Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The studies summarized in this report were intended to determine whether salmon calcitonin had direct effects on bone formation indices in vitro. The results of these investigations demonstrate acute effects of calcitonin on skeletal tissues derived from embryonic chickens to increase calvarial cell proliferation ([3H]thymidine incorporation into DNA) and bone matrix synthesis ([3H]proline incorporation into collagen, as [3H]hydroxyproline) in intact calvaria and tibiae. The effects of calcitonin on [3H]thymidine incorporation were significant at 1 mU/ml (0.08 nM; P less than 0.05), additive with respect to the action(s) of F (calcitonin increased the maximum effect of F, and F increased the effect of low dose calcitonin; P less than 0.01 for each), associated with an increase in total cell protein (r = 0.82; P less than 0.02), and inversely dependent on osteoblastic differentiation (r = -0.96; P less than 0.005). The effects of calcitonin to increase bone matrix synthesis ([3H]hydroxyproline incorporation, 139% and 155% of untreated control values for tibiae and calvaria, respectively; P less than 0.005 for each) were maximal at approximately 5 mU/ml (0.4 nM) and associated with a proportional increase in alkaline phosphatase activity in the bones (r = 0.71; P less than 0.05 for tibiae). These effects of calcitonin were not dependent on continuous exposure. [3H]Thymidine incorporation was increased in calvarial cells 16 h after a 4-h limited (inductive) exposure to calcitonin (at 3 mU/ml; P less than 0.01). [3H]Proline incorporation in embryonic chicken calvaria was also increased during 3 days of limited exposure (i.e. 4 h/day) to 10 mU/ml calcitonin (P less than 0.02). The proliferative action(s) of calcitonin was not unique to chicken osteoblastline cells. Salmon calcitonin also increased [3H]thymidine incorporation in the transformed murine calvarial cell lines MMB and MC-3T3-E1 and in primary cultures of cells prepared from newborn mouse calvaria (P less than 0.05 for each). Furthermore, these effects were observed at calcitonin doses (3-30 mU/ml) that also decreased murine bone resorption (i.e. 45Ca release from prelabeled neonatal mouse calvaria; P less than 0.01).
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PMID:The anti-bone-resorptive agent calcitonin also acts in vitro to directly increase bone formation and bone cell proliferation. 338 71

Amino-terminal amino acid sequences (42 residues) were determined for the products of the three common alleles at the human placental alkaline phosphatase [orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] gene locus. The sequences differ at position 3, which is proline in types 1 and 2 but is leucine in type 3. cDNA libraries were constructed in phage lambda gt11 and used to isolate clones covering the coding regions of types 1 and 3 cDNAs. Comparison of the deduced amino acid sequences of the types 1 and 3 proteins showed 7 differences out of 513 amino acids, each due to a single base substitution. cDNA sequence comparisons showed three silent substitutions in the coding regions and three base differences in the greater than 1 kilobase pairs of 3' untranslated sequences.
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PMID:Products of two common alleles at the locus for human placental alkaline phosphatase differ by seven amino acids. 346 52

Effects of H2O2 on bone were evaluated in an organ culture system. Tibiae from chick embryos were incubated for up to 3 days in culture medium containing 0.07 to 20 mM H2O2. Glucose metabolism was monitored by measuring lactate production and oxygen consumption, and collagen synthesis was determined by hydroxylation of proline. In addition to markedly inhibiting these parameters, H2O2 also decreased bone weight and alkaline phosphatase activity. Multiple exposures to H2O2 were somewhat more effective than a single exposure. Since H2O2 inhibits bone at low concentrations in vitro, the results suggest that the potential for harmful effects of H2O2 in the oral cavity should be investigated.
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PMID:Hydrogen peroxide inhibits glucose metabolism and collagen synthesis in bone. 347 27

The purpose of this study was to correlate the histologic and biochemical responses of the interparietal suture to a range of tensile forces. Stainless steel spring implants, calibrated to generate expansive forces from 50 to 250 g, were placed across the interparietal suture in 85 female Sprague-Dawley rats. After experimental periods from 2 hours to 14 days, the interparietal sutures were evaluated by radiography, histology, and biochemistry. An in vivo/in vitro system was used for the biochemical analysis; total protein, proline incorporated, percent collagen, and alkaline phosphatase activity were measured. The radiographs and histology showed that in vivo suture expansion was achievable with 50 to 70 g of force, but the heavier forces showed greater sutural opening, more cellular proliferation, and more bone formation. This increased biologic response by the heavier forces was substantiated by an increase in sutural protein and alkaline phosphatase activity but not in percent collagen. It was concluded that changes in the total protein content of the suture were not primarily caused by proliferation of osteogenic cells and fibroblasts but due to an influx of transudate. In contrast, the increase in incorporation of 3H-proline and alkaline phosphatase activity correlated with the observance of bone formation. This study indicated a positive correlation between the magnitude of tensile forces and osteogenic response.
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PMID:The morphologic and biochemical effects of tensile force application to the interparietal suture of the Sprague-Dawley rat. 347 67

1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3], but not 24,25-(OH)2D3 stimulates the alkaline phosphatase activity of cultured human bone cell populations. The stimulatory effect of the sterol was dose dependent (10(-10)-10(-7) M), evident by 24 h, and observed over a range of cell densities. Analysis of the radiolabeled collagens synthesised by human bone cell cultures indicated the synthesis of predominantly type I collagen. In the presence of 1,25-(OH)2D3, but not 24,25-(OH)2D3, there was a dose-dependent (10(-11)-10(-9) M) increase in radiolabeled proline incorporation into collagenase-digestible protein and in the amount of collagen synthesized, expressed as a percentage of the total protein synthesis. The effect of 1,25-(OH)2D3 was observed over a range of cell densities and appeared to be specific for the synthesis of type I collagen. The stimulatory effect of 1,25-(OH)2D3 on alkaline phosphatase activity and the increase in proline incorporation into collagenase-digestible protein were accompanied by a dose-dependent (5 X 10(-11) to 5 X 10(-8) M) inhibition of bone cell proliferation. These findings suggest that 1,25-(OH)2D3 is an important modulator of the growth and differentiation of human bone cells in vitro. They are also consistent with the possibility that 1,25-(OH)2D3 has direct effects on bone formation in vivo.
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PMID:1,25-Dihydroxyvitamin D3 and human bone-derived cells in vitro: effects on alkaline phosphatase, type I collagen and proliferation. 348 8

Rats were subjected to a two-stage subtotal nephrectomy or sham operation, and treated with aluminum (Al) or both aluminum and vitamin D3 metabolites for 5 weeks with a cumulative dose of 13.6 mg aluminum. Animals were injected with 3H-thymidine and 3H-proline. The following analyses were performed: quantitative histology of tibial metaphyses and cytomorphometric electron microscopy of osteoclasts, quantitative (ICP-spectroscopy) and qualitative determination (histochemical staining) of aluminum within organs, and serum biochemistry (Ca, P, Mg, vitamin D3 metabolites, alkaline phosphatase, urea). The following new facts of the aluminum-related bone disease became evident: (a) Application of aluminum to growing uremic rats induced rickets, whose major epiphyseal growth plate changes were 1 alpha,25(OH)2D3-dependent. Addition of 1 alpha,25(OH)2D3 prevented the formation of rachitic metaphysis, but failed to prevent osteoid accumulation on epiphyseal and metaphyseal trabecular surfaces. Moreover, calcitriol produced hyperosteoidosis and osteosclerosis in the same rats. Aluminum did not alter the function of osteoblasts, while osteoclasts seemed inactivated. (b) The development of rickets was associated with suppressed serum levels of 1,25(OH)2D3, reduced phosphorus level and the high content of aluminum in the bone, kidney, and liver. The addition of 24R,25(OH)2D3 markedly exaggerated the reduction of serum levels of calcitriol. We suggested that aluminum induces rickets in growing uremic rats, which consists of two components: vitamin D refractory osteomalacia and 1 alpha,25(OH)2D3-dependent epiphyseal growth plate changes.
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PMID:Effects of 1 alpha,25- and 24R,25-dihydroxyvitamin D3 on aluminum-induced rickets in growing uremic rats. 350 83

Brush-border microvillous plasma membrane vesicles were prepared from human full-term placental syncytiotrophoblasts and purified 33-fold from the homogenate with reference to a membrane marker enzyme, alkaline phosphatase (EC 3.1.3.1). Transport of alpha-(methylamino)isobutyrate by the membrane vesicles was stimulated in the presence of an Na+ gradient from the outside to the inside of the vesicles. The initial rate of uptake in a 10-s period was enhanced with increasing concentration of Na+ in the external medium. The level of alpha-(methylamino)isobutyrate transported into the vesicles reached a maximum 1 min after the start of incubation at 37 degrees C, and then decreased with time due to efflux. Extrapolation to infinite medium osmolarity showed no uptake, indicating transport of alpha-(methylamino)isobutyrate into membrane vesicles. The initial rate of uptake was dependent on temperature and pH: the highest rate occurred at 37 degrees C and the optimal pH was 8.0. When the alpha-(methylamino)isobutyrate concentration was varied, the initial rate of uptake dependent on an Na+ gradient (out greater than in) obeyed Michaelis-Menten kinetics with Km and Vmax values of 1.07 mM and 3.23 nmol/10 s per mg of protein, respectively. Cross-inhibition patterns indicated that at least three Na+-dependent and two Na+-independent carrier-mediated pathways existed in the human placental brush border. One Na+-dependent pathway interacted with all substrates tested. Another Na+-dependent route interacted with L-proline, alpha-(methylamino)isobutyrate, and L-methionine, while a third pathway was selective for L-methionine. One Na+-independent pathway was selective for L-cysteine, while the other pathway interacted with all substrates tested.
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PMID:Characterization of amino acid transport systems in human placental brush-border membrane vesicles. 366 75

Tumor necrosis factor (TNF) was studied for its effects on bone formation in cultured rat calvariae. TNF alpha at 100-100,000 U/ml stimulated [3H]thymidine incorporation into DNA, an effect that appeared after 24 h of treatment and lasted 96 h. Transient (24-h) treatment with TNF alpha increased [3H]proline incorporation into type I collagen 24-72 h after the factor was removed; this effect was DNA synthesis dependent and blocked by hydroxyurea. Transient treatment with TNF alpha also increased alkaline phosphatase activity. In contrast, continuous treatment with TNF alpha for 48-96 h caused a marked inhibition on [3H]proline incorporation into type I collagen and alkaline phosphatase activity. TNF alpha caused a small increase in collagen degradation. Lymphotoxin had similar effects to those of TNF alpha. In conclusion, TNF alpha stimulates calvarial DNA synthesis which causes an increased number of collagen-synthesizing cells, but TNF alpha has a direct inhibitory effect on osteoblastic function.
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PMID:Effects of tumor necrosis factor on bone formation in vitro. 366 33

1,25-dihydroxyvitamin D3 [1,25(OH)2D3] is essential for normal growth and mineralization, but its direct effects on various aspects of bone formation remain controversial. 1,25(OH)2D3 was studied for its effects on DNA, collagen and noncollagen protein synthesis, and alkaline phosphatase activity (APA) in the periosteum and periosteum-free bone from 21-day fetal rat calvariae. 1,25(OH)2D3 (0.01 to 10 nM) inhibited the incorporation of 3H-proline into collagenase-digestible protein (CDP) and the percent of collagen synthesized, and, at 10 nM, APA in the periosteum-free bone. 1,25(OH)2D3 inhibited type I collagen without affecting other collagen types. In contrast, 1,25(OH)2D3 at 10 nM caused a small but significant stimulation of the incorporation of 3H-thymidine into acid-insoluble residues (DNA) and on DNA content; both effects were exclusively observed in the periosteum. Hydroxyurea did not modify the inhibitory effect of 1,25(OH)2D3 on 3H-proline incorporation into CDP. These studies indicate that 1,25(OH)2D3 stimulates periosteal DNA synthesis but inhibits type I collagen synthesis and APA in the periosteum-free bone.
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PMID:1,25-Dihydroxyvitamin D3 effects on collagen and DNA synthesis in periosteum and periosteum-free calvaria. 384 46

The effect of varying the amino acid concentrations of the culture medium on matrix vesicle formation was studied in primary cultures of chicken epiphyseal growth plate chondrocytes grown in Dulbecco's modified Eagle's medium (DME) supplemented with 10% fetal bovine serum (FBS). Decreasing the levels of free amino acids in the culture medium to levels of one-half, one quarter, and one eighth of the values normally present in DME caused a progressive decline in matrix vesicle (MV) formation. Increasing the level in the culture medium of those amino acids that are enriched in extracellular fluid (ECF) of growth plate cartilage significantly increased formation of matrix vesicles (MV), as assayed by the alkaline phosphatase (AP) activities present in high-speed sediments from spent culture media. However, adjusting the levels of all amino acids to match those of the ECF produced the greatest stimulation of MV formation. Of the amino acids that are notably enriched in ECF, glutamate (GLU), alanine (ALA), serine (SER), asparagine (ASN), and taurine (TAU) individually enhanced MV production, whereas proline (PRO), glycine (GLY), and aspartate (ASP) had essentially no effect. The simple combination of ECF levels of ALA and GLU resulted in a stimulation of MV formation equal to that observed when the eight aforementioned amino acids were elevated to ECF levels. Other combinations of ASP and GLY, or of TAU, SER, and ASN showed some stimulation, but at a lower level. Increasing the amino acid concentrations, alone or in combination, also increased the levels of cellular AP, and to a lesser extent cellular protein. While increases in cellular AP were generally correlated with increased formation of AP-rich MV, this was not uniformly true. These results indicate that in addition to hormones and growth factors, nutritional factors such as the levels of amino acids are also critical for normal phenotypic expression, growth, and matrix formation by epiphyseal chondrocytes.
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PMID:Effect of amino acid levels on matrix vesicle formation by epiphyseal growth plate chondrocytes in primary culture. 394 89


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