterobacterial isolates . Clinical studies were conducted to examine the prevalence of such bacteria in four different populations: orthodontic patients, malodor clinic patients, complete-denture wearers, and a healthy young population . The prevalence of Enterobacteriaceae in the oral cavities of the denture-wearing population was very high (48.0%) as compared with the other groups: 27.1% in the malodor clinic patients, 16.4% in the normal population, and 13% among orthodontic patients . Isolates of Klebsiella and Enterobacter emitted foul odors in vitro which resembled bad breath, with concomitant production of volatile sulfides and cadaverine, both compounds related to bad breath . When incubated on a sterile denture, enterobacterial isolates produced typical denture foul odor . Isolates exhibited cell-surface hydrophobic properties when tested for adhesion to acryl and aggregation with ammonium sulphate . The results, taken together, suggest that Klebsiella and related Enterobacteriaceae may play a role in denture malodor. FEBS Lett, 1997 Nov 24, 418(1-2), 27 - 9 Ribosomal efficiency and growth rates of freshly isolated Escherichia coli strains originating from the gastrointestinal tract; Rang CU et al.; It has been previously reported that for natural Escherichia coli isolates from the ECOR collection, there were differences in the ribosomal efficiencies and there was a direct correlation between growth rate and the ribosome efficiency (R-factor) . The aim of this study was to determine whether strains freshly isolated (i.e . subcultured < 5 times) from the gastrointestinal tract ecosystem also exhibited this correlation . Eleven E . coli and two Enterobacter spp . isolates from either humans, pigs, rats or a mammoth were investigated . Considerable variability in the R-factor was noted using an in vitro translation assay, however no consistent correlation between the R-factor and growth rate was noted. Antibiot Khimioter, 1997, 42(9), 27 - 32 {Analysis of the etiologic structure of urinary tract infection and antibiotic-resistance of its pathogens}; Derevianko II et al.; The main pathogens of inflammatory diseases of the kidneys and upper urinary tracts in inpatients of an urological unit were gramnegative organisms of the family Enterobacteriaceae while the pathogens of the infection of the lower urinary tracts (nonspecific urethritis) and male genitalia were grampositive cocci . Pseudomonas aeruginosa, Enterobacter agglomerans and Proteus spp . (indole positive) were the chief causative agents of the hospital infections . The analysis of the materials revealed a tendency towards an increase in the microflora resistance to the most widely used antibiotics: aminoglycosides, cephalosporins and fluoroquinolones . This especially applied to the "problem" pathogen P.aeruginosa . Thus, in 1987 the portion of the P.aeruginosa gentamicin susceptible strains amounted to 52 per cent whereas in 1996 it was 13 per cent . The strains susceptible to ofloxacin equaled 79 per cent in 1988 and 44 per cent in 1995 . At present the drugs of choice in the treatment of urinary tract infections due to P.aeruginosa are ceftazidime, cefpirome and amikacin (65, 64 and 62 per cent of the susceptible strains respectively) . The importance of permanent microbiological monitoring and the respective correction of the therapy are indicated. Antibiot Khimioter, 1997, 42(8), 26 - 30 {Antimicrobial effects of medicines which are not antibiotics}; Shenderov BA; The data on antimicrobial activity of 69 different drugs not belonging to the class of typical antibiotics were examined . It was shown that many of them had antimicrobial activity against enterobacteria susceptible and resistant to antibiotics . Some of such drugs were able to eliminate the property of resistance to antibiotics and in particular that to chloramphenicol and tetracyclines . The data are indicative of the fact that many of the so called nonantibiotics have the capacity of active interference with the human microbial ecology. Med Dosw Mikrobiol, 1997, 49(1-2), 89 - 94 {Aerobic and anaerobic bacterial flora in chronic sinusitis in adults}; Radosz-Komoniewska H et al.; The aim of the study was to analyse microbiologically samples obtained from 30 patients aged from 21 to 73 years treated for chronic sinusitis . Aerobic bacteria only were isolated in 16 patients (53%), and anaerobic organisms only in 5 patients (17%) . Mixed aerobic and anaerobic isolates were recovered from 9 patients (30%) . The isolated aerobic bacteria were as follows: streptococci from the species Streptococcus salivarius, Streptococcus anginosus, Streptococcus group C, Streptococcus sanguis, Staphylococcus aureus, Gram-negative rods from the genus Haemophilus and rods from the Enterobacteriaceae family, and strains of Moraxella catarrhalis . The isolated anaerobic microorganisms Gram-negative rods from the genus Prevotella, Bacteroides, Fusobacterium, Gram-positive cocci from the genus Peptostreptococcus . Other organisms from the genus Vailonella, Eubacterium and Actinomyces were isolated less frequently . In 15 patients only one isolate was recovered, in 15 patients isolated bacteria were mixed with other microorganisms. Med Dosw Mikrobiol, 1997, 49(1-2), 75 - 81 {Analysis of aerobic and anaerobic bacterial flora colonizing drains after surgical abdominal incisions}; Michalska W et al.; Aerobic and anaerobic bacterial flora of post-operative incisions with drainage were examined . From each of 28 patients three specimens were taken; during operation (smear from peritoneal cavity), liquid from drain (taken at 3-th day after operation) and smear from drain taken at the end of drainage . Enterococci, Enterobacteriaceae spp . and anaerobes, especially Bacteroides spp . were most often isolated from specimens taken during operation . Enterococci and coagulase negative Staphylococci-often resistant to methicillin, were most often isolated from specimens taken at the end of drainage. Klin Lab Diagn, 1997 Mar, (3), 18 - 21 {Biovars and antibiotic resistance of enterobacter cloacae strains isolated from colonized newborns}; Morozova OT et al.; A total of 120 strains of E . cloacae were isolated from colonized newborns in a maternity hospital (96 strains) and from inpatients of other hospitals (23 strains) . Biovars of these strains and of 1 reference strain were determined by two methods of biochemical typing and their sensitivity to 14 antibiotics assessed . The overwhelming majority of isolates from newborns were referred to type 2 according to typing after Old (original) or to biovar 62 if typed using the modification of this method . The phenotypical profile of determinants of resistance to 10 antibacterial drugs were the same, including those to aminoglycosides (except amikacin) and third-generation cephalosporins . The strains isolated from non-obstetrical inpatients belonged to biovars with phenotypical profiles of resistance to only 3 antibiotics and sensitive to aminoglycosides and third-generation cephalosporins. Contracept Fertil Sex, 1997 Jul-Aug, 25(7-8), 572 - 5 {Acute salpingitis: current antibiotic protocols}; Judlin PG et al.; Nowadays, most patients with uncomplicated acute salpingitis undergo ambulatory treatment . The choice of medications must take features of PID into account: they are mult-microbial infections and a prolonged follow-up is necessary in order to decrease the risk of sequellae . To fulfil these goals, combination of antibiotics are prescribed that are active against C . trachomatis, enterobacteria and anaerobes. Enferm Infecc Microbiol Clin, 1997 Sep, 15 Suppl 1, 69 - 72 {Monotherapy with meropenem in febrile granulocytopenic patients}; Sanz MA et al.; Infection remains the major cause of morbidity and mortality for cancer patients who become granulocytopenic . Combinations of beta-lactams plus aminoglycosides have been the standard empiric therapy for febrile granulocytopenic patients, especially those with profound long-lasting granulocytopenia . The advent of new broad-spectrum cephalosporins and carbapenems has favoured the possibility of empiric monotherapy . Meropenem is a parenteral carbapenem antibiotic stable to renal dehydropeptidase-I which has excellent bactericidal activity against almost all clinically significant aerobic and anaerobic organisms . Meropenem hasta an antibacterial spectrum similar to that of imipenem but it is more active against Pseudomonas aeruginosa, all Enterobacteriaceae, Haemophilus influenzae, Proteus spp, Morganella spp and Providencia spp . Recently, the efficacy, safety, and tolerance of meropenem monotherapy for the empirical treatment of fever in granulocytopenic cancer patients have been compared in two large prospective randomized multicenter trials . The Meropenem Study Group compared monotherapy with meropenem versus ceftazidime and the EORTC conducted a comparative study of meropenem monotherapy versus the combination of ceftazidime plus amikacin . In both groups, success rates were similar by type of infection and infection-related mortality was low . Related adverse events were also similar in both groups . These studies confirm that monotherapy with meropenem is as effective as ceftazidime-containing regimens for the empiric treatment of fever in granulocytopenic patients. Enferm Infecc Microbiol Clin, 1997 Sep, 15 Suppl 1, 20 - 6 {in vitro activity of carbapenems against Enterobacteriaceae and Pseudomonas aeruginosa hyperproducers of group 1 chromosomal beta-lactamases}; Martinez-Beltran J et al.; Resistance to imipenem and meropenem, reported sporadically in Enterobacteriaceae and more frequently in Pseudomonas aeruginosa, can be caused, among other mechanisms, by the combination of changes in permeability and hyperproduction of inducible chromosomal beta-lactamases . In this study, the in vitro activity of imipenem and meropenem was analysed by the agar dilution method against cefotaxime, ceftazidime, and aztreonam resistant clinical strains of Enterobacteriaceae (n = 202) and P . aeruginosa (n = 90) . This phenotype is consistent with the hyperproduction of group 1 chromosomal beta-lactamases and was previously determined in stably derepressed mutants in the same species, obtained from strains with inducible beta-lactamase expression by selection with cefotaxime and ceftazidime . Likewise, the activity of imipenem and meropenem against the same number of clinical isolates susceptible to cefotaxime, ceftazidime, and aztreonam was evaluated . In general, imipenem and meropenem showed an excellent activity, which was intrinsically greater for meropenem against Enterobacteriaceae and P . aeruginosa organisms . Nevertheless, imipenem and meropenem activity was slightly affected on cefotaxime, ceftazidime, and aztreonam resistant isolates of E . cloacae (MIC90, 1 and 0.2 microgram/ml, respectively), E . aerogenes (1 and 0.2 microgram/ml), C . freundii (1 and 0.1 microgram/ml), M . morganii (1 and 0.5 microgram/ml), and S . marcescens (4 and 0.5 micrograms/ml) . On the other hand, the activity of imipenem and meropenem against ceftazidime and aztreonam resistant isolates of P . aeruginosa was more significantly affected, with MIC90 values of 64 and 16 micrograms/ml, respectively. Enferm Infecc Microbiol Clin, 1997 Sep, 15 Suppl 1, 14 - 9 {Penetration of meropenem in gram-negative bacilli . Differences in activity with imipenem}; Garcia de Lomas J et al.; The outer membrane of gramnegative bacteria cell-wall contain channels formed by proteins known as porins, which facilitate the penetration of molecules into the cell . Imipenem and meropenem possess an important intrinsic activity against most gramnegative bacteria due to their high affinity for the penicillin-binding proteins PBP-2 and/or PBP-3 . Meropenem is slightly more active against certain species of Enterobacteriaceae, Pseudomonas aeruginosa and nonfermenters bacilli . In this sense the differences in the activity between the two carbapenems may be attributable to differences in their affinity for PBPs, differences in their resistance to beta-lactamases hydrolysis, or to the differences in the capacity to employ certain porin channels . OprF is the main porin channel involved in the beta-lactam penetration of bacteria, though OprC and OprD2 may also contribute to the penetration of carbapenems into P . aeruginosa . However, evidences suggest that although impenem requires the presence of OprD2, meropenem may use other pathways for penetration . In Escherichia coli both carbapenems use ompF and ompC, and no specific porin channels have been detected . Only in the case of Enterobacter cloacae may exceptions exists among Enterobacteriaceae. Biochim Biophys Acta, 1997 Oct 9, 1354(1), 7 - 12 Sequence of a melibiose transporter gene of Enterobacter cloacae; Okazaki N et al.; We cloned a fragment of the chromosomal DNA of Enterobacter cloacae, which enabled a melibiose-negative Escherichia coli mutant lacking melB to grow on melibiose as the sole source of carbon . Transformed cells harboring the hybrid plasmid carrying the cloned DNA showed melibiose transport activity . The nucleotide sequence of the DNA region was determined . One complete open reading frame (ORF) and a part of another ORF were found in the region, and the amino acid sequences were deduced . The complete ORF was found to encode a melibiose transporter which consisted of 425 amino acid residues . Hydropathy analysis revealed that there are about 12 hydrophobic domains in this transporter . The incomplete ORF which exists in the upstream region of the transporter gene seemed to encode an alpha-galactosidase. Br J Rheumatol, 1997 Oct, 36(10), 1051 - 3 IgM, IgG and IgA class enterobacterial antibodies in serum and synovial fluid in patients with ankylosing spondylitis and rheumatoid arthritis; Maki-Ikola O et al.; IgM, IgG and IgA class antibodies against three Klebsiella pneumoniae capsular types, Escherichia coli and Proteus mirabilis, as well as total immunoglobulin concentrations, were measured by enzyme immunoassay and radial immunodiffusion technique, respectively, in paired serum and synovial fluid samples from eight patients with ankylosing spondylitis and 10 with rheumatoid arthritis . No clear evidence for intra-articular antibody production against any of the studied microbes was found. J Antimicrob Chemother, 1997 Oct, 40(4), 543 - 9 Detection of mutations in the gyrA and parC genes in quinolone-resistant clinical isolates of Enterobacter cloacae; Deguchi T et al.; We have determined partial sequences of the gyrA and parC genes of Enterobacter cloacae type strain including the regions analogous to the quinolone resistance-determining region of the Escherichia coli gyrA gene . The deduced 65- and 49-amino acid sequences of the determined regions of the E . cloacae gyrA and parC genes were identical to the corresponding regions of the E . coli GyrA and ParC proteins, respectively . We examined 40 clinical strains of E . cloacae isolated from patients with urinary tract infection for susceptibilities to nalidixic acid and ciprofloxacin . Based on the nalidixic acid and ciprofloxacin MICs, these isolates were divided into 19 quinolone-susceptible strains (MICs of nalidixic acid, 3.13-25 mg/L; MICs of ciprofloxacin, < or = 0.025 mg/L) and 21 quinolone-resistant strains (MICs of nalidixic acid, 400 to > 800 mg/L; MICs of ciprofloxacin, 0.39-100 mg/L) . We analysed five quinolone-susceptible and 21 quinolone-resistant strains for alterations in GyrA and ParC . The five quinolone-susceptible strains had amino acid sequences in GyrA and ParC identical to those of type strain . Of the 21 quinolone-resistant isolates, three (MICs of nalidixic acid, 400 to > 800 mg/L; MICs of ciprofloxacin, 0.39-3.13 mg/L) had a single amino acid change at the position equivalent to Ser-83 in the E . coli GyrA protein and no alterations in ParC; one (MIC of nalidixic acid, > 800 mg/L; MIC of ciprofloxacin, 3.13 mg/L) had a single amino acid change at Ser-83 in GyrA and a single amino acid change at the position equivalent to Glu-84 in the E . coli ParC protein; two (MIC of nalidixic acid, > 800 mg/L; MIC of ciprofloxacin, 25 mg/L) had double amino acid changes at Ser-83 and Asp-87 in GyrA and no alterations in ParC; and 15 (MICs of nalidixic acid, > 800 mg/L; MICs of ciprofloxacin, 25-100 mg/L) had double amino acid changes at Ser-83 and Asp-87 in GyrA and a single amino acid change at Ser-80 or Glu-84 in ParC . This study suggests, that in clinical isolates of E . cloacae, DNA gyrase is a primary target of quinolones, that only a single amino acid change at Ser-83 in GyrA is sufficient to generate high-level resistance to nalidixic acid and to decrease susceptibility to ciprofloxacin, and that the accumulation of amino acid changes in GyrA and the simultaneous presence of the ParC alterations play a central role in developing high-level resistance to ciprofloxacin. J Antimicrob Chemother, 1997 Oct, 40(4), 533 - 41 Bases of variation in resistance to beta-lactams in Klebsiella oxytoca isolates hyperproducing K1 beta-lactamase; Gheorghiu R et al.; Nineteen isolates of Klebsiella oxytoca were examined, representing 18 distinct strains . All were from a 1994 survey of resistance amongst klebsiellae in intensive care units in Europe, and all had reduced susceptibility, or were resistant, to cefuroxime, ceftriaxone and aztreonam, suggesting hyperproduction of the chromosomal K1 beta-lactamase . We sought to confirm this mechanism and to identify why the levels of resistance varied between isolates . Possible reasons for variation were differences in the quantity or subtype of the K1 enzyme or differences in this enzyme's interplay with permeability . Spectrophotometric assays showed that all 19 isolates had K1-like beta-lactamases and that these were present at > or = 15-fold higher levels than in beta-lactam-sensitive K . oxytoca isolates . Fourteen of the 19 isolates had the OXY-2 form of K1 enzyme, while the remaining five had the OXY-1 form, as determined by isoelectric focusing and PCR amplification . Most isolates with the OXY-2 enzyme were more resistant than those with the OXY-1 subtype, but this difference partly reflected enzyme quantity rather than subtype . More generally, and irrespective of enzyme subtype, levels of resistance were broadly related to beta-lactamase specific activity, and the degree of hyperproduction was a major determinant of the level of resistance . Nevertheless, other factors had a role too: several isolates had reduced susceptibility or were resistant to cefoxitin, which is not a substrate for K1 enzyme, and examination of outer membrane protein profiles revealed considerable strain-to-strain diversity in the molecular weight range typical of the major enterobacterial porins (40-48 kDa). Res Microbiol, 1997 Jan, 148(1), 87 - 93 Electrochemical measurement of trace concentrations of biological hydrogen produced by Enterobacteriaceae; Podesta JJ et al.; Accurate measurements of the hydrogen gas produced by Escherichia coli and Hafnia alvei pure cultures during glucose metabolism were performed under different growth conditions: stagnant, with magnetic stirring or with vibrational shaking . These measurements were carried out using an electrochemical hydrogen sensor based on a platinum-coated solid polymer electrolyte membrane (Pt-SPE) . The results obtained were dependent on the hydrodynamic conditions of the growth, with greater hydrogen production being associated with the stagnant conditions . These measurements will eventually enable us to elucidate whether the pathway used for glucose metabolism is either strictly or mainly anaerobic and to modify experimental conditions so as to influence the reaction. Mol Microbiol, 1997 Nov, 26(3), 531 - 44 The rap and hor proteins of Erwinia, Serratia and Yersinia: a novel subgroup in a growing superfamily of proteins regulating diverse physiological processes in bacterial pathogens; Thomson NR et al.; The enteric bacterium Serratia marcescens is an opportunistic human pathogen . The strain ATCC39006 makes the red pigment, prodigiosin (Pig), and the beta-lactam antibiotic carbapenem (Car) . Mutants were isolated that were concomitantly defective for Pig and Car production . These mutants were found to have a mutation in the rap gene (Regulation of Antibiotic and Pigment) . Sequence analysis of the rap gene revealed a predicted protein product showing strong homology to SlyA, originally thought to be a haemolytic virulence determinant in Salmonella typhimurium . Homologues of rap were detected in several bacterial genera, including Salmonella, Yersinia, Enterobacter, and species of the plant pathogen, Erwinia . The Erwinia hoeEr (homologue of rap) and the Yersinia horYe genes were also found to be very similar to rap and slyA . Marker exchange mutagenesis of horEr revealed that it encoded a regulatory protein controlling the production of antibiotic and exoenzyme virulence determinants in the phytopathogen, Erwinia carotovora subspecies carotovora . We have shown that these new homologues of SlyA form a highly conserved subgroup of a growing superfamily of bacterial regulatory proteins controlling diverse physiological processes in human, animal and plant pathogens. Diagn Microbiol Infect Dis, 1997 Nov, 29(3), 173 - 86 Comparative in vitro assessment of sparfloxacin activity and spectrum using results from over 14,000 pathogens isolated at 190 medical centers in the USA . SPAR Study Group; Ballow CH et al.; Sparfloxacin, a new orally administered fluoroquinolone, was tested against 14,182 clinical strains isolated (generally blood stream and respiratory tract cultures) at nearly 200 hospitals in the United States (USA) and Canada . Sparfloxacin activity was compared with 13 other compounds by Etest (AB BIODISK, Solna, Sweden), broth microdilution, or a standardized disk diffusion method . Using the Food and Drug Administration/product package insert MIC breakpoint for sparfloxacin susceptibility (< or = 0.5 microgram/ml), 94% of Streptococcus pneumoniae (2666 isolates) and 89% of the other streptococci (554 isolates) were susceptible . However, at < or = 1 microgram/ml (the breakpoint for all nonstreptococcal species) sparfloxacin susceptibility rates increased to 100% and 98%, respectively, for the two groups of streptococci . Only 50% and 65% of pneumococci were susceptible to ciprofloxacin (MIC90, 3 micrograms/ml) and penicillin (MIC90, 1.5 micrograms/ml), respectively . Although there were significant differences between regions in the USA in the frequency of penicillin-resistant pneumococcal strains, results indicate that the overall sparfloxacin MIC90 was uniformly at 0.5 microgram/ml . Nearly all (> or = 99%) Haemophilus species and Moraxella catarrhalis, including those harboring beta-lactamases, were susceptible to sparfloxacin, ciprofloxacin, and amoxicillin/clavulanic acid . Only cefprozil and macrolides demonstrated lower potency and spectrum against these two species . Sparfloxacin was active against oxacillin-susceptible Staphylococcus aureus (96 to 97%), Klebsiella spp . (95%), and other tested enteric bacilli (93%) . Comparison between broth microdilution MIC and disk diffusion interpretive results for M . catarrhalis, Staphylococcus aureus, and the Enterobacteriaceae showed an absolute intermethod categorical agreement of > 95% using current sparfloxacin breakpoints, in contrast to those of cefpodoxime for S . aureus where a conspicuous discord (98% versus 59%) between methods was discovered . These results demonstrate that sparfloxacin possesses sufficient in vitro activity and spectrum versus pathogens that cause respiratory tract infections (indications), especially strains resistant to other drug classes such as the earlier fluoroquinolones, oral cephalosporins, macrolides, and amoxicillin/clavulanic acid . The sparfloxacin susceptibility breakpoint for streptococci may require modification (< or = 1 microgram/ml) based on the MIC population analysis presented here . A modal MIC (0.38 to 0.5 microgram/ml) was observed at the current breakpoint . Regardless, sparfloxacin inhibited 89% (nonpneumococcal Streptococcus spp.) to 100% (Haemophilus spp., M . catarrhalis) of the isolates tested with a median activity of 97% against indicated species. Infect Control Hosp Epidemiol, 1997 Nov, 18(11), 769 - 71 Ribotyping and random amplification of polymorphic DNA for nosocomial Enterobacter cloacae isolates in a pediatric intensive-care unit; Hou ST et al.; Between 1987 and 1989, two sequential outbreaks of nosocomial infection caused by Enterobacter cloacae occur-red in the pediatric intensive-care unit of a tertiary-care teaching hospital . Seventeen strains retrieved from the outbreaks and two control strains identified in other wards were typed by ribotyping and random amplification of polymorphic DNA (RAPD) . The results indicated that the genomic pattern of strains identified between the first and second outbreaks was different . We conclude that both ribotyping and RAPD are highly discriminatory and reproducible methods for typing E cloacae . RAPD seems to be more efficacious and cost-effective. Chemotherapy, 1997 Nov-Dec, 43(6), 393 - 3 In vitro activity of biapenem against recent gram-negative and gram-positive clinical isolates; Bonfiglio G et al.; The in vitro activity of biapenem, a new carbapenem, against 535 clinical recent isolates was compared with those of other antibiotics . Biapenem showed broad-spectrum activity against gram-negative and gram-positive clinical isolates . The new carbapenem was more active than imipenem against members of the family Enterobacteriaceae with MIC90S ranging from 0.12 to 2 mg/l and from 0.25 to 4 mg/l, respectively . Moreover it was 2-fold more active than imipenem against Pseudomonas aeruginosa (MIC90, 8 and 16 mg/l, respectively) . Taken together, these results indicate that biapenem shares the favorable in vitro activity properties of imipenem and merits further study in the treatment of infections caused by a wide range of pathogens. Kansenshogaku Zasshi, 1997 Oct, 71(10), 1017 - 24 {Trend of bacterial meningitis in children over a 14 year period (1981 through 1994) in Japan--an analysis based on studies in 27 institutions}; Kobayashi Y et al.; We observed 266 children with purulent meningitis in 27 institutions in Japan during the 14 years from 1981 on dividing these years into 3 periods, 1981-1985, 1986-1990 and 1991-1994, and studied the trend of causative organisms identified in 254 among the 266 patients . Their ages were less than 3 months after birth in 50 children and 3 months or older in 216: there were 141 boys and 125 girls . The causative organisms were H . influenzae in 134 patients and S . pneumoniae in 50, most of them being aged 3 months or older . Next to the above bacteria ranked S . agalactiae in 29 and E . coli in 12, many of the patients were aged less than 3 months . Staphylococcus spp . was found in 7 patients and about 70% of them were aged 3 months or older . L . monocytogenes was found in 4 patients and N . meningitidis in 3 and they were aged 3 months or older in both patient groups . S . pyogenes, Enterococcus spp., Peptostreptococcus spp., P . Mirabilis and Enterobacter spp . were detected each in 1 patient . The causative organism was unknown in 21 patients and there was no double infection . H . influenzae were detected in 18 patients in 1981-1985 period (36.7%), in 56 in 1986-1990 (54.9%) and in 60 in 1991-1994 (63.8%) showing an increasing tendency, but S . pneumoniae exhibited neither an increasing nor decreasing tendency . There was a decreasing tendency with S . agalactiae and E . coli, but the details were not clear because there were few patients aged less than 3 months . Although the period of coexistence of 4 main bacterial species was not made clear in this study . Listeria is considered to develop mainly in the early childhood, and we believe that the conventional way of using a cephem preparation and ampicillin combined for patients under 6 years need not be altered . However, panipenem (phonetic) is likely to be effective for insensible S . pneumoniae for the time being. J Bacteriol, 1997 Nov, 179(22), 7063 - 71 Temperature-dependent regulation of the ribosomal small-subunit protein S21 in the cyanobacterium Anabaena variabilis M3; Sato N et al.; The rpsU gene, which encodes the ribosomal small-subunit protein S21 in Anabaena, is not a part of the macromolecular-synthesis operon as in most enterobacteria but rather is located downstream of the rbpA1 gene, which encodes an RNA-binding protein . Two types of transcripts were detected for this gene cluster . The level of the major rbpA1-rpsU transcript was about 10 times higher at 22 degrees C than at 38 degrees C, whereas the minor monocistronic rpsU transcript was more abundant at the higher temperature . The level of the S21 protein in relation to total protein was three times lower at 38 degrees C than at 22 degrees C . Analysis of isolated ribosomes indicated that S21 was present at an equimolar ratio with regard to other ribosomal proteins at 22 degrees C but that its level decreased with temperature . Conversely, the relative abundance of S5 increased with temperature . A decrease in the level of S21 at high temperature was also found in Synechocystis, in which rpsU is located downstream of the rrn operon . These results suggest that S21 is involved in the adaptation to changes in temperature in cyanobacteria. Antimicrob Agents Chemother, 1997 Nov, 41(11), 2544 - 6 Improved antimicrobial activity of DU-6859a, a new fluoroquinolone, against quinolone-resistant Klebsiella pneumoniae and Enterobacter cloacae isolates with alterations in GyrA and ParC proteins; Deguchi T et al.; MICs of DU-6859a, a novel fluoroquinolone, for 18 Klebsiella pneumoniae isolates and 21 Enterobacter cloacae isolates with altered GyrA or altered GyrA and ParC ranged from < or =0.025 to 6.25 microg/ml and from 0.1 to 3.13 microg/ml, respectively . Based on the MICs at which 90% of the isolates were inhibited for these strains of K . pneumoniae and E . cloacae, DU-6859a exhibited 16- to 256-fold-greater activity than currently available fluoroquinolones. J Med Microbiol, 1997 Nov, 46(11), 941 - 8 Comparison of Vibrio cholerae O1 isolates by polymerase chain reaction fingerprinting and ribotyping; Shangkuan YH et al.; The rRNA gene restriction patterns and the polymerase chain reaction (PCR) fingerprinting types of 53 Vibrio cholerae O1 isolates were studied . Five and eight patterns were observed from 27 toxigenic and 26 non-toxigenic O1 isolates after BglI cleavage . PCR fingerprinting with three primer sets aimed at enterobacterial repetitive intergenic consensus (ERIC) sequences, ERIC-related sequences in V . cholerae, another kind of repeated sequences in V . cholerae (VCR) and arbitrary sequences divided the same strains into seven and 10 PCR types, respectively . Eight ribotypes had unique PCR patterns . PCR fingerprinting identified more than one pattern among isolates within each of the remaining ribotypes . However, ribotyping was able to differentiate the same PCR types in one case . A single ribotype and a single PCR pattern were found in toxigenic O1 strains isolated in Taiwan from imported food and imported cases of cholera between 1993 and 1995 . Typing of V . cholerae O1 by PCR fingerprinting correlated well with ribotyping, but was more discriminating . PCR assay provides a rapid and simple means of typing these strains for epidemiological studies. J Med Microbiol, 1997 Nov, 46(11), 903 - 12 Serratia marcescens; Hejazi A et al.; Over the last 30 years, Serratia marcescens has become an important cause of nosocomial infection . There have been many reports concerning the identification, antibiotic susceptibility, pathogenicity, epidemiological investigations and typing of this organism . Accurate identification is important in defining outbreaks . The API 20E system has been used widely, but is not individually satisfactory . The growth of S . marcescens in the environment has been investigated in relation to water, disinfectants and plastics such as blood bags . Certain extracellular products are unique to S . marcescens . Pigment (prodigiosin) biosynthesis by S . marcescens has been investigated fully since the emergence of the organism as a cause of infection . Many other aspects of the pathogenicity and virulence of S . marcescens have been studied, including adherence and hydrophobicity, lipopolysaccharide (LPS) and extracellular products . Two modes of adhesion to host epithelial surfaces have been suggested . These are mannose-resistant (MR) pili and mannose-sensitive (MS) pili . LPS, which is responsible for the biological activity of endotoxin, has been investigated fully and 24 somatic antigens have been described . The production of different enzymes by S . marcescens as virulence factors has also been reported, including chitinase, lipase, chloroperoxidase and an extracellular protein, HasA . Antibiotics used to treat serratia infection include beta-lactam agents, aminoglycosides and fluoroquinolones and a variety of different resistance mechanisms have been demonstrated . Typing methods used to study the epidemiology of S . marcescens include biotyping, bacteriocin typing, phage typing, plasmid analysis, polymerase chain reaction amplification of enterobacterial repetitive intergenic consensus sequences (ERIC-PCR) and ribotyping . Serological typing has also been used and this method seems to be a suitable first-line typing method for S . marcescens, although some strains remain untypable . RAPD-PCR has also been applied to a small number of isolates and seems to be a promising method, especially for rapid monitoring of an outbreak and tracing the source of initial infection. J Immunol, 1997 Nov 15, 159(10), 4868 - 78 Bacterial lipoprotein and lipopolysaccharide act synergistically to induce lethal shock and proinflammatory cytokine production; Zhang H et al.; Septic shock is a major cause of death in the world . Although much is known about the role of LPS in septic shock, little is known about the role of other bacterial components . Lipoprotein (LP) is a major component of bacteria in the family Enterobacteriaceae . LP purified from Escherichia coli was shown to induce TNF-alpha and IL-6 production in peritoneal exudate macrophages obtained from LPS-responsive (C3H/HeOuJ) and LPS-nonresponsive (C3H/HeJ) mice . LP and LPS acted synergistically to induce cytokine production not only in C3H/HeOuJ macrophages but also in C3H/HeJ macrophages . These results suggest that LPS can induce cellular signaling in C3H/HeJ macrophages, and that LPS and LP activate macrophages via different receptors and/or signaling pathways . The role LP plays in septic shock was investigated using the mouse D-galactosamine model . LP induced lethal shock and in vivo production of TNF-alpha and IL-6 in both LPS-responsive and LPS-nonresponsive mice . LPS failed to induce lethal shock or in vivo cytokine production in C3H/HeJ mice . However, LP and LPS acted synergistically in inducing lethal shock and in vivo cytokine production in both LPS-responsive and LPS-nonresponsive mice . Finally, a heat-killed preparation of an E . coli mutant strain that lacked LP was shown to be less efficient than heat-killed wild-type E . coli at inducing lethal shock in C3H/HeJ mice . Collectively, these results suggest that LP and LPS induce cytokine production via different mechanisms and that LP plays an important role in septic shock induced by bacteria in the family Enterobacteriaceae. Mol Microbiol, 1997 Sep, 25(5), 831 - 8 Control of virulence gene expression by plant calcium in the phytopathogen Erwinia carotovora; Flego D et al.; Plant calcium can modulate a particular plant-pathogen interaction and have a decisive role in disease development . Enhanced resistance to the phytopathogenic enterobacterium Erwinia carotovora, the causal agent of bacterial soft rot disease, is observed in high-calcium plants . One of the main virulence determinants of E . carotovora, the PehA endopolygalacturonase, is specifically required in the early stages of the infection . Production of PehA was found to be dependent on the calcium concentration in the bacterial environment . An increase in extracellular calcium to mM concentrations repressed pehA gene expression without reducing or even enhancing expression of other extracellular enzyme-encoding genes of this pathogen . An increase in plant calcium levels could be correlated to enhanced resistance to E . carotovora infection and to an inhibition of in planta production of PehA . Ectopic expression of pehA from a calcium-insensitive promoter allowed E . carotovora to overcome this calcium-induced resistance . The results imply that plant calcium can constitute an important signal molecule in plant-pathogen interaction, which acts by modulating the expression of virulence genes of the pathogen. Appl Environ Microbiol, 1997 Nov, 63(11), 4331 - 9 Identification of N2-fixing plant- and fungus-associated Azoarcus species by PCR-based genomic fingerprints; Hurek T et al.; Most species of the diazotrophic Proteobacteria Azoarcus spp . occur in association with grass roots, while A . tolulyticus and A . evansii are soil bacteria not associated with a plant host . To facilitate species identification and strain comparison, we developed a protocol for PCR-generated genomic fingerprints, using an automated sequencer for fragment analysis . Commonly used primers targeted to REP (repetitive extragenic palindromic) and ERIC (enterobacterial repetitive intergenic consensus) sequence elements failed to amplify fragments from the two species tested . In contrast, the BOX-PCR assay (targeted to repetitive intergenic sequence elements of Streptococcus) yielded species-specific genomic fingerprints with some strain-specific differences . PCR profiles of an additional PCR assay using primers targeted to tRNA genes (tDNA-PCR, for tRNA(IIe)) were more discriminative, allowing differentiation at species-specific (for two species) or infraspecies-specific level . Our protocol of several consecutive PCR assays consisted of 16S ribosomal DNA (rDNA)-targeted, genus-specific PCR followed by BOX- and tDNA-PCR; it enabled us to assign new diazotrophic isolates originating from fungal resting stages (sclerotia) to known species of Azoarcus . The assignment was confirmed by phylogenetic analysis of 16S rDNA sequences . Additionally, the phylogenetic distances and the lack of monophyly suggested emendment of the genus Azoarcus: the unnamed species Azoarcus groups C and D and a new group (E) of Azoarcus, which was detected in association with fungi, are likely to have the taxonomic rank of three different genera . According to its small subunit rRNA, the sclerotium-forming basidiomycete was related to the Ustilagomycetes, facultatively biotrophic parasites of plants . Since they occurred in a field which was under cultivation with rice and wheat, these fungi might serve as a niche for survival for Azoarcus in the soil and as a source for reinfection of plants. J Infect Dis, 1997 Nov, 176(5), 1260 - 8 Antiserum against Escherichia coli J5 contains antibodies reactive with outer membrane proteins of heterologous gram-negative bacteria; Hellman J et al.; The binding of IgG in antiserum to Escherichia coli J5 to the surface of Enterobacteriaceae and to cell wall fragments released from serum-exposed bacteria was studied in a search for potentially protective epitopes other than lipopolysaccharide (LPS) . IgG titers to multiple heterologous gram-negative smooth bacteria increased following incubation of the bacteria in serum and decreased following absorption with serum-exposed heterologous bacteria . IgG eluted from absorbing bacteria bound to at least three conserved bacterial outer membrane proteins (OMPs), but not LPS, as assessed by immunoblotting . The same OMPs were present in LPS-containing macromolecular cell wall fragments released by incubation of heterologous gram-negative bacteria in human serum . Part of the protection offered by J5 antiserum could be from binding of IgG to conserved OMPs at the bacterial surface or to OMPs in cell-wall fragments released from dying bacteria. Avian Dis, 1997 Jul-Sep, 41(3), 548 - 58 Inhibition of Salmonella typhimurium attachment to chicken cecal mucus by intestinal isolates of Enterobacteriaceae and lactobacilli; Craven SE et al.; The ability of selected strains of Enterobacteriaceae or lactobacilli isolated from the intestines of adult chickens to inhibit in vitro attachment of Salmonella typhimurium 3333/O to cecal mucus in the presence or absence of D-mannose was determined . Attachment in the absence of mannose was reduced by prior exposure of mucus to cultures of two isolates of Enterobacteriaceae, an Escherichia coli and a Hafnia alvei strain, but not to a third isolate, an Enterobacter agglomerans strain . Attachment of S . typhimurium was not inhibited when mannose was present in the blocking or attachment step . Formation of fimbriae by the two inhibitory Enterobacteriaceae strains and the S . typhimurium strain, as indicated by titers of mannose-sensitive hemagglutination of guinea pig erythrocytes was optimal in Z biphasic medium (consisting of tryptone, yeast extract, dextrose, and NaCl) incubated anaerobically at 42 C . Fimbriae of each of three strains prepared from these cultures also inhibited attachment . These are characteristics consistent with attachment and inhibition of attachment mediated by a mannose-sensitive adhesin associated with type 1 fimbriae on bacterial cells of Enterobacteriaceae strains . Attachment in the presence of mannose was significantly reduced by prior exposure of mucus to cultures of a Lactobacillus salivarius strain and a Lactobacillus delbrueckii delbrueckii strain but not to a strain of Lactobacillus for which the species had not been determined . Washed cells or spent culture supernatant fluid from brain-heart infusion broth, Z broth, or Z biphasic cultures of the inhibitory strains of lactobacilli incubated at 37 or 42 C inhibited this form of attachment . Of 27 intestinal isolates of Enterobacteriaceae and 21 of lactobacilli, the lactobacilli strains were generally more hydrophobic than the Enterobacteriaceae as determined by adherence to hexadecane . The lactobacilli isolates did not agglutinate guinea pig erythrocytes . The data suggest more than one mechanism for mediating attachment of inhibitory bacterial strains and for subsequent attachment of S . typhimurium. J Biochem (Tokyo), 1997 Jun, 121(6), 1129 - 33 Chemical structure of lipid A from Helicobacter pylori strain 206-1 lipopolysaccharide; Suda Y et al.; The chemical structure of a novel lipid A, which was obtained as a major component from lipopolysaccharide of Helicobacter pylori strain 206-1, was determined to be a glucosamine beta(1-6) disaccharide 1-(2-aminoethyl)phosphate acylated by (R)-3-hydroxyoctadecanoic acid and (R)-3-(octadecanoyloxy)octadecanoic acid at the 2- and 2'-position, respectively . The absence of a phosphoryl group at the 4'-position and fatty acyl groups at the 3- and 3'-position, and the stoichiometric presence of 2-aminoethyl phosphate at the 1-position are unique features, distinguishing it from the lipid A of enterobacteria. Microbiologia, 1997 Sep, 13(3), 321 - 30 Purification of OmpU from Vibrio cholerae classical strain 569B: evidence for the formation of large cation-selective ion-permeable channels by OmpU; Benz R et al.; The outer membrane of the classical Vibrio cholerae strain 569B was isolated by sucrose density centrifugation . The simple treatment of the isolated outer membrane or the cell envelopes with different detergents allowed the purification of two outer membrane proteins, the 38 kDa OmpU and the 25 kDa OmpV . Furthermore, a 35 kDa outer membrane protein (probably the 35 kDa OmpA-like protein) was purified by two-fold treatment of the cell envelope with 2% SDS solution . A subsequent wash of the SDS-pellet with 2% Genapol buffer yielded in the 38 kDa OmpU protein, which formed SDS-resistant oligomers (66 kDa) . The Genapol pellet contained OmpV . Reconstitution experiments with lipid bilayer membranes demonstrated that OmpU was a channel-forming component, whereas OmpV had a small channel-forming ability if any . The OmpU channels appeared to be large and water-filled and had a single-channel conductance of about 2 nS in 1 M KCl for the monomer in a trimer, which means that they have a larger cross-section than enterobacterial porins . The channels showed rapid switching between open and closed configuration . They were slightly cation-selective, which suggests that they contain an excess of negatively charged amino groups. EMBO J, 1997 Nov 3, 16(21), 6394 - 406 The chaperone-assisted membrane release and folding pathway is sensed by two signal transduction systems; Jones CH et al.; The assembly of interactive protein subunits into extracellular structures, such as pilus fibers in the Enterobacteriaceae, is dependent on the activity of PapD-like periplasmic chaperones . The ability of PapD to undergo a beta zippering interaction with the hydrophobic C-terminus of pilus subunits facilitates their folding and release from the cytoplasmic membrane into the periplasm . In the absence of the chaperone, subunits remained tethered to the membrane and were driven off-pathway via non-productive interactions . These off-pathway reactions were detrimental to cell growth; wild-type growth was restored by co-expression of PapD . Subunit misfolding in the absence of PapD was sensed by two parallel pathways: the Cpx two-component signaling system and the sigma E modulatory pathway. J Appl Microbiol, 1997 Oct, 83(4), 456 - 63 A selective medium for the rapid detection by an impedance technique of Pseudomonas spp . associated with poultry meat; Salvat G et al.; A new medium for detecting and enumerating Pseudomonas spp . associated with poultry meat spoilage by a rapid impedance technique was developed, after testing potential growth promoters for eight Pseudomonas strains and inhibitors against eight competing strains (Enterobacteriaceae) able to grow on the medium of Mead and Adams (1977) . Four basal media (brain heart infusion, brucella broth, Shaedler broth and Whitley impedance broth (WIB)) and a synthetic medium were evaluated . Whitley impedance broth was the best basal medium for detecting variations in impedance in relation to Pseudomonas growth . The efficiency of WIB was improved by adding compounds which enhanced the growth of Pseudomonas on the synthetic medium . Among the incubation temperatures tested, 22 degrees C proved to be the best compromise between growth of Pseudomonas associated with poultry meat spoilage and inhibition of competitors . Among the 15 inhibitory substances evaluated against Pseudomonas competitors, five were chosen for inclusion in the final medium: metronidazole, carbenicilline, cetrimide, cycloheximide and diamide (MCCCD medium) . Preliminary results obtained from experiments with beef and pork meat showed that this medium could also be used without diamide and at an incubation temperature of 25 degrees C . The impedance technique using MCCCD medium was then compared with an official method which uses the medium of Mead and Adams (1977) on 106 samples of poultry neck skin . The linear regression coefficient between the two techniques was approximately r = 0.85 . Impedance was able to detect 10(3) Pseudomonas g-1 within less than 19 h making it a promising technique for predicting poultry meat spoilage. J Appl Microbiol, 1997 Sep, 83(3), 367 - 74 Growth of enterobacteria on fructo-oligosaccharides; Hartemink R et al.; Fructo-oligosaccharides (FOS) can be fermented by most species of enterobacteria present in the human intestine . Fermentation was confirmed by increased growth rates, low final pH and degradation patterns using high performance anion exchange chromatography (HPAEC) . Growth rates were increased when FOS was added to the growth medium . Growth rates on all substrates differed widely between strains within the same species . HPAEC analysis showed that each strain degraded the oligosaccharides differently, but a preference for the smaller oligosaccharides was observed . No differences were observed between the two commercial preparations, the inulo-oligosaccharides and neosugars . Fermentation was rapid as could be determined by acidification tests using cell suspensions . It can be concluded that enterobacteria may play a role in overall fermentation of FOS in the colon and, in addition, due to competitive exclusion, may prevent survival of ingested pathogenic enterobacteria. Trends Microbiol, 1997 Oct, 5(10), 389 - 94 Origins of the mobile gene cassettes found in integrons; Recchia GD et al.; Many of the acquired antibiotic resistance genes found in enterobacteria and pseudomonads are part of small mobile elements known as gene cassettes, and other genes are also likely to be found in cassettes . The origins of the genes and the recombination sites that make up cassettes are not known, but recent analyses of available data suggest that cassettes may be ancient structures, and some hypotheses for how they are formed can now be examined. J Clin Microbiol, 1997 Nov, 35(11), 2773 - 7 Evaluation of BBL CHROMagar orientation medium for detection and presumptive identification of urinary tract pathogens; Hengstler KA et al.; The microbiological performance of BBL CHROMagar Orientation medium and CPS ID2 agar was compared to that of Columbia agar with 5% sheep blood and MacConkey agar without crystal violet for the enumeration and presumptive identification of bacteria responsible for urinary tract infections . Of a total of 658 clinical urine specimens, 118 specimens yielded no growth, 402 specimens yielded growth with cell counts of > or = 10(5) CFU/ml, and 138 specimens yielded growth with cell counts of < 10(5) CFU/ml . Of the specimens with cell counts of > or = 10(5) CFU/ml, 163 were pure cultures and 239 were mixed cultures . A total of 266 Escherichia coli organisms were isolated on both chromogenic media, 260 were isolated on blood agar, and 248 were isolated on MacConkey agar . One strain (0.4%) failed to develop the expected pink color on CHROMagar Orientation medium, and 23 strains (8.7%) failed to develop the expected pink color on CPS ID2 agar . Enterococci (CHROMagar Orientation medium, n = 266; CPS ID2 agar, n = 265) produced small blue-green colonies on both chromogenic media . Fifty of the mixed cultures contained enterococci that were detected only on the chromogenic media . The Klebsiella-Enterobacter-Serratia (KES) and the Proteus-Morganella-Providencia (PMP) groups could be identified on both chromogenic media . Of 66 isolates of the KES group, 63 grew with the expected color on CHROMagar Orientation medium and 58 of 64 isolates grew with the expected color on CPS ID2 agar . Other microorganisms required further identification . The use of chromogenic medium formulations offers a time-saving method for the reliable detection, enumeration, and presumptive identification of urinary tract pathogens . One of the greatest advantages of these media is the easy recognition of mixed cultures. Ann Biol Clin (Paris), 1997 Sep-Oct, 55(5), 465 - 9 {Molecular typing by pulsed field gel electrophoresis of Enterobacter cloacae strains isolated from osteoarticular infections at the Nancy University Hospital 1990-1994}; Simeon D et al.; Enterobacteriaceae represent more than 11% of bacteria involved in osteoarticular infections in adult and, among them, Enterobacter cloacae is found in 12% of the cases . The increased evolution of the antibiotic resistance rate of this species is worrying . However, all isolates responsible for this type of infection at the University Hospital of Nancy from 1990 to 1994 (24 strains isolated from 22 patients presented an identical antibiotype with especially a natural resistance phenotype to beta-lactam antibiotics except one cephalosporinase-over-producing strain . The DNA of these strains was studied by pulse-field gel electrophoresis after digestion by the restriction enzyme XbuI . The great genomic diversity obtained showed that the stability of the antibiotic susceptibility during the period studied was not due to the existence of unique clone but to that of multiple clones . The analysis of the restriction profiles has permitted to achieve a better differenciation of the strains than biotyping and antibiotyping, which confirms the high discrimination power of this genotypic method. Biochem Biophys Res Commun, 1997 Oct 9, 239(1), 240 - 6 Synthetic genes specifying periodic polymers modelled on the repetitive domain of wheat gliadins: conception and expression; Elmorjani K et al.; In order to optimise new polypeptide based biomaterials, we developed a procedure for producing homoblock polypeptides using recombinant DNA technology . Synthetic genes encoding periodic polypeptides modelled on the consensus sequence of wheat gliadins (a family of wheat storage proteins) were devised to be expressed in Escherichia coli . The construction strategy followed allows the construction of three genes encoding 8, 16, and 32 copies of the PQQPY module . The optimal expression conditions in the enterobacteria were established and a convenient purification procedure was shown to be useful in recovery of sizable amounts of strictly periodic polypeptides . The identities of the synthesized polypeptides were assessed using positive cross reactions to antibodies raised against a synthetic decapeptide (PQQPYPQQPA) and amino acid composition was determined as well. J Am Vet Med Assoc, 1997 Oct 15, 211(8), 1029 - 35 Number of viable bacteria and presumptive antibiotic residues in milk fed to calves on commercial dairies; Selim SA et al.; OBJECTIVE: To assess the number of bacteria and presumptive antibiotic residues in milk fed to calves and to identify those bacteria and the antibiotic susceptibility of selected bacterial strains . DESIGN: Cross-sectional prospective study . SAMPLE POPULATION: 189 samples obtained from 12 local dairies . PROCEDURE: Samples of waste milk and milk-based fluids (eg, milk replacer, colostrum, bulk-tank milk) were obtained . Cumulative number of viable bacteria was determined . Bacteria were cultured aerobically, and antibiotic susceptibility testing of selected strains was performed . Presumptive antibiotic residues were detected by use of test kits . RESULTS: Geometric mean of the cumulative number of bacteria for waste milk samples was significantly higher than for other types of milk or milk-based products . Streptococcus sp (84/165 samples) and Enterobacteriaceae (83/165 samples) were the predominant bacteria identified, followed by Staphylococcus sp (68/165 samples) . Escherichia coli was the gram-negative species most commonly isolated (52/165 samples; 32%); however, none were strain O157 . Salmonella sp or Mycoplasma sp were not isolated . Of 189 samples, 119 (63%) were positive when tested for beta-lactams or tetracycline by use of 2 commercially available assays . In vitro, some bacteria were resistant to commonly used antibiotics . CLINICAL IMPLICATIONS: Waste milk that has not been effectively treated (eg, pasteurization) to reduce microbial load prior to use as calf feed should be used with caution, because it may contain a high number of bacteria that may be pathogenic to cattle and human beings . Antibiotic residues that would constitute violative amounts and existence of multiple antibiotic resistant bacterial strains are concerns in calf health management and dairy food safety. Zh Mikrobiol Epidemiol Immunobiol, 1997 Jul-Aug, (4), 89 - 92 {The effect of therapeutic mud on the viability and persistence properties of bacteria}; Abdrakhmanov AR et al.; The influence of therapeutic salt mud on the viability and some biological properties of bacteria, responsible for their survival in the macroorganisms, was shown . Therapeutic mud had low bactericidal properties, and enterobacteria were, on the whole, even less sensitive to these properties than staphylococci . Therapeutic mud inhibited the capacity of bacteria for inactivating complement, lysozyme and the bactericidal component of the preparation of interferon and also reduced the hydrophobic properties of bacterial cells . At the same time Escherichia were found to be more susceptible to the modifying action of the mud than staphylococci . The greatest effect on the hydrophobic properties and anticomplement activity of bacteria was observed after their incubation in mud solution. J Bacteriol, 1997 Oct, 179(19), 6192 - 5 Isolation and characterization of Enterobacter cloacae mutants which are defective in chemotaxis toward inorganic phosphate; Kusaka K et al.; Enterobacter cloacae IFO3320 is attracted to Pi when cells are starved for Pi . Two Tn1737KH-induced mutants, which were constitutive for alkaline phosphatase, failed to exhibit Pi taxis even under conditions of Pi limitation . Both of the mutant strains exhibited normal chemotactic responses to peptone, suggesting that they are specifically defective in Pi taxis . Cloning and sequence analysis showed that the TN1737KH insertions were located in either the pstA or pstB genes which encode the channel-forming proteins of the Pi-specific transport (Pst) system in E . cloacae . These results suggest that the E . cloacae Pst system is required for Pi chemoreception. J Antimicrob Chemother, 1997 Sep, 40(3), 393 - 9 Plasmid-encoded fosfomycin resistance in bacteria isolated from the urinary tract in a multicentre survey; Arca P et al.; Sixty out of 219 fosfomycin-resistant bacteria selected from more than 7400 urinary pathogens in an epidemiological multicentre survey performed in Italy were screened for plasmid genes fosA and fosB conferring fosfomycin resistance . Only five strains, three enterobacteria and two staphylococci, carried plasmids harbouring, respectively, fosA and fosB genes . Fosfomycin resistance in the other isolates was caused by an alteration of the chromosomally encoded GlpT transport system . One strain, Morganella morganii 279, incorporated alpha-glycerolphosphate and its mechanism of fosfomycin resistance needs to be further investigated . Our study showed that PCR amplification is the most accurate, simple and rapid method for epidemiological studies of plasmid-encoded fosfomycin resistance, and that fosfomycin resistance conferred by plasmid genes (both fosA and fosB) accounts for only a low percentage of the fosfomycin-resistant strains. Nature, 1997 Oct 9, 389(6651), 636 - 9 Survival of FimH-expressing enterobacteria in macrophages relies on glycolipid traffic; Baorto DM et al.; Strains of Escherichia coli persist within the human gut as normal commensals, but are frequent pathogens and can cause recurrent infection . Here we show that, in contrast to E . coli subjected to opsonic interactions stimulated by the host's immune response, E . coli that bind to the macrophage surface exclusively through the bacterial lectin FimH can survive inside the cell following phagocytosis . This viability is largely due to the attenuation of intracellular free-radical release and of phagosome acidification during FimH-mediated internalization, both of which are triggered by antibody-mediated internalization . This different processing of non-opsonized bacteria is supported by morphological evidence of tight-fitting phagosomes compared with looser, antibody-mediated phagosomes . We propose that non-opsonized FimH-expressing E . coli co-opt internalization of lipid-rich microdomains following binding to the FimH receptor, the glycosylphosphatidylinositol-linked protein CD48, because (1) the sterol-binding agents filipin, nystatin and methyl beta-cyclodextrin specifically block FimH-mediated internalization; (2) CD48 and the protein caveolin both accumulate on macrophage membranes surrounding bacteria; and (3) antibodies against CD48 inhibit FimH-mediated internalization . Our findings bring the traditionally extracellular E . coli into the realm of opportunistic intracellular parasitism and suggest how opportunistic infections with FimH-expressing enterobacteria could occur in a setting deprived of opsonizing antibodies. Electrophoresis, 1997 Aug, 18(9), 1512 - 8 Amplification of anonymous DNA fragments using pairs of long primers generates reproducible DNA fingerprints that are sensitive to genetic variation; Gillings M et al.; The reproducibility and potential applications of anonymous amplification protocols can be improved by using pairs of primers, each of 18 to 24 bases, to replace the single 8 to 10 base primers normally used in randomly amplified polymorphic DNA (RAPD) or DNA amplification fingerprinting (DAF) methods . Amplification using large primer pairs (LP-RAPD) generates 5 to 30 bands that can be resolved on standard agarose gels . Complex fingerprints can be readily generated from viruses, bacteria, fungi, plants, invertebrates and vertebrates . We also present evidence that a number of polymerase chain reaction (PCR) methods, including those based on the use of enterobacterial repetitive intergenic consensus (ERIC-PCR) or microsatellite primed (MP-PCR) sequence, may in essence operate by the same mechanism as LP-RAPD . Using standard LP-RAPD protocols, reproducible fingerprints can be generated from a single specimen using different thermocyclers, regardless of the mechanism used for thermocycling (air-cooled, Peltier effect, or robotic arm) . LP-RAPD is sensitive to intraspecific and interspecific genetic variation, demonstrated here by analysis of mites and apple cultivars . Approximately 50% of LP-RAPD products are expected to have different primers at either end . Polymorphic bands with this arrangement can be recovered from the gel and directly sequenced using the LP-RAPD primers themselves . The efficiency of sequencing is improved by the length of the LP-RAPD primers . This method has the potential to allow the production of allele-specific species markers in less than two days. Int J Syst Bacteriol, 1997 Oct, 47(4), 1253 - 4 Phylogenetic relationships of Salmonella typhi and Salmonella typhimurium based on 16S rRNA sequence analysis; Chang HR et al.; The 16S rRNA gene sequences of Salmonella typhi and Salmonella typhimurium were amplified by PCR, cloned, and sequenced . These sequences were analyzed by comparison with reference organisms from the family Enterobacteriaceae . Both S . typhi and S . typhimurium belong to the gamma subdivision of the class Proteobacteria. Rev Assoc Med Bras, 1997 Apr-Jun, 43(2), 137 - 44 {Evaluation of in vitro activity of new fluoroquinolones, cephalosporins and carbapenems against 569 gram-negative bacteria}; Gales AC et al.; OBJECTIVE: Evaluation of the in vitro activity of new fluoroquinolones, cephalosporins and carbapenems against gram-negative bacteria . MATERIAL AND METHOD: A total of 569 clinical isolates were obtained from inpatients at Sao Paulo Hospital--UNIFESP/EPM in June and July of 1992 . The species distribution was as follows: Enterobacter sp . (62), Escherichia coli (308), Klebsiella pneumoniae (27), Klebsiella sp . (9), Proteus mirabilis (23), Pseudomonas aeruginosa (88), Pseudomonas sp . (4), Serratia sp . (30) and other gram-negatives (7) . Susceptibility tests were performed by broth microdilution . The antimicrobials agents tested were: ciprofloxacin, ofloxacin, levofloxacin, grepafloxacin, DU 6859-alpha, ceftazidime, cefepime, FK 037, imipenem, meropenem and biapenem . RESULTS: DU 6859-alpha showed the highest anti-microbial activity among the fluoroquinolones . It was two- to four-fold more active than ciprofloxacin against some species . The potency and antimicrobial spectrum were similar between the fourth-generation cephalosporins against Enterobacteriaceae, except for Enterobacter sp . strains which were more susceptible to cefepime than they were to cefetazidime or FK 037 . When testing Pseudomonas aeruginosa, ceftazidime was slightly more active than the other cephalosporins . Against Enterobacteriaceae and Pseudomonas aeruginosa strains, meropenem was more active than imipenem or biapenem . In addition, the percentage of strains, susceptible to meropenem was higher than the percentage susceptible to the other cerbapenems against these species . CONCLUSION: The new antimicrobial agents demonstrated in vitro activity higher than that of agents commercially available . However, more studies are necessary to further evaluate the in vivo activity and the clinical benefit of these compounds. Antimicrob Agents Chemother, 1997 Oct, 41(10), 2312 - 6 Comparative in vitro activities of trovafloxacin (CP-99,219) against 221 aerobic and 217 anaerobic bacteria isolated from patients with intra-abdominal infections; Citron DM et al.; Four hundred thirty-eight bacteria cultured from specimens of patients with serious intra-abdominal infections were tested by agar dilution against trovafloxacin and other quinolones and antimicrobial agents . Trovafloxacin inhibited 435 strains (99.3%) at < or =2 microg/ml . All the quinolones had similar activities against Enterobacteriaceae and Pseudomonas sp., but trovafloxacin showed superior activities against streptococci, enterococci, and anaerobic organisms . Because of its excellent in vitro activities against diverse bacteria, trovafloxacin has potential use as a single agent for polymicrobial infections. Antimicrob Agents Chemother, 1997 Oct, 41(10), 2113 - 20 The signal molecule for beta-lactamase induction in Enterobacter cloacae is the anhydromuramyl-pentapeptide; Dietz H et al.; Beta-lactamase induction in Enterobacter cloacae, which is linked to peptidoglycan recycling, was investigated by high-performance liquid chromatographic analysis of cell wall fragments in genetically defined cells of Escherichia coli . After treatment of cells with beta-lactams, we detected an increase in a D-tripeptide (disaccharide-tripeptide, N-acetylglucosaminyl-1,6-anhydro-N-acetylmuramyl-L-alanyl-D-glutamyl-mes o-diaminopimelic acid), aD-tetrapeptide (disaccharide-tetrapeptide, N-acetylglucosaminyl-1,6-anhydro-N-acetylmuramyl-L-alanyl-D-glutamyl-mes o-diaminopimelic acid-D-alanine), and aD-pentapeptide (disaccharide-pentapeptide, N-acetylglucosaminyl-1,6-anhydro-N-acetylmuramyl-L-alanyl-D-glutamyl-mes o-diaminopimelic acid-D-alanyl-D-alanine)levels in the periplasms of bacterial cells . Furthermore, only the accumulation of aD-pentapeptide correlates with the beta-lactamase-inducing capacity of the beta-lactam antibiotic . The transmembrane protein AmpG transports all three aD-peptides into the cytoplasm, where they are degraded into the corresponding monosaccharide peptides . In the absence of AmpD the constitutive overproduction of beta-lactamase is accompanied by an accumulation of aM-tripeptide (monosaccharide-tripeptide, anhydro-N-acetylmuramyl-L-alanyl-D-glutamyl-meso-diaminopimelic acid) and aM-pentapeptide (L1,6-anhydro-N-acetylmuramyl-L-alanyl-D-glutamyl-meso-diaminopimelic acid-D-alanyl-D-alanine), but not aM-tetrapeptide (anhydro-N-acetylmuramyl-L-alanyl-D-glutamyl-meso-diaminopimelic acid-D-alanine), in the cytoplasm . Only the amount of aM-pentapeptide is increased upon treatment with imipenem . These findings indicate that aD-pentapeptide is the main periplasmic muropeptide, which is converted into the cytoplasmic signal molecule for beta-lactamase induction, the aM-pentapeptide. Eur J Cardiothorac Surg, 1997 Sep, 12(3), 443 - 9 Mediastinitis after aorto-coronary bypass surgery; Antunes PE et al.; OBJECTIVES: To identify risk factors in 60 cases of mediastinitis amongst 2512 patients (2.3%) subjected to isolated coronary bypass surgery from March 1988 through December 1995, treated by a closed irrigation/drainage system . PATIENTS AND METHODS: The mean age of the 60 patients was 56.9 +/- 6.8 years (45-81 years) and 55 (91.6%) were male . Early mediastinal reexploration was performed in all cases immediately after the diagnosis of mediastinitis, with debridement of necrosed tissues, followed by implantation of a closed-circuit irrigation system of the mediastinum constituted by irrigation catheter and drain, closure of the sternum and skin, and specific systemic antibiotic therapy . The mean interval between the original surgery and reexploration was 9.4 days (range 6-14 days) . No patient required more extensive procedures, namely omental or muscular flaps . Twenty potential risk factors in patients with mediastinitis, including diabetes mellitus, obesity, coexistence of peripheral vascular disease, decreased LV function, use of inotropes, mediastinal blood drainage and utilization of double IMA, were compared with the group without mediastinitis . RESULTS: Mean cardiopulmonary bypass time was 74.1 +/- 8.1 min, anesthetic time 3.5 +/- 0.8 h and postoperative mechanical ventilation 18 +/- 3 h . A total of 23 patients (38.3%) received one IMA and 35 (58.3%) two IMAs . In the postoperative period, 7 of the 60 patients (11.6%) had required inotropes because of low output . Mediastinal blood loss was 1112cc +/- 452cc and 9 patients (15%) were transfused . Cultures were positive in 40 cases (66.6%) and the most frequent infecting agent was the Staph . epidermidis in 25 cases (62.5%), followed by Candida albicans and Enterobacter and Serratia species (7.5% each); 1 patient (1.7%) died and 9 (15%) had renal failure . The irrigation/drainage was maintained for a mean of 9.1 days (5-83 days) . Patients with mediastinitis had a significantly higher prevalence of diabetes (41.6% vs . 18.8%; P < 0.01), obesity (48.3% vs . 15.2%; P < 0.001), peripheral vascular disease (11.6% vs . 4.0%; P < 0.05), but a lower incidence of poor LV function (18.3% vs . 32.7%; P < 0.05) . A double IMA was used more frequently in patients who had mediastinitis (58.3% vs . 23.5%; P < 0.001) CONCLUSIONS: Diabetes mellitus, obesity, co-existence of peripheral vascular disease and use of double IMA are risk factors for mediastinitis after coronary artery surgery . The efficacy of the closed method of treatment with a mediastinal irrigation/drainage system was increased with early diagnosis and reintervention. Clin Infect Dis, 1997 Aug, 25(2), 318 - 20 Incidence and epidemiology of nosocomial infections in patients infected with human immunodeficiency virus; Frank U et al.; In a prospective study, we investigated the incidence, characteristics, and risk factors of nosocomial infections (NIs) in patients with human immunodeficiency virus disease . There was a total of 528 admissions of 405 eligible patients; 46 NIs (8.7% per discharge) were identified in 39 patients . The proportional frequencies of NIs were as follows: 16 skin and/or soft-tissue infections (including localized catheter-associated infections), 3.0%; 14 respiratory tract infections, 2.7%; 11 bloodstream infections, 2.1%; and 5 urinary tract infections, 0.9% . The most common etiologic agents were Staphylococcus aureus (27.6%), Pseudomonas aeruginosa (13.8%), and Enterobacter cloacae (13.8%) . The duration of hospitalization was not significantly prolonged by NI in the cohort. Am J Med Sci, 1997 Oct, 314(4), 245 - 9 Urinary tract infection: an overview; Barnett BJ et al.; Urinary tract infection (UTI) remains very common . As many as 50% of women report having had at least one UTI in their lifetimes . Urinary tract infection is the most common cause of infection in nursing home residents and the most common source of bacteremia in the elderly population . Urinary tract infection occurs in patients with structurally or functionally abnormal urinary tracts (complicated UTI) and in patients with anatomically normal urinary tracts (uncomplicated UTI) . Escherichia coli (E coli) is the most common cause of uncomplicated UTI, whereas antibiotic-resistant Enterobacteriaceae, enterococci, and Candida species often are the causes of complicated UTI . In this article we review current concepts of the epidemiology, microbiology, pathophysiology, clinical manifestations, diagnosis, and treatment of urinary tract infection. Harefuah, 1997 Jul, 133(1-2), 1 - 2, 80 {Bacterial culture of chip tissue of enucleated prostates}; Ben-Zion IZ et al.; To assess the prevalence of infection and colonization of the prostate by bacteria, chip tissue samples from 166 patients undergoing retropubic prostatectomy were submitted for bacterial tissue culture . In 28 patients with an indwelling catheter before surgery, E . coli, Klebsiella, Pseudomonas and Enterobacter were the commonest species encountered, the first the most common . In only 7 patients (20%) who didn't have an indwelling catheter before operation was the culture positive . We confirmed that the longer the time the catheter was indwelling before surgery, the greater the likelihood of positive cultures . However, postoperative outcome and morbidity were not related to culture results . We conclude that even though it is worth trying to sterilize the urine and prostate before prostatectomy, the effect on the postoperative outcome is minimal when proper antimicrobial therapy is given perioperatively. Acta Microbiol Immunol Hung, 1997, 44(2), 165 - 71 In vitro antimicrobial properties of azidothymidine (AZT); Monno R et al.; In addition to the activity against a number of retroviruses, azidothymidine (AZT) has antibacterial activity against many bacteria . The effect of AZT on 224 bacterial species, including 25 strains of Salmonella spp . isolated from HIV-positive patients, was tested . AZT had no activity against all the strains of tested Gram-positive bacteria and Pseudomonas species (MIC > 128 micrograms/ml), whereas a different activity against Enterobacteriaceae (MIC range, 128 to 0.06 micrograms/ml) was found . In particular 76% of Salmonella spp . isolated from HIV-positive patients showed MICs > 1 microgram/ml, whereas similar MICs value were found in 50% of the Salmonella strains isolated from HIV-negative subjects . In addition, strains of Salmonella isolated from stools were more resistant to AZT when compared to strains isolated from blood even if this difference was not statistically significant . No correlation was found between length of therapy and Salmonella resistance to AZT in HIV-positive patients and a low incidence of Salmonella relapses in subjects treated with AZT was observed . The possibility that AZT may have an ancillary benefit in controlling some bacterial infections in AIDS patients is discussed. FEMS Immunol Med Microbiol, 1997 Sep, 19(1), 33 - 45 Tracking of clinical and environmental Vibrio cholerae O1 strains by combined analysis of the presence of toxin cassette, plasmid content and ERIC PCR; Colombo MM et al.; Clinical and environmental Vibrio cholerae O1 strains associated with the cholera epidemic in the Luanda province of Angola from 1991 to 1994 were tracked by toxin distribution, plasmid content and chromosomal polymorphism of the enterobacterial repetitive intergenic consensus (ERIC) sequences by PCR fingerprinting . To follow the distribution of ace, zot and ctxA toxin genes, 6 specific PCR tests were applied to 100 Vibrio strains, after preliminary hybridization experiments . Clinical isolates of Vibrio cholerae O1 were characterized by high stability of the toxigenic cassette and the presence of a large conjugative multi-resistant plasmid of incompatibility class C . Such characteristics were present in all isolates during the four years of the epidemic . Environmental strains, isolated from the river supplying water to the Luanda population showed three different genetic profiles: the presence of both cassette and plasmid, the presence of cassette only or absence of both . To assess the clonal relationship between the clinical isolates and the three groups of environmental strains, the strains were analyzed by PCR ERIC polymorphism . This analysis, supported by the toxin and plasmid content, suggested the stability of the epidemic strain in clinical cases during the epidemic and led to the finding that there was a strict genetic relationship of the epidemic strain with the environmental ones as characterized by the presence of the toxin cassette . The role of the water supply from Bengo River as a reservoir of the Vibrio epidemic strain is discussed. J Hosp Infect, 1997 Sep, 37(1), 25 - 37 Role of quantitative cultures and microscopic examinations of endotracheal aspirates in the diagnosis of pulmonary infections in ventilated patients; Albert S et al.; Endotracheal aspirates (EA) from 20 intubated patients in a surgical intensive care unit (mean ventilation time/patient = 16.5 days) were investigated serially by performing quantitative cultures using growth of 10(5) cfu/mL as a cut-off point . Microscopic examinations were made using Giemsa's stain for polymorphonuclear neutrophils (PMN) . The spectrum of pathogens encountered was determined and compared with clinical data to distinguish colonization from infection of the lower respiratory tract . Out of 301 EA cultures, 156 (51.8%) were positive and 145 (48.2%) were below the cut-off point . Counts of PMN were significantly higher in samples which gave positive cultures . Seventy-five different bacterial strains were isolated (64% were Gram-negative bacilli) . Seventeen patients (85%) were colonized with Gram-negative bacteria . Nine patients (45%) developed nosocomial pneumonia (NP), five (25%) had no signs of pneumonia, and six (30%) had an uncertain status . Main causative agents for NP were Pseudomonas aeruginosa, Enterobacteriaceae and Staphylococcus aureus . Quantitative EA cultures had a sensitivity of 81.5%, a specificity of 64.8%, a positive predictive value of 55% and a negative predictive value of 87% . Our results suggest that EA quantitative cultures (cut-off value 10(5) cfu/mL), species identification and microscopic examination of EA may help to differentiate tracheobronchial colonization and infection, especially when bronchoscopic techniques are not available. Infect Immun, 1997 Oct, 65(10), 4199 - 206 Specificity of the high-mannose recognition site between Enterobacter cloacae pili adhesin and HT-29 cell membranes; Pan YT et al.; Enterobacter cloacae has been implicated as one of the causative agents in neonatal infection and causes a septicemia thought to be initiated via the gastrointestinal tract . The adhesion of radiolabeled E . cloacae to HT-29 cells was concentration and temperature dependent and was effectively blocked by unlabeled bacteria or by millimolar concentrations of alpha-mannosides and micromolar concentrations of high-mannose oligosaccharides . A variety of well-characterized mannose oligosaccharides were tested as inhibitors of adhesion . The best inhibitor was the Man9(GlcNAc)2-tyrosinamide, which was considerably better than other tyrosinamide-linked oligosaccharides such as Man7(GlcNAc)2, Man6(GlcNAc)2 or Man5(GlcNAc)2 . Further evidence that the bacteria preferred Man9(GlcNAc)2 structures was obtained by growing HT-29 cells in the presence of glycoprotein processing inhibitors that block mannosidase I and increase the amount of protein-bound Man9(GlcNAc)2 at the cell surface . Such cells bound 1.5- to 2-fold more bacteria than did control cells . The adhesin involved in binding to high-mannose structures was purified from isolated pili . On sodium dodecyl sulfate-gels, a 35-kDa protein was identified by its specific binding to a mannose-containing biotinylated albumin . The amino acid sequences of several peptides from the 35-kDa subunit showed over 85% identity to FimH, the mannose-specific adhesin of Salmonella typhimurium . Pili were labeled with 125I and examined for the ability to bind to HT-29 cells . Binding showed saturation kinetics and was inhibited by the addition of Man9(GlcNAc)2-tyrosinamide but not by oligosaccharides with fewer mannose residues . Polyclonal antibody against this 35-kDa protein also effectively blocked adhesion of pili or E . cloacae, but no effect was observed with nonspecific antibody . These studies demonstrate that the 35-kDa pilus subunit is a lectin whose specificity is directed toward Man, (GlcNAc)2 oligosaccharides. J Clin Microbiol, 1997 Oct, 35(10), 2642 - 8 Evaluation of a fluorescence-labelled oligonucleotide probe targeting 23S rRNA for in situ detection of Salmonella serovars in paraffin-embedded tissue sections and their rapid identification in bacterial smears; Nordentoft S et al.; A method for the detection of Salmonella based on fluorescence in situ hybridization (FISH) has been developed and applied for the direct detection of Salmonella in pure cultures and in formalin-fixed, paraffin-embedded tissue sections . On the basis of the 23S rRNA gene sequences representing all of the S . enterica subspecies and S . bongori, an 18-mer oligonucleotide probe was selected . The specificity of the probe was tested by in situ hybridization to bacterial cell smears of pure cultures . Forty-nine of 55 tested Salmonella serovars belonging to subspecies I, II, IIIb, IV, and VI hybridized with the probe . The probe did not hybridize to serovars from subspecies IIIa (S . arizonae) or to S . bongori . No cross-reaction to 64 other strains of the family Enterobacteriaceae or 18 other bacterial strains outside this family was observed . The probe was tested with sections of formalin-fixed, paraffin-embedded tissue from experimentally infected mice or from animals with a history of clinical salmonellosis . In these tissue sections the probe hybridized specifically to Salmonella serovars, allowing for the detection of single bacterial cells . The development of a fluorescence-labelled specific oligonucleotide probe makes the FISH technique a promising tool for the rapid identification of S . enterica in bacterial smears, as well as for the detection of S . enterica in histological tissue sections. Chemosphere, 1997 Oct, 35(7), 1487 - 95 Volatile metabolites from some gram-negative bacteria; Scholler C et al.; A survey of volatile organic compounds (VOCs) excreted from various Gram-negative bacteria (Pseudomonas spp., Serratia spp . and Enterobacter spp.) was carried out . Compounds were identified by gas chromatography-mass spectrometry . VOCs identified included dimethyl disulphide, dimethyl trisulphide and isoprene. FEMS Microbiol Lett, 1997 Sep 15, 154(2), 245 - 50 Induction of gram-negative bacterial growth by neurochemical containing banana (Musa x paradisiaca) extracts; Lyte M; Bananas contain large quantities of neurochemicals . Extracts from the peel and pulp of bananas in increasing stages of ripening were prepared and evaluated for their ability to modulate the growth of non-pathogenic and pathogenic bacteria . Extracts from the peel, and to a much lesser degree the pulp, increased the growth of Gram-negative bacterial strains Escherichia coli O157:H7, Shigella flexneri, Enterobacter cloacae and Salmonella typhimurium, as well as two non-pathogenic E . coli strains, in direct relation to the content of norepinephrine and dopamine, but not serotonin . The growth of Gram-positive bacteria was not altered by any of the extracts . Supplementation of vehicle and pulp cultures with norepinephrine or dopamine yielded growth equivalent to peel cultures . Total organic analysis of extracts further demonstrated that the differential effects of peel and pulp on bacterial growth was not nutritionally based, but due to norepinephrine and dopamine . These results suggest that neurochemicals contained within foodstuffs may influence the growth of pathogenic and indigenous bacteria through direct neurochemical-bacterial interactions. Int J Food Microbiol, 1997 Jul 22, 37(2-3), 225 - 9 Effect of nitrate and nitrite curing salts on microbial changes and sensory quality of rapid ripened sausages; Sanz Y et al.; The effect of the use of either nitrite or nitrate curing salts on microbial changes and sensory quality in rapid ripened sausages inoculated with a mixed starter culture (Lactobacillus sake and Staphylococcus carnosus) was investigated . Lactic acid bacteria and Micrococcaceae were not greatly affected by the added curing salt . Conversely, the inhibition exerted by nitrite on the undesirable flora (Enterobacteriaceae and psychrotrophs) was evident from the early stages of the processing keeping highly significant differences (P < 0.01) with respect to nitrate made sausages till the end of the ripening stage . The use of nitrite in sausage processing was found to reduce hygienic risks. Behring Inst Mitt, 1997 Feb, (98), 103 - 13 The Escherichia coli hemolysin secretion apparatus--a versatile antigen delivery system in attenuated Salmonella; Gentschev I et al.; The E . coli hemolysin (HlyA) secretion apparatus represents a type I secretion system that is fully functional in Salmonella . The system which consists of the two specific membrane proteins HlyB and HlyD and the outer membrane protein TolC, recognizes on HlyA a C-terminally located signal sequence of about 60 amino acids . Fusion proteins to which this signal sequence is covalently linked at the C-terminus are also recognized by this secretion apparatus . The efficiency of secretion is dependent on the rate of folding of the reporter protein . Secretion-competent regions of a given reporter protein that is not secretable as entire protein can be screened by a recently constructed transposon TnhlyAs which allows the insertion of the secretion signal into any region of the reporter protein . The genetic information for antigens of any source ranging in size between 10 and 1000 amino acids can be easily inserted into a recently constructed secretion vector which will allow the secretion of the fused antigen(s) in attenuated Salmonella typhimurium strains and in other attenuated Enterobacteriaceae . By manipulation of the Hly secretion system the antigen can be either completely secreted into the environment, fixed on the outer membrane or arrested in the cytoplasm of the used carrier strain . By the use of appropriate attenuated Salmonella strains the antigen is delivered in isolated compartments or to the cytosolic compartment . The extracellular delivery of such antigens is also possible with the help of appropriate carrier strains . The immunological consequences of the different display of the processed antigen will be discussed in the paper by Hess et al in this volume . With a similar antigen delivery system the easy identification and molecular characterization of unknown antigens recognized by the immune system in an infection is also feasible. Can J Microbiol, 1997 Aug, 43(8), 770 - 3 Nematophin, a novel antimicrobial substance produced by Xenorhabdus nematophilus (Enterobactereaceae); Li J et al.; A new antibiotic, nematophin, was isolated from strain BC1 of Xenorhabdus nematophilus and detected in all strains of X . nematophilus studied . Its structure is fully established as 3-indoleethyl (3'-methyl-2'-oxo)pentanamide by extensive spectroscopic study . The production of nematophin is affected by the strain type and culture conditions . The compound shows strong in vitro bioactivity against a series of fungal and bacterial species. Can J Microbiol, 1997 Aug, 43(8), 729 - 33 Purification and characterization of a cytotoxin from Enterobacter cloacae; Barnes AI et al.; Leukotoxic activity was assayed in clinical isolates of Enterobacter cloacae . Two strains were selected out of 38 by their greater hemolytic activity in blood agar plates . Leukotoxin was purified by salt precipitation, dialysis, chromatography by gel filtration, and high pressure liquid chromatography (HPLC) . Human leukocytes, when incubated with purified E . cloacae toxin, showed high percentages of death and lysis, with time and dose dependence . The chromatographic profile of gel filtration presented three protein peaks and toxic activity was detected in the second peak . After HPLC, leukotoxin coeluted with the hemolytic activity and both activities were detected only after 2-mercaptoethanol treatment . Coomassie-stained sodium dodecyl sulfate--polyacrylamide gels showed a single band . This band was estimated to represent a protein of 13300 Da on the basis of both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography. Nephrologie, 1997, 18(3), 95 - 101 {Pefloxacin as a first-line treatment for nephrotic syndrome in minimal glomerular lesions in the adult . Multicenter study of 32 patients}; Pruna A et al.; Minimal change nephrotic syndrome (MCNS) is the most frequent single cause of nephrotic syndrome occurring both in adults and children . Although it appears to be a self-limiting disorder (10% spontaneous remissions within the fortnight following the initial flare), MCNS displays a high rate of complications during the nephrotic period (10 to 15% cases) and prompts one to treat patients as early as possible . Corticosteroids are currently used as first-line treatment . A 16 weeks full-dose steroid course (1 mg/kg/day) usually induces remission in 75% MCNS in adults . Nevertheless, duration of treatment (9 months) and occurrence of relapses despite a slowly tapering dosage schedule, expose patients to steroids side-effects . Immunosuppressive drugs are recommended in case of steroid resistance and their side-effects are not harmless . Therefore, an alternative to steroids or immunosuppressives would lend a serious helping hand in MCNS management . The present work is dealing with pefloxacin efficacy in 40% MCNS in adults . Thirty-two MCNS adult patients were treated in a national multicenter study . A short-duration pefloxacin course (4 to 6 weeks) allowed partial or complete remission in 13 out of 32 cases . Thus far, this effect was undescribed for this class of drugs . Pefloxacin belongs to antibacterial agents of the fluoroquinolone family and is active against Gram negative Enterobacteria species . Fluoroquinolones also act on eukaryotic cells as lymphocytes and chondrocytes and alter IL2, gamma IFN and integrin expression . Although their precise mode of action is unknown in this kind of immunological disorder, fluoroquinolones might represent an alternative to steroids in some adult form of MCNS . However, predictive criteria for sensitivity to fluoroquinolones are currently not available and further controlled studies would be helpful using fluoroquinolones as first-line treatment in all the MCNS. Res Microbiol, 1996 Nov-Dec, 147(9), 733 - 7 Biochemical identification of a lipoprotein with maltose-binding activity in the thermoacidophilic Gram-positive bacterium Alicyclobacillus acidocaldarius; Herrmann A et al.; Growth of the thermoacidophilic Gram-positive bacterium Alicyclobacillus acidocaldarius strain ATCC 27009 on maltose resulted in the increased production of a protein with apparent molecular mass of 40 kDa . By metabolic labelling with 14C-palmitic acid, the 40-kDa protein was identified as a lipoprotein . The protein exhibited mal