PubMed Health⌕ Search

Biomedical subjects

W E Finkbeiner

Publications and source records attributed to W E Finkbeiner.

At least 19 recordsLinked to original sources
PubMed1992

Characterization of immortal cystic fibrosis tracheobronchial gland epithelial cells.

Tracheobronchial glands were isolated and cultured from a patient with cystic fibrosis (CF). Cultured epithelial cells were transformed with pSVori-. All transformed cell lines express cytokeratin filaments and at early passages express the junctional complex molecule cell CAM 120/80, indicating their epithelial origin. Several gland cell lines express antigens that localize to secretory cells in vivo. Cl- transport measured by 36Cl efflux shows that CF gland epithelial cells, like CF surface airway and nasal polyp epithelial cells, are unable to respond to increases in intracellular cAMP. However, they do produce an increase in intracellular cAMP after treatment with isoproterenol or forskolin. One CF gland cell line shows increased intracellular calcium in response to a number of agents and increased Cl- efflux comparable to that observed in a non-CF airway surface epithelial cell line after addition of calcium ionophore. All cell lines express CF transmembrane conductance regulator mRNA, as measured by PCR amplification of first-strand cDNA. The CF tracheobronchial gland cell lines described here are compound heterozygotes, having a single copy of the delta F508 mutation.

Base Sequence↗
PubMed1992

Ablation of ventricular tachycardia using multiple sequential transcatheter application of radiofrequency energy.

Multiple sequential radiofrequency energy was applied in the left and right ventricles of 24 dogs to produce large ablated areas limited to endocardial and subendocardial regions. Endocardial ablation was performed in nine dogs with normal ventricles and 15 that had survived remote myocardial infarcts, three with inducible sustained monomorphic ventricular tachycardia. A quadripolar catheter was positioned either at the site of earliest ventricular activation during induced monomorphic ventricular tachycardia or at circumscribed areas of the left ventricle. Radiofrequency energy was delivered between two adjacent poles of the catheter, successively applying radiofrequency energy to the distal, middle, and proximal electrode pairs; this was repeated 9 to 11 times with the catheter in a slightly different position. A cumulative energy of 9,688 +/- 4,191 joules resulted in an ablated endocardial/subendocardial surface area of 4.7 +/- 2.2 cm2 (range 2.4-10 cm2, maximum depth 4 mm). Sustained tachycardia was not inducible by aggressive programmed ventricular stimulation in the dogs with previously inducible tachycardia, indicating successful ablation of the tachycardia foci. Only seven normal dogs were available for electrophysiological studies; three were used in acute and four in chronic studies. Ventricular tachycardia was not induced in the remaining dogs either before or after radiofrequency ablation, indicating the lack of an arrhythmogenic effect of this method. Histologic examination was performed in all nine normal dogs (five were sacrificed for acute pathological examination) as well as in the 15 with myocardial infarction. The late pathological examination of the radiofrequency lesion in these 19 animals showed homogeneous areas of coagulation necrosis and endocardial proliferation. Thus, this modified technique of radiofrequency ablation produced large homogeneous endocardial/subendocardial scars suitable for treating ventricular tachycardia and showed no evidence of an arrhythmogenic influence.

Animals↗
PubMed1992

Changes in permeability of dog tracheal epithelium in response to hydrostatic pressure.

We tested the hypothesis that, in asthma, the airway epithelial damage and leakage of blood proteins into the lumen are the result of edema and raised submucosal hydrostatic pressure. Sheets of dog tracheal epithelium were mounted in Ussing chambers, and the effects of transepithelial hydrostatic pressure differences (delta P) on conductance (G), [3H]mannitol flux (Jman), and fluorescein isothiocyanate-albumin flux (Jalb) were determined. delta P values of 20 cmH2O directed from the mucosal to submucosal side of the tissue (m----s) had no significant effects on G, Jman, Jalb, or tissue ultrastructure. delta Ps----m caused increases in conductance (G) with a maximal effect at approximately 20 cmH2O. delta Ps----m of 20 cmH2O significantly (P less than 0.05) increased G (4.3 +/- 0.6 to 10.6 +/- 1.6 mS/cm2), Jman s----m (18 +/- 5 to 411 +/- 54 nmol.cm-2.h-1), J(alb)s----m (0.3 +/- 0.1 to 6.0 +/- 2.0 micrograms.cm-2.h-1), and J(alb)m----s (0.7 +/- 0.3 to 1.8 +/- 0.4 micrograms.cm-2.h-1). Jman m----s was not affected. On removal of delta P, G and Jman s----m returned to preexposure values, though J(alb)s----m remained slightly elevated at 1.1 +/- 0.3 micrograms.cm-2.h-1. Morphologically, delta Ps----m caused dilation of lateral intercellular spaces, disruption of tight junctions, and submucosal edema. The large increases in s----m fluxes of albumin and mannitol are consistent with bulk flow of fluid toward the lumen via the areas of epithelial damage.

Animals↗
PubMed1992

Cloning of human airway 15-lipoxygenase: identity to the reticulocyte enzyme and expression in epithelium.

Lipoxygenases constitute a family of enzymes which are implicated in a variety of inflammatory disorders including asthma. Although the 15-lipoxygenase has been identified as the major route of arachidonic acid metabolism in human lung, airway epithelial cells, eosinophils, and developing red cells, the localization of the enzyme within lung has not been clearly defined. Furthermore, the existence of isoforms of 15-lipoxygenase in different tissues has recently been proposed. To address these issues, we isolated a 2.6-kb cDNA encoding human airway 15-lipoxygenase from a human bronchus cDNA library using a previously characterized reticulocyte 15-lipoxygenase cDNA as a probe. The airway 15-lipoxygenase sequence was found to be identical to that of the reticulocyte-derived clone. Immunocytochemical studies using an antibody to human recombinant 15-lipoxygenase specifically localizes the enzyme to the basal and ciliated cells of the trachea, bronchi, and bronchioles. Staining for 15-lipoxygenase is not present in the airway secretory epithelial cells, epithelial cells located in the gas-exchanging regions of lung, vascular structures, or inflammatory cells. Taken together these results suggest that the 15-lipoxygenase of human lung is identical to that of the reticulocyte enzyme and is preferentially expressed in airway epithelium.

Arachidonate 15-Lipoxygenase↗
PubMed1992

Bovine tracheal serous cell secretion: role of cAMP and cAMP-dependent protein kinase.

The role of adenosine 3',5'-cyclic monophosphate (cAMP) and protein phosphorylation during beta-adrenergic receptor stimulation of bovine tracheal gland serous cells was investigated in vitro. Isoproterenol, a beta-adrenergic agonist, increased the secretion of 35S-labeled molecules. Intracellular cAMP levels were increased within 1 min after stimulation of bovine tracheal gland serous cells with isoproterenol. The dose-response relationship for isoproterenol-stimulated generation of cAMP correlated with the dose-response relationship for isoproterenol-stimulated secretion of 35S-labeled molecules. The phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine potentiated both isoproterenol-evoked secretion of 35S-labeled molecules and the production of intracellular cAMP, and the beta-adrenergic receptor antagonist propranolol completely blocked both effects. The secretory response of the cells to isoproterenol could be mimicked by the cAMP analogues 8-bromoadenosine 3',5'-cyclic monophosphate and dibutyryl adenosine 3',5'-cyclic monophosphate. Activity of cAMP-dependent kinase was measured in soluble and particulate cell extracts. cAMP effected the state of phosphorylation of proteins associated with the soluble but not the particulate fraction. These studies are consistent with the hypothesis that beta-adrenergic stimulation of secretion from bovine tracheal gland serous cells occurs via a cAMP-mediated pathway and that one of the molecular events in this pathway is cAMP-dependent protein phosphorylation.

1-Methyl-3-isobutylxanthine↗
PubMed1992

Differentiated structure and function of cultures from human tracheal epithelium.

Here we describe the conditions which allow cultured human tracheal epithelial cells to retain the ion transport properties and ultrastructure of the original tissue. The order of potency of growth supports and media additives in elevating baseline short-circuit current (Isc) and responses to mediators were vitrogen gel (VIT) greater than extracellular matrix from bovine corneal endothelial cells (ECM) greater than human placental collagen (HPC), and 2% Ultroser G serum substitute (USG) greater than 5% fetal calf serum (FCS) greater than defined growth factors (GF). For all combinations of medium and growth supports, an air interface (AIR) gave better electrical properties than immersion feeding (IMM). As opposed to our earlier conditions (HPC/FCS/IMM), the best new combination (VIT/USG/AIR) produced higher baseline Isc (58.0 +/- 10.6 vs. 5.1 +/- 1.0 microA/cm2) and increased Isc responses to isoproterenol (6.1 +/- 1.5 vs. 0.8 +/- 0.3 microA/cm2) and bradykinin (9.6 +/- 2.0 vs. 1.0 +/- 0.2 microA/cm2), while retaining high transepithelial resistance (227 +/- 5 omega.cm2). VIT/USG/AIR led to the appearance of cilia, an increase in the depth of the cell sheets (50 vs. 10 microns), longer and more frequent apical microvilli, and increased interdigitations of the basolateral membrane. Protein and DNA content were also significantly increased. Secretory granules were present which stained with antibody to goblet cells, but not to serous or mucous gland cells. CF cells grown in VIT/USG/AIR showed high baseline Isc (69 +/- 18 microA/cm2) and a proportionately larger inhibition of Isc by amiloride (70 +/- 10 vs. 34 +/- 3%). Isc did not respond to isoproterenol, and the response to bradykinin was 22% normal.

Amiloride↗
PubMed1991

Hyaluronic acid in a cardiac myxoma: a biochemical and histological analysis.

Cardiac myxoma is the most common primary tumor of the heart. This tumor has a gelatinous stroma that is thought to be composed of glycosaminoglycans, the classical acid mucopolysaccharide ground substance. We examined both biochemically and histologically the hyaluronic acid in a case of cardiac myxoma using a newly developed hyaluronic acid-binding protein probe. We observed that hyaluronic acid was localized in the amorphous stroma and occurred at levels 30 times that found in normal atrial septum.

Heart Neoplasms↗
PubMed1991

Simple technique for culture of highly differentiated cells from dog tracheal epithelium.

Cultures of dog tracheal epithelium have proved very useful in studies of ion transport. Their short-circuit current (Isc), however, is usually much less than the original tissue. We have tested a variety of conditions in an attempt to produce large numbers of cells with electrical properties comparable with the original tissue. Of several growth supports, human placental collagen (HPC) gave the best results. When plated at 2.5 x 10(5) cells/cm2 onto HPC, cells grown in serum-free, growth factor-supplemented medium (GF medium) showed increases in cells per unit area, thickness of cell sheet, numbers of domes, numbers of apical microvilli, and degree of basolateral membrane interdigitation compared with cells grown in medium containing 5% fetal calf serum (FCS medium). Transepithelial resistance (Rte) and the increases in Isc and intracellular Ca in response to isoproterenol were also increased. However, baseline Isc and adenosine 3',5'-cyclic monophosphate levels were not changed. The improved electrical properties were maintained for up to 4 mo. GF medium combined with an air interface produced further increases in Rte, Isc, and changes in Isc in response to amiloride and isoproterenol. Ultrastructural features such as the presence of cilia, greater thickness of the cell sheet, and increased amplification of apical and basolateral membranes also indicated improved differentiation. Our results show that GF medium and an air interface can be combined with a simple growth support and a relatively low-plating density to allow the easy production of greater than 500 cm2 of cultured cells from a single trachea, with a level of differentiation similar to that of the original tissue.

Animals↗
PubMed1991

Ion transport by cultures of human tracheobronchial submucosal glands.

Acini of human tracheobronchial submucosal glands were isolated by enzymatic disaggregation, and, when plated on flasks coated with human placental collagen (HPC) in media containing Ultroser G serum substitute (USG) and a variety of growth factors (GF), they became confluent after 14-20 days. The cells were then isolated by trypsinization and replated in media containing USG and GF at 10(6) cells/cm2 on porous-bottomed inserts coated with HPC. Confluent monolayers formed on day 1 after replating and were studied on day 10. Transepithelial resistance and short-circuit current (Isc) were 578 +/- 89 omega.cm2 and 12.9 +/- 1.9 microA/cm2 (means +/- SE, n = 23 cell sheets). The potency sequence for stimulation of Isc by mediators was methacholine greater than bradykinin greater than isoproterenol approximately or equal to phenylephrine. Amiloride decreased baseline Isc by 42 +/- 9% (n = 6 cell sheets) but had little effect on the Isc response to mediators. Diphenylamine-2-carboxylic acid, however, had no effect on baseline Isc but markedly inhibited the Isc response to all mediators. These results show that submucosal gland cells from human trachea can be grown in culture to produce epithelial sheets of high resistance, which secrete Cl in response to bradykinin and alpha- and beta-adrenergic and cholinergic agents.

Adult↗
PubMed1991

Altered ion transport by tracheal glands in cystic fibrosis.

Acini of tracheal glands from 12 humans without cystic fibrosis (CF) and from two CF patients were isolated by enzymatic digestion. They were plated on flasks coated with human placental collagen (HPC) in media containing Ultroser G serum substitute and a variety of growth factors. Confluent cell sheets formed after 20 days. Cells were then isolated by trypsinization and replated at 10(6) cells/cm2 onto porous-bottomed inserts coated with HPC. Confluent sheets formed on day 1 after replating and were studied on day 10 in Ussing chambers. Transepithelial resistance (Rte) and baseline short-circuit current (Isc) of CF cultures (171 +/- 67 omega.cm2, 3.8 +/- 0.8 microA/cm2; n = 5) were significantly less than non-CF (541 +/- 116 omega.cm2, 9.9 +/- 2.3 microA/cm2; n = 14). Responses in Isc to mediators were also significantly reduced in CF: isoproterenol (10(-5)M) = 0.04 +/- 0.04 vs. 1.9 +/- 0.6; methacholine (10(-5)M) = 0.2 +/- 0.1 vs. 7.1 +/- 1.7; bradykinin (10(-6)M) = 0.4 +/- 0.1 vs. 5.0 +/- 1.0 microA/cm2; n = 5 for CF, n = 14 for non-CF. When CF and non-CF cells were matched for baseline Isc and Rte, the responses of CF cells to mediators still remained statistically lower than normal. The reduced responses of CF cells to bradykinin and methacholine, in addition to isoproterenol, suggest that both Ca-dependent and adenosine 3',5'-cyclic monophosphate-dependent regulation of Cl secretion are defective in CF tracheobronchial glands. The resulting reduction in fluid secretion by glands may contribute to the accumulation of airway mucus in CF.

Adult↗
PubMed1990

Denervation-induced inflammation in the rat.

We report that section of the sciatic and saphenous nerves, in the hindlimb of the rat, evokes an inflammatory response in the denervated tissue that can be distinguished from the previously described peptide-mediated neurogenic inflammation. This novel form of neurogenic inflammation has a very delayed onset (9.75 +/- 2.1 h, mean +/- S.E.M., n = 8), persists for more than 30 h, and is characterized by a marked neutrophilic cellular infiltrate. These features cannot be mimicked by electrical stimulation of the peripheral nerve and are not prevented by either prior application of local anesthetics to the nerve lesion site or by neonatal treatment with capsaicin.

Animals↗
PubMed1990

Effect of repetitive brief episodes of cardiac ischemia on 31P magnetic resonance spectroscopy in the cat.

Angina is characterized by brief periods of ischemia followed by reperfusion; the cumulative effect of these episodes on energetics of the myocardium has not been fully elucidated. This study used an in vivo feline model for the assessment of high-energy phosphate compounds during brief sequential periods of ischemia and reperfusion. Nine adult, open-chest, anesthetized cats were prepared with a reversible occluder around the proximal left anterior descending artery and a 1.2-cm-inside diameter coil sutured on the myocardial surface in the distribution of the left anterior descending coronary artery. Levels of PCr, Pi, and ATP (beta-phosphate signal) were measured by 31P MRS in a GE CSI 2-T NMR spectrometer/imager. Measurements were obtained during a control period and during three successive occlusion-deocclusion periods of roughly 12 and 20 min' duration, respectively. The last deocclusion period was observed for 60 min. Electron microscopy was performed in two animals. PCr declined (P less than 0.01) rapidly following each occlusion to 51 +/- 5.2% (occlusion 1), 53 +/- 5.8% (occlusion 2), and 48 +/- 5.7% (occlusion 3) of the control value by 6 min. Pi rose (P less than 0.01) with the three sequential occlusions to 253 +/- 46, 288 +/- 57, and 277 +/- 46%, respectively. PCr and Pi returned to baseline promptly with reperfusion, while ATP showed a gradual decline throughout the experiment, decreasing to 77 +/- 7.2% of control at the end of the last reperfusion (P less than 0.05). Although PCr returned to baseline during reperfusion, ATP did not, suggesting a reduction in the nucleotide pool. These findings indicate that the repeated episodes of ischemia, which are insufficient to produce necrosis, can have an effect on myocardial high-energy phosphate metabolism as evidenced by mild depletion of ATP.

Adenosine Triphosphate↗
PubMed1990

Etiology of intestinal damage in gastroschisis, II. Timing and reversibility of histological changes, mucosal function, and contractility.

Previous work in the fetal lamb examined the relative effects of amniotic fluid and bowel constriction in the etiology of bowel damage in gastroschisis. The present study used the same model to assess the timing and reversibility of these changes during gestation. Gastroschisis was created at 80 days' gestation, and a tape was placed around the bowel to cause gradual constriction with growth. Lambs were killed at 100 days, 120 days, and term. Bowel damage was assessed using histology, mucosal enzyme activity, and in vitro motility. In an additional "repaired" group, the constrictor was removed at 120 days, a silastic pouch placed over the bowel, and bowel damage assessed at term. Normal fetuses at each gestational age were used as controls. A fibrous peel was observed at all gestational ages. Mucosal villous atrophy and mesenteric venous and lymphatic dilation were mild at 100 and 120 days, but severe at term. These changes were present but mild in repaired animals at term. Mucosal enzyme activity decreased gradually with gestational age; inhibition of maltase activity was maximal at term, and was significantly reversed by repair, whereas inhibition of aminooligopeptidase activity was maximal at 120 days, and was not affected by repair. Protein/DNA, DNA/weight, and protein/weight ratios showed that repaired mucosal cells were significantly more proliferative, smaller, and less mature than control or gastroschisis cells. In vitro motility studies demonstrated a mild decrease in contractility at 100 and 120 days, and a large decrease at term. This deleterious effect at the end of gestation was only partially reversed by repair in utero.(ABSTRACT TRUNCATED AT 250 WORDS)

Abdominal Muscles↗
PubMed1990

The serous cell.

Explore the source record for details and available documents.

Animals↗
PubMed1990

Receptors on airway gland cells.

Airway submucosal glands are by volume the most important source of macromolecules in airway secretions. These secretions, containing gel-forming mucins, antibacterial proteins, and antiproteases, comprise the major defensive barrier protecting the host against airborne pathogens. The identification of the mechanisms regulating secretion from the submucosal glands is key to understanding the genesis of this barrier and how it is altered by disease processes. Using a variety of methods, we and others have identified on the gland cells of several species receptors specific for ACh, norepinephrine, substance P, VIP, PGE1, PGE2, PGA1, PGD2, histamine and bradykinin. These receptors all participate in modulating the secretory activity of the airway submucosal glands. Studies of homogeneous cultures of bovine airway serous cells have yielded detailed information regarding the beta-adrenergic receptor on these cells. Using radioligand binding techniques, we found evidence for the presence of a single high affinity beta receptor of beta-2 subtype. Occupancy of this receptor by isoproterenol causes an elevation in the concentration of intracellular cAMP, which in turn stimulates the phosphorylation of a subset of cytoplasmic and membrane proteins. Based on the kinetics and pharmacology of these effects, it is likely that cAMP functions as a second messenger in the serous cell secretory pathway, probably acting through protein kinases. Current efforts are directed at identification of those phosphoproteins whose phosphorylation and dephosphorylation times are consistent with their possible roles in secretion.

Animals↗
PubMed1990

Human tracheobronchial submucosal gland cells in culture.

Cellular mechanisms regulating airway secretion, secretory products of individual airway cell types, and control of airway cell growth and differentiation are poorly understood. In order to aid studies of these questions, we have established a system for culturing human tracheobronchial submucosal gland cells. Gland acini were isolated by enzymatic disaggregation from submucosal tissue obtained postmortem from patients without pulmonary diseases and from patients with cystic fibrosis. In culture, acini attached to a collagen substratum, and gland cells proliferated and formed confluent monolayers which were homogeneous by phase microscopy. In contrast to cells of freshly disaggregated acini which expressed either serous or mucous gland cell secretory antigens, in culture virtually all cells (greater than or equal to 95%) concurrently expressed both antigens as assessed by immunocytochemical staining with serous and mucous cell-specific antibodies. Similarly, electron microscopy revealed cells with serous- or mucous-type secretory granules, and cells containing both types of granules. Cultures incorporated 35S into high (greater than 10(6) D) and lower (greater than 700 kD; 150 kD) molecular weight molecules. Cholinergic and adrenergic agonists increased release of radio-labeled secretions. These findings demonstrate that human tracheal gland cells in culture retain immunocytochemical, ultrastructural, and functional features of both differentiated serous and mucous gland cells. This culture system will be useful for studying the biology and pathology of human tracheobronchial submucosal gland cells.

Adolescent↗