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G Aubert

Publications and source records attributed to G Aubert.

At least 37 records · Page 2Linked to original sources

Sensitivity and rapidity of blood culture bottles in the detection of cornea organ culture media contamination by bacteria and fungi.

AIMS: To test the bactericidal activity of standard organ culture medium, and to compare the sensitivity and rapidity of blood culture bottles with conventional microbiological methods for detection of bacteria and fungi inoculated in a standard cornea organ culture medium. METHODS: The bactericidal activity of contaminated standard organ culture medium containing 100 IU/ml penicillin, 0.1 mg/ml streptomycin, and 0.25 micro g/ml amphotericin B was evaluated after 48 hours of incubation at 31 degrees C with five inocula of 14 bacteria. Two yeasts (Candida spp) and one Aspergillus were also tested. Contaminated media were then inoculated in three blood bottles (aerobic, anaerobic, fungal) placed in a Bactec 9240 automat; three conventional microbiological broths were the control. Changes in colour of organ culture medium and growth on conventional broth were screened daily by visual inspection. The sensitivity and rapidity of detection of contamination were compared between the three methods: blood bottle, conventional, and visual. RESULTS: Organ culture medium eradicated five bacteria irrespective of the starting inoculums: Streptococcus pneumoniae, Branhamella catarrhalis, Escherichia coli, Propionibacterium acnes, and Haemophilus influenzae. For micro-organisms where the medium was ineffective or bactericidal only (methicillin resistant Staphylococcus aureus, methicillin sensitive Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Pseudomonas aeruginosa, Acinetobacter baumannii, Bacillus subtilis, Klebsiella pneumoniae, Enterococcus faecalis, Candida albicans, Candida kruzei, Aspergillus fumigatus), the blood bottle, conventional, and visual methods detected microbial growth in 100%, 76.5%, and 70% of cases respectively. Mean detection time using blood bottles was 15.1 hours (SD 13.8, range 2-52). In cases of detection by the blood bottle method and the conventional method, the former was always faster: 95.5% against 65.2% detection within 24 hours (p=0.022) respectively. CONCLUSIONS: Blood bottles detect more efficiently and more rapidly a wider range of bacteria and fungi than the conventional microbiological method and the visual inspection of organ culture media.

Bacteria↗

Development of an AFLP-based linkage map and localization of QTLs for seed fatty acid content in condiment mustard (Brassica juncea).

A genetic linkage map of Brassica juncea based on AFLP and RAPD markers was constructed using 131 F1-derived doubled-haploid (DH) plants from a cross between two mustard lines. The map included 273 markers (264 AFLP, 9 RAPD) arranged on 18 linkage groups, and covered a total genetic distance of 1641 cM; 18.3% of the AFLP markers showed a segregation distortion (P < 0.01). The markers with biased segregation were clustered on seven linkage groups. QTLs for oil contents, palmitic acid (16:0), stearic acid (18:0), oleic acid (18:1), linoleic acid (18:2), linolenic acid (18:3), eicosenoic acid (20:1), and erucic acid (22:1), were mapped on the AFLP linkage map. Correlation studies among fatty acids in the DH population and the localization of QTLs involved in their control indicated that a major gene located on linkage group (LG) 2 controlled the elongation step of erucic acid.

Brassica↗

[Use of a pair of blood culture bottles for sterility testing of corneal organ culture media].

PURPOSE: To test the effectiveness and rapidity of a pair of blood culture bottles in the diagnosis of bacterial and fungal contamination of corneal organ culture media. MATERIAL: and methods: Seven hundred and sixty one microbiological analysis of storage media (Inosol(R) and Exosol(R), Opsia, Toulouse, France), sampled in all phases of the organ culture at 31 degrees C of 410 consecutive corneas, were analyzed. Each medium was inoculated in a pair of Bactec Plus Aerobic/F(R) and Bactec Lytic/10 Anaerobic/F(R) blood bottles (Becton Dickinson, Cockeysville, MD) and placed in a Bactec 9240 incubator for 14 days at 37 degrees C and in a Sabouraud broth at 20 degrees C. Changes in color or turbidity of storage media were evaluated daily at the corneal bank. Recipients were screened after graft for signs of infection. RESULTS: The overall contamination rate was 2.4% (18/761). Contamination was detected in less than 1 day in 78% (14/18) and in less than 2 days in 94% (17/18). Positivity of the microbiological controls of starting media preceded medium color changes in 10 out of 14 cases. Bactec blood bottles allowed detection of bacteria as well as Candida sp. yeasts. DISCUSSION: The use of a pair of aerobic and anaerobic blood culture bottles is a simple, effective and rapid method for the diagnosis of a wide range of microbiological contaminations of organ-cultured corneas during banking. CONCLUSION: The validation of this protocol will require a prospective study to compare it with the conventional microbiological method.

Aerobiosis↗

Direct evidence that leukemic cells present HLA-associated immunogenic peptides derived from the BCR-ABL b3a2 fusion protein.

The BCR-ABL oncogene is central in the pathogenesis of chronic myeloid leukemia (CML). Here, tandem nanospray mass spectrometry was used to demonstrate cell surface HLA-associated expression of the BCR-ABL peptide KQSSKALQR on class I-negative CML cells transfected with HLA-A*0301, and on primary CML cells from HLA-A3-positive patients. These patients mounted a cytotoxic T-lymphocyte response to KQSSKALQR that also killed autologous CML cells, and tetramer staining demonstrated the presence of circulating KQSSKALQR-specific T cells. The findings are the first demonstration that CML cells express HLA-associated leukemia-specific immunogenic peptides and provide a sound basis for immunization studies against BCR-ABL.

Adult↗

Cytomegalovirus-specific cellular immune responses and viremia in recipients of allogeneic stem cell transplants.

The immune suppression inherent in allogeneic stem cell transplantation (SCT) offers a favorable environment for infection by opportunistic agents, such as human cytomegalovirus (CMV). Despite the application of potent antiviral prophylaxis, patients remain at risk for CMV infection until adequate immunity is restored. CMV-specific CD8(+) T cell counts were monitored, using HLA-A2 tetrameric complexes, to establish the level of immune response to the viral phosphoprotein UL83 in patients after allogeneic SCT. Correlating this with viral replication and clinical status shows that the level of tetramer-positive T cells provides an assessment of CMV immune reconstitution after stem cell transplantation. Most patients with seropositive donors did reconstitute long-term CMV immunity, unless prolonged immunosuppression to control graft-versus-host disease was induced. Together with polymerase chain reaction testing, this technique provides measurable parameters that can be a guide to therapeutic decision making and can form the basis of CMV immunotherapy.

Adolescent↗

Isolation and expansion of cytomegalovirus-specific cytotoxic T lymphocytes to clinical scale from a single blood draw using dendritic cells and HLA-tetramers.

Cytomegalovirus (CMV) reactivation in immunocompromised recipients of allogeneic stem cell transplantation is a cause of morbidity and mortality from viral pneumonitis. Antiviral drugs given to reactivating patients have reduced the mortality from CMV but have toxic side effects and do not always prevent late CMV disease. Cellular immunotherapy to prevent CMV disease is less toxic and could provide prolonged protection. However, a practical approach to generating sufficient quantities of CMV-specific cytotoxic T cells (CTLs) is required. This study describes a system for generating sufficient CMV-specific CTLs for adoptive immunotherapy of HLA-A*0201 bone marrow transplant recipients from 200 mL donor blood. Donor monocytes are used to generate dendritic cells (DCs) in medium with autologous plasma, interleukin 4, granulocyte-macrophage colony-stimulating factor, and CD40 ligand. The DCs are pulsed with the immunodominant HLA-A*0201-restricted CMV peptide pp65(495-503), and incubated with donor T cells. These cultures are restimulated twice with peptide-pulsed lymphoblastoid cell lines (LCLs) or CD40-ligated B cells and purified with phycoerythrin (PE)-labeled pp65(495-503)/HLA-A*0201 tetramers by flow sorting, or with anti-PE paramagnetic beads. The pure tetramer-positive population is then rapidly expanded to obtain sufficient cells for clinical immunotherapy. The expanded CTLs are more than 80% pure, of memory phenotype, with a Tc1 cytokine profile. They efficiently kill CMV-infected fibroblasts and express the integrin VLA-4, suggesting that the CTLs could cross endothelial barriers. This technique is reproducible and could be used for generating CMV-specific CTLs to prevent CMV disease after allogeneic blood and marrow transplantation. (Blood. 2001;98:505-512)

Antigen-Presenting Cells↗

[Isolation of Vibrio strains in French coastal waters and infection with Vibrio cholerae non-O1/non-O139].

OBJECTIVE: Although the incidence in France of V. cholerae non-O1/non-O139 infection in man has increased since 1996, it remains low (7 cases in 1999). After the death in 1994 of an immunodepressed patient presenting a skin lesion showing superinfection by a strain of non-O1/non-O139 V. cholerae following exposure to seawater, we examined 22 samples of sea-water collected from 20 French coastal areas (Mediterranean coast). METHODS: The sea-water samples were filtered and enriched with alkaline peptone water (APW), and the strains of Vibrio were isolated on TCBS, SS and BCP media and identified using the API 20 E system (bioMérieux, France). RESULTS: We isolated 6 strains belonging to 3 species of Vibrio: 2 V. cholerae (non-O1/non-O139), 3 V. parahaemolyticus and 1 V. alginolyticus. One of the V. cholerae strains was isolated from sea-water sampled at the coastal town in which the patient had been staying. The seawater strains exhibited high sensitivity (MIC determined by agar dilution) to the following antibiotics: aminoglycosides, tetracyclines, azithromycin, cotrimoxazole, rifampicin and fluoroquinolones. The beta-lactams were very active against strains of V. cholerae isolated from seawater, while the strain isolated from this patient presented a new carbenicillinase (CARB-6) recently described. CONCLUSION: The presence of Vibrio in seawater along the French coast-line constitutes a risk for immunocompromised patients, and the severity of Vibrio infections warrants improved monitoring both of these organisms and of the marine environment. In addition, awareness on the part of doctors would allow patients at risk to be warned against these dangers.

Adult↗

Prospective study of nosocomial colonization and infection due to Pseudomonas aeruginosa in mechanically ventilated patients.

OBJECTIVE: To investigate the respective contribution of endogenous and exogenous transmission of Pseudomonas aeruginosa in the colonization of lungs in the mechanically ventilated patient, to estimate the role of P. aeruginosa colonization in the occurrence of severe infections, and to extrapolate appropriate control measures for the prevention of P. aeruginosa ventilator-associated pneumonia. DESIGN: Prospective study of the presence of P. aeruginosa (in stomach fluid, throat specimens, stool, and sputum) on admission, twice a week throughout the patient's stay, and in their environment. O-serotyping, pulsed-field gel electrophoresis, and arbitrarily-primed polymerase chain reaction were used to characterize the strains. SETTING: The two intensive care units (ICUs 1 and 2) of a university hospital. PATIENTS: During a 6-month period, 59 patients were included (21 in ICU 1 and 38 in ICU 2). RESULTS: P. aeruginosa was isolated in 26 patients, including ten pneumonia cases and seven colonizations on admission. The incidence of acquired colonization was statistically different between the two ICUs: 5.5 and 20.5 per 1000 days of mechanical ventilation, in ICUs 1 and 2, respectively. Endogenous acquisition was the main origin of P. aeruginosa colonization (21 of 26 patients) and the upper respiratory tract was the main bacterial reservoir in broncho-pulmonary colonization and infection. However, during the 6-month period of the study, a multidrug-resistant strain of P. aeruginosa O:11, isolated in the sink of the room of 12 patients, was found responsible for two colonizations (1 digestive, 1 throat/lungs) and one pneumonia. As a whole, from 26 cases of colonization/infection with P. aeruginosa, 5 were related to an exogenous contamination (environmental reservoir in 4 patients and cross-contamination in one patient). CONCLUSIONS: These results emphasize the need for applying various infection control measures to prevent colonization of patients with P. aeruginosa, including strategies to limit the potential of sinks from acting as a source or reservoir for this bacterium.

Aged↗

Evaluation of a new rapid urine screening analyser: CellFacts.

The aim of this study was to evaluate a new urine screening analyser in a Hospital Bacteriology Laboratory: the Cellfacts Urine Screening Analyser (Microbial Systems Limited (MSL), Coventry, England). A cohort of 1036 urine specimens were analysed by both the CellFacts and routine traditional methods. Using the CDC urinary tract infection decision levels, and compared to reference methods, the sensitivity and specificity were respectively 88.9% and 76.2%, and the predictive negative value and predictive positive value respectively 93.3% and 64.8%. Compared to the microscopy, the correlations of white blood cell count and red blood cell count were good, respectively (r = 0.8, p < 0.0001) and (r = 0.6, p < 0.0001). These results indicate that, although several positive samples were not reported, CellFacts facilitates efficient, rapid screening of infected urine specimens. This leads to perform a cytobacteriological analysis only on those results screened as positive by the automatic analysis system.

Adolescent↗

Space-time segmentation using level set active contours applied to myocardial gated SPECT.

This paper presents a new variational method for the segmentation of a moving object against a still background, over a sequence of [two-dimensional or three-dimensional (3-D)] image frames. The method is illustrated in application to myocardial gated single photon emission computed tomography (SPECT) data, and incorporates a level set framework to handle topological changes while providing closed boundaries. The key innovation is the introduction of a geometrical constraint into the derivation of the Euler-Lagrange equations, such that the segmentation of each individual frame can be interpreted as a closed boundary of an object (an isolevel of a set of hyper-surfaces) while integrating information over the entire sequence. This results in the definition of an evolution velocity normal to the object boundary. Applying this method to 3-D myocardial gated SPECT sequences, the left ventricle endocardial and epicardial limits can be computed in each frame. This space-time segmentation method was tested on simulated and clinical 3-D myocardial gated SPECT sequences and the corresponding ejection fractions were computed.

Algorithms↗

Evaluation of BacT/Alert 3D liquid culture system for recovery of mycobacteria from clinical specimens using sodium dodecyl (lauryl) sulfate-NaOH decontamination.

A total of 52 mycobacterial isolates were recovered from 1,197 clinical specimens decontaminated by a sodium dodecyl (lauryl) sulfate (SDS)-NaOH protocol. Of these, 94% were recovered with the BacT/Alert 3D system (Organon Teknika, Durham, N.C.) and 79% were recovered on Löwenstein-Jensen (LJ) medium. Mean times to detection of organisms of the Mycobacterium tuberculosis complex (n = 47) were 22.8 days with LJ medium and 16.2 days with the system. The BacT/Alert 3D system is a rapid and efficient detection system which can be used with an SDS-NaOH decontamination procedure.

Bacteriological Techniques↗

Performance of the chromogenic medium CHROMagar Staph Aureus and the Staphychrom coagulase test in the detection and identification of Staphylococcus aureus in clinical specimens.

CHROMagar Staph aureus (CSAM) (CHROMagar Microbiology, Paris, France) is a new chromogenic medium designed to enable detection of colonies of Staphylococcus aureus by their pink color. A total of 775 specimens were cultured in parallel on CHROMagar Staph aureus and conventional media. Among the 267 S. aureus strains recovered on at least one medium, 263 were isolated on CSAM medium (sensitivity, 98.5%), and 245 (sensitivity, 91.8%) were isolated on conventional media. The specificity of presumptive identification of S. aureus on the basis of pink colony color on CSAM medium was 97% (493 of 508). This specificity increased to 100% when coagulase detection with the Staphychrom coagulase test was added and to 98.8% when S. aureus surface components were detected by agglutination in the Pastorex Staph Plus test. Susceptibility testing of 67 S. aureus strains, performed in parallel on pink CSAM colonies and on colonies grown on blood agar, gave similar results. Thus, rapid and accurate recognition and identification of S. aureus isolates were achieved with CSAM as the primary isolation medium, followed by the staphylocoagulase Staphychrom test. Antimicrobial susceptibility testing (disk-diffusion method or ATB STAPH System) can be performed directly on pink CSAM colonies.

Chromogenic Compounds↗

Use of a pair of blood culture bottles for sterility testing of corneal organ culture media.

AIMS: To test the effectiveness and rapidity of a pair of blood culture bottles in the diagnosis of bacterial and fungal contamination of corneal organ culture media. METHODS: 761 microbiological analyses of storage media (Inosol and Exosol, Opsia, Toulouse, France), sampled in all phases of the organ culture at 31 degrees C of 410 consecutive corneas, were analysed. Each medium was inoculated in a pair of Bactec Plus Aerobic/F and Bactec Lytic/10 Anaerobic/F blood bottles and placed in a Bactec 9240 incubator for 14 days at 37 degrees C and in a Sabouraud broth at 20 degrees C. Changes in colour or turbidity of storage media were evaluated daily at the corneal bank. Recipients were screened post-graft for infectious signs. RESULTS: Overall contamination rate was 2.4% (18/761). Contamination was detected in less than 1 day in 78% (14/18) and less than 2 days in 94% (17/18). Positivity of the microbiological controls of starting media preceded changes medium colour in 10 out of 14 cases. Bactec blood bottles allowed detection of bacteria as well as yeasts. CONCLUSION: The use of a pair of Bactec blood culture bottles appears reliable for the rapid diagnosis of a wide range of microbiological contaminations of organ cultured corneas during banking.

Bacteria↗

Microsleep during a simplified maintenance of wakefulness test. A validation study of the OSLER test.

Daytime somnolence is both a symptom in many patients and a prevalent complaint in the general population, but its objective assessment remains elusive. The current available tests are technically complex and thus inadequate for routine clinical use or epidemiological studies. A simplified behavioral maintenance of wakefulness test (OSLER test) has been recently described that could allow for widespread availability of objective measurements of this symptom. We verified the occurrence of (micro)sleep (episodes of sleep of > or = 3 s duration) during the performance of the OSLER test in 10 normal subjects after a non-sleep-deprived night and a sleep-deprived night in randomized order. Sleep was assessed electrophysiologically according to standard methods. The OSLER test (mean of four measurements) was significantly shorter after the sleep-deprived night (25 min versus 38 min). Single missed stimuli were frequent with or without (micro)sleep, but (micro)sleep was almost always present when four or more consecutive stimuli were missed. The sensitivity and specificity of the test in detecting sleep (of > or = 3 s duration) are 85% and 94%, respectively. The total number of missed stimuli per minute duration of the test could add valuable information to the simpler mean test result. Sensitivity and specificity are not altered when only three measurements are performed. We conclude that the OSLER test appears as a simple, easy, and reliable method to objectively assess daytime somnolence.

Female↗

Sleep fragmentation: comparison of two definitions of short arousals during sleep in OSAS patients.

The measurement of arousals during sleep is useful to quantify sleep fragmentation. The criteria for electroencephalography (EEG) arousals defined by the American Sleep Disorders Association (ASDA) have recently been criticized because of lack of interobserver agreement. The authors have adopted a scoring method that associates the increase in chin electromyography (EMG) with the occurrence of an alpha-rhythm in all sleep stages (Université Catholique de Louvain (UCL) definition of arousals). The aim of the present study was to compare the two scoring definitions in terms of agreement and repeatability and the time taken for scoring in patients with obstructive sleep apnoea syndrome (OSAS) of varying severity. Two readers using both ASDA and UCL definitions scored twenty polysomnographies (PSGs) each on two occasions. The PSGs were chosen retrospectively to represent a wide range of arousal index (from 6-82) in OSAS patients. There was no difference in the arousal indices between readers and between scoring methods. The mean+/-SD difference between the two definitions (the bias) was 1.1+/-3.76 (95% confidence interval: -0.66-2.86). There was a strong linear relationship between the arousal index scored with the two definitions (r=0.981, p<0.001). Mean+/-SD scoring duration was significantly shorter for UCL than for ASDA definitions (18.5+/-5.4 versus 25.3+/-6.6 min, p<0.001). In conclusion, it has been found that in obstructive sleep apnoea syndrome patients, the American Sleep Disorders Association and Université Catholique de Louvain definitions were comparable in terms of agreement and repeatability.

Adult↗

Serum and ascitic procalcitonin levels in cirrhotic patients with spontaneous bacterial peritonitis: diagnostic value and relationship to pro-inflammatory cytokines.

OBJECTIVE: To assess the potential role of procalcitonin and pro-inflammatory cytokines, TNF-alpha, and IL-6, in the diagnosis of spontaneous bacterial peritonitis (SBP). DESIGN: Prospective study. SETTING: The emergency unit of a teaching hospital. PATIENTS: We included 21 patients with SBP and 40 patients with sterile ascitic fluid. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: For the diagnosis of SBP, the best markers were serum levels of procalcitonin with a cut-off value of 0.75 ng/ml, a sensitivity of 95%, a specificity of 98%, and ascitic fluid levels of IL-6 with a cut-off value of 5,000 ng/ml, a sensitivity of 100%, and a specificity of 88%. C-reactive protein and serum polymorphonuclear count have low sensitivity/specificity at 62/92% and 57/90%, respectively. From 21 patients with SBP, ascitic fluid to serum ratio of TNF-alpha and IL-6 was greater than to 2 in all cases with a mean at 6.2 +/- 6.5 and 34 +/- 31, respectively. By contrast, this ratio for procalcitonin was less than 1 in all cases with a mean at 0.31 +/- 0.25. We found no correlation between procalcitonin levels and cytokine levels in either ascitic fluid or serum. CONCLUSIONS: Serum procalcitonin level may become a useful marker for the diagnosis of SBP in cirrhotic patients. The low ratio of ascitic fluid to serum procalcitonin supports the hypothesis that procalcitonin is not produced intraperitoneally.

Ascitic Fluid↗