scholarly journals In vitro modulation of alkaline phosphatase activity in neutrophils from patients with chronic myelogenous leukemia by monocyte-derived activity

Blood ◽  
1986 ◽  
Vol 67 (2) ◽  
pp. 492-497 ◽  
Author(s):  
T Matsuo
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Chronic Myelogenous Leukemia ◽  
Alkaline Phosphatase Activity ◽  
Culture Medium ◽  
Underlying Mechanism ◽  
Myelogenous Leukemia ◽  
Neutrophil Alkaline Phosphatase

Abstract To clarify the underlying mechanism of low neutrophil alkaline phosphatase (NAP) activity in chronic myelogenous leukemia (CML), CML neutrophils were cultured in liquid medium with different numbers of monocytes. Alkaline phosphatase activity in CML neutrophils, assessed cytochemically, increased with the numbers of monocytes. NAP activity was not induced by the interaction between neutrophils and monocytes, but by the presence of a monocyte-derived soluble activity. NAP activity in normal neutrophils was also lowered by depletion of monocytes from culture medium. Under such monocyte-depleted conditions, both CML and normal neutrophils proliferated and differentiated to produce mature neutrophils. Thus induction of NAP activity can be modified in vitro by changing the amount of NAP-inducing activity released from monocytes. However, whether a reduction of NAP-inducing activity in CML neutrophil is the cause of low NAP activity in vivo remains uncertain.

scholarly journals In vitro modulation of alkaline phosphatase activity in neutrophils from patients with chronic myelogenous leukemia by monocyte-derived activity

Blood ◽  
1986 ◽  
Vol 67 (2) ◽  
pp. 492-497
Author(s):  
T Matsuo
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Chronic Myelogenous Leukemia ◽  
Alkaline Phosphatase Activity ◽  
Culture Medium ◽  
Underlying Mechanism ◽  
Myelogenous Leukemia ◽  
Neutrophil Alkaline Phosphatase

To clarify the underlying mechanism of low neutrophil alkaline phosphatase (NAP) activity in chronic myelogenous leukemia (CML), CML neutrophils were cultured in liquid medium with different numbers of monocytes. Alkaline phosphatase activity in CML neutrophils, assessed cytochemically, increased with the numbers of monocytes. NAP activity was not induced by the interaction between neutrophils and monocytes, but by the presence of a monocyte-derived soluble activity. NAP activity in normal neutrophils was also lowered by depletion of monocytes from culture medium. Under such monocyte-depleted conditions, both CML and normal neutrophils proliferated and differentiated to produce mature neutrophils. Thus induction of NAP activity can be modified in vitro by changing the amount of NAP-inducing activity released from monocytes. However, whether a reduction of NAP-inducing activity in CML neutrophil is the cause of low NAP activity in vivo remains uncertain.

scholarly journals Neutrophil alkaline phosphatase: comparison of enzymes from normal subjects and patients with polycythemia vera and chronic myelogenous leukemia

Blood ◽  
1975 ◽  
Vol 45 (3) ◽  
pp. 335-343
Author(s):  
D Rosenblum ◽  
SJ Petzold
Keyword(s):  
Alkaline Phosphatase ◽  
Polycythemia Vera ◽  
Phosphatase Activity ◽  
Chronic Myelogenous Leukemia ◽  
Alkaline Phosphatase Activity ◽  
Normal Subjects ◽  
Myelogenous Leukemia ◽  
Precipitin Line ◽  
Neutrophil Alkaline Phosphatase ◽  
Sepharose 4B

To determine whether decreased alkaline phosphatase activity in the granules from neutrophils of patients with chronic myelogenous leukemia (CML) was due to an absence of enzyme or the production of defective enzyme, we compared the immunologic properties of granule alkaline phosphatase derived from patients with CML with that of normal subjects and patients with polycythemia vera (PRV). Antisera prepared in rabbits against granule alkaline phosphatase purified from the neutrophils of a patient with PRV produced a single precipitin line of antigenic identity when reacted with extracts of normal, PRV, and CML neutrophil granules. A histochemical stain for alkaline phosphatase activity (alpha-naphthyl acid phosphate coupled with Fast Blue RR) specifically stained the precipitin line. A variety of quantitative precipitin techniques failed to produce satisfactory precipitation of alkaline phosphatase activity. Comparative analyses were therefore performed by affinity chromatography using goat antirabbit-gammaglobulin linked to Sepharose 4B to adsorb alkaline phosphatase complexed with rabbit gamma globulin. With this method, 100% of CML, normal, and PRV alkaline phosphatase could be adsorbed. Using limiting concentrations of antibody, a proportionally smaller fraction of enzyme activity was absorbed as the concentration of PRV alkaline phosphatase or normal alkaline phosphatase was increased. Extracts of CML granules containing comparable amounts of protein but 200-fold less alkaline phosphatase activity per milligram did not specifically reduce adsorption. Thus, in CML, we found no evidence that the granulocytes contained a large amount of antigenically normal but enzymatically defective alkaline phosphatase. Examination of electron micrographs revealed no significant differences in the number or distribution of granules in the granulocytes of normal subjects or patients with PRV or CML. This suggests that the low level of neutrophil alkaline phosphatase in CML granulocytes is the result of decreased enzyme content and not a consequence of synthesis of catalytically defective enzyme.

scholarly journals Neutrophil alkaline phosphatase: comparison of enzymes from normal subjects and patients with polycythemia vera and chronic myelogenous leukemia

Blood ◽  
1975 ◽  
Vol 45 (3) ◽  
pp. 335-343 ◽  
Author(s):  
D Rosenblum ◽  
SJ Petzold
Keyword(s):  
Alkaline Phosphatase ◽  
Polycythemia Vera ◽  
Phosphatase Activity ◽  
Chronic Myelogenous Leukemia ◽  
Alkaline Phosphatase Activity ◽  
Normal Subjects ◽  
Myelogenous Leukemia ◽  
Precipitin Line ◽  
Neutrophil Alkaline Phosphatase ◽  
Sepharose 4B

Abstract To determine whether decreased alkaline phosphatase activity in the granules from neutrophils of patients with chronic myelogenous leukemia (CML) was due to an absence of enzyme or the production of defective enzyme, we compared the immunologic properties of granule alkaline phosphatase derived from patients with CML with that of normal subjects and patients with polycythemia vera (PRV). Antisera prepared in rabbits against granule alkaline phosphatase purified from the neutrophils of a patient with PRV produced a single precipitin line of antigenic identity when reacted with extracts of normal, PRV, and CML neutrophil granules. A histochemical stain for alkaline phosphatase activity (alpha-naphthyl acid phosphate coupled with Fast Blue RR) specifically stained the precipitin line. A variety of quantitative precipitin techniques failed to produce satisfactory precipitation of alkaline phosphatase activity. Comparative analyses were therefore performed by affinity chromatography using goat antirabbit-gammaglobulin linked to Sepharose 4B to adsorb alkaline phosphatase complexed with rabbit gamma globulin. With this method, 100% of CML, normal, and PRV alkaline phosphatase could be adsorbed. Using limiting concentrations of antibody, a proportionally smaller fraction of enzyme activity was absorbed as the concentration of PRV alkaline phosphatase or normal alkaline phosphatase was increased. Extracts of CML granules containing comparable amounts of protein but 200-fold less alkaline phosphatase activity per milligram did not specifically reduce adsorption. Thus, in CML, we found no evidence that the granulocytes contained a large amount of antigenically normal but enzymatically defective alkaline phosphatase. Examination of electron micrographs revealed no significant differences in the number or distribution of granules in the granulocytes of normal subjects or patients with PRV or CML. This suggests that the low level of neutrophil alkaline phosphatase in CML granulocytes is the result of decreased enzyme content and not a consequence of synthesis of catalytically defective enzyme.

scholarly journals A biodegradable porous composite scaffold of PCL/BCP containing Ang-(1-7) for bone tissue engineering

Cerâmica ◽  
2012 ◽  
Vol 58 (348) ◽  
pp. 481-488 ◽  
Author(s):  
F. A. Macedo ◽  
E. H. M. Nunes ◽  
W. L. Vasconcelos ◽  
R. A. Santos ◽  
R. D. Sinisterra ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Composite Scaffold ◽  
Weight Ratio ◽  
High Porosity ◽  
X Rays ◽  
Solvent Casting

Highly porous three-dimensional biodegradable scaffolds was obtained from beta-tricalcium phosphate-hydroxyapatite bioceramic (BCP), PCL, and Angiotensin-(1-7). We used the solvent casting and particulate leaching methods (SC/PL). The processed scaffolds were characterized by X-ray microtomography (µ-CT). Biocompatibility tests in vitro were performed during three and seven days using MTT and Alkaline Phosphatase Activity (APA) assays. Both the MTT activity and APA were evaluated using a one-way ANOVA test. The µ-CT results showed that the increase of the PCL:BCP weight ratio leads to structures with lower pore sizes. The pore interconnectivity of the processed scaffolds was evaluated in terms of the fragmentation index (FI). We observed that the obtained composites present poorly connected structures, with close values of FI. However, as the polymer phase is almost transparent to the X-rays, it was not taken into consideration in the µ-CT tests. The MTT activity assay revealed that scaffolds obtained with and without Angiotensin-(1-7) present mild and moderate cytotoxic effects, respectively. The APA assay showed that the rat osteoblasts, when in contact for three days with the PCL composites, presented an APA similar to that observed for the control cells. Nevertheless, for an incubation time of seven days we observed a remarkable decrease in the alkaline phosphatase activity. In conclusion, using the solvent casting and salt leaching method we obtained 3D porous that are composites of PCL, BC and Ang-(1-7), which have suitable shapes for the bone defects, a high porosity and interconnect pores. Furthermore, the viability in vitro showed that the scaffolds have potential for drug delivery system and could be used in future in vivo tests.

scholarly journals Simultaneous autoradiographic and histochemical analysis of bone formed in vitro.

1986 ◽  
Vol 34 (6) ◽  
pp. 769-773 ◽  
Author(s):  
H C Tenenbaum ◽  
C A McCulloch ◽  
K Palangio
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Time Course ◽  
Bone Cell ◽  
Histochemical Demonstration ◽  
Thymidine Uptake ◽  
Kinetics In Vivo

The simultaneous histochemical demonstration of alkaline phosphatase activity and autoradiographic demonstration of [3H]-thymidine uptake is valuable for study of bone cell kinetics in vivo or in vitro. By use of this technique, it has been possible to detect changes induced by a single dose of dexamethasone (10(-7) M) in the time course of alkaline phosphatase activity, the number of alkaline phosphatase-positive cells, and [3H]-thymidine labeling in bone formed in vitro.

Experimental Thrombosis in Rabbits: Evidence that Alkaline Phosphatase Enhances the Thrombogenic Effect of the Injected Ellagic Acid

1966 ◽  
Vol 16 (03/04) ◽  
pp. 645-656 ◽  
Author(s):  
P. G Iatridis ◽  
J. H Ferguson
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Ellagic Acid ◽  
Plasma Samples ◽  
Electrophoretic Study ◽  
Negative Results ◽  
Thrombotic Tendency

SummaryIntravenous injection of ellagic acid plus alkaline phosphatase in rabbits, induces a thrombotic tendency, which correlates with a hypercoagulability of the blood revealed by thrombelastography. Either ellagic acid (which suboptimally activates Hageman factor) or alkaline phosphatase, separately, gave negative results concerning the incidence of thrombosis. This clearly indicates that alkaline phosphatase in vivo, as was previously shown in vitro, acts only in the presence of an active SF. The electrophoretic study of the distribution of alkaline phosphatase, in platelet-poor plasma samples secured from rabbits, with and without injection of an alkaline phosphatase preparation, suggests the probability that it is the alkaline phosphatase activity which is located in the “beta” fraction that contributes the responsible co-factor for the thrombogenesis.

Osteogenic protein-1 enhances phenotypic expression in ROS 17/2.8 cells

1995 ◽  
Vol 269 (5) ◽  
pp. E918-E926 ◽  
Author(s):  
A. M. Kitten ◽  
J. C. Lee ◽  
M. S. Olson
Keyword(s):  
Alkaline Phosphatase ◽  
Parathyroid Hormone ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Thymidine Incorporation ◽  
Phenotypic Expression ◽  
Proliferating Cells ◽  
Osteogenic Protein

Osteogenic protein-1 (OP-1) stimulates bone morphogenesis in vivo and modulates osteoblast growth and differentiation in vitro. Treatment of ROS 17/2.8 cells with OP-1 resulted in a time- and concentration-dependent inhibition of [3H]thymidine incorporation. In contrast, OP-1 treatment stimulated phenotypic differentiation in ROS 17/2.8 cells, as indicated by enhanced 1) alkaline phosphatase activity (4-fold); 2) alkaline phosphatase mRNA (5-fold); 3) parathyroid hormone receptor mRNA (2-fold), and 4) parathyroid hormone-stimulated adenosine 3',5'-cyclic monophosphate accumulation (2-fold). OP-1-induced changes in cell growth and gene expression were sensitive to cycloheximide and actinomycin D. Measurement of [3H]thymidine incorporation and alkaline phosphatase activity in situ revealed heterogeneity in the cellular responses to OP-1. Proliferating cells exhibited less alkaline phosphatase activity than nonproliferating cells, whereas cells expressing high levels of alkaline phosphatase incorporated little [3H]thymidine. Our data delineating the responses of mature differentiated osteoblasts to OP-1 suggest that potentiation of osteoblast differentiated function is an important component of bone morphogenesis in vivo.

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