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A Collignon

Publications and source records attributed to A Collignon.

At least 19 recordsLinked to original sources

Report of a case of congenital malaria Plasmodium malariae in France.

Congenital malaria (CM) has been considered to be rare, even in malaria-endemic areas but the disease can result in significant neonatal morbidity. Because of its rarity, the disease may go undiagnosed for a prolonged period in a seriously ill infant. We report the first case of Plasmodium malariae CM from a HIV mother. HIV could have facilitated the transfer of erythrocytic persistent P. malariae through the placenta to the fetus.

Adult↗

[Evaluation of four immunoenzymatic tests for detecting Clostridium difficile toxins A and B].

Four immunoenzymatic tests for detecting Clostridium difficile toxins A and B were studied: two rapid tests (Tox A/B QUIK CHEK-Techlab and NoviView Toxine-A-Hiss diagnostics) and two Elisa tests (C. difficile TOX A/B II -Techlab and Toxin A+B Elisa Test, Novitec-Hiss diagnostics). The results were compared to those obtained with ImmunoCard Tox A+B -ICTAB (Meridian), C. difficile Toxine A (Oxoid) for rapid test and Elisa Premier A+B Meridian for Elisa. A total of 41 stools and 16 isolates were studied with rapid tests. On stools, the sensitivity and specificity of QUIK CHEK test was 94.1% and 100% respectively compared to the test ICTAB. On the isolates, sensitivity and specificity was 100%. With the Noviview test, the sensitivity on stools and isolates was respectively 88.2 and 85.7% and the specificity was 100% compared to Oxoid. A total of 38 stools were studied with Elisa tests. With Techlab test compared to the test Premier, sensitivity and specificity was 100%. The Novitec test gave five false negative reactions with consequently a sensitivity of 70.6%.

Bacterial Proteins↗

[Schistosoma haematobium among immigrants consulting in municipal free clinics in Paris in 2003].

The "Consultations de Diagnostic et d'Orientation" (CDO) are free medical consultations for precarious populations, proposed by the Health Department of Paris. More than two-thirds of the CDO patients come from Sub-Saharan Africa. Schistosoma haematobium (SH) is one of the most frequent infectious diseases detected within CDO. More than a thousand people have consulted for the first time in CDO in 2003 in one of the municipal free Clinics which proposes this service. Parasitologic test of urine has been performed among 220 patients and found 24 positive results: viable eggs of SH (10.8%). All 24 patients are male, most of them are under 35 years-old and come from the region of the Senegal River that lies in the junction of Mali, Senegal and Mauritania. We want to remind physicians in non-tropical setting to think of SH when they see a patient originating from Africa. To ask him if he presents haematuria and if not, to prescribe a parasitologic test of urine. If all patients from endemic regions had undergone this screening in 2003, we would have detected about 20 more cases of SH. If treated early enough, it could avoid severe uronephrological complications, which are rare but represent a high health care cost (bladder tumor, renal failure).

Adolescent↗

[Characterization of an extracellular protease from Clostridium difficile].

Clostridium difficile is an intestinal pathogen, which produces two main virulence factors, the exotoxins A and B. Other bacterial structures have been implicated in the colonization of the gastrointestinal tract, which is the first step of the pathogenic process. C. difficile expresses adherence factors and also, displays some surface-associated proteolytic activity, which could play a role in the physiopathology of this bacterium. The aim of this work was to study the protein named Cwp84 which displays significant homologies with many cysteine proteases. The coding catalytic domain of this protein has been cloned in the expression system pGEX-6P-1, as an in-frame fusion with the gluthatione S-transferase, and subsequently purified. The purified fraction showed proteolytic activity on gelatine and BAPNA, but not on azocoll, suggesting a highly selective substrate specificity. The results obtained from inhibition experiments confirmed that Cwp84 belongs to the cysteine protease family. Cwp84 could play a role in degrading some specific host proteins or in the maturation of surface-associated bacterial proteins.

Amino Acid Sequence↗

[Alcohol consumption patterns and behaviour among people using the health care system in Lower-Normandy].

A national study on the prevalence of excessive alcohol consumption behaviour was undertaken throughout France. The results presented in this paper are those from Lower-Normandy. Two surveys were carried out: one among patients of general practitioners, and another among those in hospitals. The following data were specifically examined: the CAGE test score, the use of alcohol, the level of dependence, the conclusion of the care provider and the patient's profile. The results show that the frequency of excessive alcohol consumption in Lower-Normandy remains slightly higher than the national average. However, a significant reduction, which has been confirmed by the decrease in mortality rates, has been obtained within the last two decades. But nevertheless, the fight must continue and with on-going determination, and this aspect must rely more heavily upon general practitioners.

Adult↗

Multicentre study of the molecular epidemiology, serotypes and antimicrobial susceptibility patterns of invasive Streptococcus pneumoniae invasive isolated from children in the Ille de France area.

Between January 1997 and April 2002, 73 consecutive invasive strains of Streptococcus pneumoniae were isolated from children under 16 years of age in four hospitals in suburban Paris. Their genetic diversity was investigated by serotyping and analysis of pulsed-field gel electrophoresis restriction patterns. Antibiotic susceptibility patterns were analysed by disk susceptibility testing and determination of minimal inhibitory concentrations. The genetic basis of macrolide resistance was investigated by polymerase chain reaction. Studies of penicillin and vancomycin tolerance were performed for each strain. Despite the high prevalence (45.2%) of penicillin-nonsusceptible Streptococcus pneumoniae, resistance to amoxicillin (1.4%) was rare, and no strain was resistant to cefotaxime. Overall, 4.1% of pneumococcal strains were resistant to penicillin. Penicillin or vancomycin tolerance was not detected in any of the 73 strains studied. Of the erythromycin-resistant strains (48%), all but one carried the ermB gene. No strains showing a decreased susceptibility to ciprofloxacin (MIC, >4 mg/l) or overexpressing an efflux pump inhibited by reserpine were isolated. The serotypes found, in order of frequency, were as follows: 18C, 14, 6B, 19F, 19A, 9V, 23F, 1, 7F, 9A, 38. Strains of penicillin-nonsusceptible Streptococcus pneumoniae belonged predominantly to serotypes 14, 6B, 9V, 9A, 23F, 19F and 19A. The seven-valent conjugated vaccine covered 85.5% of the serogroups isolated in children under 2 years of age and 65.6% of the serogroups identified in children over 2 years of age. The genetic analysis showed a high identity for some serotypes, such as 14/9V, 6B and 23F. The use of the seven-valent conjugated vaccine is a critical measure to prevent invasive pneumococci infections in children in the Ille de France area.

Adolescent↗

[Regional evaluation of long-term care facilities for the elderly].

The increase in the total number of very elderly people demands the precision and specification of the needs of long-term care structures in the coming years. A survey conducted in Lower-Normandy is presented and described. Questionnaires were sent to directors of residence homes and nursing homes in order to investigate their operations and their problems. The increase in the number of spaces to foresee seems moderate if the progressive trend remains unchanged (+1% per year until the year 2010), but within the same timeframe, the demographic decline in the number of potential family helpers and home health workers to take care of the elderly in their homes, coupled with the establishment of a new state allowance for dependent people, could alter the situation. Furthermore, more than 50% of nursing homes and residence homes for the elderly are in need of significant improvements: a reduction in the number of shared rooms (52.5% of nursing homes) and the development of equipment to meet the needs of the handicapped and disabled. The means in personnel and staff qualifications are particularly heterogeneous and difficulties in coping with dependency are reported most everywhere. The application of the 1999 decree stipulating the approval of these structures based upon thorough evaluations of available services is urgently needed.

Aged↗

Analysis of expression of GroEL (Hsp60) of Clostridium difficile in response to stress.

Our laboratory has previously shown that adherence of Clostridium difficile to tissue culture cells is augmented by various stresses and that GroEL, a heat shock protein, serves an adhesive function in this bacterium. In this communication, RT-PCR, SDS-PAGE and immunoblotting were used to study the stress response in C. difficile following heat, acid or osmotic shock, iron deprivation or presence of a subinhibitory concentration of ampicillin in the culture medium. All these stresses increased transcription of groEL and production of GroEL to various degrees. Furthermore, the protein was found in membrane fractions and in the extracellular space after heat stress.

Ampicillin↗

GroEL (Hsp60) of Clostridium difficile is involved in cell adherence.

Previous results have demonstrated that adherence of Clostridium difficile to tissue culture cells is augmented by various stresses; this study focussed on whether the GroEL heat shock protein is implicated in this process. The 1940 bp groESL operon of C. difficile was isolated by PCR. The 1623 bp groEL gene is highly conserved between various C. difficile isolates as determined by RFLP-PCR and DNA sequencing, and the operon is present in one copy on the bacterial chromosome. The 58 kDa GroEL protein was expressed in Escherichia coli in fusion with glutathione S:-transferase and the fusion protein was purified from IPTG-induced bacterial lysates by affinity chromatography on glutathione-Sepharose. A polyclonal, monospecific antiserum was obtained for GroEL which established by immunoelectron microscopy, indirect immunofluorescence and immunoblot analysis that GroEL is released extracellularly after heat shock and can be surface associated. Cell fractionation experiments suggest that GroEL is predominantly cytoplasmic and membrane bound. GroEL-specific antibodies as well as the purified protein partially inhibited C. difficile cell attachment and expression of the protein was induced by cell contact, suggesting a role for GroEL in cell adherence.

Amino Acid Sequence↗

Role of FliC and FliD flagellar proteins of Clostridium difficile in adherence and gut colonization.

In vitro and in vivo adhesive properties of flagella and recombinant flagellin FliC and flagellar cap FliD proteins of Clostridium difficile were analyzed. FliC, FliD, and crude flagella adhered in vitro to axenic mouse cecal mucus. Radiolabeled cultured cells bound to a high degree to FliD and weakly to flagella deposited on a membrane. The tissue association in the mouse cecum of a nonflagellated strain was 10-fold lower than that of a flagellated strain belonging to the same serogroup, confirming the role of flagella in adherence.

Animals↗

Characterization of a cell surface protein of Clostridium difficile with adhesive properties.

Our laboratory has previously shown that Clostridium difficile adherence to cultured cells is enhanced after heat shock at 60 degrees C and that it is mediated by a proteinaceous surface component. The present study was undertaken to identify the surface molecules of this bacterium that could play a role in its adherence to the intestine. The cwp66 gene, encoding a cell surface-associated protein of C. difficile 79-685, was isolated by immunoscreening of a C. difficile gene library with polyclonal antibodies against C. difficile heated at 60 degrees C. The Cwp66 protein (66 kDa) contains two domains, each carrying three imperfect repeats and one presenting homologies to the autolysin CwlB of Bacillus subtilis. A survey of 36 strains of C. difficile representing 11 serogroups showed that the 3' portion of the cwp66 gene is variable; this was confirmed by sequencing of cwp66 from another strain, C-253. Two recombinant protein fragments corresponding to the two domains of Cwp66 were expressed in fusion with glutathione S-transferase in Escherichia coli and purified by affinity chromatography using gluthatione-Sepharose 4B. Antibodies raised against the two domains recognized Cwp66 in bacterial surface extracts. By immunoelectron microscopy, the C-terminal domain was found to be cell surface exposed. When used as inhibitors in cell binding studies, the antibodies and protein fragments partially inhibited adherence of C. difficile to cultured cells, confirming that Cwp66 is an adhesin, the first to be identified in clostridia.

Adhesins, Bacterial↗

Molecular characterization of fliD gene encoding flagellar cap and its expression among Clostridium difficile isolates from different serogroups.

The fliD gene encoding the flagellar cap protein (FliD) of Clostridium difficile was studied in 46 isolates belonging to serogroups A, B, C, D, F, G, H, I, K, X, and S3, including 30 flagellated strains and 16 nonflagellated strains. In all but three isolates, amplification by PCR and reverse transcription-PCR demonstrated that the fliD gene is present and transcribed in both flagellated and nonflagellated strains. PCR-restriction fragment length polymorphism (RFLP) analysis of amplified fliD gene products revealed interstrain homogeneity, with one of two major patterns (a and b) found in all but one of the strains, which had pattern c. A polyclonal monospecific antiserum raised to the recombinant FliD protein reacted in immunoblots with crude flagellar preparations from 28 of 30 flagellated strains but did not recognize FliD from nonflagellated strains. The fliD genes from five strains representative of the three different RFLP groups were sequenced, and sequencing revealed 100% identity between the strains with the same pattern and 88% identity among strains with different patterns. Our results show that even though FliD is a structure exposed to the outer environment, the flagellar cap protein is very well conserved, and this high degree of conservation suggests that it has a very specific function in attachment to cell or mucus receptors.

Bacterial Proteins↗

Phenotypic and genotypic diversity of the flagellin gene (fliC) among Clostridium difficile isolates from different serogroups.

Phenotypic and genotypic diversity of the flagellin gene (fliC) of Clostridium difficile was studied in 47 isolates from various origins belonging to the serogroups A, B, C, D, F, G, H, I, K, X, and S3. Electron microscopy revealed 17 nonflagellated strains and 30 flagellated strains. PCR and reverse transcription-PCR demonstrated that the flagellin gene was present in all strains and that the fliC gene was expressed in both flagellated and nonflagellated strains. Southern blotting showed the presence of only one copy of the gene and three different hybridization patterns. DNA sequence analysis of fliC from the strains belonging to serogroups C, D, and X, representative of each profile, disclosed great variability in the central domain, whereas the N- and C-terminal domains were conserved. The variability of the flagellin gene fliC was further studied in the isolates by PCR-restriction fragment length polymorphism (RFLP) analysis. Nine different RFLP groups were identified (I to IX), among which three (I, VII, and VIII) corresponded to numerous serogroups whereas the six others (II, III, IV, V, VI, and IX) belonged to a single serogroup. Flagellin gene RFLP analysis could constitute an additional typing method employable in conjunction with other typing methods currently available.

Amino Acid Sequence↗

Clostridium difficile cell attachment is modified by environmental factors.

Adherence of Clostridium difficile to Vero cells under anaerobic conditions was increased by a high sodium concentration, calcium-rich medium, an acidic pH, and iron starvation. The level of adhesion of nontoxigenic strains was comparable to that of toxigenic strains. Depending on the bacterial culture conditions, Vero cells could bind to one, two, or three bacterial surface proteins with molecular masses of 70, 50, and 40 kDa.

Adhesins, Bacterial↗

[Susceptibility of Clostridium difficile to metronidazole using the E-test: effect of the culture medium].

The treatment of intestinal Clostridium difficile infections rests on administration of either a glycopeptide or metronidazole. Given the current shifts in resistance patterns of anaerobes to antimicrobials, a study of the susceptibility of C. difficile to metronidazole was timely. The objective of this study was to evaluate the influence of the culture medium on the Minimal Inhibitory Concentration (MIC) of metronidazole as determined using the E-test. Thirty-one strains were grown on three different media supplemented with 5% horse blood, namely Columbia agar, Wilkens Chalgren agar, and Brucella agar. Results were compared to those obtained using the reference agar dilution method (ADM). As recommended by the French Society for Microbiology, susceptibility was defined as an MIC < or = 4 mg/L. When used on strains susceptible by the ADM, the E-test yielded lower values than the ADM with all three media. Furthermore, findings suggest that E-test results obtained with strains whose MIC is in the 4 to 8 mg/L range by the ADM should be interpreted with caution and, in some cases, tested using the ADM.

Animals↗

Protease activity of Clostridium difficile strains.

The production of proteolytic enzymes by 10 Clostridium difficile isolates of varying toxigenicity and clinical origin was studied to determine if all isolates secreted proteases. Different protease substrates were studied: gelatin, collagen, phenylazobenzyloxycarbonyl-leucyl-glycyl-L-prolyl-D-arginine (Pz-peptide), casein, azocasein, and azocoll. All isolates degraded gelatin, collagen, and azocoll. The supernatants of all isolates contained an enzyme capable of attacking gelatin incorporated in a polyacrylamide gel (zymograms) and forming two closely spaced lytic bands with an estimated molecular mass of 35-40 kDa. Polyclonal antibodies, produced against the C. difficile gelatinase, revealed in Western blots a 35-kDa protein in the culture supernatants of all C. difficile isolates. In the same manner, Clostridium perfringens collagenase polyclonal antibodies detected a 120-kDa protein in the culture supernatants of all isolates; this suggests that at least two proteases may exist in C. difficile. The protease activities of the 10 strains examined did not seem strikingly different quantitatively but were in general weak and their role in pathogenicity is suspect.

Caseins↗