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Biomedical subjects

F Lebargy

Publications and source records attributed to F Lebargy.

53 records · Page 3Linked to original sources

Tissue-specific expression of two gamma-glutamyl transpeptidase mRNAs with alternative 5' ends encoded by a single copy gene in the rat.

Two different cDNAs have been isolated and characterized from a rat kidney cDNA library. The two cDNA sequences are identical in the coding region and in the 144 bases upstream from the initiation codon but have alternate sequences (154 and 138 bases) at their 5' ends. Primer extension analysis on kidney mRNA reveals that both cDNAs are full-length and correspond to two mRNAs of nearly the same size (2142 and 2127 bases). Synthesis of two mRNAs with alternative 5' ends can be explained only by initiation at two separate promoters on the single rat gamma-glutamyl transpeptidase (GGT) gene. The alternate 5' end nucleotide sequences were used as probes to detect the corresponding mRNAs in several rat tissues. In the kidney, the expression of both RNAs was detected by in situ hybridization in the distal part of the proximal convolutions of the renal tubule. Northern blot analysis of kidney mRNAs reveals that the expression of both mRNAs increases from birth to the adult stage. Neither of these two transcripts is expressed in the liver or in seminal vesicles in which a larger mRNA (2.4 kilobase pairs) is transcribed from the same gene. Thus, two GGT mRNAs, initiated on two separate promoters on the single GGT gene, are expressed in the rat in a tissue-specific manner and coordinately regulated.

Animals↗

Expression of Tau protein and Tau mRNA in the cerebellum during axonal outgrowth.

UNLABELLED: "In situ" hybridization and immunohistochemical analysis of the expression of Tau mRNAs and Tau proteins in the developing cerebellum showed that: 1. At early postnatal stages Tau mRNAs are expressed in the deeper region of the external granular layer (EGL II) i.e. in the cells that begin to migrate from the proliferative zone. Little labeling was seen in the upper layer (EGL I) where the cerebellar interneurons actively proliferate during the first two postnatal weeks. Anti-Tau antibodies failed to detect Tau proteins both in EGL I and II. 2. Tau transcripts were also clearly detected in the migrating cells present in the molecular layer; no Tau immunoreactivity was seen in this layer. This suggests that Tau mRNAs remain very poorly translated in the migrating granule cells and in the other interneurons. 3. Tau proteins begin to be detected at postnatal day 8 in the molecular layer but only at the level of the parallel fibers that are present in the Purkinje cell dendritic field. This suggests that the Tau mRNAs transcribed in the migrating cells are not actively translated for several days and that Tau proteins accumulate only in the more mature sections of their axons, the parallel fibers. IN CONCLUSION: Tau mRNAs are transcribed in the migrating cells several days before Tau proteins are actively translated and transported to their axons. Tau proteins accumulation occurs only at the end of granule cell migration i.e. when the parallel fibers interact with their post-synaptic counterparts, the dendrites of the Purkinje cells. Thus, axonal outgrowth and differentiation seem to be a multistep process.

Aging↗

HIV-1 infection of lung alveolar fibroblasts and macrophages in humans.

We have studied the infected cell populations in the lungs of four human immunodeficiency virus type 1 (HIV-1) seropositive patients suffering from lymphocytic alveolitis or lymphocytic interstitial pneumonitis. Adherent cells were obtained by bronchoalveolar lavage (BAL) and were analyzed by various technical approaches. The cells considered here were alveolar macrophages and fibroblasts, and could be clearly identified morphologically and by the expression of specific cell-surface markers using monoclonal antibodies. The presence of HIV-1 in both of these cell types was established by serological, virological, and molecular procedures. Our results show that alveolar macrophages and fibroblasts are naturally infected in the lungs of HIV+ patients. Both cell types express the CD4 receptor molecule, in contrast to skin fibroblasts which are negative. Alveolar macrophages and fibroblasts thus may act as eventual HIV-1 reservoirs in vivo, and are probably involved in the induction of inflammatory reactions because they are targets for CD8 cytotoxic T lymphocytes (CTL).

AIDS-Related Complex↗

Localization by in situ hybridization of gamma-glutamyl transpeptidase mRNA in the rat kidney using 35S-labeled RNA probes.

gamma-Glutamyl transpeptidase (GGT) is involved in the extraction of plasma glutathione in the postglomerular compartment of the kidney. The enzyme is distributed in the proximal tubule associated with the apical brush border. Using in situ hybridization with 35S-labeled anti-sense and sense RNA GGT probes on rat kidney cryostat sections, the present study demonstrates that GGT mRNA transcripts are detected in proximal tubules localized in the inner cortex, outer medulla, and medullary rays of the cortex. Such a distribution suggests that the GGT gene is mainly expressed in the more distal part of the proximal tubule, i.e., the pars recta. As a control, in situ hybridization has also been performed using a 35S-labeled beta-actin anti-sense RNA probe showing a diffuse pattern of distribution of beta-actin RNA transcripts particularly in the outer cortex. This highly sensitive method using single strand asymetric RNA probes, with a markedly reduced nonspecific binding is promising for the study of gene expression patterns in the kidney in order to clarify their heterogeneity in the various segments of the nephron.

Animals↗

Intraepithelial T-lymphocyte subsets in the airways of normal subjects and of patients with chronic bronchitis.

Lymphocyte infiltration of central airway epithelium was evaluated in 13 normal nonsmoking subjects (Group 1), in 11 smokers without clinical signs of chronic bronchitis (Group 2), and in 34 patients who were smokers with chronic bronchitis and mild airflow limitation (Group 3). Bronchial samples were obtained through fiberoptic bronchoscopy. Murine monoclonal antibodies directed against cell-surface antigens and an immunoperoxidase technique were used on cryostat sections to label in situ the following lymphocyte populations: T-lymphocytes (CD3+), helper/inducer T-cells (CD4+), suppressor/cytotoxic T-cells (CD8+) and B-lymphocytes (leu 12+). Virtually no B-cells were found in central airway epithelium from subjects of any group. Conversely, consistent infiltration of epithelial layers with T-lymphocytes of both subsets was observed in all subjects, with a constant predominance of CD8+ over CD4+ cells. For any T-cell marker, differences between mean scores from Group 1 and Group 2 subjects were not statistically significant. On the other hand, mean lymphocyte numbers of both subsets were found increased in patients from Group 3 compared with subjects from the two other groups: statistically significant differences were observed for CD3+, CD4+, and CD8+ cells (p less than 0.001). Furthermore, lymphocyte scores at two different airway generation were compared in some patients from Groups 2 and 3, and a significant positive correlation was observed. These results suggest that T-lymphocyte infiltration of central airway epithelium (1) may be a naturally occurring phenomenon that is amplified in the airways of smokers with chronic bronchitis, and (2) may represent the counterpart to the intraepithelial population of the intestine.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Effects of digoxin on diaphragmatic strength generation in patients with chronic obstructive pulmonary disease during acute respiratory failure.

We studied the effects of digoxin, a compound that has an inotropic effect on the myocardium, on diaphragmatic function in 8 patients with chronic obstructive pulmonary disease. All the patients were in acute respiratory failure and were artificially ventilated. Diaphragmatic strength was assessed by measuring the transdiaphragmatic pressure generated at functional residual capacity during bilateral supramaximal electrical stimulation of the phrenic nerves. The latter were stimulated before and at 45 and 90 min after administration of digoxin (0.02 mg/kg infused for 10 min). In all the patients, cardiac output was measured by the thermodilution technique using a Swan-Ganz catheter placed in the pulmonary artery. Arterial blood gases and pH were maintained within normal range by mechanical ventilation. In all the patients, digoxin plasma levels reached the therapeutic range (mean values, 2.82 +/- 0.17 and 2.90 +/- 0.20 nmol/L at 45 and 90 min, respectively) after digoxin administration. Diaphragmatic strength improves significantly after digoxin administration, the transdiaphragmatic pressure for an identical phrenic stimulation increasing by 19.5% (p less than 0.001) on the average. This increase was noted 45 and 90 min after digoxin administration. We conclude that digoxin has a potent effect on diaphragmatic strength generation that may be beneficial in patients with chronic obstructive pulmonary disease during acute respiratory failure. Furthermore, this inotropic positive effect of digoxin on the diaphragm, as previously observed for the myocardium, emphasizes the similarities between these 2 contractile tissues.

Action Potentials↗

[Is bronchial fiberscopy always indicated in hemoptysis? Apropos of 427 tests].

The authors report the results of a retrospective study of 427 fiberoptic bronchoscopies in patients with haemoptysis. The patients were classified into 4 groups according to the results: Group I: a specific diagnosis was made (cancer, tuberculosis, infection); 151 cases (35 p. 100); Group II: inflammatory mucosa, 172 cases (40 p. 100); Group III: normal bronchoscopy, 84 cases (20 p. 100); Group IV, only blood, 20 cases (5 p. 100). Of the 151 patients in Group I, 12 had a normal chest X-ray (8 p. 100); seven of them had bronchopulmonary carcinoma (7 out of 65 cases of cancer: 10.7 p. 100). These results show that bronchoscopy is essential in cases of haemoptysis even when the chest X-ray is normal.

Bronchial Neoplasms↗

Morphological changes in rat tracheal mucosa immediately after antigen challenge.

We have investigated the morphological effects of an intratracheal challenge with 50 micrograms ovalbumin (OA) on sensitized rat tracheas, in vivo. Female Sprague-Dawley rats were primed ten days before challenge with a single i.t. injection of 100 micrograms OA plus Bordetella pertussis (OA-BP). Two additional groups of animals served as controls: primed animals challenged with saline only and non-primed but OA-challenged animals. Sacrifices--and subsequent morphological studies--were performed prior to and 5, 15 and 60 min after challenge. At each time, the total numbers of epithelial nuclei, subepithelial mast cells (SEMC) and intraepithelial mast cells (IEMC) were scored in six non-adjacent cross sections per trachea. We found that: 1) priming with OA-BP alone did not induce any change in the tracheal mucosa with respect to morphological structure and mast cell counts; 2) no morphological change nor significant modification of the cell counts occurred at any time in tracheas from either of the control groups; 3) in contrast, a luminal heterogeneous exudate and a subepithelial oedema developed in 9 of the 15 tracheas of primed animals within 60 min of OA challenge. In those nine tracheas, the scores of intraepithelial nuclei, of IEMC and of SEMC were found to decrease significantly 15 min after challenge as compared with starting values (p less than 0.05 for each score). The decrease in the number of mucosal mast cells is probably related to the damages of the epithelial cells and to the difficulty with which depleted mast cells can be seen by toluidine blue staining.

Animals↗

[Nicotine: a psychoactive drug, a doping agent?].

A 43-year-old high-level athlete used nicotine gum to stop his smoking habit. Later, use of nicotine gum was directly related to improved sports performance. It is suggested that nicotine might be added to the official list of prohibited drugs in athletes.

Adult↗

CFTR gene mutations in adults with disseminated bronchiectasis.

The severity and type of clinical manifestations are variable in patients with cystic fibrosis (CF). The respiratory syndromes in these patients consist of lung infections associated with disseminated bronchiectasis (DB), asthma, and chronic obstructive pulmonary disease. To investigate the possible involvement of the cystic fibrosis transmembrane conductance regulator (CFTR) gene in chronic pulmonary disease in adults, we studied 32 DB patients with a clinically isolated respiratory syndrome. Careful analysis of all the CFTR gene exons and their flanking regions revealed a significantly increased frequency of CFTR gene mutations in these patients. Thirteen CFTR gene mutations were identified in sixteen different alleles. Six of these mutations, which have previously been reported as CF defects, were found on nine alleles. A further four, two of which had not previously been described (D192N and 406-2 AdeltaC), are potentially disease-causing mutations. We also identified three rare substitutions (R31C, L997F, T1220I), which could be involved in mild CFTR gene disease. Four patients were compound heterozygotes, one carried two CFTR gene mutations (possibly allelic) and six were heterozygous for a mutation. These results indicate that CFTR gene mutations may play a role in bronchiectatic lung disease, possibly in a multifactorial context. These findings have implications for genetic counselling of DB patients and their families.

Adult↗