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Biomedical subjects

C Carlier

Publications and source records attributed to C Carlier.

At least 37 records · Page 2Linked to original sources

Shuttle vectors containing a multiple cloning site and a lacZ alpha gene for conjugal transfer of DNA from Escherichia coli to gram-positive bacteria.

The mobilizable shuttle cloning vectors, pAT18 and pAT19, are composed of: (i) the replication origins of pUC and of the broad-host-range enterococcal plasmid pAM beta 1; (ii) an erythromycin-resistance-encoding gene expressed in Gram- and Gram+ bacteria; (iii) the transfer origin of the IncP plasmid RK2; and (iv) the multiple cloning site and the lacZ alpha reporter gene of pUC18 (pAT18) and pUC19 (pAT19). These 6.6-kb plasmids contain ten unique cloning sites that allow screening of derivatives containing DNA inserts by alpha-complementation in Escherichia coli carrying the lacZ delta M15 deletion, and can be efficiently mobilized by self-transferable IncP plasmids co-resident in the E. coli donors. Plasmids pAT18, pAT19 and recombinant derivatives have been successfully transferred by conjugation from E. coli to Bacillus subtilis, Bacillus thuringiensis, Listeria monocytogenes, Enterococcus faecalis, Lactococcus lactis, and Staphylococcus aureus at frequencies ranging from 10(-6) to 10(-9). The presence of a restriction system in the recipient dramatically affects (by three orders of magnitude) the efficiency of conjugal transfer of these vectors from E. coli to Gram+ bacteria.

Cloning, Molecular↗

Prevalence of malnutrition and vitamin A deficiency in the Diourbel, Fatick, and Kaolack regions of Senegal: feasibility of the method of impression cytology with transfer.

The evaluation of the prevalence of ophthalmological diseases and vitamin A deficiency in Senegalese preschool-aged children enabled us to confirm the method of impression cytology with transfer (ICT) and to assess the impact of ophthalmological diseases on the cytological appearance of conjunctival cells. A simplification of the ICT method consisted of transferring cells present on a strip of paper to a glass slide by finger pressure. Harris-Schorr staining of the sample papers confirmed the results obtained by the transfer. Inflammatory trachoma influenced the results of the test. This simple method will prove very helpful for detection of vitamin A deficiency in children in underdeveloped areas.

Child↗

Prevalence of malnutrition and vitamin A deficiency in the Diourbel, Fatick, and Kaolack regions of Senegal: epidemiological study.

The purpose of our study was to determine the prevalence of protein-calorie malnutrition (PCM), ocular diseases, and vitamin A deficiency in preschool children selected at random in a rural zone of the groundnut belt of Senegal. The prevalence of PCM was 37.1% (95% CI 33.8-40.2%) according to the Waterlow classification, with a majority of stunting, and prevalence of hypovitaminosis A was estimated to be 11.4% (95% CI 9.3-13.5%) by using impression cytology. Furthermore, 19.4% (95% CI 15.8-22.0%) of the children might be defined at risk of deficiency. The prevalence of Bitot's spots was equal to 0.2% (95% CI 0.03-0.9%). A problem of PCM associated with a health-endangering vitamin A deficiency existed.

Anthropometry↗

Prevalence of malnutrition and vitamin A deficiency in the Diourbel, Fatick, and Kaolack regions of Senegal: a controlled study.

Two hundred and six Senegalese preschool children included in an epidemiological study were selected by their results from impression cytology with transfer (ICT) for assessment of their nutritional state by means of biological variables and for assessment of the diagnostic values of the ICT. A problem of protein-calorie malnutrition existed (transthyretin and retinol-binding protein concentrations were low) associated with vitamin A deficiency (retinol concentrations were low). The sensitivity and specificity of the ICT defined with respect to retinol (threshold fixed at 0.35 mumol/L) varied with the classification criteria of ICT and seemed to be fairly insensitive but specific.

C-Reactive Protein↗

Nucleotide sequences specific for Tn1545-like conjugative transposons in pneumococci and staphylococci resistant to tetracycline.

The distributions of tet(M) and conjugative transposons related to Tn1545 were studied by hybridization in 47 clinical isolates of Streptococcus pneumoniae resistant to tetracycline. Resistance to tetracycline was always associated with resistance to minocycline and was due to the presence of the tet(M) gene. Association of tet(M) with int-Tn, the gene encoding the protein required for the movements of Tn1545-like transposons, was found in all but one strain of S. pneumoniae. In contrast, int-Tn was detected in only 2 of 37 strains of Staphylococcus spp. harboring tet(M).

Base Sequence↗

Transferable plasmid-mediated antibiotic resistance in Listeria monocytogenes.

A strain of Listeria monocytogenes, isolated from a patient with meningoencephalitis, was resistant to chloramphenicol, erythromycin, streptomycin, and tetracycline. The genes conferring resistance to these antibiotics were carried by a 37-kb plasmid, pIP811, that was self-transferable to other L monocytogenes cells, to enterococci-streptococci, and to Staphylococcus aureus. The efficacy of transfer and the stability of pIP811 were higher in enterococci-streptococci than in the other gram-positive bacteria. As indicated by nucleic acid hybridisation, the genes in pIP811 conferring resistance to chloramphenicol, erythromycin, and streptomycin were closely related to plasmid-borne determinants that are common in enterococci-streptococci. Plasmid pIP811 shared extensive sequence homology with pAM beta 1, the prototype broad host range resistance plasmid in these two groups of gram-positive cocci. These results suggest that emergence of multiple antibiotic resistance in Listeria spp is due to acquisition of a replicon originating in enterococci-streptococci. The dissemination of resistance to other strains of L monocytogenes is likely.

Chloramphenicol Resistance↗

The integration-excision system of the conjugative transposon Tn 1545 is structurally and functionally related to those of lambdoid phages.

Excision of Tn1545 and related conjugative transposons of Gram-positive bacteria occurs by reciprocal site-specific recombination between non-homologous regions of the transposon-target junctions. Excisive recombination requires two transposon-encoded proteins designated Xis-Tn and Int-Tn. We have shown that, following excision, Tn1545 is a circular structure with ends separated by either of the two hexanucleotides that were present at the transposon-target junctions. Using a trans-complementation assay, we have demonstrated that Int-Tn is able to catalyse in vivo integration of a circular intermediate of Tn1545 defective for integration and excision. comparison of integration sites suggests that limited sequence homology at the vicinity of the recombining sites is required for integration of the element. These data support the hypothesis that the integration/excision systems of conjugative transposons from Gram-positive cocci and of lambdoid phages from Gram-negative bacilli have evolved from a common ancestor.

Bacteriophage lambda↗

Gene homogeneity for aminoglycoside-modifying enzymes in gram-positive cocci.

Aminoglycoside-resistant strains of Staphylococcus and Enterococcus, approximately 500 of each, were screened by dot blot hybridization for the presence of genes encoding aminoglycoside-modifying enzymes. The MICs of various aminoglycosides for the strains were determined, and the enzyme contents of the cells were inferred from the resistance phenotypes. The agreements (in percent) of the hybridization results with the deduced enzyme contents for Staphylococcus and Enterococcus species were, respectively, 80 and 87.6 for ANT(6) (aminoglycoside nucleotidyltransferase), 99.8 and 100 for both APH(3') (aminoglycoside phosphotransferase) and APH(2")-AAC(6') (aminoglycoside acetyltransferase), and 100 and 100 for ANT(4'). The weak correlation obtained with the probe for ANT(6) was due to the fact that gram-positive cocci can also be streptomycin resistant by synthesis of APH(3") or ANT(3")(9) and by ribosomal mutation. The remaining probes appeared to be specific: they hybridized with all the resistant clinical isolates but not with the susceptible strains. These results indicate that, except for streptomycin, nucleic acid hybridization is a valid approach for the detection and characterization of aminoglycoside resistance in gram-positive cocci.

Acetyltransferases↗

Emergence of 4',4"-aminoglycoside nucleotidyltransferase in enterococci.

Enterococcus faecium BM4102 was resistant to macrolide-lincosamide-streptogramin B-type (MLS) antibiotics; tetracycline-minocycline; and high levels of kanamycin, neomycin, tobramycin, and dibekacin but not gentamicin. This aminoglycoside resistance phenotype is new in enterococci. The genes conferring resistance to aminoglycosides and MLS antibiotics in this strain were carried on a plasmid, pIP810, that was self-transferable to to other Enterococcus strains. Resistance to tobramycin and structurally related aminoglycosides, kanamycin, neomycin, and dibekacin, was due to synthesis of a 4',4"-aminoglycoside nucleotidyltransferase. Homology was detected by hybridization between pIP810 DNA and a probe specific for a gene encoding an enzyme with identical site specificity in staphylococci. The bacteriostatic activity of amikacin apparently was not affected by the presence of the enzyme, although it was modified in vitro. However, the bactericidal activity of amikacin and the synergism of this aminoglycoside with penicillin were abolished.

Anti-Bacterial Agents↗

[Long-term results of percutaneous transluminal angioplasty of the iliac arteries].

A well-mastered technique will cause percutaneous transluminal angioplasty of iliac arteries to achieve long-term results comparable to those obtained surgically. In isolated iliac stenosis, a 95% good result rate is attained (81% cured, 14% improved). More complex cases of iliac stenosis yield 90% good results (72% cure), with a mean Doppler ankle/arm pressure index gain of 0.5 point in a series including 673 patients at 5 years follow-up. Only the presence of combined distal femoral lesions may cause the good result score to drop to 85%, including merely 35% cure. General complications are practically lacking and local complications are minimal, which incites one to propose this technique as a first-line treatment of iliac stenosis whenever distal blood circulation is maintained.

Angioplasty, Balloon↗

[Femoral angioplasty. Long-term results].

A study on the long-term efficacy of femoral-popliteal angioplasty was carried out on 185 angioplasty cases over a 5 year follow-up period. A classification of data according to the type of lesion treated, revealed that results were favorable in case of stenosis (87%), short obstruction (70%) and long obstruction (35%). A special study of the outcome of treatments for stage IV arteritis was carried out. After comparing results with those obtained by other teams, the authors list the complications encountered, hematomas, and thromboses, and show their current tendency for regression. Lastly, the authors stress the advantages of angioscopy, which permits to identify the nature of the treated lesions and to predict possible complications, which are usually underrated by angiography.

Angioplasty, Balloon↗

[Angioscopy and angioplasty of the iliac and femoral arteries].

Angioscopy is a technique of endoscopic investigation of the vascular lumen and its contents, which we have coupled with iliac and femoral angioplasty. The material comprises 3 main elements: the angioscope, the TV-monitoring assembly and the infusion system. No complication was recorded in a series of 94 angioscopies (47 iliac, 44 distal femoral and 3 grafts) performed over a period of 18 months. The technical quality of the images obtained was good in 90% of cases (85/94). Angioscopy revealed lesions that had either been nonvisualized or underrated by angiography, and played a determinant role in 16% of cases (15/94), whereby the therapeutical procedure could be modified accordingly.

Angioplasty, Balloon↗

[Percutaneous angioscopy in the diagnosis of arteriopathies of the lower limbs].

Because of the poor specificity of the arteriography, the authors have performed a percutaneous angioscopy before every angioplasty of the arteries of lower limbs for nearly 3 years. They define the angioscopy and describe materials and technique. The percutaneous route without surgery and without anesthesia underlines the originality of the technique. The great difficulty is to counter-pulse the arterial inflow. The rate of saline irrigation and all the means of arterial blockade are described. In the second part, the "elementary images" are classified. The authors insist on the specificity of the angioscopy in the interpretation of arterial obstructions. Because of this specificity, the right treatment can be applied to any type of lesion. Angioscopy permits to foresee endoluminal surgery technique.

Angiography↗

Molecular characterization of two proteins involved in the excision of the conjugative transposon Tn1545: homologies with other site-specific recombinases.

Excision is probably the initial and rate-limiting step of the movements of conjugative transposons of Gram-positive bacteria such as Tn916 and Tn1545. We have shown, by molecular cloning and DNA sequencing, that a 2058 bp Sau3A right-junction fragment of transposon Tn1545 specifies two gene products that are involved in the excision of the element. The DNA sequence of these genes, designated orf1 and orf2, has been determined and the corresponding proteins, ORF1 and ORF2, have been identified in a bacterial cell-free coupled transcription-translation system. These proteins are freely diffusible since they are able to trans-complement in vivo a deletion derivative of Tn1545 defective for excision. Using an in vivo complementation assay, we have demonstrated that ORF2 alone is able to catalyse excision and that ORF1 strongly stimulates the activity of ORF2. We also found that ORF1 and ORF2 display local homology with, respectively, proteins Xis and Int from lamboid phages, which suggests that these excision systems have a common origin. Based on the functional properties of the integrase of bacteriophage lambda, on the analysis of the nucleotide sequence of the junction fragments and of the target before insertion and after excision, a model is proposed for ORF2-catalysed excision of Tn1545 and related conjugative transposons.

Amino Acid Sequence↗

Assessment of vitamin A status in an elderly French population using impression cytology with transfer.

The vitamin A status of 105 elderly French people was assessed by ocular impression cytology with transfer (ICT). 5 patients (4.8%) had a negative ICT defined by the absence of goblet cells and the enlargement of the epithelial cells (deficient vitamin A status). 100 patients had a positive ICT with goblet cells and small, numerous epithelial cells (sufficient vitamin A status). ICT results were compared to serum biochemical parameters. Mean serum levels of retinol, retinol-binding protein (RBP) and transthyretin (TTR) were significantly lower while C-reactive protein (CRP) and orosomucoid were significantly higher in negative ICT than in positive one. The etiology of vitamin A deficiency is difficult to conclude (malnutrition, vitamin A deficiency, inflammation). ICT is a good indicator of peripheral vitamin A deficiency in our experience.

Aged↗