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 SAPONIFICATION-INDUCED INCREASE IN THE PERIODIC ACID-SCHIFF REACTION IN THE GASTROINTESTINAL TRACT MECHANISM AND DISTRIBUTION OF THE REACTIVE SUBSTANCE

1973 ◽  
Vol 21 (5) ◽  
pp. 473-482 ◽  
Author(s):  
P. E. REID ◽  
C. F. A. CULLING ◽  
W. L. DUNN
Keyword(s):  
Alternative Hypothesis ◽  
Periodic Acid ◽  
Periodate Oxidation ◽  
Periodic Acid Schiff ◽  
Initial Oxidation ◽  
Vicinal Diol ◽  
Sodium Borohydride Reduction ◽  
Vicinal Diols ◽  
Schiff Reaction ◽  
Carbonyl Function

The mechanism and distribution of the KOH-periodic acid-Schiff (PAS) effect in rat large intestine, human sigmoid colon, rabbit rectum and rabbit Brunner's glands have been investigated following the abolition of normally occurring PAS-positive vicinal diols by means of a periodate oxidation-sodium borohydride reduction sequence. The KOH-PAS effect was shown to be independent of the initial oxidation time and could be abolished by pretreatment with either methanolic-HCl or alcoholic KOH. The evidence suggests that the KOH-PAS effect is due to the removal of an o-acyl-ester substituent on a potential vicinal diol. The alternative hypothesis, that the effect is due to an ester involving the carbonyl function of a macromolecule or a lactone, is less likely. The difficulty of interpretation of histochemical results following methylation is discussed.

scholarly journals THE HISTOCHEMICAL DEMONSTRATION OF O-ACYLATED SIALIC ACID IN GASTROINTESTINAL MUCINS THEIR ASSOCIATION WITH THE POTASSIUM HYDROXIDE-PERIODIC ACID-SCHIFF EFFECT

1974 ◽  
Vol 22 (8) ◽  
pp. 826-831 ◽  
Author(s):  
C. F. A. CULLING ◽  
P. E. REID ◽  
M. G. CLAY ◽  
W. L. DUNN
Keyword(s):  
Sialic Acid ◽  
Gastrointestinal Tract ◽  
Acid Hydrolysis ◽  
Potassium Hydroxide ◽  
Periodic Acid ◽  
Histochemical Demonstration ◽  
Sialic Acids ◽  
Side Chains ◽  
Periodic Acid Schiff ◽  
Alcoholic Potassium Hydroxide

O-Acylated sialic acids have been demonstrated histochemically in the epithelial mucins of the lower end of the gastrointestinal tract. The potassium hydroxide-periodic acid-Schiff effect has been shown to be due, in most areas, to the presence of these O-acyl esters on the polyhydroxy side chains of sialic acid. The esters have been shown to hinder the removal of the sialic acids by either neuraminidase or acid hydrolysis unless tissues were pretreated with alcoholic potassium hydroxide. It is suggested that substitution at the C4 position is responsible for their neuraminidase resistance.

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.

Sign in / Sign up

Export Citation Format

Share Document