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P Rodriguez

Publications and source records attributed to P Rodriguez.

At least 55 records · Page 3Linked to original sources
PubMed1996

ATP-regulated activity of the plasmin-streptokinase complex: a novel mechanism involving phosphorylation of streptokinase.

Streptokinase, an extracellular protein produced by Streptococci, is capable of activating the human fibrinolytic zymogen plasminogen. The rate of amidolytic activity of the plasminogen-streptokinase complex is greatly diminished by micromolar concentrations of ATP and heparin oligosaccharides. In addition, the plasminogen activator activity of the plasminogen-streptokinase complex is also inhibited by these effectors. ATP and heparin oligosaccharides show structural similarity, suggesting that the inhibition is caused by binding of these molecules to a common newly formed binding pocket in streptokinase, which appears after interaction with plasminogen. Addition of the bivalent cations Ca2+ and Mg2+ reverses the inhibition caused by ATP and heparin. In the presence of ATP and bivalent cations, the complex between plasminogen and streptokinase develops an autophosphorylating activity whose target is the sequence LTSRPAHG in the 4.5 kDa streptokinase N-terminal peptide, which is an early autolysis peptide. This streptokinase N-terminal peptide, which is essential for streptokinase activating activity, may serve, once phosphorylated, in mechanisms related to the pathogenicity of Streptococci. These studies suggest a critical role for plasminogen in regulating the activity of the streptokinase molecule.

Adenosine Triphosphate↗
PubMed1996

A Tn1545-like transposon carries the tet(M) gene in tetracycline resistant strains of Bacteroides ureolyticus as well as Ureaplasma urealyticum but not Neisseria gonorrhoeae.

The presence of the tet(M) determinant and conjugative transposons related to Tn1545 in urogenital pathogens, Bacteroides ureolyticus (10 strains), Neisseria gonorrhoeae (37 strains) and Ureaplasma urealyticum (81 strains), was studied by PCR analysis and hybridization assay. All tetracycline-resistant strains that hybridized with a probe specific for tet(M) gave the expected fragment when their DNA's were subject to PCR. The tet(M) gene and int-Tn, the gene encoding the protein required for the movement of Tn1545-like transposons, were always found together in tetracycline resistant strains of B. ureolyticus and U. urealyticum. In contrast, int-Tn was not detected in tet(M)-mediated tetracycline-resistant N. gonorrhoeae strains. These data suggest carriage of tet(M) by a Tn1545-like transposon in some tetracycline-resistant B. ureolyticus.

Bacteroides↗
PubMed1996

Intestinal paracellular permeability during malnutrition in guinea pigs: effect of high dietary zinc.

BACKGROUND: Zinc has been shown to have beneficial effects in vitro on epithelial barrier function, and in vivo to reduce intestinal permeability in malnourished children with diarrhoea. AIMS: To determine whether malnutrition alters intestinal paracellular permeability, and whether zinc prevents such alterations. METHODS: Guinea pigs were fed a normal protein diet (NP group), a low protein diet (LP group), or a low protein diet enriched with 1800 ppm zinc (LPZn group) for three weeks. Intestinal permeability was measured on jejunal segments mounted in Ussing chambers by measuring ionic conductance and mucosal to serosal fluxes of 14C-mannitol, 22Na, and horseradish peroxidase. Tight junction morphology was assessed on cryofracture replicas. RESULTS: Mannitol and Na fluxes and ionic conductance increased in the LP group compared with the NP group but remained normal in the LPZn group. Accordingly, jejunal epithelia from the LP group, but not from the LPZn group, showed a small decrease in number of tight junctional strands compared with epithelia from the NP group. Neither malnutrition nor zinc treatment modified horseradish peroxidase fluxes. CONCLUSIONS: Malnutrition is associated with increased intestinal paracellular permeability to small molecules, and pharmacological doses of zinc prevent such functional abnormality.

Animals↗
PubMed1996

Establishment of hepatic cell polarity in the rat hepatoma-human fibroblast hybrid WIF-B9. A biphasic phenomenon going from a simple epithelial polarized phenotype to an hepatic polarized one.

By immunofluorescence and freeze fracture methods, we have studied the establishment of hepatic cell polarity in WIF-B9 cells, a subclone of the WIF-B rat hepatoma-derived hybrid cell line. As previously shown (Ihrke et al. (1993) J. Cell Biol. 123, 1761-1775; Shanks et al. (1994) J. Cell Sci. 107, 813-825), these cells are a suitable model for in vitro studies of various hepatic functions, particularly polarity: in confluent cultures, the majority of cells form bile canaliculus-like structures; membrane domains are settled, according to plasma membrane protein localization similar to rat hepatocytes in situ. We here report that the establishment of WIF-B9 cell polarity is a slow progressive biphasic phenomenon. During the first days of culture, the majority of cells do not make bile canaliculus-like structures. However, they display a polarity similar to that of simple epithelial cells: apical membrane proteins and villin are found at the cell apex; basolateral ones, excluded from this area, are expressed in the remaining membrane area; the tight junction-associated protein ZO-1 and actin are concentrated at the boundary of these two poles, whereas E-cadherin is present at the lateral pole just under the apex. With time in culture, the number of cells expressing this simple epithelial polarized phenotype decreases progressively and, after 10-15 days, depending on the plating density, nearly all the cells express the typical hepatic polarized phenotype. The expression of these two phenotypes is mutually exclusive. Freeze-fracture replicas of both types of polarized cells show either macula occludens, fascia occludens (simple epithelial polarity) or zonula occludens (hepatic polarity), associated with gap junctions. In this last case, two or three continuous strands are generally present all around the bile canaliculus-like structures.

Actins↗
PubMed1996

[Molecular typing by pulsed field gel electrophoresis of Stenotrophomonas maltophilia isolated in a department of hematology].

Stenotrophomonas maltophilia is an important nosocomial pathogen. The increased isolates of S. maltophilia among hematology unit patients led to an epidemiological survey. Over 26 months, 24 strains isolated from 23 patients and an environmental isolate from blood pressure armband have been identified. The isolated were first analysed by the use of phenotypical markers: biotype, antibiotic susceptibility, but the minor differences observed justified a genotypic analysis. Pulsed field gel electrophoresis of genomic DNA was carried with XbaI and DraI restriction endonucleases by contour-clamped homogeneous electric field method (CHEF). The data obtained showed a great genomic diversity within the species S. maltophilia. Nevertheless, the same restriction profile was found for 3 patients and 3 other profiles were obtained for 3 couples of patients hospitalized at the same time. All the other strains isolated from subjects hospitalized during the same period exhibited pulsotypes independent from each other. Compared to biotypes and antibiotic susceptibility, these results indicate field only with non modified primers, and the absence of 460 mutations was confirmed by sequencing. Two isolates P1 and P2, from a transplanted patient were amplified with both primers MCMM and MCMW: sequencing analysis shown the presence of a mixture of strains, one of them harbouring A- > G 1378 mutation. One resistant strain was amplified neither with MCMM nor with MCMW: a C- > T silent mutation at nt 1368 was present. As sequencing analysis confirmed PCR results, discriminative PCR enables isolates to be rapidly assessed for the presence or absence of 460 mutations. Moreover, it can distinguish Met to Val from Met to Ile mutations, and allows the analysis of mixtures of sensitive and resistant strains.

Anti-Bacterial Agents↗
PubMed1995

Tumour necrosis factor-alpha induces morphological and functional alterations of intestinal HT29 cl.19A cell monolayers.

TNF-alpha is a widely distributed proinflammatory cytokine, involved in many disease states. Although it has widely distributed effects, a precise mechanism of action has never been described, in particular at the epithelial level. Morpho-functional changes of the intestinal epithelial monolayer HT29 cl.19A exposed to TNF-alpha were therefore assessed, using electron microscopy (including freeze-fracture replica analysis), as well as measurement of mannitol, Na+ and horseradish peroxidase fluxes across intestinal HT29 cl.19A cell monolayers using Ussing chambers. TNF-alpha receptors were induced on HT29 cl.19A cells by a small non-toxic dose of IFN-gamma (5 U/ml). After 4 h of the combined presence of TNF-alpha (10 ng/ml) and IFN-gamma (5 U/ml), the tight junction structure was altered as shown by a significant decrease in the average strand number measured in the apico-basal direction (5.50 +/- 2.70 vs 3.73 +/- 1.39 in control and treated cells respectively, P < 0.0001) and by a significant decrease in junctional depth (0.27 +/- 0.14 and 0.17 +/- 0.10 microns in control and treated cells respectively, P < 0.0001). These results are in agreement with a decrease in number of 'kiss' sites between contiguous membranes of TNF-alpha treated cells observed in ultrathin sections.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Membrane Permeability↗
PubMed1995

Detection of Chlamydia trachomatis by ligase chain reaction compared with polymerase chain reaction and cell culture in urogenital specimens.

OBJECTIVE: The aim of this study was to evaluate the newly developed ligase chain reaction (LCR) assay for the detection of Chlamydia trachomatis in urogenital specimens using cell culture and Amplicor PCR for comparison. SUBJECTS: Two hundred and eighty patients attending hospital or urban STD clinics (high-risk population, 62 men and 84 women) and obstetric/gynaecology clinics (low-risk population, 134 women) in Bordeaux, France. METHODS: Specimens from men were tested with LCR on urethral swabs and urine, with Amplicor or urine, with cell culture on urethral swabs. Specimens from women were tested with LCR, Amplicor and cell culture on endocervical swabs and with LCR on urine. When the three methods generated different results, the LCR and Amplicor tests were repeated on the remaining samples. Samples with discordant LCR and Amplicor results and a negative culture were further analysed by major outer membrane protein gene omp1-PCR. RESULTS: After analysis of discrepant results, the overall prevalence was 7.5% (21/280) calculated on the basis of an expanded "gold standard" defined as culture positive or LCR plus Amplicor positive or omp1-PCR positive for discrepant results between LCR and Amplicor tests. Of the 21, 20 were detected by LCR, 17 by Amplicor and culture. The specificity of LCR and Amplicor was 99.6%. CONCLUSION: The LCR Chlamydia trachomatis test is a highly sensitive nonculture technique and a good alternative test for the detection of chlamydial infections.

Bacteriological Techniques↗
PubMed1995

[Molecular typing by pulsed field gel electrophoresis of Pseudomonas (Burkholderia) cepacia isolated from a nosocomial infection].

Because of the emergence of Pseudomonas (Burkholderia) cepacia, isolated among patients from an intensive care unit of the Hôpital Pellegrin (Bordeaux), strains were studied in the aim to determine the source and the mode of transmission of the bacteria. The study was performed on 44 isolates of P. cepacia from 21 patients (23 strains) or from the environment (16 isolates from ventilators and five from the distilled water used for these ventilators). Susceptibility to 23 antibiotics of these strains and endonuclease restriction polymorphism on their total DNA were determined. Pulsed field gel electrophoresis was performed after digestion of the genomic DNA, using three restriction enzymes. Results obtained by analysis of genotypic characteristics demonstrated the clonal origin of the P. cepacia epidemy in the different intensive care units, but did not localize the source of the infection.

Aminoglycosides↗
PubMed1994

Homogeneity of the major outer membrane protein gene of feline Chlamydia psittaci.

Thirteen feline Chlamydia psittaci strains isolated in various countries over a 50-year period were examined by restriction mapping of the major outer membrane protein (MOMP) gene obtained after DNA amplification by the polymerase chain reaction and compared with avian, ovine and guinea pig inclusion conjunctivitis (GPIC) C psittaci strains. All the feline isolates produced a unique AluI pattern distinct from the other C psittaci strains and were characterised by a typical fragment doublet of 185/180 base pairs. Such a homogeneity argues for their genetic relatedness and suggests their clonal origin. These data demonstrate the usefulness of MOMP-gene restriction mapping in C psittaci typing.

Animals↗
PubMed1994

A physical map and candidate genes in the BRCA1 region on chromosome 17q12-21.

We have constructed a physical map of a 4 cM region on chromosome 17q12-21 that contains the hereditary breast and ovarian cancer gene BRCA1. The map comprises a contig of 137 overlapping yeast artificial chromosomes and P1 clones, onto which we have placed 112 PCR markers. We have localized more than 20 genes on this map, ten of which had not been mapped to the region previously, and have isolated 30 cDNA clones representing partial sequences of as yet unidentified genes. Two genes that lie within a narrow region defined by meiotic breakpoints in BRCA1 patients have been sequenced in breast cancer patients without revealing any deleterious mutations. These new reagents should facilitate the identification of BRCA1.

Autoantigens↗
PubMed1994

Life quality of patients undergoing liver transplantation.

The aim of this study was to assess the life quality of a group of patients who had undergone liver transplantation using (1) a psychological test to evaluate family relations, work activity, emotional state and social relationships; (2) the quantification of hospital dependence and degree of fitness for work. Included in the study were 32 patients using the following criteria: diagnosis of hepatic cirrhosis and minimum posttransplant follow-up of 6 months. The average age of the study population was 44.8 +/- 10.5 years; there were 23 males and 9 females, with an average follow-up of 15 months. The psychological test used was the Quality of Life Scale (QLS) which consists of 21 items each scoring from 1 to 6 points. The questionnaire was completed before transplantation by all the patients, and after transplantation by 32 patients at 6 months, 20 at 12 months and 12 at 24 months. Hospital dependence was evaluated by the number of admissions and number of days per admission. Lastly, we compared fitness for work before transplantation and at 1 and 2 years after transplantation. The QLS test showed a post-transplant improvement in the four aspects assessed, particularly in the personal aspects (emotions and family) (P < 0.001). Hospital dependence following liver transplantation decreased significantly compared with the pretransplant situation (P < 0.01). Finally, the post-transplant percentage of unfitness for work decreased with time, reaching a significant differences 2 years after transplantation (P < 0.05).

Adult↗
PubMed1994

Genetic variability among Chlamydia trachomatis reference and clinical strains analyzed by pulsed-field gel electrophoresis.

Pulsed-field gel electrophoresis (PFGE) was applied to Chlamydia trachomatis reference strains representing each of the 18 serovars and to 29 clinical isolates from genital specimens collected in Bordeaux, France, or Malmö, Sweden. Comparison of the fingerprint patterns of the reference strains revealed a high level of polymorphism of the total DNA when SmaI was used (14 profiles), whereas the other enzymes, Sse8387I and ApaI, showed fewer differences. Some serovars, considered to be closely related on the basis of their antigenic determinants located on the major outer membrane protein (MOMP), such as D and Da or I and Ia, were shown to be different after PFGE of their genomic DNAs. However, serovars B and Ba and serovars L2 and L2a had identical patterns after analysis with the three endonucleases. When applied to clinical isolates, which were typed by restriction fragment length polymorphism analysis of the MOMP gene, PFGE allowed the detection of intragenotype polymorphisms and showed the identity of two strains successively isolated from the same patient. This technique seems to be an efficient tool for epidemiological studies when used in addition to serotyping or genotyping by restriction fragment length polymorphism analysis of the MOMP gene.

Bacterial Outer Membrane Proteins↗
PubMed1994

Genetic mapping of the BRCA1 region on chromosome 17q21.

Chromosome 17q21 harbors a gene (BRCA1) associated with a hereditary form of breast cancer. As a step toward identification of this gene itself we developed a number of simple-sequence-repeat (SSR) markers for chromosome 17 and constructed a high-resolution genetic map of a 40-cM region around 17q21. As part of this effort we captured genotypes from five of the markers by using an ABI sequencing instrument and stored them in a locally developed database, as a step toward automated genotyping. In addition, YACs that physically link some of the SSR markers were identified. The results provided by this study should facilitate physical mapping of the BRCA1 region and isolation of the BRCA1 gene.

Base Sequence↗
PubMed1993

Localization of rap1 and rap2 proteins in the gelatinase-containing granules of human neutrophils.

The subcellular localization of rap proteins in resting human neutrophils was investigated by immunoblot analysis with specific anti-rap2 and anti-rap1 antibodies of the membrane proteins obtained from distinct subcellular fractions. Rap2 protein was mainly located in gelatinase-containing granules, whereas rap1 protein was detected both in gelatinase-containing granules and in fractions enriched in plasma membrane. Neither rap1 nor rap2 proteins were found in the cytosol or in azurophilic granules. Rap2B, not rap2A, appeared to be the major rap2 protein in human neutrophils. The identification and subcellular localization of rap1 and rap2 proteins at the membranes of gelatinase-rich granules suggest that these proteins could play a role in the regulation of the rapid and selective mobilization of gelatinase-containing granules in human neutrophils.

Amino Acid Sequence↗