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Biomedical subjects

S G Kelsen

Publications and source records attributed to S G Kelsen.

At least 37 records · Page 2Linked to original sources
PubMed1994

Expression of the beta-adrenergic receptor-adenylylcyclase system in basal and columnar airway epithelial cells.

Catecholamines acting through the beta-adrenergic receptor (beta AR) coupled-adenylylcyclase system stimulate a variety of responses by airway epithelial cells which affect airway caliber and the response to inflammatory stimuli. Although the tracheobronchial epithelium (TBE) is composed of several phenotypically differentiated cell types, surprisingly little is known about the expression of the beta AR system by the major subpopulations of TBE cells (i.e., basal and columnar). We, therefore, examined the function of the beta AR system in columnar and basal cell-enriched populations of rabbit tracheocytes. Cells were collected from 35 rabbits in 17 separate experiments and separated into basal and columnar cell-enriched fractions by centrifugal elutriation. The columnar fraction demonstrated a significantly greater (P < 0.005) adenosine 3',5'-cyclic monophosphate (cAMP) response to isoproterenol (10(-9)-10(-5) M) than the basal cell-enriched fraction (i.e., 74.7 +/- 5.1 and 49.4 +/- 2.8 pmol/10(6) cells, in columnar and basal cell-enriched fractions, respectively, P < 0.0001) as well as a higher beta AR density (i.e., 8,678 +/- 840 and 4,754 +/- 406 beta AR sites/cell, respectively, P < 0.0001). However, when corrected for differences in cell size assessed from measurements of total cell protein, cAMP production per milligram protein and beta AR density per milligram protein were similar in the two cell fractions (P > 0.50 for both comparisons). beta AR subtype assessed by beta 1AR and beta 2AR subtype selective antagonists demonstrated that the beta 2AR subtype predominated (i.e., > 90%) in both cell populations (P > 0.5).(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclases↗
PubMed1994

The comparative effects of elastase-induced emphysema on costal and crural diaphragm and parasternal intercostal muscle contractility.

Chronically, hyperinflated human subjects with chronic obstructive pulmonary disease and animals with experimentally induced emphysema generate greater than expected levels of transdiaphragmatic pressure at high lung volume because of adaptive changes in the length-tension relationship of the costal diaphragm. The ability to lower intrathoracic pressure during inspiration depends on the mechanical action of all the inspiratory muscles acting in concert. However, the effect of chronic hyperinflation on the mechanical action of inspiratory muscles other than the costal diaphragm remains unknown. This study compares the effect of chronic hyperinflation in the form of elastase-induced emphysema on the contractile properties of the three major inspiratory muscles of the hamster (i.e., the costal and crural diaphragm and parasternal intercostals). Muscles were studied in vitro 6 months after emphysema was induced by intratracheal injection of elastase. Saline-injected animals served as controls. TLC in the elastase-injected hamsters was significantly greater than in controls (12.5 +/- 0.8 ml versus 9.0 +/- 0.3 ml, p < 0.002). Maximal tetanic tension, time to peak tension, maximal velocity of shortening, and the force-velocity relationship differed among the three muscles but for any given muscle were similar in control and emphysematous animals. In contrast, the fiber length optimal for tension generation (Lo) not only differed across muscles but was significantly shorter in the costal diaphragm of emphysematous animals compared with control animals. However, Lo of the parasternal intercostal and crural diaphragm was similar in both groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adaptation, Physiological↗
PubMed1994

Relaxation of rabbit tracheal smooth muscle by adenine nucleotides: mediation by P2-purinoceptors.

The relaxant action of adenine nucleotides was studied in isolated rabbit trachealis to assess the presence of P2-purinoceptors in the airways, their cellular location, and pharmacologic properties. Strips of tracheal smooth muscle with intact epithelium were incubated in tissue baths and contracted with 1 microM acetylcholine. Over a dose range of 0.1 microM to 1 mM, ATP and ADP were significantly more potent than adenosine in relaxing tracheal smooth muscle. Significant relaxations were also elicited by AMP-PCP, AMP-CPP, and AMP-PNP, three ATP analogs stable to enzymatic hydrolysis to adenosine. In the absence of acetylcholine, neither ATP nor AMP-CPP exerted any contractile effect on the tracheal strips. In tissues selectively denuded of epithelium, ATP-, ADP-, and AMP-PCP-induced relaxations were markedly reduced. ATP-induced relaxation was also inhibited by the P2y-purinoceptor antagonist Reactive Blue 2 (RB2) (50 to 300 microM) and partially reduced by the cyclooxygenase inhibitor indomethacin (10 microM), whereas adenosine-induced relaxation was not significantly affected by these agents. These results suggest that ATP can induce smooth muscle relaxation in acetylcholine-contracted tracheal strips through a distinct P2-purinoceptor. This receptor appears to be located on the epithelium where its relaxant effect is mediated in part by release of one or more cyclooxygenase products. Additional relaxation at high ATP concentrations may occur through enzymatic hydrolysis of ATP to adenosine and interaction at P1-purinoceptors.

Adenine Nucleotides↗
PubMed1994

Functional behavior of the beta-adrenergic receptor-adenylyl cyclase system in rabbit airway epithelium.

Stimulation of adenylyl cyclase mediates the effects of beta-adrenergic agonists and prostaglandin E2 (PGE2) on tracheobronchial epithelial cell function by increasing intracellular cyclic adenosine monophosphate (cAMP). In turn, increases in cAMP affect airway function by modulating ciliary beating, chloride and water transport, mucus secretion, and release of bronchoactive substances. This study examined the function and regulation of the beta-adrenergic receptor-adenylyl cyclase system (beta AR-AC) in tracheal epithelial cells isolated from the rabbit, a frequently used animal model of airway reactivity, inflammation, and electrolyte transport. beta AR number, assessed by ligand binding using the non-subtype-specific beta-antagonist [125I]iodopindolol, averaged approximately 10,700 beta AR/cell (400 fmol/mg membrane protein). Greater than 85% of the receptors were of the beta 2 subtype as determined by competitive antagonist displacement of iodopindolol by selective beta 1- (betaxolol) and beta 2- (ICI 118,551) antagonists. cAMP synthesis was stimulated with isoproterenol, PGE2, and forskolin in a time- and concentration-dependent fashion. Preincubation of epithelial cells for 30 min with either isoproterenol (10 microM) or the peptide inflammatory mediator, bradykinin (100 microM), markedly depressed subsequent isoproterenol-stimulated cAMP synthesis. Isoproterenol-induced beta AR-AC desensitization appeared to be homologous since cAMP responses to PGE2 and forskolin, a direct activator of adenylyl cyclase, were not reduced. The effect of bradykinin on isoproterenol-stimulated cAMP response was mimicked by preincubation with either dioctanoyl glyceride or phorbol myristate acetate, activators of protein kinase C.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclases↗
PubMed1994

Regulation of beta-agonist- and prostaglandin E2-mediated adenylyl cyclase activity in human airway epithelial cells.

Despite the importance of beta 2-adrenergic receptor (beta 2AR) stimulation in mediating airway epithelial cell function, little is known regarding its regulation in airway epithelium. Perturbations of the airway environment associated with disease states, including the management of bronchomotor tone with beta-agonists, expose airways to putative regulators of beta 2AR signal transduction. In this communication, we describe the desensitization of beta 2AR signal transduction in the human airway epithelial cell line BEAS-2B. Examination of both beta-agonist- and prostaglandin E2 (PGE2)-mediated cAMP generation in BEAS-2B cells reveals both agonist-specific (homologous) and non-agonist-specific (heterologous) desensitization of these G protein-coupled receptor pathways. Short-term homologous desensitization of beta 2AR-mediated cAMP generation was characterized by an approximately 60% loss of maximal responsiveness to isoproterenol (ISO) when cells were pretreated 30 min with 10 microM ISO. A reduced sensitivity to ISO was also evidenced by an approximately 4-fold increase in the EC50 for ISO stimulation of adenylyl cyclase (AC). Short-term heterologous desensitization was characterized by an increase in EC50 (approximately 2- to 3-fold) with no change in maximal responsiveness to ISO in cells pretreated with either forskolin or PGE2. Qualitatively similar findings characterized short-term homologous and heterologous desensitization of PGE2-mediated AC activity. Short-term agonist-specific desensitization of the beta 2AR was associated with, but not dependent upon, rapid beta 2AR sequestration. Long-term pretreatment of cells with 10 nM ISO and 1 microM PGE2 eliminated AC responsiveness to subsequent ISO and PGE2 stimulation, respectively. Exposure of BEAS-2B cells to ISO for 24 h resulted in an approximately 70% loss of beta 2ARs, whereas chronic forskolin or PGE2 pretreatment had no effect on beta 2AR number. Long-term pretreatment of cells designed to elicit heterologous desensitization was associated with reductions in maximal responsiveness to ISO and PGE2 that appear to be related to a loss in inherent AC activity. These findings hold strong implications regarding the effect of beta 2AR desensitization on epithelial cell function and the role of beta-agonists in the management of airway disease.

Adenylyl Cyclases↗
PubMed1994

Asthma and diving.

BACKGROUND: Approximately 10 to 15 million Americans are scuba divers. The prevalence of scuba diving and asthma makes it likely some asthmatics will be interested in scuba diving and some scuba divers will have asthma. Conditions present during scuba diving may provoke airway obstruction in asthmatic patients. Further, asthmatic patients may, in theory, face a greater than normal risk of pulmonary barotrauma from lung overdistension on ascent through the water column. OBJECTIVE: The purpose of this paper is to review the theoretical issues underlying the prohibition against scuba diving for asthmatic patients as advanced by most major diving organizations in the United States and critically examine the relevant accident data. METHODS: All reports that dealt with asthma and diving, and all available American accident data including both fatal and nonfatal accidents were reviewed. RESULTS: Actuarial data on the risk of scuba accidents attributable to asthma do not define several important variables likely to affect accident risk during scuba diving. Despite these limitations, careful review indicates the risks of serious morbidity or mortality during scuba diving appears to be inconsequentially elevated in subjects whose asthma was not characterized. CONCLUSIONS: Additional data are needed to define accurately risks of diving in subjects with different forms of asthma, however, the available data suggest asthmatic patients with normal airway function at rest, and with little airway reactivity in response to exercise or cold air inhalation, have a risk of pulmonary barotrauma similar to that of normal subjects.

Airway Obstruction↗
PubMed1993

Structure of parasternal intercostal muscles in the adult hamster: topographic effects.

The parasternal intercostals are primary inspiratory muscles like the costal and crural diaphragm. However, the structure of the rib cage and its impedance to inspiration and expiration varies regionally. We questioned whether topographic differences in rib cage structure and impedance were associated with regional differences in parasternal intercostal muscle structure. Therefore, we examined the size and percentage of histochemically stained fibers in the parasternal intercostal muscles in the first, second, third, fourth, and sixth interspaces in the hamster. We observed a rostrocaudal gradient in the percentage and size of slow oxidative (SO), fast oxidative-glycolytic, and fast glycolytic (FG) fibers in the parasternal intercostal muscles. In particular, the percentage of SO decreased while the percentage of FG increased in a rostrocaudal direction in the first through sixth interspaces. In addition, the size of SO and FG fibers increased from the first to sixth interspace. Furthermore, changes in the size and percent of the three fiber types produced, in a rostrocaudal direction, significant reductions in the relative mass of the parasternal intercostal muscle made up of SO fibers and increases in the mass of fast fibers. We speculate that topographical differences in the size and percentage of fast and slow twitch fibers in the parasternal intercostal are likely to alter force-generating capacity of the parasternal muscles in a rostrocaudal direction and likely reflect regional differences in muscle load and/or activity.

Analysis of Variance↗
PubMed1993

Explant culture of rabbit tracheobronchial epithelium: structure and prostaglandin metabolism.

This study examines the potential usefulness of explant culture of rabbit tracheal epithelium as a model for the study of epithelial function under normal and potentially pathologic conditions. Accordingly, we assessed the structure and prostaglandin E2 (PGE2) release of tracheal epithelial explants obtained from adult pathogen-free rabbits. Epithelial cells attached to their native connective tissue substratum were maintained in culture for 5 days in serum-free medium, under bipolar conditions (air-liquid interface) on a permeable membrane (pore size, 0.2 mm), and nourished from the basolateral surface. At 5 days in culture, scanning and transmission electron microscopy and light microscopy demonstrated a pseudostratified, ciliated columnar epithelium with prominent folds and mucus secretion identical in appearance to the mucosa before culture. On the day of dissection (day 0) and after 4 days in culture (day 4), explants released PGE2 into the medium spontaneously. However, day 4 explants produced 3- to 4-fold greater amounts of PGE2 than day 0 explants. Moreover, day 4 explants demonstrated increased PGE2 release in response to bradykinin, a receptor-dependent agonist, and ionomycin, a calcium ionophore, while day 0 explants did not. Primary tracheal epithelial cell cultures grown to confluence (day 9) on a collagen substrate demonstrated PGE2 responses to bradykinin and ionomycin that qualitatively resembled those of day 4 explants. We conclude that rabbit tracheal explants cultured in vitro under the above conditions maintain cellular differentiation, in situ three-dimensional organization, and PGE2 synthetic pathways over several days in culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Effect of aging on respiratory skeletal muscles.

Although it has been recognized that limb skeletal muscles undergo changes during aging, it has been unclear to what extent age-related changes occur in respiratory muscles. This article reviews the structural organization of human skeletal muscles, including those of the respiratory system. Evidence that changes do occur in the structure and function of these muscles in elderly individuals is presented. The potential clinical significance of aging on respiratory muscle strength and endurance is discussed.

Adult↗
PubMed1992

Inspiratory muscle activity during pulmonary edema in anesthetized dogs.

Pulmonary edema is known to induce a rapid and shallow breathing pattern. However, its effects on the level and pattern of distribution of motor activity to the respiratory muscles is unclear. In the present study we evaluated the effect of oleic acid induced pulmonary edema on the electrical activity of the inspiratory muscles (costal and crural diaphragm and parasternal and external intercostal muscles) in the dog, and related it to the transdiaphragmatic pressure and ventilatory parameters over the course of CO2 rebreathing. Pulmonary edema, reflected by a 7.1 +/- 0.6 wet to dry ratio, decreased lung compliance by 57%, increased pulmonary shunt to 35%, and was associated with a rapid and shallow breathing pattern. When compared at equal levels of PCO2 during CO2 rebreathing before and during edema, ventilation and mean inspiratory flow were increased only at lower levels of hypercapnia and their responses to increasing levels of PCO2 were significantly diminished during edema. Transdiaphragmatic pressures were elevated during edema as compared to control values. The rate of rise of the electrical activity of all inspiratory muscles increased significantly during edema at all levels of PCO2. Peak activity, however, remained unchanged, due to shortening of the inspiratory duration. The EMG responses to progressive hypercapnia were not affected by edema. Pulmonary edema did not change the pattern of breathing and neural output to the inspiratory muscles in vagotomized dogs. We conclude that stimulation of pulmonary proprioreceptors during edema increases neural output to all inspiratory muscles. The neural response to hypercapnia is not altered by edema, and is additive to the vagal input. The ventilatory response to CO2 is blunted during severe edema, due to alterations in lung mechanics.

Anesthesia↗
PubMed1992

Expiratory muscle activity during pulmonary edema in the anesthetized dog.

The present study evaluated the reflex response of the expiratory muscles to pulmonary congestion and edema. The electromyograms of two thoracic and four abdominal expiratory muscles were recorded in 12 anesthetized dogs. Pulmonary edema was induced by rapid saline infusion in six dogs and injection of oleic acid into the pulmonary circulation in the remaining six dogs. Both forms of pulmonary edema reduced pulmonary compliance, interfered with gas exchange, and induced a rapid and shallow breathing pattern. The electrical activity of all abdominal muscles was suppressed during both forms of pulmonary edema. In contrast, the electromyogram activity of the thoracic expiratory muscles was not significantly affected by pulmonary edema. Acute pulmonary arterial hypertension produced in two dogs by inflating a balloon in the left atrium had no effect on ventilation or expiratory muscle electrical activity. In two vagotomized dogs, pulmonary edema did not inhibit the expiratory muscles. We conclude that pulmonary edema suppresses abdominal but not thoracic expiratory muscle activity by vagal reflex pathway(s). Extravasation of fluid into the lung appears to be more important than an increase in pulmonary vascular pressure in eliciting this response.

Anesthesia↗
PubMed1992

Effect of lung volume on the respiratory action of the canine pectoral muscles.

Lung volume influences the mechanical action of the primary inspiratory and expiratory muscles by affecting their precontraction length, alignment with the rib cage, and mechanical coupling to agonistic and antagonistic muscles. We have previously shown that the canine pectoral muscles exert an expiratory action on the rib cage when the forelimbs are at the torso's side and an inspiratory action when the forelimbs are held elevated. To determine the effect of lung volume on intrathoracic pressure changes produced by the canine pectoral muscles, we performed isolated bilateral supramaximal electrical stimulation of the deep pectoral and superficial pectoralis (descending and transverse heads) muscles in 15 adult supine anesthetized dogs during hyperventilation-induced apnea. Lung volume was altered by application of a negative or positive pressure (+/- 30 cmH2O) to the airway. In all animals, selective electrical stimulation of the descending, transverse, and deep pectoral muscles with the forelimbs held elevated produced negative intrathoracic pressure changes (i.e., an inspiratory action). Moreover, with the forelimbs elevated, increasing lung volume decreased both pectoral muscle fiber precontraction length and the negative intrathoracic pressure changes generated by contraction of each of these muscles. Conversely, with the forelimbs along the torso, increasing lung volume lengthened pectoral muscle precontraction length and augmented the positive intrathoracic pressure changes produced by muscle contraction (i.e., an expiratory action). These results indicate that lung volume significantly affects the length of the canine pectoral muscles and their mechanical actions on the rib cage.

Animals↗
PubMed1992

Rabbit trachealis tension responses to receptor-mediated agonists are diminished by elastase.

The present study examined the effects of elastase, in concentrations present in respiratory secretions, on airway smooth muscle contractile responses in vitro and the magnitude of the airway epithelial inhibition of smooth muscle tension. Experiments were performed on 126 full-thickness tracheal strips from 25 rabbits. Isometric tension responses to acetylcholine (10(-8) to 10(-4) M) and potassium chloride (10 to 110 mM) were examined before and after a 5-min exposure to either porcine pancreatic elastase (PPE) or human neutrophil elastase (HNE). PPE (5 to 40 micrograms/100 microliters) reduced the tension response to acetylcholine but had no effect on the tension response to potassium chloride. PPE and HNE (20 micrograms/100 microliters) produced similar effects. Mechanical removal of the epithelium per se significantly (P less than 0.005) decreased the ED50 response to acetylcholine but did not affect maximal tension. However, the airway epithelial inhibitory effect on the acetylcholine tension response was similar in the presence and absence of PPE (20 micrograms/100 microliters). These data suggest that the diminution of tracheal smooth muscle tension responses to receptor-mediated agonists induced by elastase is a direct effect on the muscle and is not mediated by an effect of elastase on the respiratory epithelium.

Acetylcholine↗
PubMed1992

A technique to harvest viable tracheobronchial epithelial cells from living human donors.

The ability to obtain airway epithelial cells from the lower respiratory tract in living human donors will facilitate study of the biologic properties of these cells. We report our experience harvesting tracheobronchial epithelial cells from living human donors by brushing the mucosal surface of the trachea and mainstem bronchi. Cells were obtained on 21 occasions from 18 healthy adult subjects under direct vision with a brush-tipped catheter during fiberoptic bronchoscopy. The average number of cells harvested per subject was 14 +/- 2 x 10(6), and cell viability determined by trypan blue exclusion averaged 36 +/- 4%. Of note, cell viability was significantly enhanced when lidocaine was confined to the nares. Lidocaine was also observed to diminish cell viability in vitro in a dose-dependent fashion. Morphologic and staining properties were used to classify harvested cells into the three major cell types present in the mucosa (i.e., ciliated, secretory, and basal cells). All three subtypes were obtained. The percentage of ciliated, secretory, and basal-like cells was 24 +/- 2%, 11 +/- 1%, 29 +/- 1%, respectively, while the remaining 36% were difficult to type. In one subject in whom brushing was performed on three occasions over a 7-wk period, the percentage of each of the three subtypes was similar across procedures. Harvested cells could be successfully placed in primary culture with a plating efficiency of 50 to 60% and could be subcultured for up to seven passages. Acutely dissociated cells could be used to study the beta-adrenergic receptor adenylyl cyclase system since they produced cAMP in response to isoproterenol.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1990

Effects of branched-chain amino acids on nitrogen metabolism in patients with cirrhosis.

This study was conducted to determine whether an amino acid solution enriched with branched-chain amino acids altered protein catabolic rates and plasma ammonia in patients with cirrhosis. Nine stable subjects were given two peripheral intravenous infusions: a standard amino acid solution (solution A) and a branched-chain-enriched solution containing 97% more leucine (solution B). Each solution was given for separate 9-day (group 1, n = 6) or 3-day (group 2, n = 3) periods. Amino acid solutions delivered 0.7 gm protein.kg-1.day-1. Diets provided an additional 0.3 gm protein plus maintenance calories. Protein turnover was assessed by a primed continuous infusion of [1-14C] leucine in six patients (three patients in group 1 and three patients in group 2). Nitrogen balance and urinary 3-methyl histidine excretion were determined in group 1 patients. Compared with solution A, solution B increased leucine flux and leucine oxidation but had no significant effect on protein synthesis or catabolism based on the plasma specific activity of either leucine or alpha-ketoisocaproic acid. The additional leucine infused with solution B was quantitatively oxidized. Nitrogen balance did not differ with the two solutions and there was also no difference in the urinary excretion of 3-methyl histidine, suggesting that muscle protein catabolism was unchanged. Plasma ammonia concentration decreased significantly during the infusion of solution B and was associated with a slight fall in plasma glucagon concentration. The results indicated that a branched-chain-enriched amino acid solution did not alter protein synthesis or catabolism although it did lower the plasma ammonia when compared with a standard amino acid formula in stable cirrhotic patients.

Amino Acids↗
PubMed1990

Distribution of motor activity to expiratory muscles during sciatic nerve stimulation in the dog.

The present study examined the effect of increasing sensory input from the lower limbs (assessed from the response to electrical stimulation of the sciatic nerve) on the distribution of electrical activity to the expiratory muscles. Expiratory muscle response to sciatic nerve stimulation (SNS) was compared to the response of the inspiratory muscles, and to their response to hypercapnia. In 16 anesthetized dogs afferent SNS increased ventilation and augmented the integrated EMG of all six expiratory muscles studied. Increases in abdominal muscle electrical activity were not uniform, being greater in the transversus abdominis and external oblique as compared to the internal oblique and the rectus abdominis. Increases in thoracic expiratory and inspiratory muscle activity during SNS were similar in magnitude. SNS performed while dogs were breathing 7% CO2, produced increased neural activity similar to those observed during O2 breathing. During CO2 rebreathing, at equal levels of minute ventilation, expiratory muscle responses to SNS and to CO2 were similar. In contrast, the rate of rise of the inspiratory muscle EMGs was greater during SNS. The present study indicates that the abdominal muscles participate in the respiratory response to afferent neural drive from skeletal muscles. The magnitude of their response is independent of their pre-stimulation level of activity and is similar to that observed during CO2 stimulated breathing.

Abdominal Muscles↗