A New Fluorometric Method for the Estimation or Detection of Total and Fractionated Alkaline Phosphatase

1969 ◽  
Vol 15 (2) ◽  
pp. 108-123 ◽  
Author(s):  
Roy B Johnson
Keyword(s):  
Alkaline Phosphatase ◽  
Relative Activity ◽  
Normal Serum ◽  
Starch Gel Electrophoresis ◽  
Total Alkaline Phosphatase ◽  
Fluorometric Method ◽  
Highly Sensitive ◽  
Starch Gel ◽  
Alpha Beta ◽  
Total Alkaline

Abstract A highly sensitive fluorometric method for the assay of total alkaline phosphatase and the detection of its components is described. The commercially available substrate naphthyl AS-MX phosphate, combined in 1 M 2-amino-2-methyl-1-propanol buffer, pH 9.8, is cleaved to the highly fluorescent naphthol AS-MX. Fluorometry requires filters passing 405 mµ primary and 505 mµ secondary. As little as 20 µl. of normal serum per 3 ml. of reaction mixture can be assayed. Alkaline phosphatase components, separated by vertical starch gel (or other methods of) electrophoresis, produce yellow fluorescence under ultraviolet light when incubated at 37° with the same buffer-substrate. After vertical starch gel electrophoresis, all normal serums exhibit at least one component (β-globulin region), but six distinct areas of activity have been located. These correspond to Taswell and Jeffers’ origin, beta-lipoprotein, alpha-2, alpha-beta, and beta (5). Few serums contain all of these; rather there appears to be a correlation between the ones present, their relative activity, and the disease state.

scholarly journals Bone mineral density and serum levels of 25 OH vitamin D in chronic users of antiepileptic drugs

2004 ◽  
Vol 62 (4) ◽  
pp. 940-948 ◽  
Author(s):  
Carolina A.M. Kulak ◽  
Victória Z.C. Borba ◽  
John P. Bilezikian ◽  
Carlos E. Silvado ◽  
Luciano de Paola ◽  
...  
Keyword(s):  
Bone Mineral Density ◽  
Vitamin D ◽  
Alkaline Phosphatase ◽  
Lumbar Spine ◽  
Antiepileptic Drugs ◽  
Serum Levels ◽  
Mineral Density ◽  
Total Alkaline Phosphatase ◽  
Blood Samples ◽  
Total Alkaline

The aim of this cross sectional study was to evaluate bone mineral density (BMD) and serum levels of 25-hydroxy vitamin D (25OHD) in a group of patients taking antiepileptic drugs (AED) for a seizure disorder. Between May-2001 and January-2003, we evaluated 58 patients (40 women/18 men), 34.4±6 years old living in Curitiba or in its metropolitan area, on antiepileptic therapy for 2 to 38 years (10 on monotherapy /48 on multiple drugs regime). The group was matched by age, gender, and bone mass index to 29 healthy subjects (20 women/ 9 men); 34.2±5.9 years old. Medical history and physical exam were performed on all subjects with particular information sought about fractures and risks factors for osteoporosis. Blood samples were collected for total serum calcium, albumin, phosphorus, creatinine, total alkaline phosphatase, and liver function tests. BMD of the lumbar spine, femur and forearm was determined by dual energy X-ray absorptiometry (DXA, Hologic QDR 1000). Between February and April-2003, other blood samples were collected to measure 25OHD, intact paratohormone (PTH) and calcium. Unemployment and smoking history were more frequent among patients than among controls (p<0.05). Fifteen patients had a fracture history, all of which occurred during a seizure. The BMD of the lumbar spine (0.975±0. 13 g/cm² vs. 1.058±0.1 g/cm²; p<0.03) and of the total femur (0.930±0.1 g/cm² vs. 0.988±0.12 g/cm²; p<0.02) was lower in patients than in controls. In 63.5% of patients and in 24.1 % of controls a T-score < -1.0 in at least one site was seen. The AED users had higher total alkaline phosphatase and lower 25OHD (p<0.02). No correlations between BMD and 25OHD were found. The use of phenytoin was correlated with a greater incidence of fractures (RR: 2.38). We conclude that patients on chronic use of AED have alterations in bone metabolism characterized in this study by lower BMD of the lumbar spine and total femur and lower serum concentrations of 25OHD.

scholarly journals The distribution of some hydrolases in glomeruli and tubular fragments prepared from rat kidney by zonal centrifugation

1971 ◽  
Vol 122 (5) ◽  
pp. 641-645 ◽  
Author(s):  
D. G. Taylor ◽  
R. G. Price ◽  
D. Robinson
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Gel Electrophoresis ◽  
Leucine Aminopeptidase ◽  
Rat Kidney ◽  
Kidney Cortex ◽  
Starch Gel Electrophoresis ◽  
Multiple Forms ◽  
Rat Kidney Cortex ◽  
Starch Gel

1. A collagenase digest of rat kidney cortex was separated into four bands by zonal centrifugation. 2. Two of these bands were shown by light-microscopy to contain glomeruli and tubular fragments, which were free from each other and well separated from other renal material. 3. Protein, N-acetyl-β-glucosaminidase, 5′-nucleotidase, l-leucine β-naphthylamidase, leucine aminopeptidase, acid phosphatase and alkaline phosphatase were assayed across the gradient. 4. The greater proportion of these enzyme activities was recovered in the tubular fragments and acid phosphatase was the only enzyme detected in significant amounts in the glomeruli. 5. Tubular fragments and glomeruli were sedimented and multiple forms of β-naphthylamidase, N-acetyl-β-glucosaminidase, acid phosphatase and alkaline phosphatase were investigated by starch-gel electrophoresis.

scholarly journals Release of alkaline phosphatase from membranes by a phosphatidylinositol-specific phospholipase C.

1977 ◽  
Vol 167 (1) ◽  
pp. 281-284 ◽  
Author(s):  
M G Low ◽  
J B Finean
Keyword(s):  
Staphylococcus Aureus ◽  
Alkaline Phosphatase ◽  
Phospholipase C ◽  
Mammalian Species ◽  
Salt Solutions ◽  
Total Alkaline Phosphatase ◽  
Total Alkaline Phosphatase Activity ◽  
Wide Range ◽  
Releasing Effect ◽  
Total Alkaline

Purified phosphatidylinositol-specific phospholipase C from Staphylococcus aureus released a substantial proportion of the total alkaline phosphatase activity from a wide range of tissues from several mammalian species. Co-purification of the phospholipase C and alkaline phosphatase-releasing activities and the inhibition of both these activities by iso-osmotic salt solutions suggested that the releasing effect was unlikely to be due to a contaminant.

scholarly journals Race, Mineral Homeostasis and Mortality in Patients with End-Stage Renal Disease on Dialysis

2015 ◽  
Vol 42 (1) ◽  
pp. 25-34 ◽  
Author(s):  
Julia J. Scialla ◽  
Rulan S. Parekh ◽  
Joseph A. Eustace ◽  
Brad C. Astor ◽  
Laura Plantinga ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
African American ◽  
African Americans ◽  
Serum Calcium ◽  
Mortality Risk ◽  
Fibroblast Growth Factor 23 ◽  
Total Alkaline Phosphatase ◽  
Mineral Homeostasis ◽  
Calcium Phosphorus ◽  
Total Alkaline

Background: Abnormalities in mineral homeostasis are ubiquitous in patients on dialysis, and influenced by race. In this study, we determine the race-specific relationship between mineral parameters and mortality in patients initiating hemodialysis. Methods: We measured the levels of fibroblast growth factor 23 (FGF23) and 25-hydroxyvitamin D (25 D) in 184 African American and 327 non-African American hemodialysis patients who enrolled between 1995 and 1998 in the Choices for Healthy Outcomes in Caring for ESRD Study. Serum calcium, phosphorus, parathyroid hormone (PTH) and total alkaline phosphatase levels were averaged from clinical measurements during the first 4.5 months of dialysis. We evaluated the associated prospective risk of mortality using multivariable Cox proportional hazards models stratified by race. Results: PTH and total alkaline phosphatase levels were higher, whereas calcium, phosphorus, FGF23 and 25 D levels were lower in African Americans compared to those of non-African Americans. Higher serum phosphorus and FGF23 levels were associated with greater mortality risk overall; however, phosphorus was only associated with risk among African Americans (HR 5.38, 95% CI 2.14-13.55 for quartile 4 vs. 1), but not among non-African Americans (p-interaction = 0.04). FGF23 was associated with mortality in both groups, but more strongly in African Americans (HR 3.91, 95% CI 1.74-8.82 for quartiles 4 vs. 1; p-interaction = 0.09). Serum calcium, PTH, and 25 D levels were not consistently associated with mortality. The lowest and highest quartiles of total alkaline phosphatase were associated with higher mortality risk, but this did not differ by race (p-interaction = 0.97). Conclusions: Aberrant phosphorus homeostasis, reflected by higher phosphorus and FGF23, may be a risk factor for mortality in patients initiating hemodialysis, particularly among African Americans.

scholarly journals Activity of Total Alkaline Phosphatase in Water of the Barlinek Lake of the during Stagnation Time of 2008

2013 ◽  
Vol 2 ◽  
pp. 42-45
Author(s):  
Piotr Daniszewski
Keyword(s):  
Alkaline Phosphatase ◽  
Phosphatase Activity ◽  
Total Alkaline Phosphatase ◽  
Activity Maximum ◽  
Seasonal Oscillations ◽  
Total Alkaline

To estimate differentiation of activity of total alkaline phosphatase in water of the Barlinek Lake, of the during stagnation time of 2008. Studies upon water revealed that, among the zones analyzed, upper sublittoral layer was characterized by the highest phosphatase activity. From the analysis of seasonal oscillations it follows that activity maximum for alkaline phosphatase, both in water, occurred July 2008.

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