A theoretical study of concentration of profiles of primary cytochemical-enzyme reaction products in membrane-bound cell organelles and its application to lysosomal acid phosphatase

1976 ◽  
Vol 8 (6) ◽  
pp. 609-624 ◽  
Author(s):  
C. J. Cornelisse ◽  
W. Th. Hermens ◽  
M. Tjok Joe ◽  
W. A. L. Duijndam ◽  
P. Van Duijn
Keyword(s):  
Acid Phosphatase ◽  
Theoretical Study ◽  
Enzyme Reaction ◽  
Reaction Products ◽  
Cell Organelles ◽  
Lysosomal Acid ◽  
Membrane Bound

scholarly journals Acid-beta-glycerophosphatase reaction products in the central nervous system mitochondria following x-ray irradiation.

1975 ◽  
Vol 23 (6) ◽  
pp. 402-410 ◽  
Author(s):  
L Roizin ◽  
D Orlovskaja ◽  
J C Liu ◽  
A L Carsten
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Acid Phosphatase ◽  
Enzyme Reaction ◽  
Reaction Products ◽  
Huntington's Chorea ◽  
X Ray ◽  
System A ◽  
Chemical Lesion ◽  
The Central Nervous System

A survey of the literature to date on the enzyme histochemistry of intracellular organelles has not yielded any reference to the presence of acid phosphatase reaction products in the mammalian mitochondria of the central nervous system. A combination of Gomori's acid phosphatase mehtod, however, with standard electron microscopy has disclosed the presence of enzyme reaction products in the mitochondria of the central nervous system of rats from 2 hr to 22 weeks after x-ray irradiation, as well as in a cerebral biopsy performed on a patient affected by Huntington's chorea. No enzyme reaction products, on the other hand, were observed in serial sections that had been incubated in substrates either containing sodium fluoride or lacking in beta-glycerophosphate. The abnormal mitochondrial enzyme reaction (chemical lesion) is considered to be the consequenco of the pathologic process affecting the ultrastructural-chemical organization of the organelle.

scholarly journals Further observations on the activation of lysosomal acid α-glucosidase by cortisone derivatives

1973 ◽  
Vol 132 (3) ◽  
pp. 435-438 ◽  
Author(s):  
E. J. Bourne ◽  
K. Clarke ◽  
J. B. Pridham ◽  
J. J. M. Rowe
Keyword(s):  
Acid Phosphatase ◽  
Rat Liver ◽  
Lysosomal Membrane ◽  
Cortisone Acetate ◽  
Lysosomal Acid ◽  
Liver Slices ◽  
Liver Lysosomes ◽  
Membrane Bound ◽  
Isolated Liver

1. Cortisone acetate activates the acid α-glucosidase in rat liver slices and in isolated liver lysosomes. 2. The reaction is steroid specific and moreover does not occur with lysosomal acid phosphatase or β-galactosidase. 3. After pretreatment of the lysosomes with cortisone, substrate (maltose) binding to the soluble lysosomal acid α-glucosidase is not affected, but the steroid does increase the Vmax. value. Membrane-bound enzyme is not activated by cortisone. 4. 4-[14C]Cortisone is preferentially bound to the lysosomal membrane and the possible involvement of this structure in the activation phenomenon is discussed.

scholarly journals A STUDY OF ARYL SULFATASE AND ACID PHOSPHATASE IN THE DEVELOPING NERVOUS SYSTEM OF THE RAT

1968 ◽  
Vol 16 (9) ◽  
pp. 582-589 ◽  
Author(s):  
A. E. CHOY ◽  
H. CRAVIOTO
Keyword(s):  
Nervous System ◽  
Cerebral Cortex ◽  
Acid Phosphatase ◽  
Enzyme Reaction ◽  
The Other ◽  
Reaction Products ◽  
Aryl Sulfatase ◽  
Essential Enzyme ◽  
Increased Activity ◽  
Developing Nervous System

Aryl sulfatases A and B and acid phosphatase activities were studied in the nervous system of rats aged 5, 10, 15, 20 and greater than 45 days. The enzyme reaction products appeared as granules localized in neurons of the cerebral cortex, brain stem, choroid plexus and ependyma and in blood vessels. The distribution and intensity of enzyme reaction product varied with the age of the animals. Aryl sulfatase B activity appeared earlier and reached a maximum of intensity sooner than the other enzymes. Acid phosphatase and aryl sulfatase A activities appeared at the same time. However, acid phosphatase had a greater intensity of activity than aryl sulfatase A. The increased activity of aryl sulfatase B was parallel with the maturation of neurons. It is suggested that aryl sulfatase B may be an essential enzyme for myelination.

Lead aspartate: a new en bloc stain for electron microscopy

1978 ◽  
Vol 36 (2) ◽  
pp. 34-35
Author(s):  
J.R. Walton
Keyword(s):  
Electron Microscopy ◽  
Calcium Ion ◽  
Enzyme Reaction ◽  
Lead Citrate ◽  
Reaction Products ◽  
En Bloc ◽  
Thin Sections ◽  
Lead Salts ◽  
Thin Sectioning ◽  
High Alkalinity

In electron microscopy, lead is the metal most widely used for enhancing specimen contrast. Lead citrate requires a pH of 12 to stain thin sections of epoxy-embedded material rapidly and intensively. However, this high alkalinity tends to leach out enzyme reaction products, making lead citrate unsuitable for many cytochemical studies. Substitution of the chelator aspartate for citrate allows staining to be carried out at pH 6 or 7 without apparent effect on cytochemical products. Moreover, due to the low, controlled level of free lead ions, contamination-free staining can be carried out en bloc, prior to dehydration and embedding. En bloc use of lead aspartate permits the grid-staining step to be bypassed, allowing samples to be examined immediately after thin-sectioning.Procedures. To prevent precipitation of lead salts, double- or glass-distilled H20 used in the stain and rinses should be boiled to drive off carbon dioxide and glassware should be carefully rinsed to remove any persisting traces of calcium ion.

The Demonstration of Human Prostatic Acid Phosphatase (Pap) With Phosphorylcholine

1976 ◽  
Vol 34 ◽  
pp. 88-89
Author(s):  
José A. Serrano ◽  
Hannah L. Wasserkrug ◽  
Anna A. Serrano ◽  
Arnold M. Seligman
Keyword(s):  
Acid Phosphatase ◽  
Prostate Gland ◽  
Prostatic Acid Phosphatase ◽  
Human Prostate ◽  
Rat Tissues ◽  
Specific Substrate ◽  
Acid Phosphatases ◽  
Lysosomal Acid ◽  
Cacodylate Buffer

As previously reported (1, 2) phosphorylcholine (PC) is a specific substrate for prostatatic acid phosphatase (PAP) as opposed to other acid phosphatases, e.g., lysosomal acid phosphatase. The specificity of PC for PAP is due to the pentavalent nitrogen in PC, a feature that renders PC resistant to hydrolysis by all other acid phosphatases. Detailed comparative cytochemical results in rat tissues are in press. This report deals with ultracytochemical results applying the method to normal and pathological human prostate gland.Fresh human prostate was obtained from 7 patients having transurethral resections or radical prostatectomies. The tissue was fixed in 3% glutaraldehyde- 0.1 M cacodylate buffer (pH 7.4) for 15 min, sectioned at 50 μm on a Sorvall TC-2 tissue sectioner, refixed for a total of 2 hr, and rinsed overnight in 0.1 M cacodylate buffer (pH 7.4)-7.5% sucrose.

Prostatic Acid Phosphatase (PAP) Differentiated Ultracytochemically from Lysosomal Acid Phosphate in the Rat with a PAP/Specific Substrate, Phosphorylcholine

1975 ◽  
Vol 33 ◽  
pp. 450-451
Author(s):  
W. Allen Shannon ◽  
José A. Serrano ◽  
Hannah L. Wasserkrug ◽  
Anna A. Serrano ◽  
Arnold M. Seligman
Keyword(s):  
Acid Phosphatase ◽  
Phosphate Buffer ◽  
Prostatic Carcinoma ◽  
Prostatic Acid Phosphatase ◽  
Chemotherapeutic Agents ◽  
Specific Substrate ◽  
Acid Phosphate ◽  
Lysosomal Acid ◽  
Design And Synthesis ◽  
New Chemotherapeutic Agents

During the design and synthesis of new chemotherapeutic agents for prostatic carcinoma based on phosphorylated agents which might be enzyme-activated to cytotoxicity, phosphorylcholine, [(CH3)3+NCH2CH2OPO3Ca]Cl-, has been indicated to be a very specific substrate for prostatic acid phosphatase (PAP). This phenomenon has led to the development of specific histochemical and ultracytochemical methods for PAP using modifications of the Gomori lead method for acid phosphatase. Comparative histochemical results in prostate and kidney of the rat have been published earlier with phosphorylcholine (PC) and β-glycerophosphate (βGP). We now report the ultracytochemical results.Minced tissues were fixed in 3% glutaraldehyde-0.1 M phosphate buffered (pH 7.4) for 1.5 hr and rinsed overnight in several changes of 0.05 M phosphate buffer (pH 7.0) containing 7.5% sucrose. Tissues were incubated 30 min to 2 hr in Gomori acid phosphatase medium (2) containing 0.1 M substrate, either PC or βGP.

High Degree of Homology Between Primary Structure of Human Lysosomal Acid Phosphatase and Human Prostatic Acid Phosphatase

1989 ◽  
Vol 370 (1) ◽  
pp. 177-182 ◽  
Author(s):  
Christoph PETERS ◽  
Carola GEIER ◽  
Regina POHLMANN ◽  
Abdul WAHEED ◽  
Kurt VON FIGURA ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Primary Structure ◽  
Prostatic Acid Phosphatase ◽  
Lysosomal Acid ◽  
High Degree

scholarly journals THE UNLABELED ANTIBODY ENZYME METHOD OF IMMUNOCYTOCHEMISTRY QUANTITATIVE COMPARISON OF SENSITIVITIES WITH AND WITHOUT PEROXIDASE-ANTIPEROXIDASE COMPLEX

1974 ◽  
Vol 22 (8) ◽  
pp. 782-801 ◽  
Author(s):  
JOHN P. PETRALI ◽  
DENNIS M. HINTON ◽  
GWEN C. MORIARTY ◽  
LUDWIG A. STERNBERGER
Keyword(s):  
Secretory Granules ◽  
Enzyme Reaction ◽  
Fold Increase ◽  
Immunocytochemical Staining ◽  
Intermediate Lobe ◽  
Reaction Products ◽  
Electron Microscopic ◽  
Specific Staining ◽  
Enzyme Method ◽  
Unlabeled Antibody

Araldite sections of rat pituitary intermediate lobe were used with anti-17-39 adrenocorticotropin in the unlabeled antibody enzyme method to compare electron microscopic immunocytochemical staining by peroxidase-antiperoxidase complex (PAP) with antiserum or purified antibody to peroxidase followed by peroxidase (PO). Quantitation of normalized optical densities of secretory granules offered high significance comparison (P < 0.0001) of experimental with control values and of experimental values with each other. Use of purified antibody (prepared by a new density gradient method) yielded four times higher sensitivity than antiserum to PO, while a 6.5-fold increase would have been expected from the degree of contamination of anti-PO in the serum by nonanti-PO immunoglobulin. Use of PAP was four to five times more sensitive than purified anti-PO and 20 times more sensitive than antiserum to PO. The increased sensitivity of PAP is explained by the high over-all binding affinity of PO for anti-PO in the cyclic PAP molecule, thus preventing the losses of PO that occur during washing when anti-PO and PO have been applied in sequence. Identification of the characteristic, cyclic PAP molecules affords confirmation of specific staining at high resolution. In the sequential application of anti-PO and PO, no PAP molecules are formed, thus making distinction of specific from nonspecific deposition of enzyme reaction products ambiguous. With the use of anti-17-39 ACTH and the intermediate lobe, the unlabeled antibody enzyme method was 16,000-100,000 times more sensitive than radioimmunoassay.

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