A technique for the isolation of submucosal gland cells from cat trachea

1983 ◽  
Vol 55 (3) ◽  
pp. 1035-1041 ◽  
Author(s):  
D. J. Culp ◽  
D. P. Penney ◽  
M. G. Marin
Keyword(s):  
Trypan Blue ◽  
Mechanical Treatment ◽  
Average Rate ◽  
Periodic Acid ◽  
Surface Epithelium ◽  
Submucosal Gland ◽  
Isolated Cells ◽  
Periodic Acid Schiff ◽  
Gland Cells ◽  
Glandular Cells

We have developed a procedure to isolate submucosal gland cells from cat trachea. The excised trachea was stripped of surface epithelium by stroking the luminal surface with a nylon brush. The remaining submucosa was scraped free from underlying cartilage and minced into small fragments. To disperse glandular cells from these fragments, we subjected the minced tissue to both enzymatic (collagenase and elastase) and mechanical treatment. In 23 preparations of cells we yielded an average (+/- SE) of 8.4 +/- 0.9 (X 10(6] cells. In eight cell preparations 95 +/- 1% of the cells stained with periodic acid-Schiff stain, suggesting that the cells are of glandular origin. We used the following criteria to assess cell viability. The dye trypan blue was excluded by 92 +/- 1% of the cells (n = 23). Under the electron microscope, cellular membranes and organelles appeared normal. The isolated cells consumed oxygen at an average rate of 1.34 +/- 0.05 microliters O2 X h -1 X (10(6) cell) -1, (n = 65). Oxygen consumption was constant for at least 4 h after cell isolation, was inhibited 21% by 10(-4) M ouabain, and was subsequently stimulated to 135% above basal levels by 4 X 10(-5) M dinitrophenol.

Mucus glycoconjugate secretion in cool and hypertonic solutions

1997 ◽  
Vol 272 (6) ◽  
pp. L1121-L1125 ◽  
Author(s):  
T. M. Dwyer ◽  
J. M. Farley
Keyword(s):  
Average Rate ◽  
Transient Increase ◽  
Productive Cough ◽  
Submucosal Gland ◽  
Basal Secretion ◽  
Cold Air ◽  
Dry Air ◽  
Gland Cells ◽  
Hypertonic Solutions ◽  
Exercise Induced

For sensitive individuals, exercise-induced asthma is triggered by cold and dry air and is often accompanied by a productive cough. In this study, we determined whether cold solutions and/or solutions of increased tonicity directly caused an increase in glycoconjugate (GC) secretion. To test this, we used isolated swine tracheal submucosal gland cells (TSGCs) and measured the rate of GC secretion at 37 and 32 degrees C in isotonic solutions and in solutions made hypertonic by 30 mosM. TSGCs were isolated under conditions that minimized the rate of GC secretion and were perfused with medium 199 equilibrated with 5% CO2 to a pH of 7.4. A lectin-based assay was used to specifically detect GC present in each 2-min fraction of the perfusate. Basal secretion was 3.1-fold greater at 32 degrees C (n = 3) than at 37 degrees C (n = 4; P < 0.05). At 37 degrees C, increasing perfusate osmolarity by 30 mosM increased the average rate of secretion by 41 +/- 11% (n = 4; P < 0.05); return to isotonic perfusate caused a 4.5 +/- 1.8-fold transient increase in secretion (n = 4; P < 0.05) that was complete within 10 min. At 32 degrees C, changing tonicity of the perfusate had no significant effect but returning to isotonic perfusate caused a 2.3 +/- 0.7-fold transient increase in secretion (n = 3; P < 0.05). Thus key stimuli that trigger obstruction of airflow (cold and increased osmolarity) can also directly stimulate GC secretion in the airway. Such increased secretions may contribute to the productive cough observed in some individuals in response to cold air.

scholarly journals Histological observations on the dendretic organ of the farmed adult African catfish (Clarias Gariepinus) from eastern Nigeria

2013 ◽  
Vol 58 (2) ◽  
pp. 139-146
Author(s):  
Ekele Ikpegbu ◽  
Uchenna Nlebedum ◽  
Okechukwu Nnadozie ◽  
Okezie Agbakwuru
Keyword(s):  
Periodic Acid ◽  
Clarias Gariepinus ◽  
Surface Epithelium ◽  
African Catfish ◽  
Dense Layer ◽  
Mucous Cells ◽  
Respiratory Organ ◽  
Periodic Acid Schiff ◽  
Stratified Squamous Epithelium ◽  
Subepithelial Layer

The histology of the accessory respiratory organ of the African catfish - dendretic organ was investigated to reveal its microanatomy. The data obtained will provide baseline data for further investigative research and assist fish pathologists. The histology showed that the tubular shaped dendretic organ was covered by stratified squamous epithelium containing periodic acid-Schiff (PAS) and alcian blue (AB) positive mucous cells. On most surfaces, the epithelial cells were organized into columns with mucous cells placed in-between the epithelial cell columns. At the tip of the surface, capillaries were lined by endothelium at the surface epithelium/air contact area. The subepithelial layer was of loose connective tissue containing adipose tissue and occasional blood vessels. The core of the dendretic organ contained elastic cartilage surrounded by a dense layer of perichondrium. Elastic fibres were observed the territorial and inter-territorial spaces.

Stimulation of secretion into human and feline airways by Pseudomonas aeruginosa proteases

1991 ◽  
Vol 70 (5) ◽  
pp. 2259-2267 ◽  
Author(s):  
M. Somerville ◽  
P. S. Richardson ◽  
A. Rutman ◽  
R. Wilson ◽  
P. J. Cole
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Alkaline Protease ◽  
Periodic Acid ◽  
Surface Epithelium ◽  
Dependent Manner ◽  
Periodic Acid Schiff ◽  
Concentration Dependent Manner ◽  
Human Bronchus ◽  
Secretion Inhibition

We have investigated the effect of elastase and alkaline protease from Pseudomonas aeruginosa on airway secretion into the trachea of anesthetized cats and from human bronchial mucosa in vitro. Secretory macromolecules were radiolabeled biosynthetically with two precursors in the cat, [3H]glucose and [35S]sulfate, and with [35S]-sulfate only in human tissue. Both enzymes (2.6 x 10(-9) to 1.3 x 10(-6)M elastase and 8 x 10(-9) to 2.4 x 10(-6)M alkaline protease) released radiolabeled macromolecules in a concentration-dependent manner from the two preparations. Purified elastase, 1.3 x 10(-6)M, released radiolabeled macromolecules (delta 3H = +397 +/- 72%, delta 35S 225 +/- 40% over control, P less than 0.001) and periodic acid-Schiff- (PAS) reactive glycoconjugates (delta PAS = +4.1 +/- 0.96 micrograms/min or +102 +/- 20%; P less than 0.01) from cat trachea, as did alkaline protease, 2.4 x 10(-6)M (delta 3H = +356 +/- 57%, delta 35S = +176 +/- 25%, delta PAS = +7.5 +/- 1.3 micrograms/min or 194 +/- 36%, P less than 0.001). Increases in 3H exceeded those of 35S, suggesting surface epithelium as the main source of secretion. Inhibition of enzyme activity abolished secretory effects. Both enzymes also stimulated secretion from human bronchus (e.g., with elastase, 1.3 x 10(-6)M: delta 35S = +331 +/- 67%, delta PAS = +4.3 +/- 0.92 micrograms/min or +131 +/- 24%, P less than 0.001; with alkaline protease, 2.4 x 10(-6)M: delta 35S = +220 +/- 67%, delta PAS = +12.7 +/- 3.2 micrograms/min or +575 +/- 245%, P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of Cytokeratins 7 and 20 in Helicobacter pylori-associated Chronic Gastritis in Children

2004 ◽  
Vol 7 (2) ◽  
pp. 180-186 ◽  
Author(s):  
M. Cohen ◽  
E. Cueto Rúa ◽  
N. Balcarce ◽  
R. Drut
Keyword(s):  
Helicobacter Pylori ◽  
Chronic Gastritis ◽  
Structural Changes ◽  
Periodic Acid ◽  
Gastric Epithelium ◽  
Surface Epithelium ◽  
Ki 67 ◽  
Periodic Acid Schiff ◽  
Group A ◽  
H Pylori

Helicobacter pylori gastric infection induces structural changes in the gastric epithelium. Among them, variations in the expression of cytokeratins have been reported in adult patients. In the present study, we describe the expression of CK7 and CK20 in gastric samples taken from the antrum in three groups of pediatric patients: (A) Helicobacter pylori-associated chronic gastritis (mean age: 11.4 years); (B) previous H. pylori chronic gastritis patients (mean age: 9.4 years); and (C) controls (mean age: 8.8 years). In all, the presence of sulfomucins was assessed with Alcian blue-periodic acid-Schiff pH 1.0. Immunoreactivity was graded as absent (0), weak (1 +), moderate (2+), or intense (3+), in accordance with the intensity of the staining, and its distribution as focal or diffuse. CK7 reactivity was 2 + either focal or diffuse in all group A biopsies. The reactivity was more evident in the cells at the neck of the glands, in the areas with more inflammatory infiltrates, decorating long vertical segments of epithelium. In groups B and C, CK7 reactivity was also focal and 1 + at the cells of the necks of the glands. However, group B presented longer vertical segments of positive cells as compared to group C, and shorter than those of group A. The deeper glandular structures were focally 1 + in both groups. CK20 expression was comparable in all three groups, depicting a 2+ diffuse reactivity at the surface epithelium and interposed pits with absence or focal reactivity at the neck and coiled gland areas. Ki-67 immunostaining paralleled that of the CK7. Staining for sulfated mucosubstances was positive in two of five cases of groups A and B, and in none of the cases of group C. We conclude that: (1) the long segments of CK7-positive glandular necks in H. pylori cases most probably indicate intense regenerative activity during active inflammation; (2) eradication of H. pylori does not warrant ad integrum restitution since long segments of Ki-67+, CK7+ cells at the germinative compartment of the glands (as well as cells with sulfomucins) were still recognizable in ex- H. pylori patients; (3) finally, differing from what happens in adults, children somehow manage to maintain fully differentiated CK20+ superficial epithelium while the H. pylori is in action.

Muscarinic stimulation of submucosal glands in swine trachea

1988 ◽  
Vol 64 (1) ◽  
pp. 200-209 ◽  
Author(s):  
C. M. Yang ◽  
J. M. Farley ◽  
T. M. Dwyer
Keyword(s):  
Acetylcholine Receptors ◽  
Relative Proportion ◽  
Muscarinic Acetylcholine Receptors ◽  
Mucus Secretion ◽  
Submucosal Gland ◽  
Isolated Cells ◽  
Gland Cells ◽  
Submucosal Glands ◽  
Tracheal Explants

The properties of muscarinic acetylcholine receptors (mAChR) on tracheal explants and isolated submucosal gland cells were determined using [3H]quinuclidinyl benzilate ([3H]QNB) and N-[3H]methylscopolamine ([3H]NMS) as ligands. Analysis of competitive displacement of ([3H]NMS binding by pirenzepine demonstrated the presence of M1- (27 +/- 2%) and M2G- (73 +/- 2%) receptors on isolated tracheal submucosal gland cells (TSGC's) in control. Daily administration of diisopropylfluorophosphate (DFP) inhibited cholinesterase activity by greater than 95%. After 7 days of DFP treatment, [3H]QNB binding to intact TSGC's decreased from 14.2 +/- 0.6 to 6.3 +/- 0.8 fmol/10(6) cells; similarly, [3H]NMS binding fell from 8.1 +/- 1.9 to 2.0 +/- 0.8 fmol/10(6) cells. The loss of mAChR's was predominantly of the M2G subtype with the relative proportion dropping to 33%. In addition, 90% of the receptors assumed the high-affinity state for carbachol displacement of [3H]NMS. Mucus secretion was quantitated by measuring the release of 3H-labeled mucus macromolecules from explants of tracheal submucosal glands and isolated cells. Acetylcholine (ACh), 2 X 10(-5) M, stimulated mucus secretion by 2.5 and 2.3 times the basal rate, respectively. Elimination of acetylcholinesterase (AChe) by DFP increased the ACh sensitivity by 18- and 5-fold. Tracheal explants or TSGC's obtained 2 h after an in vivo DFP treatment showed a 6- and 3-fold ACh stimulation. This ACh sensitivity decreased during the continued daily dosing with DFP such that only a 1.3- and 1.1-fold ACh stimulation was apparent after 7 days of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals Histochemical reactivity of peanut lectin-horseradish peroxidase conjugate.

1980 ◽  
Vol 28 (9) ◽  
pp. 979-990 ◽  
Author(s):  
P J Stoward ◽  
S S Spicer ◽  
R L Miller
Keyword(s):  
Sialic Acid ◽  
Horseradish Peroxidase ◽  
Periodic Acid ◽  
Apical Cell ◽  
Goblet Cells ◽  
Surface Epithelium ◽  
Renal Tubules ◽  
Peanut Lectin ◽  
Periodic Acid Schiff ◽  
Terminal Galactose

A peanut lectin-horseradish peroxidase (PL-HRP) conjugate has been applied to histochemical staining of paraffin sections of various mouse organs. The PL-HRP conjugate has selectively reacted with secretory bodies, the Golgi zone, and the apical cell surface in various cell types. Some positive sites, including lingual and tracheal serous glands, Brunner's glands, and the brush border of the proximal straight nephron, contained periodic acid-Schiff (PAS)-positive glycoconjugate with no affinity for basic reagents. The stored secretion in these sites was interpreted as containing neutral glycoprotein with terminal galactose residues which could, in part at least, account for the PAS reactivity. Duodenal goblet cells, which exhibited basophilia attributable to sulfate esters, also bound PL-HRP. As the binding was affected by prior sialidase digestion, the secretory glycoprotein in the duodenal goblet cells was judged to contain oligosaccharides with sulfate esters and terminal galactose uncapped by sialic acid. All sites known from their basophilia to form sialomucin failed to stain with the PL-HRP conjugate, but consistently gained reactivity following sialidase digestion and were inferred, therefore, to possess glycoproteins with oligosaccharide side chains containing subterminal galactose and terminal sialic acid. Lingual mucous glands, known to secrete a mucosubstance with basophilic properties indicative of the presence of sulfate esters but not sialic acid, stained with PL-HRP only after sialidase digestion and, accordingly, were reinterpreted as containing both sulfate esters and terminal galactose-sialic acid dimers. Staining of gastric surface epithelium demonstrated a srongly PAS-reactive neutral glycoprotein, and that of goblet cells in the cecum disclosed PAS-positive sulfated glycoprotein. The latter two sites lacked PL-HRP affinity without or with prior sialidase treatment and apparently possessed neither terminal galactose residues nor galactose-sialic acid dimers. PL-HRP affinity was observed exclusively in the Golgi cisternae of some epithelial cells, thus indicating that galactose occurs transiently as a terminal residue in this site. A few histologic sites, such as pancreatic and gastric zymogen cells and renal tubules, were devoid of both PAS reactivity and basophilia indicative of the presence of complex carbohydrate but stained strongly with the PL-HRP conjugate by means which are not understood. Galactose in the PL-HRP solution blocked or reversed the PL-HRP binding in most of the structures with an affinity for the conjugate, supporting the conclusions that the reagent is specific for galactosyl residues.

Some Secretory Products of the Invasive Stage of a Digenetic Trematode

1968 ◽  
Vol 26 ◽  
pp. 156-157
Author(s):  
Paul L. Krupa ◽  
Arya K. Bal ◽  
Gilles H. Cousineau
Keyword(s):  
Secretory Granules ◽  
Periodic Acid ◽  
Cell Types ◽  
Digenetic Trematode ◽  
Width Ratio ◽  
Uniform Density ◽  
Acid Hydrolases ◽  
Periodic Acid Schiff ◽  
Gland Cells ◽  
Secretory Products

The fine structure of various gland cells and their secretory products was studied in the invasive stage (cercaria) of the platyhelminth parasite, Cryptocotyle lingua. Secretory granules or droplets occur in several different specialized cell types, but those that we call attention to here are found in the (1) surface cytoplasmic tegument or “cuticle”, (2) ducts of cephalic (penetration) glands, and (3) epithelial lining of the “excretory bladder”.The tegumental granules appear as numerous, membrane-bounded circular or oval profiles of uniform density (Fig. 1). They are scattered more or less randomly among mitochondria and other inclusion bodies of the tegument. Some of the longer granules, with a length to width ratio of about 7 to 1, have their long axes oriented perpendicularly to the surface plasma membrane of the parasite. In cercariae tested for acid hydrolases with sodium β-glycerophosphate in Barka and Anderson's modification of Gomori's medium, clumps of reaction product appear in the vicinity of the granules and elsewhere within the tegument, but not within the granules themselves. As granules that stain with periodic acid-Schiff, they are seen in certain subsurface gland cells as well as in the tegument under the light microscope.

scholarly journals The Distributions of Glycogen in the Stomach on Long Tailed Macaque (Macaca fascicularis) during Pre and Postnatal Period

2008 ◽  
Vol 2 (2) ◽  
Author(s):  
Erdiansyah Rahmi
Keyword(s):  
Macaca Fascicularis ◽  
Periodic Acid ◽  
Postnatal Period ◽  
Periodic Acid Schiff ◽  
Fetal Period ◽  
Glandular Cells

The study was done to investigate the glycogen distribution in the stomach of long tailed macaques (Macaca fascicularis). The study used eight individuals of long tailed macaques consist of 6 fetuses (55, 70, 85, 100, 120, and 150 days pregnancy) and 2 infants (10 and 105 days). The gastrict tract were histologically stained using Hematoxyllin-Eosin (HE), and histochemically stained using Periodic Acid Schiff (PAS), to detect glycogen. The results revealed that the histogenesis pattern of stomach glandullar cells are developed in line with the age of the fetuses and infant. We also found that glycogen existance are increase in line with the age in the glandullar stomach. From the present study we concluded that the glycogen distribution in gastrict glandular cells were influenced by the age in the fetal period, while in the infant was under influence of the digestion activities, and the activities of the gastrict glandular cells was already seen at 55 days gestation.

Infection of human respiratory submucosal glands with rhinovirus: effects on cytokine and ICAM-1 production

1999 ◽  
Vol 277 (2) ◽  
pp. L362-L371 ◽  
Author(s):  
Mutsuo Yamaya ◽  
Kiyohisa Sekizawa ◽  
Tomoko Suzuki ◽  
Norihiro Yamada ◽  
Masayuki Furukawa ◽  
...  
Keyword(s):  
Tumor Necrosis Factor ◽  
Mrna Expression ◽  
Tumor Necrosis ◽  
Tumor Necrosis Factor Α ◽  
Surface Epithelium ◽  
Submucosal Gland ◽  
Gland Cells ◽  
Factor Α ◽  
Necrosis Factor ◽  
Viral Titers

To further understand the early biochemical events that occur in infected surface epithelium, we developed for the first time a model in which a respiratory submucosal gland cell population can be infected with rhinovirus (RV). Viral infection was confirmed by demonstrating with PCR that viral titers in supernatants and lysates from infected cells increased with time. Infection by RV14 upregulated the expression of intercellular adhesion molecule-1 (ICAM-1) mRNA, the major RV receptor, on submucosal gland cells, and it increased production of interleukin (IL)-1α, IL-1β, IL-6, IL-8, tumor necrosis factor-α, and granulocyte-macrophage colony-stimulating factor in supernatants. Antibodies to ICAM-1 inhibited RV infection of submucosal gland cells and decreased the production of cytokines after RV infection. Both IL-1α and IL-1β upregulated ICAM-1 mRNA expression and increased susceptibility to RV infection, whereas other cytokines failed to alter ICAM-1 mRNA expression. Furthermore, neutralizing antibodies to IL-1α and IL-1β significantly decreased the viral titers in supernatants and ICAM-1 mRNA expression after RV infection, but a neutralizing antibody to tumor necrosis factor-α was without effect. These findings suggest that respiratory submucosal gland cells play an important role in the initial stages of inflammation and provide useful insights into the pathogenesis of RV infection.

scholarly journals STRUCTURE OF SOME STRIGEATAS' ORGAN OF BRANDES

2019 ◽  
pp. 399-403
Author(s):  
Nacheva ◽  
Manikovskaya
Keyword(s):  
Periodic Acid ◽  
Secretory Activity ◽  
Bromphenol Blue ◽  
Main Function ◽  
Periodic Acid Schiff ◽  
Alaria Alata ◽  
Sudan Black B ◽  
Glandular Cells ◽  
Secondary Function ◽  
Schiff Reaction

The purpose of the research: the analysis of histology and histochemistry of the Brandes organ of Strigea strigis and Alaria alata. The trematode marites of Strigea striges and Alaria alata were the material. Fixation of the material was carried out in 10% neutral formalin. The treatment of the specimens was carried out using the conventional histological and histochemical methods: with hematoxylin-eosin according to the method of Van Gizon, Mallory, with Sudan black B, sulema-Bromphenol blue according to Bonhage, with periodic acid Schiff reaction by Mac-Manus, alcian blue according to Stedman and Mowry and with toluidine blue. Histochemical reactions were performed with appropriate controls. The studies have shown that the structure of the Brandes organ of Strigea strigis and Alaria alata differs by their constituents and morphology of glandular cells. The histochemical reactions are similar. The cells of the glandular complex show bromophenolophilia, toluudinophilia and fuchsinophilia in periodic acid Schiff reaction speaks about the glycoprotein nature of the secreted substances. Bromphenophilia and sudanophilia of glandular cells cytoplasm indicate the presence of lipoprotein substances in them. The Brandes of S. strigis and A. alata is a morphofunctional unit, to which the principle of multi-functionality is inherent. It performs the main function – digestion of food components by means of developed glandular structures and specialized secretory activity. Its ability to fix the helminth tightly in the endostatin can be considered a secondary function of the organ.

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