Alkaline Phosphatase Isoenzymes In Plasma Research Articles

Novel methodology for the determination of alkaline phosphatase isoenzymes in plasma circulating

dp er os+1.5μg/kg/w sc. Eight rats were sham operated. Histomorphometric and histopathological studies were performed on tibias obtained after 30 days of treatment. OVX decreased trabecular bone (%BV/TV), and Z prevented and raised it (Sham: 35.3±4.7, control: 21.5±1.5*, MFP: 17.1±1.0*, Z: 64.3±3.5*, MFP±Z: 52.2±3.2*, * p<0.05 vs sham, mean±SE, ANOVA). Histological examination showed, in the sham group, an adequate bone-hemopoietic (HT) tissue relationship and mature bone trabeculae (Tb). The control group showed higher HT, immature Tb, fewer osteoblasts (Ob) and higher osteoclasts (Oc) compared with the sham group. The MFP group showed abundant HT and thin Tb with abundant lining cells (LC) and Ob. In Z, anastomosed and immature Tb with invasive tendency on the marrow space were observed. A decrease in Ob and HT was also observed. In contrast, the MFP+Z group showed mature Tb with well defined marrow spaces. The trabecular edge was covered by LC and Ob. We conclude that Z treatment, alone or combined with MFP, prevented the bone loss caused by OVX. Its combination with MFP normalized the histological features of bone. The MFP, by stimulating osteoblast activity, could modulate bone remodeling by producing a better quality bone. This article is part of a Special Issue entitled AAOMM 2010 Abstracts.

Alkaline phosphatase isoenzymes in mouse plasma detected by polyacrylamide-gel disk electrophoresis

Plasma alkaline phosphatase (ALP) activity is frequently measured in toxicity studies. In the present study, we assessed the usefulness of a commercially available polyacrylamide-gel (PAG) disk electrophoresis kit used in humans (AlkPhor System, Jokoh Co., Ltd., Tokyo, Japan) for identifying plasma ALP isoenzymes in mice of the Crlj:CD1 strain (ICR mice), which are commonly used in toxicity studies. We also examined age-related changes in plasma ALP isoenzymes in ICR mice. Electrophoresis was performed according to the manufacturer's instructions. In order to identify the origin of each ALP isoenzyme, in addition to plasma samples, tissue ALP extracts from the liver, bone and small intestine were treated with neuraminidase, anti-small intestinal ALP antibody, ALP inhibitor levamisole and/or wheat germ agglutinin (WGA). The kit revealed that main plasma ALP isoenzyme in intact ICR mice was bone-derived one, and it tended to decrease with age. On the other hand, liver-derived ALP isoenzyme greatly increased in plasma of cholestasis model mice induced by bile duct ligation. This model mouse had also a large molecular ALP detected in the stacking gel. This ALP was thought to be of intestinal origin because its activity remained even after levamisole inhibition. In addition, a minimum sample volume for sufficient resolution of plasma ALP isoenzymes was only 14µl. The results of this study suggest that the present method is a useful tool for detecting plasma ALP isoenzymes in mice and that pre-treatment of plasma with neuraminidase and concomitant levamisole inhibition with another gel is applicable for the evaluation of organ toxicity.

Alkaline phosphatase isoenzymes in plasma of chagasic and healthy pregnant women.

Catedra de Quimica Biologica, Facultad de CienciasMedicas **Catedra de Estadistica y Biometria,Facultad de Ciencias Exactas, Fisicas y Naturales,Universidad Nacional de Cordoba, Pabellon Argentina2° Piso, Ciudad Universitaria, Cordoba, Argentina*Servicio Nacional de Chagas, Cordoba, ArgentinaKey words: alkaline phosphatase - pregnancy -Chagas disease

A simple electrophoretic method for separating elevated liver and bone alkaline phosphatase isoenzymes in plasma after neuraminidase treatment

A simple electrophoretic method for separating elevated liver and bone alkaline phosphatase isoenzymes in plasma after neuraminidase treatment

Alkaline phosphatase isoenzymes in plasma in hyperthyroidism.

Alkaline phosphatase (ALP; EC 3.1.3.1) isoenzymes were measured in the plasma of 63 untreated hyperthyroid patients (the hyperthyroid group), 58 treated hyperthyroid patients, and 100 blood donors. Total, liver, and bone ALP activities were significantly higher in the hyperthyroid group than in the treated hyperthyroid group or the blood donors. Bone ALP was more frequently and more markedly abnormal than liver ALP. Intestinal ALP did not differ significantly between the groups. The hyperthyroid patient group had significantly higher plasma calcium concentrations and lower serum parathyrin concentrations than those of the treated hyperthyroid group.

Properties of alkaline phosphatase isoenzymes in plasma of preterm and term neonates.

The isoenzymes of alkaline phosphatase (EC 3.1.3.1) in plasma of 37 preterm and 21 term neonates two weeks postpartum have been studied with regard to electrophoretic mobility, sialic acid content, inhibition properties, heat lability, molecular mass, and binding to lectins. Term infants generally had a single form of alkaline phosphatase present in significant amounts, identified by the above criteria as originating in bone. All the preterm neonates had two alkaline phosphatase isoenzymes in plasma, namely, bone- and fetal-type intestinal alkaline phosphatase. Liver-, placental-, and adult-type intestinal alkaline phosphatase were not detected in any of the plasma samples.

Incubation with neuraminidase and affinity electrophoresis with wheat-germ lectin compared for separating and quantifying alkaline phosphatase isoenzymes in plasma.

Of 98 patients' specimens examined for alkaline phosphatase (EC 3.1.3.1) isoenzymes by electrophoresis on cellulose acetate membrane after incubation with neuraminidase, 50 showed only a single liver or bone isoenzyme staining band; in 15 of these, the tissue origin of the fraction could not be accurately identified from its electrophoretic location. In the remaining 48 specimens, both liver and bone fractions were identifiable, but in only 25 of these was the electrophoretic resolution sufficient to yield separate peaks on densitometry. In contrast, both liver and bone alkaline phosphatase isoenzymes were identified in 95 of the 98 specimens by affinity electrophoresis involving wheat-germ lectin, the detection of both fractions being in agreement with the results of sequential heat inactivation. The tissue origin of the enzyme bands was readily ascertainable from their consistent electrophoretic location in this medium, and in 89 of the specimens the isoenzyme fractions could be resolved into separate peaks on densitometry. We conclude that resolution of liver and bone alkaline phosphatase by incubation with neuraminidase followed by cellulose acetate electrophoresis is greatly inferior to that obtained by wheat-germ lectin affinity electrophoresis.

Two new methods for separating and quantifying bone and liver alkaline phosphatase isoenzymes in plasma.

We describe two new methods for the separation and quantification of the bone and liver isoenzymes of alkaline phosphatase (EC 3.1.3.1) in plasma. In the first, we use wheat-germ lectin to precipitate the bone isoenzyme. About 80% of this, but minimal liver isoenzyme, is precipitated. The activity of the bone isoenzyme is calculated from measuring the alkaline phosphatase activity in the precipitate, that of liver alkaline phosphatase by subtracting the activity of the bone isoenzyme from total alkaline phosphatase activity. The liver fraction will also contain biliary, intestinal, and placental alkaline phosphatase if these are present in the original plasma, but correction for such activity is readily made. In the second method, samples are separated on cellulose acetate membranes that, before electrophoresis, have been soaked in buffer containing wheat-germ lectin. The bone isoenzyme is retarded and clearly separated from the liver fraction, allowing these isoenzymes to be quantified by densitometry. Both methods are rapid, reproducible, and suitable for use in the diagnostic laboratory.

Isoenzymes of canine plasma alkaline phosphatase: an investigation using isoelectric focusing and related to diagnosis

The isoenzymes of canine alkaline phosphatase have been separated by isoelectric focusing on agarose gels. Alkaline phosphatase from normal tissue extracts gave heterogeneous focusing patterns with the following isoelectric points: liver pH 4.3, bone pH 4.0 to 4.9, kidney pH 4.4 to 4.7 and intestine pH 3.6 to 4.6. Plasma alkaline phosphatase isoenzymes were examined in 123 dogs with activities at least twice the normal maximum. The isoenzymes from these plasma samples had multiple bands at pH 4.3 to 4.6 but in 67 per cent of all cases, the predominant isoenzyme, named isoenzyme A, was a single band of isoelectric point pH 3.5. Isoenzyme A occurred in all dogs with diabetes mellitus or Cushing's syndrome and in 93 per cent of animals receiving steroid therapy. Increased activities of the pH 4.3 to 4.6 isoenzymes were more likely to occur in cases of hepatic degeneration.

Infectious stunting and leg weakness em broilers ii. Studies on alkaline phosphatase isoenzymes in blood plasma1

Total alkaline phosphatase (ALP) activity in blood plasma of broilers suffering from infectious stunting after artificial infection was constantly higher than that of uninoculated control birds. While total ALP activity in blood plasma from inoculated broilers of various ages decreased after incubation at 56 degrees C, it increased in similarly treated plasma collected from broilers at 3, 21 and 28 days after inoculation. Investigations into the organ origin of the plasma ALP isoenzymes with agarose electrophoresis, column chromatography, L phenylalanine inhibition and heat treatment showed that the main part of ALP activity in blood plasma of both, inoculated and uninoculated birds was most likely of intestinal origin. In 3-day-old broilers no heat sensitive part of total ALP could be measured.

Plasma alkaline phosphatase isoenzymes in hepatobiliary disease.

BY CELLULOSE ACETATE OR ACRYLAMIDE GEL ELECTROPHORESIS IT IS POSSIBLE TO SEPARATE THESE ALKALINE PHOSPHATASE ISOENZYMES FROM SERUM: [anode] fast liver, slow liver, placenta/Regan, bone, intestine, bile [cathode]. Heat or chemical inhibition can confirm the differentiation. Normal adult serum always contains slow-liver isoenzyme, and sometimes bone isoenzyme: the latter is always present in serum of children. In hepatobiliary disease slow-liver isoenzyme was always increased: intestinal isoenzyme appeared in many cases of cirrhosis (of blood groups B and 0) but fast-liver and bile isoenzymes were occasionally seen in miscellaneous cases. The findings in other diseases included Regan isoenzyme in six out of 45 cases of malignant disease.