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Biomedical subjects

C Mougin

Publications and source records attributed to C Mougin.

At least 73 records · Page 4Linked to original sources
PubMed1991

Detection of human papillomavirus and human cytomegalovirus in cervical lesions by in situ hybridization using biotinylated probes.

Infections with specific types of human papillomavirus (HPV) have emerged as necessary but not sufficient factors in the development of the majority of cervical cancers. The infection by human cytomegalovirus (HCMV) has also been implicated in both cervical intraepithelial neoplasia (CIN) and cancer. In order to test prevalence of these viral pathogens in genital lesions with suspect cytopathic changes after observation of smears, cervical biopsies from 131 patients were obtained under colposcopic guidance. The biopsies were tested for the presence of HPV and HCMV by the in situ hybridization technique using biotinylated DNA probes on paraffin-embedded sections. Presence of HCMV is twice more frequent in women with HPV-induced cervical lesions (40%) than in women with any detectable HPV (20%). It may be concluded that HCMV might contribute as one synergistic factor in the development of cervical dysplasia.

Cervix Uteri↗
PubMed1991

Optimization of in situ hybridization for detection of viral genomes in cultured cells on 96-microwell plates: a cytomegalovirus model.

In situ hybridization (ISH) for identification of infectious replicative cytomegalovirus (CMV) in cell culture microplates (96 microwells) infected by clinical specimens was tested by using a biotin-labeled DNA probe and an avidin-alkaline phosphatase conjugate. A total of 395 specimens were examined by using ISH and a monoclonal antibody (MAb) specific for an early antigen of CMV. Of 47 specimens that gave a positive signal for CMV by ISH, 33 were confirmed virus positive by MAb staining. Of 141 blood samples tested, 4.96% were positive by ISH, and 0.7% were positive by the MAb technique. ISH shows 40% more sensitivity than MAb staining. This technique should be widely applicable for the specific identification of viral isolates (e.g., herpesvirus, myxovirus, paramyxovirus, and enterovirus) in cell culture 96-microwell microplates, thereby making it feasible to screen a larger number of samples than is possible with classical methods using conventional culture tubes, shell vials, or 24-well plates.

Antibodies, Monoclonal↗
PubMed1990

Histochemical detection of the messenger RNAs coding for calcitonin and calcitonin gene-related peptide in medullary thyroid carcinomas with radioactive and biotinylated oligonucleotide probes.

The present study has been undertaken to investigate the efficiency of biotinylated synthetic oligonucleotide probes in detection by in situ hybridization of the mRNAs coding for calcitonin (CT) or calcitonin gene-related peptide (CGRP) in human medullary thyroid carcinomas (MTCs). Tissue sections fixed with formaldehyde were hybridized with 45-base long oligonucleotides, specific for CT or CGRP mRNA. Recombinant DNA probe or synthetic oligonucleotides radioactively labelled with 32P or 35S were used as controls to detect by autoradiography the corresponding mRNAs in the tumour cells. Oligonucleotide probes labelled by fixation of one biotin molecule at their 5'-end, or by incorporation of a tail of biotin-11-dUTP at their 3'-end, were used and were revealed by incubation with streptavidin-alkaline phosphatase associated with the corresponding substrate. Each biotinylated probe stained exclusively the cytoplasm of the tumour cells, the CT probe giving a much higher level of staining than the CGRP probe. The same cells were found to contain CT and CGRP mRNAs. Controls performed with either radioactive or biotinylated probes confirmed the specificity of the staining. These results demonstrate that biotinylated synthetic oligonucleotides can be used as efficient tools to investigate gene expression in tissue sections, thus avoiding the various inconveniences connected with the use of radioactive probes, especially bio-hazards, the use of autoradiography, the limited histological resolution, and the delay in obtaining results.

Autoradiography↗
PubMed1990

Topography and ontogeny of the neurons expressing vasopressin, oxytocin, and somatostatin genes in the rat brain: an analysis using radioactive and biotinylated oligonucleotides.

1. The use of radioactive and biotinylated oligonucleotide probes has been optimized to detect and analyze by in situ hybridization, neurons expressing neuropeptide genes (vasopressin, oxytocin, somatostatin). 2. In situ hybridization was performed on cryostat-cut sections obtained from tissues perfused with 1% formaldehyde. Radioactive probes were labeled by tailing with 35S-dATP and revealed with autoradiography. Biotinylated probes were obtained either by the incorporation of 11-biotin dUTP or by the addition of biotinylated nucleotides to the oligonucleotide during its synthesis. Biotin was revealed with streptavidin alkaline phosphatase and the appropriate substrate. 3. In the adult rat brain, radioactive and biotinylated probes revealed peptidergic neurons. The biotinylated probes provided an optimal cellular and subcellular resolution with a sensitivity similar to that observed with radioactive probes. Staining was selectively restricted to the cytoplasm and to the proximal part of processes. 4. Biotinylated vasopressin probes with 10 biotins added demonstrated magnocellular neurons and parvocellular neurons in the suprachiasmatic nucleus and the bed nucleus stria terminalis. 5. Vasopressin gene expression was studied during ontogeny in the rat fetus and neonate. Vasopressin mRNA was first detectable at gestational day 16 in the supraoptic nucleus in neurons of neuroblastic appearance. An aspect similar to the one present in adult was found at gestational day 19 in magnocellular neurons and at day 3 postnatal in parvocellular neurons. 6. The results confirm that radioactive oligonucleotide probes are efficient tools to investigate neuropeptide gene expression by in situ hybridization and demonstrate that biotinylated oligonucleotides are very efficient and provide a much higher resolution than radioactive probes with a reasonable sensitivity.

Animals↗
PubMed1988

Plasma levels of beta-endorphin, prolactin and gonadotropins in male athletes after an international nordic ski race.

Plasma beta-endorphin, prolactin (PRL), FSH and LH were measured in 17 volunteer male subjects at rest and under the stress caused by a long-distance nordic ski race. The race induced increased levels of beta-endorphin and PRL in all skiers. The changes in PRL with exercise were significantly related to the changes in beta-endorphin (r = 0.69, p less than 0.001). Furthermore, the highly trained skiers training over 150 km.week-1 of nordic ski showed consistently higher post-exercise beta-endorphin and PRL levels than the moderately trained skiers who trained for 20 km.week-1. In addition the race induced slight falls in FSH and LH; however plasma gonadotropin levels did not show any correlation with plasma beta-endorphin concentrations and did not differ between the two groups of skiers. These results suggest that endogenous opioid peptides may modulate PRL secretion in heavy exercise, since they are of minor importance in the release of FSH and LH in such a situation. The observations also suggest that the degree of previous training and the exercise intensity do seem to be responsible for the hormonal changes.

Gonadotropins↗
PubMed1988

Histological detection of messenger RNAs with biotinylated synthetic oligonucleotide probes.

We achieved histological detection of the messenger RNAs coding for vasopressin, calcitonin, or calcitonin gene-related peptide by using biotinylated synthetic oligonucleotides, and defined the technical parameters enabling optimal detection of these mRNAs. Oligonucleotides labeled by fixation of one biotin at their 5' end or by addition of a biotin-11-dUTP tail at their 3' end can be used to detect mRNAs, although the latter are more sensitive. Streptavidin-alkaline phosphatase revealed with nitroblue tetrazolium-bromo-chloro-indolyl phosphate as substrate makes possible detection of the biotinylated oligonucleotides. Increasing formaldehyde concentration in the fixative decreases the signal intensity; 1% formaldehyde fixation provides the most intense signal. Several controls, including those with addition of unlabeled oligonucleotides to the hybridization buffer, confirm the specificity of mRNA detection. The sensitivity of the biotinylated probes is identical or lower as compared to the corresponding radiolabeled oligonucleotides. Histological and subcellular resolution is greatly enhanced with biotinylated probes. The rat vasopressin probes stain magnocellular neurons in the supraoptic and paraventricular nuclei and, under optimal conditions, parvocellular neurons in the suprachiasmatic nucleus. Vasopressin mRNA is present in the cytoplasm of the cell bodies and in the roots of certain processes. Calcitonin and calcitonin gene-related peptide mRNA are found co-localized in the cytoplasm of the same tumor cells in human medullary thyroid carcinoma.

Animals↗
PubMed1987

Assessment of plasma opioid peptides, beta-endorphin and met-enkephalin, at the end of an international nordic ski race.

Plasma met-enkephalin, beta-endorphin, cortisol and lactic acid concentrations were measured in seventeen volunteer male subjects at rest and after a long-distance nordic ski race. Immediately after the race, mean plasma met-enkephalin did not show any significant change, but significant rises in beta-endorphin, cortisol and lactic acid were noted in all skiers. The change in beta-endorphin with exercise was significantly related to the change in cortisol (r = 0.68; p less than 0.001) and to the change in plasma lactic acid (r = 0.60; p less than 0.001). Furthermore, the experienced skiers training over 150 km X week-1 of nordic ski had significantly faster skiing times in this event and showed greater beta-endorphin, cortisol and lactic acid levels than the recreational skiers who trained for 20 km X week-1. Our results imply that the changes in plasma beta-endorphin depend on the intensity of exercise. However the significance of higher levels of skiing training or previous nordic ski experience in the release of beta-endorphin is expected and cannot be excluded.

Adult↗
PubMed1987

Molecular hybridization versus isoelectric focusing to determine TEM-type beta-lactamases in gram-negative bacteria.

Isoelectric focusing and molecular hybridization with a TEM DNA probe were used to screen for TEM beta-lactamase in 328 bacterial isolates representing 11 gram-negative genera. The TEM enzyme was detected in 50% of isolates, and nine additional types of beta-lactamase could be identified in 36.9% of isolates. The TEM gene was detected in 53.6% of isolates. The results obtained by both methods were concordant in 92.7% of the entire sample. In situ colony hybridization with a specific probe therefore appears to be a convenient method to screen rapidly for the presence of homologous genetic sequences among a large number of isolates. Positive hybridization was observed for 16 isolates in which no TEM beta-lactamase was detected by isoelectric focusing. The significance of this hybridization remains to be determined.

DNA, Bacterial↗
PubMed1985

[Intervention of the calcium-calmodulin system in the release of growth hormone induced by somatocrinin].

This study was carried out to evaluate the role of the calcium-calmodulin system in cellular mediation of somatocrinin action by cells producing growth hormone. We compared GH release to intra- and extracellular cAMP levels, after incubation of pituitary cells in monolayer culture with somatocrinin alone or somatocrinin associated with exogenous calmodulin, or specific calmodulin inhibitor (W13). Calmodulin (10(-6) M) decreases somatocrinin - induced cAMP levels, maintaining elevated GH levels. W13 (10(-5) M) increases somatocrinin - induced cAMP and GH levels. Thus GH release induced by somatocrinin is modulated by two interrelated systems, adenylcyclase - cAMP system and Ca++-calmodulin system.

Animals↗
PubMed1984

Effects of forskolin and cholera toxin on cyclic AMP release in a neurotensin-secreting rat C-cell line.

The effects of forskolin and cholera toxin on the regulation of cAMP release were studied in a neurotensin-secreting rat C-cell line. The interaction of these agents with norepinephrine, a potent neurotensin secretagogue, was also investigated. Forskolin stimulated cAMP release 10(2)-10(3) fold while it increased neurotensin release 2-3 fold. Cholera toxin caused a 10(2)-10(3) fold increase in cAMP release and had no effect on neurotensin release. We conclude that the 44-2 C-cells provide a new model for studying the regulation of the concomitant (via forskolin) or independent (via cholera toxin) secretion of cyclic AMP and/or neurotensin.

Adenylyl Cyclases↗
PubMed1984

[Roles of cyclic AMP and calcium in the mechanism of the release of growth hormone by somatocrinin].

Growth hormone (GH) release from Rat pituitary cell monolayers in response to synthetic somatocrinin (7.8 to 1,000 pmol/l) is paralleled with an increase in cellular cyclic AMP (cAMP) content and efflux of cAMP in the extracellular medium. Somatostatin and blockers of calcium-dependent cellular mechanisms inhibit somatocrinin-induced GH release but only partially decrease (somatostatin, CoCl2) or even increase (trifluoperazine) cAMP levels. Thus, calcium is required for somatocrinin action and GH release is not simply dependent on stimulation of cAMP metabolism.

Animals↗
PubMed1984

[Comparative study of urinary calcium, oxalate and uric acid in calcium oxalate lithiasis].

The purpose of this study was to determine and to compare the frequency of hypercalciuria, hyperoxaluria and hyperuricosuria in 49 patients with a pure or mixed calcium oxalate urolithiasis. During this study, all patients were on a normal diet and had no special disease or medication. We noted that hypercalciuria (35%) and hyperoxaluria (24%) were more often associated (18%) than isolated (8 and 2%). They were more frequent in mixed stones (44% et 39%) than in pure one (29% and 16%). Hyperuricuria (39%) had a same frequency wathever the renal stone type may be. A same frequency was noted when hyperuricuria was associated with hypercalciuria and hyperoxaluria (12%) or isolated (14%). None of these three biologic disturbances was observed in 45% of our patients.

Adult↗
PubMed1983

[Transcutaneous wirsungography under echoscopic control].

Two cases of contrast radiography of Wirsung's canal performed by direct percutaneous puncture of the canal under ultrasonic control are reported. The exploration was well tolerated by the patients. This method appears to be of considerable interest in liver and pancreas diseases, being reliable and providing valuable information.

Adult↗
PubMed1983

Regulation of neurotensin secretion in a mammalian C cell line: effect of dexamethasone.

We established in culture two colony clones of rMTC 44-2 cells, rMTC 44-2B and 44-2C which secrete substantially greater quantities of neurotensin (NT) than the parent cell line. We describe here the effects of the synthetic glucocorticoid, dexamethasone, on NT and cAMP release. Medium and intracellular levels of NT and cAMP were measured by specific RIAs. Long-term release experiments were performed in Dulbecco's Modified Eagle's Medium supplemented with 15% horse serum (DMEM). Short-term release experiments were performed in Krebs-Ringer-bicarbonate-glucose buffer (KRBG) supplemented with 1.0 mm Ca2+. Dexamethasone stimulated NT release and increased intracellular NT levels. The ED50 values for stimulation of NT release following 24 or 48 h incubation of cells in DMEM with dexamethasone were 5 X 10(-9) and 7 X 10(-9) M, respectively. Dexamethasone markedly enhanced intracellular levels of NT in rMTC 44-2 cells while it decreased cell growth. Cells pretreated with dexamethasone for 48 h released greater amounts of NT in response to Ca2+ (1.0 mM) with or without K+ (50 mM) or NE (10(-6) M) following a 10 min incubation with these substances in KRBG. This experimental paradigm was also used to measure the efflux of cAMP following a brief (10 min) exposure of cells to NE. We conclude that the rMTC 44-2B and 44-2C cells are useful tools for studying the effects of dexamethasone on the regulation of cell growth, as well as the secretion of NT and cAMP.

Animals↗
PubMed1982

[Urinary excretion of calcium and cyclic adenosine monophosphate in children. Comparison between the two variables].

The urinary excretion of calcium (mg/g creatinine) and cyclic adenosine 3'5' monophosphate (cyclic AMP) (mumol/g creatinine) was determined in 79 healthy children aged 2 to 12 years. An ag--related pattern to the urinary excretion of calcium was not seen. But urinary cyclic AMP levels are inversely correlated with age (r = -0,66; p less than 0,01) and there is a significant correlation between urinary calcium and cAMP levels (r = 0,2; p less than 0,05).

Aging↗