Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Growth-plate cartilage is organized into four cellular zones containing resting, proliferating, maturing, and hypertrophic cells. Rabbit chondrocytes were isolated from growth-plate costal cartilage of 4-week-old New Zealand rabbits, the cells (15 x 10(4)) were suspended in 1 ml of Iscove's modified Dulbecco's medium (IMDM) with 10% fetal bovine serum, 50 micrograms ascorbic acid, and 60 micrograms kanamycin (medium A), then transferred to a 15 ml of plastic centrifuge tube, and centrifuged at 1500 rpm for 5 min. The cell pellet was incubated at 37 degrees C under 5% CO2 in air. The cultures reorganized into growth plate-like tissue which could be seen 7-14 days after cell seeding. This growth-plate, histologically, was organized longitudinally into cellular columns and horizontally into four cellular zones containing resting, proliferating, maturing and hypertrophic cells. The hypertrophic cells in the upper were large in size and round or oval in shape, the proliferating and the mature chondrocytes in the lower were small in size and spherical or elongated in shape. These chondrocytes were surrounded by an extensive matrix. Biochemically, DNA content of cultures began to rise on the 2nd day after cell seeding and reached a plateau after 10 days later. The uronic acid content increased from day 4 and reached the maximum on day 15. In contrast in the early culture, alkaline phosphatase activity was extremely low, it began to rise on day 9 and was the highest on day 20. The sequential increase of DNA, uronic acid and alkaline phosphatase contents was analogous to the in vivo changes of growth-plate chondrocytes.
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PMID:[Reorganization of growth-plate-like tissue by isolated chondrocytes in culture]. 797 58

The effect of beta-alanyl-L-histidinato zinc (AHZ) on bone cell function was investigated in osteoblastic MC3T3-E1 cells. Cells were cultured for 3 days at 37 degrees C in a CO2 incubator in plastic dishes containing alpha-modified minimum essential medium supplemented with 10% fetal bovine serum. After the cultures, the medium was exchanged for that containing 0.1% bovine serum albumin plus AHZ (10(-7)-10(-5) M) or other reagents, and the cells were cultured further for appropriate periods of time. The presence of AHZ (10(-7)-10(-5) M) produced a remarkable increase of alkaline phosphatase activity and protein concentration in osteoblastic cells. Thus increase were seen with the prolonged cultivation (12-21 days). With the culture of 1, 3 and 12 days, the effect of AHZ (10(-6) M) to increase alkaline phosphatase activity and protein concentration was more intensive than the effect of zinc sulfate (10(-6) M). The AHZ effects were completely abolished by the presence of cycloheximide (10(-6) M), indicating that AHZ stimulates protein synthesis in the cells. The present study suggests that AHZ has a stimulatory effect on cell differentiation, and that this effect is partly involved on protein synthesis in osteoblastic cells.
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PMID:Effect of beta-alanyl-L-histidinato zinc on differentiation of osteoblastic MC3T3-E1 cells: increases in alkaline phosphatase activity and protein concentration. 804 61

Abrupt changes in oxygen availability within the periodontium have been suggested to have a regulatory role in alveolar bone remodeling during tooth movement; arguably, similar to that seen in bone growth or fracture healing. The purpose of this investigation was therefore to study the effects of ambient hypoxia and hyperoxia on osteoblast function in vitro. Osteoblast-enriched cultures from fetal rat calvariae were exposed to atmospheres of hyperoxia (90% O2) and hypoxia (10% O2) and assayed for media pH, pO2, pCO2, cellular proliferation, alkaline phosphatase (AP) activity, and collagen synthesis. Results of this study show that in low ambient oxygen tension cellular proliferation increases, whereas the AP activity, collagen synthesis, media pO2, PCO2 decreases. In contrast, in hyperoxic conditions cellular proliferation is suppressed with concomitant increases in: AP activity, collagen synthesis, and partial pressures for oxygen and carbon dioxide. Media pH remained unaffected. In crossover experiments, where cells were initially grown in hypoxic conditions and were switched to hyperoxic conditions, their metabolic activities were abruptly reversed. These findings in conjunction with earlier reports, suggest a triggering role for oxygen tension (an environmental factor) in bone remodeling.
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PMID:Oxygen tension regulates osteoblast function. 816 95

Periodontal ligament (PDL) cells, osteoblasts (OB), and gingival (GIN) cells originating from human periodontium were co-cultured indirectly with human peripheral blood lymphocytes (PBL). The formation of osteoclasts (OC) from each the co-cultured PBL was compared with a standard PBL culture. A marked suppression of OC formation was observed in PBL co-cultured with PDL cells, and an enhanced OC formation was observed in PBL co-cultured with OB and GIN cells, when compared with the standard PBL culture. The suppressing activity of PDL cells and the enhancing activity of OB and GIN cells on the formation of OC derived from PBL were also found, when the co-culture fluids of PDL/PBL, OB/PBL, and GIN/PBL were added to PBL, and the numbers of OC were counted after 7 days' incubation. Furthermore, the alkaline phosphatase (ALPase) activity of PDL cells was stimulated by co-culturing them with PBL, and the ALPase activity of OB and GIN cells was inhibited by co-culturing them with PBL. When PDL cells were seeded on the surfaces of titanium discs and incubated at 37 degrees C in a 5% CO2 incubator, PDL cells could adhere faster onto titanium surfaces that were coated with a cell-and-tissue-adhesive substance than onto non-coated titanium surfaces. These cultures formed a confluent monolayer on the surfaces of titanium discs by means of an autologous serum containing alpha MEM. These results clearly suggest that the periodontal ligament is a specifically differentiated tissue whose function is to protect alveolar bone from bone resorption due to biting force.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:The artificial restoration of a dental root. 824 90

The effect of a 2 hour exposure to adriamycin (1 mg/litre) on alkaline phosphatase (ALPase) activity of the golden hamster 4-5 day old second maxillary molars (M2) was investigated in vitro. The molars were grown in BGJb medium containing 15% fetal bovine serum, glutamine (200 micrograms/ml), vitamin C (250 micrograms/ml), penicillin G (50 micrograms/ml), and streptomycin sulphate (30 micrograms/ml). The gas phase contained 50% O2 + 5% CO2 + 45% N2. The molars were supported on cellulosic membrane filters and grown for 3, 5, and 7 days at the medium-gas interface in a closed humidified chamber. Biochemical analysis indicated a steady increase in ALPase activity throughout this study in the control samples. However, after adriamycin treatment no increase in ALPase activity could be observed. The histochemical data showed that the increased activity in the control was confined to the peripheral pulp, sub-odontoblastic layer, stratum intermedium, ameloblasts and odontoblasts. Although these layers showed a decreased activity after adriamycin treatment, the ameloblasts showed an increase in activity over the control. The data has shown that adriamycin caused a reduction in total ALPase activity in developing molars in vitro; osteodentin production by pulp cells; and appeared to produce an acceleration in the differentiation of ameloblasts.
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PMID:Adriamycin alters the alkaline phosphatase activity in hamster molars during development in vitro. 832 61

The effect of biomechanical force on growth of skeletal tissue was studied in monolayer cultures of mouse osteoblastic MC3T3-E1 cells which were centrifuged at 320 g for 15 min to 72 h in a CO2 incubator. Centrifugation of the cells for 30 min in low concentrations (0.3 or 1%) of fetal bovine serum (FBS) caused a two-fold increase of [3H]thymidine incorporation at 20 h from the start of centrifugation. However, centrifugation under 10% FBS caused no increase in [3H]thymidine incorporation into DNA. Under 0.3% FBS, [3H]thymidine incorporation increased in a manner dependent on the period of centrifugation and reached a maximum when the cells were centrifuged for 3 h. Stimulation of DNA synthesis by centrifugation was abolished in the presence of H-7, an inhibitor of protein kinase C. Moreover, conditioned medium collected from the centrifuged cultures increased [3H]thymidine incorporation by two-fold over the basal when added to a quiescent culture of MC3T3-E1 cells. These results suggest that centrifugal force stimulates growth of osteoblastic cells through autocrine secretion of some diffusible growth-promoting activity. On the other hand, centrifugation of the cells inhibited induction by FBS of alkaline phosphatase activity and calcium-uptake, two indices of the differentiated phenotype of osteoblasts.
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PMID:Effect of centrifugal force on growth of mouse osteoblastic MC3T3-E1 cells in vitro. 836 Mar 84

Rats were tested in the forced swim test to evaluate the effects of duration of exposure (0, 5, 15, or 25 min), and water temperature (0, 35, 30, 25, or 20 degrees C), on a variety of physiological measures. Serum corticosterone, glucose, lactate, phosphorus levels, and the anion gap (a measure of acid-base status) were increased significantly, whereas carbon dioxide and potassium levels were consistently decreased by testing, as was the potassium/phosphorus ratio; creatinine, triglycerides, and magnesium were not altered significantly in any study. Effects on prolactin, amylase, lipase, cholesterol, alkaline phosphatase, sodium, and chloride were inconsistent. Levels of serum corticosterone were increased at each duration of testing, and the increments were significantly higher than the previous duration. Corticosterone levels were also increased in proportion to decreasing water temperature, but the increments were not significantly different from the previous or following temperatures. Glucose levels were increased at every duration and at every water temperature with the exception of the coldest water temperature. Lactate and phosphorus levels and the anion gap were all increased, whereas carbon dioxide levels decreased after 5 min of immersion. Potassium levels did not decrease until some time after 5 min of testing. Immobility times were marginally correlated with corticosterone levels (r = -0.38) but were highly correlated with serum carbon dioxide (r = 0.59), potassium (r = 0.67), and phosphorus levels (r = -0.73); the correlation between immobility times and the ratio of potassium/phosphorus was 0.82. The potassium/phosphorus ratio accounted for 67% of the variance in immobility. These high correlations were interpreted in terms of acid-base changes associated with testing.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Physiological correlates of the forced swim test in rats. 837 26

After direct photoaffinity cross-linking of [3H]GTP to the beta-subunit of tubulin, followed by tryptic digestion and alkaline phosphatase treatment, we employed cis-diol-specific boronate gel chromatography and reversed-phase high-pressure liquid chromatography to purify a peptide containing most of the covalently bound radioactivity. The sequence of this peptide corresponded to that of residues 3-19 of beta-tubulin. Residue 10 of the peptide, which is Cys-12 in beta-tubulin, could not be identified. The fast atom bombardment mass spectrum of this peptide showed the presence of a predominant species with a molecular mass of 2022 kDa (2021 kDa for the 12C variant), which is 255 Da greater than the molecular mass of the peptide. Fast atom bombardment collision-activated decomposition mass spectrometry analysis produced fragments which are consistent with the beta(3-19) peptide but having a unit of mass of 358 at position 12. Thermolysin digestion of the tryptic peptide restricted the cross-linking site to the 9-amino acid sequence, I(L)QAGQXGNQ. The molecular mass of this peptide was 1174 kDa, which is equal to the mass of the beta(7-15) peptide containing an extra group of mass 255. To explain the molecular masses of the two labeled peptides, which are 26 atomic mass units less than expected, a mechanism of photolabeling is proposed that involves opening of the guanine ring and loss of the C-6 carbonyl function as CO2.
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PMID:Exchangeable GTP binding site of beta-tubulin. Identification of cysteine 12 as the major site of cross-linking by direct photoaffinity labeling. 841 20

Milk replacer formulas based on cow's milk and egg yolks are frequently recommended for use in neonatal puppies. These formulas are lower in protein, kilocalories, calcium, and phosphorus than bitch's milk. In addition, the cholesterol content is greater than bitch's milk. The effect of feeding these formulas on serum chemistry profiles, lipid profiles, and alkaline phosphatase isoenzyme profiles of 5-week-old beagle puppies was studied. Three groups of beagle puppies were fed bitch's milk (control) (n = 18), a homemade milk-egg-oil formula (Formula 1) (n = 18), or a homemade milk-egg-oil formula supplemented with additional calcium and phosphorous (Formula 2) (n = 18). Concentrations of serum urea nitrogen, albumin, and total CO2 were lower (P < 0.05), and concentrations of serum phosphorus, globulins, sodium, chloride, and cholesterol were higher (P < 0.05) in formula-fed puppies than bitch-fed puppies. Serum potassium concentration was lower in the puppies fed Formula 1 than in the control puppies (P < 0.05), and serum potassium concentration in the puppies fed Formula 2 was not significantly different from that in puppies fed Formula 1 or the control puppies. Total triglyceride (TG) and high density lipoprotein2 cholesterol (HDL2) concentrations were similar in all three groups of puppies but the combined high density lipoprotein1 (HDL1) plus low density lipoprotein (LDL) cholesterol fraction was higher (P < 0.05) in the formula-fed puppies and accounted for the majority of the increase in cholesterol. There were no differences (P < 0.05) in total serum alkaline phosphatase (ALP) or bone-derived ALP (BALP) concentrations among the groups, however there was a higher (P < 0.05) serum concentration of liver-derived ALP (LALP) in the Formula 1-fed puppies. Feeding homemade egg and cow's milk-based puppy replacement formulas is not recommended for long term use.
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PMID:Serum chemistry and lipid profiles in neonatal beagle puppies fed homemade milk replacer formulas. 846 96

Metabolic acidosis has been shown to alter vitamin D metabolism. There is also evidence that calcium may modulate 1,25(OH)2D3 by a parathyroid hormone (PTH)-independent mechanism. To investigate the effect of rapid correction of chronic metabolic acidosis on serum 1,25(OH)2D3 levels by free calcium clamp in chronic renal failure, 20 patients with mild to moderate metabolic acidosis (mean pH 7.31 +/- 0.04) and secondary hyperparathyroidism (mean intact PTH 156.47 +/- 84.20 ng/l) were enrolled in this study. None had yet received any dialysis therapy. Metabolic acidosis was corrected by continuous bicarbonate infusion for 3-4 h until plasma pH was around 7.4, while plasma ionized calcium was held at the preinfusion level by calcium solution infusion during the entire procedure. The plasma pH, bicarbonate, total CO2, sodium, and serum total calcium levels were significantly increased while serum concentrations of alkaline phosphatase and albumin were significantly decreased after bicarbonate infusion. The plasma ionized calcium, potassium, serum magnesium, inorganic phosphorus, and 25(OH)D levels showed no significant change before and after bicarbonate infusion. The serum 1,25(OH)2D3 levels were significantly increased (38.66 +/- 11.77 vs. 47.04 +/- 16.56 pmol/l, p < 0.05) after correction of metabolic acidosis. These results demonstrate that rapid correction of metabolic acidosis raises serum 1,25(OH)2D3 levels in vitamin D-deficient chronic renal failure patients, and may underline the importance of maintaining normal acid-base homeostasis in the presence of secondary hyperparathyroidism in chronic renal failure.
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PMID:Influence of metabolic acidosis on serum 1,25(OH)2D3 levels in chronic renal failure. 859 83


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