[Emergence of extended-spectrum beta-lactamases in Pseudomonas aeruginosa: about 24 cases at Rouen University Hospital].

Aim Of The Study: This retrospective study was conducted in order to characterize mechanisms of resistance to beta-lactams and clonality of 24 isolates of Pseudomonas aeruginosa. These strains were isolated from patients hospitalized in Rouen University Hospital between August 2004 and December 2006 and were resistant to cefepime and/or ceftazidime (PMR) by a mechanism not only related to overproduction of the cephalosporinase.

Patients And Methods: Clinical strains of PMR were characterized by conventional biochemical methods, antibiotic susceptibility testing by disk diffusion in agar with or without cloxacillin and RAPD for genetic comparison.

Proteomic analysis of Staphylococcus aureus biofilms grown in vitro on mechanical heart valve leaflets.

The in vitro colonization of three commercial heart valve leaflets by Staphylococcus aureus was investigated. The leaflets, made of pyrolytic carbon alloyed with or without silicon, displayed similar surface properties (wettability, roughness) and were readily colonized by S. aureus that formed patchy biofilms on the three supports.

Biofilm formation on pyrolytic carbon heart valves: influence of surface free energy, roughness, and bacterial species.

Objective: The aim of this study was to analyze the interaction of surface free energy and roughness characteristics of different pyrolytic carbon heart valves with three bacterial species on biofilm formation.

Methods: Three pyrolytic carbon heart valves (St Jude Medical [St Jude Medical Inc, Minneapolis, Minn], Sulzer Carbomedics [CarboMedics Inc, Austin, Tex], and MedicalCV [Medical Incorporated, Inver Grove Heights, Minn]) were tested. Roughness was measured by interferential microscopy and surface free energy by contact angle technique.

Effects of glutamine supplementation on gut barrier, glutathione content and acute phase response in malnourished rats during inflammatory shock.

Aim: To evaluate the effect of glutamine on intestinal mucosa integrity, glutathione stores and acute phase response in protein-depleted rats during an inflammatory shock.

Methods: Plasma acute phase proteins (APP), jejunal APP mRNA levels, liver and jejunal glutathione concentrations were measured before and one, three and seven days after turpentine injection in 4 groups of control, protein-restricted, protein-restricted rats supplemented with glutamine or protein powder. Bacterial translocation in mesenteric lymph nodes and intestinal morphology were also assessed.

Detection of Neisseria meningitidis DNA from skin lesion biopsy using real-time PCR: usefulness in the aetiological diagnosis of purpura fulminans.

Objective: The present study evaluated the usefulness of a real-time polymerase chain reaction (rtPCR) assay for the detection of Neisseria meningitidis (Nm) and genogrouping on skin lesion biopsies in patients with purpura fulminans (PF).

Design: Retrospective single-centre study.

Setting: Adult and paediatric intensive care units at the University Hospital of Rouen.

Tolerance to the glycopeptides vancomycin and teicoplanin in coagulase-negative staphylococci.

Tolerance to vancomycin and teicoplanin in 90 clinical isolates of coagulase-negative staphylococci (CoNS) was investigated by time-kill curve methodology. Only six strains, belonging to the Staphylococcus lugdunensis species, exhibited tolerance. The seven other S.

[Helicobacter pylori prevalence in Raynaud's disease].

Purpose: Recent studies have suggested that the prevalence of Helicobacter pylori may be more frequent in patients with primary Raynaud's phenomenon (PRP) compared to healthy subjects. These data prompted us to conduct this prospective study, in order to assess the prevalence of H. pylori infection in a large series of patients with PRP.

Multilocus sequence analysis and comparative evolution of virulence-associated genes and housekeeping genes of Clostridium difficile.

A multilocus sequence analysis of ten virulence-associated genes was performed to study the genetic relationships between 29 Clostridium difficile isolates of various origins, hosts and clinical presentations, and selected from the main lineages previously defined by multilocus sequence typing (MLST) of housekeeping genes. Colonization-factor-encoding genes (cwp66, cwp84, fbp68, fliC, fliD, groEL and slpA), toxin A and B genes (tcdA and tcdB), and the toxin A and B positive regulator gene (tcdD) were investigated. Binary toxin genes (cdtA and cdtB) were also detected, and internal fragments were sequenced for positive isolates.

[Choice of a strategy for screening for methicillin-resistant Staphylococcus aureus on admission to rehabilitation units].

Objective: To identify a strategy of MRSA screening (methicillin-resistant Staphylococcus aureus) on admission to geriatric rehabilitation units, which would lead to acceptable efficacy and cost compared with a reference maximaliste strategy combining all six sampling sites. Method MRSA screening was conducted prospectively for 3 months in all the patients admitted to a geriatric follow-up and rehabilitation unit, using samples from the nostrils, armpits, urine scars cutaneous ulcers and sores. Six strategies were defined combing different sampling sites.

Multiplex PCR targeting tpi (triose phosphate isomerase), tcdA (Toxin A), and tcdB (Toxin B) genes for toxigenic culture of Clostridium difficile.

A multiplex PCR toxigenic culture approach was designed for simultaneous identification and toxigenic type characterization of Clostridium difficile isolates. Three pairs of primers were designed for the amplification of (i) a species-specific internal fragment of the tpi (triose phosphate isomerase) gene, (ii) an internal fragment of the tcdB (toxin B) gene, and (iii) an internal fragment of the tcdA (toxin A) gene allowing distinction between toxin A-positive, toxin B-positive (A+B+) strains and toxin A-negative, toxin B-positive (A-B+) variant strains. The reliability of the multiplex PCR was established by using a panel of 72 C.

Multilocus sequence typing analysis of human and animal Clostridium difficile isolates of various toxigenic types.

A multilocus sequence typing (MLST) scheme was developed to study the genetic relationships and population structure of 72 Clostridium difficile isolates from various hosts, geographic sources, PCR ribotypes, and toxigenic types (determined by PCR targeting tcdA and tcdB genes). MLST was performed by DNA sequence analysis of seven housekeeping genes (aroE, ddl, dutA, tpi, recA, gmk, and sodA). The number of alleles ranged from five (dutA and ddl) to eleven (recA).

Comparative in vitro and in vivo study of nine alcohol-based handrubs.

Background: Hygienic hand disinfection using alcohol-based handrubs (AHRs) is an alternative method to conventional handwashing in hospital wards. Because a documented choice of such products would consider data from in-care evaluation, we designed a comparative study of 9 AHRs both in vitro and in vivo in actual care conditions.

Methods: The bactericidal activity of AHRs was first measured in vitro against 4 hospital strains exhibiting multiple antibiotic resistance: Acinetobacter baumannii, Staphylococcus aureus, Pseudomonas aeruginosa, and Enterobacter aerogenes.

Genotypic differentiation of twelve Clostridium species by polymorphism analysis of the triosephosphate isomerase (tpi) gene.

Housekeeping genes encoding metabolic enzymes may provide alternative markers to 16S ribosomal DNA (rDNA) for genotypic and phylogenetic characterization of bacterial species. We have developed a PCR-restriction fragment length polymorphism (PCR-RFLP) assay, targeting the triosephosphate isomerase (tpi) gene, which allows the differentiation of twelve pathogenic Clostridium species. Degenerate primers constructed from alignments of tpi sequences of various gram-positive bacteria allowed the amplification of a 501 bp target region in the twelve Clostridium type strains.

Nosocomial urinary tract infections in urologic patients: assessment of a prospective surveillance program including 10,000 patients.

Objective: Hospital-acquired urinary tract infections (HUTI) represent a significant impairment in the quality of health care. Incidence in catheterized patients has been estimated at approximately 20%, however few data are available in urologic patients. We report a prospective surveillance program over 6 years in our urologic department and evaluate its evolution.

Septicemia due to Pasteurella pneumotropica: 16S rRNA sequencing for diagnosis confirmation.

Bacteremia due to Pasteurella pneumotropica occurs infrequently. We report a case of septicemia in a 72-year-old woman who had no underlying illness. The microorganism was isolated from 10 blood cultures and identified by conventional and molecular methods.

In vitro synergistic effects of double and triple combinations of beta-lactams, vancomycin, and netilmicin against methicillin-resistant Staphylococcus aureus strains.

Several studies have previously reported synergistic effects between vancomycin and a given beta-lactam or a given aminoglycoside against methicillin-resistant Staphylococcus aureus (MRSA) strains. The aim of our study was to exhaustively compare the effects of different combinations of a beta-lactam, vancomycin, and/or an aminoglycoside against 32 clinical MRSA strains with different aminoglycoside susceptibility patterns. The effects of 26 different beta-lactam-vancomycin and 8 different aminoglycoside-vancomycin combinations were first studied using a disk diffusion screening method.

Microbiologic contamination study of nebulizers after aerosol therapy in patients with cystic fibrosis.

Background: To evaluate the contamination of delivery systems after an aerosol therapy session in patients with cystic fibrosis who have chronic Pseudomonas aeruginosa infection.

Methods: Fifty-three patients with cystic fibrosis were enrolled in the study from March 1996 to June 1997. All patients were age 7 years or older and had P aeruginosa infection.

Multilocus enzyme analysis in aerobic and anaerobic bacteria using gel electrophoresis-nitrocellulose blotting.

An optimized multilocus enzyme electrophoresis method, which involves polyacrylamide-agarose gel electrophoresis followed by electrophoretic transfers on nitrocellulose sheets, was developed for the analysis of enzyme polymorphism in several aerobic and anaerobic bacterial species including Staphylococcus aureus, Streptococcus pneumoniae, S. agalactiae, Klebsiella pneumoniae and K. oxytoca, Clostridium bifermentans and C.

PCR-Based assay for discrimination between invasive and contaminating Staphylococcus epidermidis strains.

The discrimination between Staphylococcus epidermidis strains that contaminate and infect blood cultures is a daily challenge for clinical laboratories. The results of PCR detection of putative virulence genes were compared for contaminating strains, sepsis-related strains, catheter strains, and saprophytic strains. Multiplex PCR was used to explore the atlE gene, which is involved in initial adherence, the intercellular adhesion gene cluster (ica), which mediates the formation of the biofilm, and the agrA, sarA, and mecA genes, which might contribute to the pathogenicity of S.

Usefulness of pulsed-field gel electrophoresis in assessing nosocomial transmission of pertussis.

During a 2-week period, three infants with a cough lasting at least 8 days with whoops, were admitted to the pediatric unit; Bordetella pertussis was isolated from nasopharyngeal aspirates collected from the three infants. Approximately 1 week later, a nurse working on the same unit developed influenza-like symptoms followed by whooping cough; B pertussis was isolated. Isolates from the nurse and from one of the infants were shown to be indistinguishable by pulsed-field gel electrophoresis.

Motility in the Roux-Y limb after distal gastrectomy: relation to the length of the limb and the afferent duodenojejunal segment--an experimental study.

Following gastrectomy, the longer is a Roux-Y limb constructed to restore digestive continuity the higher the frequency of postoperative symptoms. The aim of this experimental study was to test how the level of the jejunal transection and the length of the Roux limb affect the motility of the constructed limb and in particular the onset and the propagation of activity fronts (AFs). Three months after a distal Roux-en-Y gastrectomy, electromyographic tracings were recorded in six groups of rats grouped according to the level of the transection (20 or 40 cm from the pylorus) and the length of the limb (10, 20 or 30 cm).

Nosocomial meningococcemia in a physician.

We report the case of a pediatrician who developed meningococcal meningitis after performing endotracheal intubation without protection on a child who was suspected of having meningoencephalitis. This case emphasizes the necessity for healthcare workers who perform high-risk procedures to use personal protection devices (i.e.

Molecular analysis of Mycobacterium avium isolates by using pulsed-field gel electrophoresis and PCR.

Genetic relationships among 46 isolates of Mycobacterium avium recovered from 37 patients in a 2,500-bed hospital from 1993 to 1998 were assessed by pulsed-field gel electrophoresis (PFGE) and PCR amplification of genomic sequences located between the repetitive elements IS1245 and IS1311. Each technique enabled the identification of 27 to 32 different patterns among the 46 isolates, confirming that the genetic heterogeneity of M. avium strains is high in a given community.