scholarly journals Histochemical and biometric study of the gastrointestinal system of Hyla orientalis (Bedriaga, 1890) (Anura, Hylidae)

2014 ◽  
Author(s):  
E. Akat ◽  
H. Arıkan ◽  
B. Göçmen
Keyword(s):  
Hyaluronic Acid ◽  
Sialic Acid ◽  
Large Intestine ◽  
Periodic Acid ◽  
Fluorescein Isothiocyanate ◽  
Gastrointestinal System ◽  
Periodic Acid Schiff ◽  
System Integrity ◽  
Aldehyde Fuchsin ◽  
Small And Large Intestine

This study was carried out to assess the localization of hyaluronic acid (HA) and the distribution of glycoproteins in the gastrointestinal system of adult Hyla orientalis. Histochemical analysis of the gastrointestinal system in H. orientalis showed that mucous content included glycogene and/or oxidable dioles [periodic acid/Schiff (PAS)+], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS+) and acid sulphate [Aldehyde fuchsin (AF)+] glycoproteins. However the mucus content was not the same in stomach, small and large intestine. The mucus content of stomach included only glycogene and/or oxidable dioles and sialic acid residues. Besides these histochemical methods, the localization of HA was detected using biotinylated hyaluronic acid binding protein labeled with streptavidin-fluorescein isothiocyanate (FITC). In the extracellular matrix of the submucosa, the reaction for HA was evident. Since HA was located in submucosa beneath the epithelial layer of gastrointestinal system, it has a significant role in hydric balance, and essential to provide the gastrointestinal system integrity and functionality. According to biometric results, there were statistical differences between small and large intestine in terms of the amount of material stained positive with PAS/AB, PAS, KOH/PAS and AF/AB. Additionally, number of goblet cells in the small and large intestine was significantly different.

scholarly journals THE EFFECT OF DIGESTION WITH STREPTOMYCES HYALURONIDASE UPON CERTAIN HISTOCHEMICAL REACTIONS OF HYALURONIC ACID-CONTAINING TISSUES

1973 ◽  
Vol 21 (9) ◽  
pp. 794-803 ◽  
Author(s):  
KAZUYORI YAMADA
Keyword(s):  
Hyaluronic Acid ◽  
Alcian Blue ◽  
Periodic Acid ◽  
Colloidal Iron ◽  
Periodic Acid Schiff ◽  
High Iron ◽  
Azure A ◽  
Aldehyde Fuchsin ◽  
Testicular Hyaluronidase

The effect of digestion with Streptomyces hyaluronidase upon certain histochemical reactions of hyaluronic acid-containing tissues has been studied in a series of human, mammalian and avian specimens. These histochemical reactions are those for the demonstration of neutral and sulfated and nonsulfated acid mucosaccharides such as periodic acid-Schiff, Alcian Blue (pH 1.0), azure A (pH 1.5), high iron diamine, aldehyde fuchsin, Alcian Blue (pH 2.5), Alcian Blue (pH 2.5)-periodic acid Schiff, azure A (pH 4.5), low iron diamine and colloidal iron. In addition, the effect of digestion with testicular hyaluronidase upon the same reactions of the same tissues was observed for comparison. Digestion with Streptomyces hyaluronidase diminishes the staining reactions due to hyaluronic acid but fails to affect those for neutral and sulfated acid mucosaccharides. These results indicate that digestion with Streptomyces hyaluronidase is a method of choice for the identification of hyaluronic acid in mucosaccharide histochemistry.

scholarly journals DISTRIBUTION OF MUCOSUBSTANCES IN THE DEVELOPING RAT HEART

1972 ◽  
Vol 20 (11) ◽  
pp. 896-907 ◽  
Author(s):  
ROGER R. MARKWALD ◽  
WILLIAM N. ADAMS SMITH
Keyword(s):  
Electrolyte Concentration ◽  
Cetylpyridinium Chloride ◽  
Periodic Acid ◽  
Frozen Sections ◽  
Concentrated Solutions ◽  
Periodic Acid Schiff ◽  
Rat Hearts ◽  
Aldehyde Fuchsin ◽  
Developing Rat ◽  
Fibroblastic Cells

Mucosubstances (MS) were examined in 10½-14½-day embryonic rat hearts utilizing nonaqueous fixatives or formaldehyde vapor-fixed frozen sections hydrated in concentrated solutions of cetylpyridinium chloride. Ribonuclease-resistant, polyanionic sites were limited to the extracellular cardiac jelly, endocardium and fibroblastic cells (cushion tissue) associated with the endocardium. The cardiac jelly and endocardium of day 10½ embryos principally contained a hyaluronic acid-like carboxylated mucosubstance whose alcianophilia at pH 2.5 was abolished by hyaluronidase but was resistant to NaOH extraction and neuraminidase and trypsin digestion. A critical electrolyte concentration of 0.2 M MgCl2 abolished alcianophilia. On days 13½-14½ carboxylated MS were restricted to cushion tissue and partially resisted mild methylation. Sulfated MS were limited to primitive endocardial cells which gave origin to cushion tissue. Dye deposits of aldehyde fuchsin, high iron diamine or Alcian Blue (pH 1.0) were localized on cell surfaces and such staining was prevented by strong (60°C) methylation. Hyaluronidase sensitivity of sulfated MS decreased with gestation. The critical electrolyte concentration varied from 0.5-0.7 M MgCl2 on days 11½-12½ to 0.8-0.9 M MgCl2 after day 12½. The sulfated MS of endocardial cells were preceded by a transitory accumulation of diastase-resistant, periodic acid-Schiff-positive material. Possible roles of MS in normal and abnormal cardiac septation processes are discussed.

scholarly journals THE HISTOCHEMICAL INTERPRETATION OF THE COMPLEX RESULTS OF METHYLATION UPON GASTROINTESTINAL TRACT MUCINS, WITH SPECIAL REFERENCE TO THE PERIODIC ACID-SCHIFF REACTIVITY

1974 ◽  
Vol 22 (10) ◽  
pp. 986-991 ◽  
Author(s):  
P. E. REID ◽  
C. F. A. CULLING ◽  
W. L. DUNN
Keyword(s):  
Sialic Acid ◽  
Gastrointestinal Tract ◽  
Special Reference ◽  
Periodic Acid ◽  
Separate Study ◽  
Sulfate Ester ◽  
Periodic Acid Schiff ◽  
Smith Degradation ◽  
Vicinal Diol ◽  
Schiff Reaction

The histochemical use of methylation has complex results; particularly in respect of the periodic acid-Schiff reaction, these are analyzed and discussed. Methods are described which allow the separate study of the following effects: (a) the removal of the KOH/periodic acid-Schiff effect; (b) removal of sialic acid from a potential vicinal diol; and (c) the removal of O-sulfate ester from a potential vicinal diol. The use of the Smith degradation technique, in addition to the above, also allows inferences to be drawn in respect of the structure of the mucins (glycoproteins) being investigated.

Lectins as Cytochemical Probes of the Developing Wheat Grain. V. Demonstration of Separate Polysaccharides Containing N-Acetyl-D-Glucosamine and D-Galactose in Nuclear Epidermal Cell Walls

1984 ◽  
Vol 11 (3) ◽  
pp. 179 ◽  
Author(s):  
BA Baldo ◽  
D Barnett ◽  
JW Lee
Keyword(s):  
Epidermal Cell ◽  
Cell Walls ◽  
Wheat Germ ◽  
Periodic Acid ◽  
Fluorescein Isothiocyanate ◽  
Wheat Grain ◽  
Periodic Acid Schiff ◽  
Thin Sections ◽  
Lectin Staining ◽  
Wheat Germ Lectin

Fluorescein isothiocyanate-labelled lectin from wheat-gem, which binds N-acetyl-D-glucosamine, and Griffonia simplicifolia, Arachis hypogaea and Glycine max lectins, each of which binds D-galactose, react with nucellar epidermal cell walls in thin sections of plastic-embedded developing wheat grain. Reactivity of these cell walls with periodic acid-Schiff reagent, the absence of staining with protein stains and the failure of a number of proteases and the endoglycosidases D and H to prevent the binding suggested that the lectin-reactive wall components are neither proteins nor N-glycosidically linked glycoproteins. Morphological differences in lectin staining patterns and treatment of sections with chitinase and α-galactosidase, prior to the reaction with the lectins, indicated that two separate polysaccharides are probably involved in the binding. Chitinase removed the reactivity of the nucellar epidermal cell walls for wheat-germ lectin but the binding of D-galactose-specific lectins was unimpaired. Conversely, α-galactosidase did not affect the binding of wheat-germ lectin but reactivity with the galactose-specific lectins was abolished. From the available evidence we conclude that one polysaccharide in the nucellar epidermal cell wall reacts with wheat-germ lectin and contains N-acetyl-D-glucosamine in a chitin-like structure. The other polysaccharide reacts with D-galactose- specific lectins by virtue of terminal α-D-galactose residues. Hydrolysis and subsequent chromatographic analysis of nucellar epidermal cell walls peeled from immature grains revealed the presence of D-glucosamine, D-glucose, D-galactose, D-xylose, L-arabinose and a trace of D-mannose.

A physiological, biochemical and histological study of goose tracheal mucin and its secretion

1977 ◽  
Vol 279 (969) ◽  
pp. 513-540 ◽  
Keyword(s):  
Sialic Acid ◽  
Nerve Stimulation ◽  
Histological Study ◽  
Periodic Acid ◽  
Gel Filtration ◽  
Goblet Cells ◽  
Mucin Secretion ◽  
Electron Microscopic ◽  
Periodic Acid Schiff ◽  
Tracheal Mucus

Tracheal mucin secretion has been measured from a segment of trachea, isolated in situ , in anaesthetized geese by a method that involves radioactive labelling of tracheal mucus glycoproteins (Gallagher et al. 1975). Goose tracheal mucus comes entirely from goblet cells, since the goose trachea does not contain submucosal mucous or serous glands, and this method has been used to investigate the nervous and pharmacological control of the mucin secretion from these epithelial goblet cells. The mucins secreted have been collected, fractionated, and chemically analysed. Intracellular mucin has been examined histochemically, and the results of electron microscopic observations of epithelial cells and nerves are presented. Acetylcholine increased tracheal mucin secretion, and this effect was completely blocked by atropine. Neither α- nor β-stimulant sympathomimetic amines affected tracheal mucin secretion. Stimulation of the peripheral cut ends of the descending oesophageal nerves increased tracheal mucin secretion and the majority of this response, approximately three-quarters, appeared to be cholinergic since this proportion was blocked by atropine. The mediator for the atropine-resistant part of the response is not known, but it appears not to be a β-adrenoreceptor stimulant since the response to nerve stimulation was unaffected by propranolol given at 34 μm intrasegmentally. Other possibilities are discussed. Atropine itself decreased the resting level of tracheal mucin secretion. The local anaesthetic, lignocaine, increased tracheal mucin secretion, while at the same time blocking the responses to acetylcholine and descending oesophageal nerve stimulation. The implications of this are discussed. The electrophoretic, gel filtration and ion-exchange properties of goose tracheal mucins showed that they represented high molecular mass, negatively charged glycoproteins which could be labelled biosynthetically with [ 35 S]sulphate, [ 3 H]- and [ 14 C]glucose. These mucins could be stained with Alcian blue or periodic acid Schiff reagent. The carbohydrate composition was unusual for an epithelial glycoprotein in that fucose was absent and mannose was present in small quantities. The monosaccharides present in larger quantity were galactose, N -acetylglucosamine, N -acetylgalactosamine and sialic acid. Histochemical analysis of tissue sections of gosling tracheas demonstrated that nearly all of the glycoprotein in epithelial goblet cells contained both sialic acid and sulphate residues. Sialated mucin was present also, but to a lesser extent, and many cells contained a mixture of sialated and sulphated mucins. The adult goose trachea had a high proportion of sialated glycoprotein. Electron microscopy showed a range of epithelial cell types and intra-epithelial nerves also. Many of the nerves had neurosecretory vesicles suggestive of motor function and some were near to goblet cells.

scholarly journals Detection in milk of a 60,000 Mr mouse and a 68,000 Mr human phosphorylated glycoprotein by histochemical analysis on polyacrylamide gels.

1983 ◽  
Vol 31 (6) ◽  
pp. 709-716 ◽  
Author(s):  
M R Green
Keyword(s):  
Molecular Weight ◽  
Sialic Acid ◽  
Periodic Acid ◽  
Sodium Dodecyl ◽  
Apparent Molecular Weight ◽  
Blue Staining ◽  
Polyacrylamide Gels ◽  
Periodic Acid Schiff ◽  
Coomassie Blue Staining ◽  
Coomassie Blue

Proteins in colostrum and skimmed milk from humans and mice were separated by electrophoresis on polyacrylamide gels and stained with Coomassie blue (CB), Ethyl-Stains-all (ESA), and periodic acid-Schiff (PAS) to investigate changes that may occur in milks throughout lactation. In mouse colostrum but not in mature mouse milk, a PAS-positive protein of apparent molecular weight of 60,000 stained prominently blue with ESA. A protein in human milk with a molecular weight of 68,000 stained similarly but was present throughout lactation. The intensity of blue staining of these minor proteins in milk approached that obtained with casein phosphoproteins. The metachromatic dye ESA stains phosphoproteins and sialic acid-rich glycoproteins blue to blue-green. Removal of phosphorus from the former and sialic acid from the latter results in those proteins staining red with ESA. The intensity of blue staining of the 60,000 and 68,000 Mr proteins was diminished but not lost following treatment with phosphatase. It was eliminated following neuraminidase digestion of the mouse protein and mild acid hydrolysis of the human protein. Coomassie blue staining of the proteins was not affected by these procedures. Following electrophoresis of milk and milk fractions in a non-sodium dodecyl sulfate-containing system, the proteins were identified by their characteristic staining properties with ESA and isolated.

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.

scholarly journals Functional anatomy of the lacrimal gland in African black ostrich Struthio camelus domesticus in the embryonic and postnatal period

2015 ◽  
Vol 82 (1) ◽  
Author(s):  
Joanna Klećkowska-Nawrot ◽  
Karolina Goździewska-Harłajczuk ◽  
Renata Nowaczyk ◽  
Krzysztof Krasucki
Keyword(s):  
Morphological Changes ◽  
Periodic Acid ◽  
Age Groups ◽  
Functional Anatomy ◽  
Postnatal Period ◽  
Age Group ◽  
Periodic Acid Schiff ◽  
Struthio Camelus ◽  
Aldehyde Fuchsin ◽  
African Black

The aim of the present study was morphological and histochemical analysis of the lacrimalgland (LG) in African black ostrich Struthio camelus domesticus in the embryonic and postnatalperiod. Studies were conducted on 50 ostriches aged between the 28th day of incubation until7 months old. Tissue sections were stained with haematoxylin and eosin, Azan trichrome,periodic acid-Schiff, Alcian blue pH 2.5, aldehyde fuchsin and Hale’s dialysed iron. The LGin ostrich was classified as a tubulo-acinar type. The primordia of the lobes were determinedin the LG structure on the 28th day of incubation, whilst the weakly visible lobes with aciniand tubules were observed on the 40th day of incubation. Morphometric studies of the LGshowed steady growth, characterised by an increase in both length and width. Histometricmeasurements of lobe size showed little difference between the first, second and third agegroups, whilst in the fourth age group a marked increase in size of lobes was observed.The study showed that, apart from morphological changes, during the growth of the LGthe character of acid mucopolysaccharides changed. Sulphated acid mucopolysaccharideswere indicated, particularly with aldehyde fuchsin (AF) staining in the fourth age group.The Hale’s dialysed iron (HDI) staining showed a low concentration of carboxylated acidmucopolysaccharides in the first and second age groups and a higher concentration in thethird and fourth age groups. Periodic acid-Schiff staining (PAS)-positive cells were observedin each age group, but only a small number of cells with a weakly PAS-positive reaction weredemonstrated in the first age group.

scholarly journals Factor VIII/von Willebrand factor protein: sensitivity of periodic acid Schiff stain to carbohydrate deficiency

Blood ◽  
1982 ◽  
Vol 59 (6) ◽  
pp. 1310-1316 ◽  
Author(s):  
HR Gralnick ◽  
GM Jackson ◽  
SB Williams ◽  
MC Cregger
Keyword(s):  
Sialic Acid ◽  
Factor Viii ◽  
Von Willebrand Factor ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Human Factor Viii ◽  
Acid Deficiency ◽  
Or Organization ◽  
Von Willebrand ◽  
Willebrand Factor

Abstract We have investigated the periodic acid Schiff (PAS) Coomassie staining ratio of the human factor VIII/von Willebrand factor (fVIII/vWf) protein. The PAS-Coomassie staining ratio is consistent over 8 days. The PAS-Coomassie ratio of fVIII/vWf protein purified from different starting materials does not appear to be significantly different. The PAS stain can detect as little as 300 ng of carbohydrate in the fVIII/vWf protein. Desialation did not affect the PAS-Coomassie ratio, while removal of penultimate galactose resulted in a marked reduction in the PAS-Coomassie ratio. This reduction was further accentuated with the removal of N-acetylglucosamine. The smaller multimers of the fVIII/vWf protein have a reduced sialic acid and PAS-Coomassie staining ratio. This difference does not appear to be related to the sialic acid deficiency but may be related to the distribution or organization of the carbohydrate moieties on the smaller fVIII/vWf multimers.

scholarly journals Platelet membrane studies in the May-Hegglin anomaly

Blood ◽  
1981 ◽  
Vol 58 (2) ◽  
pp. 279-284 ◽  
Author(s):  
BS Coller ◽  
MH Zarrabi
Keyword(s):  
Sialic Acid ◽  
Periodic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Surface Glycoprotein ◽  
Galactose Oxidase ◽  
Periodic Acid Schiff ◽  
Giant Platelets ◽  
Total Sialic Acid ◽  
Bernard Soulier Syndrome

Abstract Since studies of the giant platelets in the Bernard-Soulier syndrome have shown decreased electrophoretic mobility, decreased sialic acid, and an abnormality in a membrane glycoprotein, we performed similar studies on the giant platelets from two patients with the May-Hegglin anomaly. The patients' platelet electrophoretic mobilities did not differ from control. Although the total sialic acid contents of the patients' platelets were greater than control when calculated per platelet, they were very similar to control when normalized for differences in platelet volume and surface area. When platelet proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis there were no differences between the glycoproteins of control and patient platelets as judged by the patterns of periodic acid Schiff staining and fluorescein-labeled concanavalin A binding. Similarly, patterns of surface glycoprotein labeling by neuraminidase/galactose oxidase/KB3H4 were identical. We conclude that unlike the giant platelets in the Bernard-Soulier syndrome, those of the May-Hegglin anomaly are not associated with a membrane abnormality detectable by these techniques.

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