PHOSPHODIESTERASE AND ACID PHOSPHATASE ACTIVITIES OF AVIAN BONE MARROW CELLS

1965 ◽  
Vol 43 (8) ◽  
pp. 1319-1328 ◽  
Author(s):  
Gerald Kingsley Bristow ◽  
Esther W. Yamada
Keyword(s):  
Bone Marrow ◽  
Acid Phosphatase ◽  
Bone Marrow Cells ◽  
Intracellular Distribution ◽  
Specific Substrate ◽  
Magnesium Ions ◽  
Differential Centrifugation ◽  
Alkaline Ph ◽  
Ph Optimum ◽  
Rna And Dna

Avian bone marrow has been found to contain a phosphodiesterase as well as an acid phosphatase. Some properties of these enzymes have been described. Because the phosphodiesterase of this tissue has an alkaline pH optimum, is activated by magnesium ions, and acts on the specific substrate p-nitrophenyl thymidine 5′-phosphate, it is probably a phosphodiesterase I such as is present in snake venom and other tissues.The intracellular distribution of these two enzymes in normal and regenerating bone marrow was studied. Subcellular fractions were prepared by differential centrifugation or by centrifugation through sucrose density gradients. The RNA and DNA content of each fraction was determined. By the methods used no differences in the properties or intracellular distribution of the two enzymes in normal and regenerating bone marrow were found.

The Ubiquitin Ligase Triad1 Is Involved in Myelopoiesis and Interacts with the Transcriptional Repressors Gfi1 and Gfi1b.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 64-64 ◽  
Author(s):  
Bert A. Van der Reijden ◽  
Jurgen A.F. Marteijn ◽  
Liesbeth Van Emst ◽  
Theo De Witte ◽  
Joop H. Jansen
Keyword(s):  
Bone Marrow ◽  
Zinc Finger ◽  
Colony Formation ◽  
Bone Marrow Cells ◽  
26S Proteasome ◽  
Specific Substrate ◽  
Granulocytic Differentiation ◽  
Substrate Proteins ◽  
Zinc Finger Region ◽  
Marrow Cells

Abstract We identified Triad1 as a gene that is upregulated by retinoic acid during the granulocytic differentiation of acute promyelocytic leukemia cells. In normal hematopoiesis, we show that Triad1 is weakly expressed in immature CD34+ bone marrow cells, and highly expressed in mature monocytes and granulocytes. Together, this suggests that Triad1 plays a role in the differentiation of hematopoietic cells. Triad1 contains a tripartite domain including two RING fingers, indicating that this protein might function as a ubiquitin E3 ligase, catalyzing the the conjugation of ubiquitin to substrate proteins thereby marking them for targeted degradation by the 26S proteasome. Using GST pull down experiments, we show that Triad1 binds to the ubiquitin conjugating (E2) enzymes UbcH6 and 7. In addition, immunoprecipitation of Triad1 in cells that were transfected with FLAG-tagged ubiquitin shows that Triad1 binds to ubiquitinated proteins, and that Triad1 is capable of self-ubiquitination, further corroborating the assumption that Triad1 acts as a E3 ubiquitin ligating enzyme. To study the role of Triad1 in hematopoiesis we overexpressed the gene in primary murine bone marrow cells using a retroviral vector that contains Triad1 in front of an IRES-GFP cassette. GFP positive cells were FACS sorted and used in colony assays (CFU-GM). Compared to empty vector controls (GFP alone), Triad1 expression resulted in more than 80% inhibition of clonogenic growth. Importantly, addition of the proteasome inhibitor MG132 (10E-8 M) reversed the Triad1-induced suppression of colony formation. Furthermore, three Triad1 expression constructs in which one of the conserved cys/his residues of the TRIAD domain (essential for function) were mutated did not show the suppressive effect on colony formation. Together, these data show that Triad1 is involved in myelopoiesis and acts through the ubiquitination of specific substrate proteins. To identify these substrates, a yeast-two-hybrid screen of a human bone marrow cDNA library was performed using the Triad1 protein as a bait. Interestingly, the transcriptional repressor Gfi1b was found to bind to Triad1. The interaction was confirmed by immunoprecipitation using GFP-Triad1 and FLAG-tagged Gfi1b transfections in mammalian cells. We show that Triad1 binds to the zinc finger region of Gfi1b. This region is very (>98%) homologous to the paralogue Gfi1. Further immunoprecipitation analyses showed that Triad1 also binds to the zinc finger region of Gfi1. Gfi1 plays an essential role in neutrophil development and Gfi1 pointmutations result in neutropenia in man. Currently, we are studying the direct ubiquitination of Gfi and Gfi1b by Triad1 in in vitro ubiquitination assays. In addition, we are studying the effect of Triad1 on the transcriptional repression of the ELA2 and other promoters by Gfi1.

scholarly journals A HISTOCHEMICAL STUDY OF "ACID" AND "ALKALINE" PHOSPHATASE DISTRIBUTION IN NORMAL HUMAN BONE MARROW SMEARS

Blood ◽  
1949 ◽  
Vol 4 (5) ◽  
pp. 580-594 ◽  
Author(s):  
M. RABINOVITCH ◽  
D. ANDREUCCI
Keyword(s):  
Bone Marrow ◽  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Bone Marrow Cells ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Mitotic Chromosomes ◽  
Acid And Alkaline Phosphatase ◽  
Normal Human ◽  
Normal Human Bone Marrow

Abstract 1. Three minute fixation in formol vapor at 44 C, followed by 15 minute washing proved to be the most satisfactory fixation procedure for both "acid" and "alkaline" phosphatase technics as applied to bone marrow smears. 2. For both technics a relation between staining intensity and cellular richness was found. 3. The reaction of normal human bone marrow cells to both phosphatase technics is described. Both are predominantly nuclear in location. Nuclear pattern approached that observed with common staining methods and Feulgen’s reaction. Cytoplasmic reaction was nearly negative. Nonspecific and specific granules do not stain after the "alkaline" technic. Nonspecific granules are negative for "acid" phosphatase, while specific neutrophilic are variable, and eosinophilic, constantly positive. Nucleoli are negative after the "acid" technic, being positive for the "alkaline" enzyme. Mitotic chromosomes are positive for both technics. "Acid" phosphatase reaction in cytoplasmic zones of lymphocytes, erythroblasts, plasmacytes and megakaryocytes, is described.

scholarly journals Enzymatic provision of bone marrow cells after autotransplantation

2018 ◽  
Vol 7 (3) ◽  
pp. 9-12
Author(s):  
O. V. Vorob'eva ◽  
L. A. Lyubovtseva ◽  
N. E. Gimaldinova
Keyword(s):  
Bone Marrow ◽  
Acid Phosphatase ◽  
Mast Cells ◽  
Monoamine Oxidase ◽  
Bone Marrow Cells ◽  
Statistical Processing ◽  
Ether Anesthesia ◽  
Elevated Activity ◽  
Activity Of Enzymes ◽  
Marrow Cells

The purpose of the study is assessment of enzymes’ activity and localization in bone marrow cells after autotransplantation. Material and methods. Experiments were performed on 40 male mice that underwent bone marrow autotransplantation. Under ether anesthesia 1 ml of bone marrow was extracted from mice epiphyses, then were diluted in 2 ml of isotonic solution, after that 1 ml of obtained suspension was injected into the caudal vein of the same mice. Smears and imprints were produced and studied using Glenner’s histochemical method to identify monoamine oxidase, using Gomory’s method for acid phosphatase and for succinate reductase in bone marrow cells using H. Laborit method. Statistical processing of the material was carried out in the program «Statistica 6.0». Results. In 40 min after autotransplantation an increase in enzymatic activity in mast cells, granular, some hematopoietic bone marrow cells is noted, with significant increase observed in mast cells and granular cells. Two types of cells are identified: endocrine-like and one of macrophagal types. There is an increase in acid phosphatase in some hematopoietic cells with macrophagal properties. The activity of succinate dehydrogenase increases, indicating elevated activity of oxidative phosphorylation, resulting in increased production of ATP, the energy supply of bone marrow cells is carried out. Conclusion. It is revealed that in some bone marrow cells autotransplantation results in increased activity of enzymes: monoamine oxidase, succinate reductase and acid phosphatase.

Platinum as a tracer for intracellular distribution of a drug (CIS-PLATIN) : x-ray microanalysis

1992 ◽  
Vol 50 (2) ◽  
pp. 1590-1591
Author(s):  
R. Gary Kirk ◽  
Ping Lee
Keyword(s):  
Bone Marrow ◽  
Bone Marrow Cells ◽  
Chemotherapeutic Agent ◽  
Intracellular Distribution ◽  
Cross Link ◽  
Bladder Tumors ◽  
Cellular Resistance ◽  
X Ray ◽  
Kinase C ◽  
Insight Into

Cisplatin, cis-diamminedichloroplatinum (II) , contains a platinum atom in each of its molecule. It is an effective chemotherapeutic agent, used to treat ovarian, testicular and bladder tumors. The antitumor activity and cytoxity of this drug is generally attributed to its ability to cross-link DNA in the chromosome. Recent investigators show that there are also membrane and cytosolic components in the biological activity of cisplatin. Although the mechanism for the membrane transport of cisplatin is not known, it is thought that decrease in uptake of the drug is contributory to the cellular resistance to cisplatin. In addition cytosolic concentrations of glutathione, metallothionein or protein kinase C have been shown to affect the cytotoxicity of cisplatin. A frequent toxicity seen in patients that have received cisplatin treatment is the depression of the bone marrow cells. In order to gain some insight into this toxicity event, we have studied the uptake and intracellular distribution of cisplatin using the technique of cryomicrotomy and x-ray microanalysis.

scholarly journals Observations on the Phosphatase Content of Blood and Bone Marrow Cells in Normal and Pathologic Hemopoiesis

Blood ◽  
1951 ◽  
Vol 6 (5) ◽  
pp. 454-465 ◽  
Author(s):  
WILLIAM KERPPOLA
Keyword(s):  
Rheumatoid Arthritis ◽  
Bone Marrow ◽  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Polymorphonuclear Leukocytes ◽  
Bone Marrow Cells ◽  
Malignant Tumors ◽  
Normal Individuals ◽  
Active Regeneration ◽  
Alkaline And Acid Phosphatase

Abstract The alkaline and acid phosphatase content of blood and bone marrow has been studied by the methods of Gomori-Takamatsu and Gomori in normal individuals and in a great number of different diseases. The presence of a particular phosphatase is related to cellular reaction. Alkaline phosphatase was found in orthochromic normoblasts, polymorphonuclear leukocytes, metamyelocytes and older megakaryocytes. Acid phosphatase was demonstrated in basophilic normoblasts, polymorphonuclear leukocytes and their juvenile forms and in young megakaryocytes. In red blood cells and young megakaryocytes phosphatase was present chiefly in the nuclei, in leukocytes and older megakaryocytes in the cytoplasm. By studying erythropoiesis, especially during active regeneration of blood, it has been shown that alkaline phosphatase plays a decisive part in the disappearance of the normoblast nucleus. In certain diseases abnormality is observed in the disappearance of the normoblast nucleus (effort syndrome, rheumatoid arthritis). In others acid phosphatase occurs in large amounts in the youngest normoblasts (polycythemia vera, rheumatoid arthritis, nephritis). At the same time phosphatase may also be found in polymorphonuclear leukocytes and their juvenile forms. In rare cases of severe anemia alkaline phosphatase (panmyelophthisis) or acid phosphatase (achrestic anemia) is noted in large amounts in nucleated red cells and in leukocytes. Only alkaline phosphatase is present in polymorphonuclear leukocytes and metamyelocytes in leukemias, myeloma, Hodgkin’s and Paget’s disease, and in some malignant tumors of the skeleton and the liver. Several circumstances indicate that phosphatases might destroy cells or their nuclei. In some cases phosphatase activity is physiologic, in others phosphatase concentration is related to accelerated or malignant growth of bone marrow or tissue associated with blood formation. Finally there are cases in which there is as yet no explanation for the appearance of phosphatases.

scholarly journals A comparative study of new substrates for the histochemical demonstration of acid phosphomonoesterase activity in tissues which secrete acid phosphatase.

1980 ◽  
Vol 28 (4) ◽  
pp. 316-322 ◽  
Author(s):  
D F McDonald ◽  
B H Schofield ◽  
M A Geffert ◽  
R A Coleman
Keyword(s):  
Bone Marrow ◽  
Enzyme Activity ◽  
Acid Phosphatase ◽  
Sebaceous Gland ◽  
Bone Marrow Cells ◽  
Prostate Gland ◽  
Histochemical Demonstration ◽  
Rat Tissues ◽  
Hydrolysis Of ◽  
Acid Phosphomonoesterase

The histochemical demonstration of acid phosphatase activities against phosphoethanolamine (PEA), phosphorylcholine (PC), and D-ephedrine phosphate (DEP) are reported for a variety of rat tissues and are compared to acid beta-glycerophosphatase (beta GPase) activity. Intense acid beta GPase activity was demonstrated in all tissues examined. However, liver, kidney, intestine, spleen and bone marrow cells failed to exhibit any enzyme activity against PEA, PC, or DEP. In addition, significant differences in the hydrolysis of these substrates were noted among the tissues that did demonstrate activity (bone, tooth, oral mucosa, sebaceous gland, and prostate gland). These observations suggest that PEA, PC, and DEP are more specific substrates for acid phosphatase than beta GP and permit the differential localization of several distinct acid phosphatase isoenzymes.

scholarly journals Elastase-like enzymes in human neutrophils localized by ultrastructural cytochemistry.

1980 ◽  
Vol 84 (1) ◽  
pp. 102-119 ◽  
Author(s):  
J M Clark ◽  
D W Vaughan ◽  
B M Aiken ◽  
H M Kagan
Keyword(s):  
Bone Marrow ◽  
Endoplasmic Reticulum ◽  
Enzyme Activity ◽  
Nuclear Membrane ◽  
Active Site ◽  
Blood Cells ◽  
Human Neutrophils ◽  
Alkaline Ph ◽  
Ph Optimum ◽  
Ultrastructural Studies

The thiol ester N-t-Boc-L-alanine-p-nitrothiophenyl ester (Boc-Ala-SNp) was synthesized and applied as an ultrastructural cytochemical substrate for intracellular elastase-like enzymes. Mature human neutrophils incubated with Boc-Ala-SNp and gold ions generate an electron-dense reaction product, gold p-nitrothiophenolate, which is found in the nuclear membrane, Golgi complex, endoplasmic reticulum, mitochondria, and granules of these cells. Enzyme activity against Boc-Ala-SNp is also observed in developing monkey bone marrow neutrophils and in other blood cells. The intracellular neutrophil enzyme activity is elastase-like because it is characterized by a slightly alkaline pH optimum and is inactivated by exposure of the cells to general and specific active site inhibitors of neutrophil elastase. This substrate appears to have important potential for use in ultrastructural studies of intracellular elastase-like enzymes.

The structure of pre-mRNA and mRNA from erythroid cells

1978 ◽  
Vol 36 (2) ◽  
pp. 234-235
Author(s):  
A.-M. Ladhoff ◽  
B.J. Thiele ◽  
Ch. Coutelle ◽  
S. Rosenthal
Keyword(s):  
Bone Marrow ◽  
Structural Transformation ◽  
Electron Micrographs ◽  
Ammonium Acetate ◽  
Bone Marrow Cells ◽  
Erythroid Cells ◽  
Globin Mrna ◽  
Ordered Structures ◽  
Rabbit Bone ◽  
Rabbit Reticulocytes

The suggested precursor-product relationship between the nuclear pre-mRNA and the cytoplasmic mRNA has created increased interest also in the structure of these RNA species. Previously we have been published electron micrographs of individual pre-mRNA molecules from erythroid cells. An intersting observation was the appearance of a contour, probably corresponding to higher ordered structures, on one end of 10 % of the pre-mRNA molecules from erythroid rabbit bone marrow cells (Fig. 1A). A virtual similar contour was observed in molecules of 9S globin mRNA from rabbit reticulocytes (Fig. 1B). A structural transformation in a linear contour occurs if the RNA is heated for 10 min to 90°C in the presence of 80 % formamide. This structural transformation is reversible when the denatured RNA is precipitated and redissolved in 0.2 M ammonium acetate.

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.

Sign in / Sign up

Export Citation Format

Share Document