Histochemical procedures for the simultaneous visualization of neutral sugars and either sialic acid and itsO-acyl variants orO-sulphate ester. II. Methods based upon the periodic acid—phenylhydrazine—Schiff reaction

1987 ◽  
Vol 19 (5) ◽  
pp. 257-263 ◽  
Author(s):  
C. M. Park ◽  
P. E. Reid ◽  
D. A. Owen ◽  
W. L. Dunn ◽  
D. Volz
Keyword(s):  
Sialic Acid ◽  
Periodic Acid ◽  
Neutral Sugars ◽  
Schiff Reaction ◽  
Simultaneous Visualization

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.

scholarly journals Effect of activity on performance and morphology in ischaemic rat slow muscles

1990 ◽  
Vol 152 (1) ◽  
pp. 265-279
Author(s):  
A. Corsi ◽  
A. L. Granata ◽  
O. Hudlicka
Keyword(s):  
Blood Supply ◽  
Periodic Acid ◽  
Capillary Density ◽  
Muscle Performance ◽  
Similar Proportion ◽  
Periodic Acid Schiff ◽  
Rat Soleus Muscle ◽  
Improved Performance ◽  
Isometric Twitch ◽  
Schiff Reaction

Muscle performance and structure was studied in rat soleus muscle with limited blood supply in combination with chronic muscle stimulation. Blood supply to the lower leg was restricted by ligation of the common iliac artery, electrodes were implanted in the vicinity of the sciatic nerve and ankle flexors were denervated. Three days later, soleus and gastrocnemius muscles were stimulated at 4 Hz four times a day for a period of 20 min with 2 h intervals between stimulations; this procedure was continued for 4 days. Muscle performance, histochemistry and ultrastructure were studied on the eighth day after operation in these muscles and in ischaemic unstimulated muscles with denervated ankle flexors. Both were compared with control animals. Muscles with limited blood supply developed less isometric twitch tension than control muscles (peak twitch tension in ischaemic muscle was 60.3 +/− 4.8 g g-1 muscle, mean +/− S.E.M., compared to 79.7 +/− 6.9 g g-1 in control muscle; tensions after 5 min contraction were 54.5 +/− 5.5 g g-1 in ischaemic muscle compared to 70.6 +/− 6 g g-1 in controls). Stimulated muscles with limited blood supply had higher peak (85 +/− 16.6 g g-1) and final (87 +/− 12 g g-1) tensions, and also fatigued less than muscles with limited blood supply but no stimulation. Histochemical estimation of capillary density (by staining for alkaline phosphatase) and slow (SO) and fast (FOG) fibres (by myosin ATPase staining) revealed similar capillary to fibre ratios (2.5) and a similar proportion of FOG fibres (around 18%) in all muscles. The proportion of glycogen-depleted fibres (estimated from the periodic acid Schiff reaction, PAS) in muscles removed from animals 10 min after a 5 min period of isometric twitches was significantly lower in ischaemic muscles (45.1 +/− 1.9%) than in control (80.5 +/− 1.5%) or chronically stimulated ischaemic muscles (67.3 +/− 4.0%). Electron microscopy showed disorganised myofibrils with Z-line streaming in 7.48 +/− 3.04% of fibres in muscles with limited blood supply. Swollen and degenerated mitochondria, dilated sarcoplasmic reticulum and areas of disrupted sarcolemma were also observed. Stimulated ligated muscles showed a significantly lower proportion of fibres with disorganised filaments (0.65 +/− 0.32%) and other signs of damage were much less frequent. The reduced damage and improved performance of chronically stimulated slow muscle may be the result of improved microcirculation, preventing accumulation of lactate.

scholarly journals Activation and activity of glycosylated KLKs 3, 4 and 11

2018 ◽  
Vol 399 (9) ◽  
pp. 1009-1022 ◽  
Author(s):  
Shihui Guo ◽  
Peter Briza ◽  
Viktor Magdolen ◽  
Hans Brandstetter ◽  
Peter Goettig
Keyword(s):  
Physiological Function ◽  
Periodic Acid ◽  
Hek293 Cells ◽  
Secretory Expression ◽  
Detailed Characterization ◽  
Periodic Acid Schiff ◽  
Clinical Biomarkers ◽  
Schiff Reaction

Abstract Human kallikrein-related peptidases 3, 4, 11, and KLK2, the activator of KLK3/PSA, belong to the prostatic group of the KLKs, whose major physiological function is semen liquefaction during the fertilization process. Notably, these KLKs are upregulated in prostate cancer and are used as clinical biomarkers or have been proposed as therapeutic targets. However, this potential awaits a detailed characterization of these proteases. In order to study glycosylated prostatic KLKs resembling the natural proteases, we used Leishmania (LEXSY) and HEK293 cells for secretory expression. Both systems allowed the subsequent purification of soluble pro-KLK zymogens with correct propeptides and of the mature forms. Periodic acid-Schiff reaction, enzymatic deglycosylation assays, and mass spectrometry confirmed the glycosylation of these KLKs. Activation of glycosylated pro-KLKs 4 and 11 turned out to be most efficient by glycosylated KLK2 and KLK4, respectively. By comparing the glycosylated prostatic KLKs with their non-glycosylated counterparts from Escherichia coli, it was observed that the N-glycans stabilize the KLK proteases and change their activation profiles and their enzymatic activity to some extent. The functional role of glycosylation in prostate-specific KLKs could pave the way to a deeper understanding of their biology and to medical applications.

scholarly journals Acute lymphoblastic leukemia with unusual cytoplasmic granulation: a morphologic, cytochemical, and ultrastructural study

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 406-411 ◽  
Author(s):  
J Fradera ◽  
E Velez-Garcia ◽  
JG White
Keyword(s):  
Acute Lymphoblastic Leukemia ◽  
Lymphoblastic Leukemia ◽  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Acute Leukemias ◽  
Ultrastructural Studies ◽  
Sudan Black B ◽  
Blast Cells ◽  
Chediak Higashi Syndrome ◽  
Schiff Reaction

Abstract The classification of the acute leukemias depends mainly on the morphologic and cytochemical evaluation of the blast forms. One of the main accepted morphologic criteria in the differentiation between acute lymphoblastic leukemia (ALL) and acute myeloblastic leukemia (AML) is the absence of granules in the blast cells of ALL. We evaluated a patient with ALL in whom granules were present in the cytoplasm of 35% of the blast cells, as seen in AML. Cytochemical evaluation was performed, including periodic acid-Schiff reaction, Sudan black B, alpha-naphthyl acetate, alpha-naphthyl butyrate, naphthol AS-D chloroacetate, and acid phosphatase stains. The results of these studies confirmed the morphologic impression and diagnosis of ALL. Ultrastructural evaluation revealed that the granules consisted of many tiny vesicles closely packed together in a proteinaceous matrix, resembling to some extent the inclusions described in lymphocytes in the Chediak-Higashi syndrome, but clearly different. The morphologic, cytochemical, and ultrastructural studies of this unique case are presented in detail. To our knowledge, this is the first time that such granules have been described in blast cells of ALL.

The periodic acid-Schiff reaction in the cornea of the developing chick

1960 ◽  
Vol 121 (5) ◽  
pp. 351-368 ◽  
Author(s):  
Ronan O'Rahilly ◽  
David B. Meyer
Keyword(s):  
Periodic Acid ◽  
Periodic Acid Schiff ◽  
Schiff Reaction

scholarly journals A VERTICAL MICROSYSTEM FOR DISCONTINUOUS ELECTROPHORESIS OF INSECT TISSUE PROTEINS USING THIN SHEETS OF POLYACRYLAMIDE GEL

1972 ◽  
Vol 20 (5) ◽  
pp. 368-384 ◽  
Author(s):  
ANITA C. BEEN ◽  
ELLEN M. RASCH
Keyword(s):  
Periodic Acid ◽  
Polyacrylamide Gel ◽  
Thin Sheets ◽  
Periodic Acid Schiff ◽  
Sudan Black B ◽  
Tissue Extracts ◽  
Salivary Gland Cells ◽  
Schiff Reaction ◽  
Coomassie Brilliant ◽  
Nonspecific Esterases

Proteins extracted from individual pairs of salivary glands or other larval tissues of Sciara coprophila (Diptera) were separated in a vertical microsystem for discontinuous electrophoresis using thin sheets of polyacrylamide gel cast in multiple layers of varying pore size. After electrophoresis at 150 volts for 40 min, gels were stained ( a) for total proteins with Coomassie brilliant blue, ( b) for glycoproteins with the periodic acid-Schiff reaction, ( c) for lipoproteins with Sudan black B or ( d) for nonspecific esterases with fast blue RR as coupler and α-naphthol acetate as substrate. Sequential application of these reactions to individual gel sectors permitted direct comparisons of protein profiles for 15-20 different samples of tissue extracts carried on a single gel sheet in adjacent lanes and thus subjected to identical conditions of electrophoresis. Representative photographs and densitometric scans are presented to show the suitability of thin gel sheets for autoradiography and for both qualitative and quantitative evaluation of tissue-specific differences in patterns of protein banding found for salivary gland cells, the gastric ceca, or the hemolymph of individual Sciara larvae sacrificed at particular stages of fourth instar development. Innovative details of methodology are presented, including the use of a microspectrophotometer to scan electropherograms of insect proteins and several types of human blood serum.

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 Histochemical and Autoradiographic Studies on the Effects of Aging on the Mucopolysaccharides of the Periosteum

1959 ◽  
Vol 6 (2) ◽  
pp. 171-178 ◽  
Author(s):  
Edgar A. Tonna ◽  
Eugene P. Cronkite
Keyword(s):  
Chondroitin Sulfate ◽  
Toluidine Blue ◽  
Bone Growth ◽  
Periodic Acid ◽  
Neutral Type ◽  
Autoradiographic Study ◽  
Colloidal Iron ◽  
Periodic Acid Schiff ◽  
Schiff Reaction ◽  
Effects Of Aging

An autoradiographic study was made using S35-sulfate for the localization, distribution, and variation in the mucopolysaccharide content of the femoral periosteum of rats from birth to old age. The mucopolysaccharides were also studied histochemically, using toluidine blue O, Rinehart and Abu'l-Haj's colloidal iron method, and the periodic acid-Schiff reaction, before and after hyaluronidase treatment. Autoradiograms revealed the uptake of S35 particularly in the vicinity of the preosseous zone and adjacent osteoblasts. This labelling was highest at the period of rapid bone growth. With increasing age, the S35 uptake became progressively less. The preosseous zone showed γ-metachromatic staining at all ages after treatment with toluidine blue. Active osteoblasts were mostly orthochromatic, however, ß-metachromasia was exhibited at a later age. Abundant amounts of intra- and extracellular mucopolysaccharides of both the acid and neutral type were demonstrated in the periosteum. S35 uptake and γ-metachromasia show the presence of sulfated mucopolysaccharides, of which chondroitin sulfate predominates in the preosseous zone. Since S35 uptake is high in active osteoblasts, the inability to demonstrate metachromasia in osteoblasts may indicate either that chondroitin sulfate is liberated as fast as it is being produced, or that it may be present within the cells in a precursor form not detectable by histochemical methods.

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.

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