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.

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.

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

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.

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.

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 Purification, characterization and radioimmunoassay of adenosine deaminase from human leukaemic granulocytes

1981 ◽  
Vol 195 (2) ◽  
pp. 389-397 ◽  
Author(s):  
D A Wiginton ◽  
M S Coleman ◽  
J J Hutton
Keyword(s):  
Molecular Weight ◽  
Adenosine Deaminase ◽  
Gel Electrophoresis ◽  
Specific Activity ◽  
Periodic Acid ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Polyacrylamide Gel ◽  
Periodic Acid Schiff ◽  
Apparent Homogeneity

Adenosine deaminase was purified 3038-fold to apparent homogeneity from human leukaemic granulocytes by adenosine affinity chromatography. The purified enzyme has a specific activity of 486 mumol/min per mg of protein at 35 degrees C. It exhibits a single band when subjected to sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, non-denaturing polyacrylamide-gel electrophoresis and isoelectric focusing. The pI is 4.4. The enzyme is a monomeric protein of molecular weight 44000. Both electrophoretic behaviour and molecular weight differ from those of the low-molecular-weight adenosine deaminase purified from human erythrocytes. Its amino acid composition is reported. Tests with periodic acid-Schiff reagent for associated carbohydrate are negative. Of the large group of physiological compounds tested as potential effectors, none has a significant effect. The enzyme is specific for adenosine and deoxyadenosine, with Km values of 48 microM and 34 microM respectively. There are no significant differences in enzyme function on the two substrates. erythro-9-(2-Hydroxy non-3-yl) adenine is a competitive inhibitor, with Ki 15 nM. Deoxycoformycin inhibits deamination of both adenosine and deoxyadenosine, with an apparent Ki of 60-90 pM. A specific antibody was developed against the purified enzyme, and a sensitive radioimmunoassay for adenosine deaminase protein is described.

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.

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

Concanavalin A-horseradish peroxidase bridge staining of alpha-1 glycoproteins separated by isoelectric focusing on polyacrylamide gel.

1976 ◽  
Vol 24 (8) ◽  
pp. 908-914 ◽  
Author(s):  
R C Allen ◽  
S S Spicer ◽  
D Zehr
Keyword(s):  
Horseradish Peroxidase ◽  
Isoelectric Focusing ◽  
Concanavalin A ◽  
Periodic Acid ◽  
Polyacrylamide Gel ◽  
New Method ◽  
Polyacrylamide Gels ◽  
Periodic Acid Schiff ◽  
Coomassie Blue

The Coomassie Blue protein stain and the periodic acid-Schiff stain for glycoproteins are compared to a new method of staining glycoproteins resolved electrophoretically. The method utilizes a Concanavalin A-horseradish peroxidase sequence to visualize selectively glycoproteins with terminal or internal mannose or terminal N-acetylglucosamine. The method applied to characterization of M and Z allele products of alpha-l-antitrypsins separated by isoelectric focusing of polyacrylamide gels slabs have revealed differences in carbohydrate content of various components that were previously undetected.

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