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J Jacquot

Publications and source records attributed to J Jacquot.

At least 37 records · Page 2Linked to original sources
PubMed1993

Vectorial delivery of newly-synthesized secretory proteins by human tracheal gland cells in culture.

The polarized secretion of newly-synthesized proteins of human tracheal submucosal gland cells was studied. Human tracheal gland cells were cultured on permeable filter supports allowing a separate biochemical analysis of apical and basal secretion. By transmission electron microscopy, confluent filter-grown cells were seen to form a continuous sheet of both multilayer and monolayer epithelial cells. Junctional complexes between adjacent cells were observed. On immunofluorescence microscopy, human tracheal gland cells in cultured exhibited characteristics of epithelial and secretory cells, including cytoplasmic staining for cytokeratin and for two secretory protein markers specific to the glandular serous type cell: lysozyme and antileucoprotease. [35S]methionine metabolic labelling experiments demonstrated that at least 90% of newly-synthesized secretory proteins were recovered in the apical medium. Moreover, lysozyme secretion was strongly polarized since 85% was released into the apical medium. Conversely, antileucoprotease secretion was more bidirectional since nearly 40% of released antileucoprotease was present in the basal medium. The fact that these two secretory proteins are released with differing relative polarity emphasizes that human tracheal gland cells exhibit at least two different exocytotic routing operations.

Cell Polarity↗
PubMed1992

Differential localization of the cystic fibrosis transmembrane conductance regulator in normal and cystic fibrosis airway epithelium.

Deletion of the amino acid residue Phe 508 of the cystic fibrosis transmembrane conductance regulator (CFTR) protein represents the most common mutation identified in cystic fibrosis (CF) patients. A monoclonal and a polyclonal antibody directed against different regions of CFTR were used to localize the CFTR protein in normal and CF airway epithelium derived from polyps of non-CF and CF subjects homozygous for the delta Phe 508 CFTR mutation. To identify the cellular and subcellular localization of CFTR, immunofluorescent light microscopy, confocal scanning microscopy, and immunogold transmission electron microscopy were performed on cryofixed tissue. A markedly different subcellular distribution was identified between normal and CF airway epithelial cells. In normal epithelium, labeling was restricted to the surface apical compartment of the ciliated cells. In contrast, in the epithelium from homozygous delta Phe 508 CF patients, CFTR markedly accumulated in the cytosol of all the epithelial cells. These findings are consistent with the concept that the CFTR delta Phe 508 mutation modifies the intracellular maturation and trafficking of the protein, leading to an altered subcellular distribution of the delta Phe 508 mutant CFTR.

Cystic Fibrosis↗
PubMed1992

Functions of proteins and lipids in airway secretions.

Proteins and lipids synthesized by airway secretory cells or transudated are active components in the protection of respiratory epithelium. Proteins and ions are involved in the control of mucus hydration. Secretory proteins, such as secretory immunoglobulin A (IgA), transferrin and lysozyme, participate in the airway antibacterial defence. Other biochemical components found in secretions, such as anti-inflammatory and antioxidant agents as well as antiproteases, contribute significantly to the protection of the underlying epithelium.

Bronchi↗
PubMed1991

Modulations of the epithelial phenotype and functional activity of cultured bovine tracheal gland cells: dependence on the culture medium and passage number.

Bovine tracheal gland (BTG) cells in culture show an epithelial-fibroblastoid transition after several passages. To investigate these BTG cell phenotype changes, we studied the effects of both the culture medium and passage number on the expression of epithelial cytoskeletal proteins and glandular serous cell markers. We also analyzed the intracellular cAMP level in the basal state and after adrenergic stimulation. Three culture media were used: 1) serum-free defined medium (SFDM); 2) medium supplemented with 2% Ultroser G; and 3) medium supplemented with 10% fetal calf serum (FCS). Using immunofluorescence microscopy, we showed that, in the first 4 passages whatever the culture conditions, BTG cells expressed immunoreactivities to cytokeratin filaments and desmoplakins I and II, whereas vimentin filaments were not detected. After four passages, BTG cells cultured in 10% FCS or 2% Ultroser G became progressively fibroblastoid and showed immunoreactivities to both vimentin and cytokeratin intermediate filaments. No immunoreactivity to vimentin filaments was observed on BTG cells cultured in a SFDM. Using biochemical analysis, we showed that basal levels of cAMP in cultured BTG cells and lysozyme secretion by these cells vary according to the culture medium and passage number. It was higher in BTG cells cultured in a SFDM compared to that recovered from cells cultured in medium supplemented with Ultroser G or FCS. Whatever the culture medium, BTG cells responded to stimulation by isoproterenol. However, the results of stimulation in a SFDM were higher than in Ultroser G or FCS supplemented medium. We conclude that the BTG epithelial cell organization and the regulation of biosynthesis of secretory proteins by these cells in culture depend on both the culture medium and passage number.(ABSTRACT TRUNCATED AT 250 WORDS)

1-Methyl-3-isobutylxanthine↗
PubMed1991

COR3224--a new antidepressant: pharmacokinetics and metabolism in rat.

COR3224 is a new 2-amino-2 oxazoline derivative with antidepressant properties. The distribution, excretion and metabolism of radiochemically labelled 14C-COR3224 has been investigated in the rat after intravenous injection. The compound was widely distributed and rapidly excreted. Hydroxylation and sulphate conjugation are involved in the COR3224 elimination.

Animals↗
PubMed1990

Production of lipocortin-like proteins by cultured human tracheal submucosal gland cells.

Evidence is obtained for the presence of lipocortin-like proteins in human tracheal gland cells in culture. Using polyclonal antibodies to lipocortin I, indirect immunofluorescence studies demonstrate that lipocortin I is mainly confined to the tracheal gland cell surface. From cell membranes, four Ca2(+)-dependent proteins (35, 40, 45 and 67 kDa) were identified as lipocortin related proteins by using immunoblotting and fluorography following [35S]methionine metabolic labeling experiments. A strong immunoreactivity for the 35 kDa protein was observed. In addition, lipocortin-like proteins with apparent Mr33, 35, 37 and 67 kDa, respectively, were released in the apical culture medium by tracheal gland cells cultured on microporous membrane of a double chamber culture system.

Annexins↗
PubMed1990

Modulation of albumin-like protein and lysozyme production by bovine tracheal gland serous cells. Dependence on culture conditions.

Bovine tracheal submucosal gland serous cells were cultured in medium supplemented with either 10% fetal calf serum or 2% Ultroser G, a commercial serum substitute for cell culture. The proteins synthesized and secreted into the culture medium during [35S]methionine pulse, chase and isoproterenol-stimulated periods were analyzed. Marked differences in the patterns of secretory radiolabeled proteins with Mr values ranging from 15,000 to 95,000 were observed between pulse and chase media of cells cultured in fetal calf serum and Ultroser G. In the presence of Ultroser G, albumin-like protein production was inhibited 95% as compared to cultures incubated with fetal calf serum. A bovine lysozyme-type enzymatic activity was detected only in medium from stimulated cells cultured in Ultroser G. The results suggest that bovine tracheal serous cells synthesize different proteins according to the composition of culture medium and release certain proteins when adrenergically stimulated.

Albumins↗
PubMed1989

The frog palate mucosa as a model for studying bacterial adhesion to mucus-coated respiratory epithelium.

Most of the methods proposed to quantify bacterial adherence to respiratory mucosa differ mainly from in vivo conditions in the absence of the mucus blanket and in the exposure of the sub-mucosal connective tissue (SMCT) to the micro-organisms. We propose the frog palate as a model to study bacterial adhesion to the respiratory mucosa, with a system which allows the mucus to be preserved and the bacterial adhesion to be quantified in a standardized mucosal area, where mucociliary transport is still active. In order to evaluate the role of respiratory mucus in bacteria-mucosa interaction, we compared the adhesion of radiolabelled pneumococci to 12 mucus-coated and 10 non-mucus-coated frog palate mucosae. The presence or absence of mucus was controlled by scanning electron microscopy (SEM). After a 10 min incubation period, the bacterial adhesion to mucus-coated palate mucosa was five times greater (P less than 0.01) than that to uncoated mucosa. By SEM, bacteria were never seen attached to ciliated cells but could be detected on small areas where mucus was not totally eliminated. Even after a 120 min contact of bacteria to uncoated mucosa, bacterial adhesion remained only half that to mucus-coated epithelium. In order to ascertain whether the exposure of the SMCT represented a means of attraction to bacteria, we incubated the frog palate mucosa face-down with radiolabelled Pseudomonas aeruginosa. As much as 44 per cent of added bacteria adhered to exposed SMCT and, by SEM, numerous micro-organisms were seen attached to connective tissue. In contrast, only a few bacteria were observed adhering to the mucosa, mainly to granules of mucus.

Animals↗
PubMed1989

Effect of air humidity on spinability and transport capacity of canine airway secretions.

The effect of varying the inspired air humidity on a rheological property (spinability) and transport capacity of airway mucus has been analyzed in 10 mongrel dogs. Tracheal mucus was collected in anesthetized dogs inspiring through an endotracheal tube the air of a climate chamber maintained at constant temperature (T degrees:20 degrees C). In one test, the dogs inspired air at an absolute humidity (AH) of 9 g water/m3 air directly through the endotracheal tube. In the other test, the dogs inspired through an artificial nose connected to the endotracheal tube giving a AH of 30 g water/m3 air. Tracheal mucus was collected at the external distal end of the endotracheal tube. The spinability (Sp) or thread-forming properties of mucus was measured. The relative mucociliary transport rate (TR) of mucus was analyzed on a frog palate epithelium preparation. The transport rate was significantly (p less than 0.01) lower (range: 0.59-0.80) when the AH of the inspired air was low in comparison to that obtained with high AH (range: 0.70-1.13). The variations in mucus Sp due to changing AH were positively and significantly correlated (r = 0.80, p less than 0.01) with the corresponding variations in TR. These results suggest that lowering the AH of air induces a decrease in the transport capacity which appears to be dependent on the change of spinability that occurs in the mucus.

Air↗
PubMed1989

Growth and characterization of isolated bovine tracheal gland cells in culture. Influence of a reconstituted basement membrane matrix.

We describe a method for establishing the culture of bovine tracheal submucosal gland (BTG) cells, in which we have also examined the influence of a reconstituted basement membrane matrix derived from the Engelbreth-Holm-Swarm tumor (EHS) on the growth and morphological differentiation of these cells. BTG cells have been isolated by tissue enzymatic digestion using trypsin, deoxyribonuclease I, elastase, hyaluronidase and EGTA for 1 hr at 37 degrees C. Afterwards, cells and tissue were collected by centrifugation and were incubated for 15 min with 15% newborn calf serum to inactivate the proteolytic enzymes. Enzymatic digestion using only trypsin, centrifugation and inactivation steps were repeated three times. Using this protocol, we obtained 15 +/- 4 (X 10(6] cells per g of tracheal submucosa with 72 +/- 2% (n = 5) cell viability. On microscopic observation, isolated cells were mainly composed of serous type glandular cells. Cells were cultured in a 1:1 medium of Dulbecco's Modified Eagle's/Ham's F12 supplemented with 10% fetal calf serum and subcultured in either plastic flasks or flasks coated with EHS matrix. On the plastic, the BTG cells exhibited at confluency an epithelioid appearance. They stained positively with the immunofluorescent anticytokeratin antibody and contained PAS-staining granules. By electron microscopy, lactoferrin, a protein marker specific to the serous cells, was demonstrated immunocytochemically in small secretory vesicles. BTG cells cultured on EHS matrix revealed a significantly increased growth in comparison to those cultured on plastic. In post-confluent culture of BTG cells on EHS matrix, we observed numerous dome-like structures formed by differentiated cells which were joined together around luminal spaces.

Animals↗
PubMed1988

Synthesis and secretion of an albumin-like protein by cultured bovine tracheal gland serous cells.

Bovine tracheal submucosal gland serous cells were incubated with (35S) methionine. The proteins synthesized and secreted into the culture medium during pulse and chase periods were analyzed. Three major protein bands with apparent molecular weight values of 67 kD, 47 kD and 32 kD were detected by fluorography following SDS-polyacrylamide gel electrophoresis. Based on the molecular weight, immuno-reactive cross reactivity and surface molecular change characteristics, the labeled Mr 67 kD protein was identified as an albumin-like protein. Our results suggest that albumin found in airway secretions can originate, at least in part, from tracheal gland serous cells.

Albumins↗
PubMed1987

Relationships between the lipid content and the rheological properties of airway secretions in cystic fibrosis.

The lipid composition and rheological properties of expectorated airway secretions have been analyzed in 16 patients with cystic fibrosis (CF), separated into two groups according to whether their secretions were superinfected or not. The total lipid content was higher in the superinfected CF secretions. The content in cholesterol and GL1, GL3 and GL4 glycosphingolipid fractions were significantly higher in the superinfected in comparison to the non-superinfected group. The viscosity was two-fold higher in the CF superinfected group. A significant correlation (r = 0.72, p less than 0.01) was observed between the apparent viscosity and the total lipid content of CF secretions. Cholesterol, glycosphingolipids and sphingomyelin were the lipid components which were the most closely and positively correlated to the viscosity. On the opposite, phosphoglycerol was negatively correlated (r = -0.72, p less than 0.05) to the viscoelastic properties of CF airway secretions.

Adolescent↗
PubMed1987

Identification of different molecular forms of human airway lysozyme.

Human airway lysozyme (HAL) was separated into fractions of distinct molecular forms using a Mono S cation-exchange column on a fast-protein liquid chromatography system. This new and rapid (30 min) purification procedure of human lysozyme enabled the preparation of fractions, highly enriched in different isoenzymes of HAL. Purified HAL from pathological purulent airway secretions, nonpurulent airway secretions, and normal tracheobronchial tissue culture medium was characterized by four, three, and only one enzymatically active molecular forms, respectively. All charge forms (separated or combined) recovered from either purulent or nonpurulent airway secretions or tracheobronchial culture medium exhibited the same apparent molecular weight of 15,000.

Chromatography, Ion Exchange↗
PubMed1987

Effect of human airway lysozyme on the in vitro growth of type I Streptococcus pneumoniae.

The effects of purified human airway lysozyme and hen egg-white lysozyme on growth rate and viability of growing type I Streptococcus pneumoniae were studied. Exposure of bacteria to human and hen lysozyme at the same final concentration (100 micrograms/ml) for 1.5-4.5 h resulted in a marked reduction of the number of colony-forming units per ml compared to control cultures. After a 1.5-h exposure to human or hen lysozyme, the remaining percentage of colony forming units per ml was 54% and 69%, respectively. The onset of growth only appeared after a 3.5-h exposure period for human lysozyme whereas it began at 2.5 h for hen lysozyme. After 3.5 h and 4.5 h of exposure, the number of colony-forming units was significantly lower (p less than 0.05) in human lysozyme-treated bacteria cultures compared to control cultures. Parallel electron microscopic observations of Streptococcus pneumoniae cultures confirmed that the density of pneumococci was less in the presence of either human lysozyme or hen lysozyme in comparison to control cultures, and showed the presence of numerous long, ribbon-like material and cytoplasmic condensations liberated in the culture medium.

Bronchi↗
PubMed1986

Characterization and conformational analysis by Raman spectroscopy of human airway lysozyme.

Human airway lysozyme, purified from pathological bronchial secretions, is characterized by a specific activity 3-fold higher than that of hen egg-white lysozyme. The amino acid composition of human airway lysozyme is identical to that of other human lysozymes. The laser Raman spectra of human airway lysozyme and hen egg-white lysozyme in phosphate buffer solution (pH 7.2) are recorded in the range 300-1900 cm-1 at 488 nm. Drastic intensity differences are observed between the spectra analyzed in the ranges characteristic of the peptide backbone (e.g., beta-sheet; C alpha-C, C alpha-N), and of the aromatic side-chain vibrations (tyrosine, tryptophan). The deconvolution of the Raman amide I band gives secondary structures of 38% and 39% alpha-helix, 25% and 20% beta-sheet, and 37% and 41% undefined structure for the human and hen lysozymes, respectively.

Amino Acids↗
PubMed1986

Adherence of type I Streptococcus pneumoniae to tracheal epithelium of mice infected with influenza A/PR8 virus.

Bacterial adherence to virus-infected respiratory tract cells may be one of the several mechanisms whereby virus predisposes to bacterial pneumonia. To evaluate the effect of influenza virus infection on pneumococcus adhesion, 39 mice were infected with PR8/A influenza virus. The adherence of radiolabeled pneumococcus to mice tracheal cells was determined 2, 4, and 6 days after viral inoculation. The pneumococcal adhesion to infected tracheas was significantly enhanced on Day 6 (p less than 0.001). Scanning and transmission electron microscopy revealed that by the fourth and sixth days after virus inoculation, the ciliated and the secretory cells of the tracheal epithelium had desquamated and the mucosa were coated with a continuous layer of basal cells. In a few cases, a desquamation of the basal layer was observed and the exposed basement membrane appeared as a pole of attraction for bacteria. Pneumococci were never seen attached to control tracheas. In contrast, they were observed adhered to the microvilli of the basal cells and, to a greater extent, to the exposed basement membrane.

Animals↗
PubMed1985

Rheological and transport properties of airway secretions in cystic fibrosis--relationships with the degree of infection and severity of the disease.

It has recently been suggested that in cystic fibrosis (CF), there is no rheological abnormality of airway secretions other than that associated with purulence, and that the apparent inhibition in the mucociliary transport rate might be partly due to a ciliary inhibitor present in these secretions. In order to ascertain this assumption, expectorated airway secretions were collected without salivary contamination in twenty-four CF patients and the rheological properties were measured. Using a photometric method, the effects of CF sputum samples were analysed on the ciliary beat frequency (Fm) of the frog palate, and we measured their mucociliary transport rate (TR). In all but one CF sputum, TR and Fm were lower than that of the control frog mucus (median TR: 18.7 and 11.6 mm min-1; median Fm: 12.3 and 11.3 Hz, respectively). In the eighteen patients in whom the rheological properties were outside the range for optimal mucociliary transport, the clinical Shwachman score was significantly (P less than 0.05) lower (median score: 66.2 points) than in the six patients with optimal rheologic properties (median score: 73 points). In the eleven CF patients with superinfection, the apparent viscosity (eta o) was significantly higher (P less than 0.01; median eta o: 24.4 Pa. s) and TR, expressed as a percentage of the reference value, was significantly lower (P less than 0.05; median Tr: 54.5%) in comparison with the values obtained for the thirteen non-superinfected CF patients (median eta o: 15 Pa. s and median TR: 66% respectively). The CF patients with markedly hyperviscous sputum (eta o higher than 30 Pa. s) exhibited a low Shwachman score.

Adolescent↗