Introduction: This study was performed to establish a complementary method to the diagnosis of complete hydatidiform mole and its differentiation from the other cases of gestational trophoblastic disease by measuring the total (ALP) and placental ALKALINE PHOSPHATASE (PLAP) specific activity. Materials and Methods: Serum and tissue extracts from 13 patients, 13 normal non-pregnant and 30 pregnant females were compared for these enzymes activities. Paranitrophenyl phosphate was used as substrate. Placental ALKALINE PHOSPHATASE was simply distingushed from its isoenzymes by its heat stability (65°c for 15min).Results: According to our findings from serum and tissue extract examination the activity of total ALKALINE PHOSPHATASE and placental ALKALINE PHOSPHATASE activities were reduced in hydatidiform mole patients in comparsion with pregnant females (P<0.05) but there was no significant difference in non - pregnant subject. Conclusion: These results show a severe hypophosphatasia in serum and tissue extracts of the patients. The reduction in activity of enzymes may be caused by low synthesis of enzyme by trophoblast cells or some post transcriptional changes that causse the reduction of enzyme activity. Since the ALP is probably important in active transport of suger and phosphate across the trophoblast cell membreanes, so the hydropic villi of hydatidiform mole placenta may due to reduction of enzyme activity in placental villi. Our results suggest that measurment of ALP and PLAP activity in serum and tissue extract can be a complementary method in diagnosis of hydatidiform mole.

Introduction: Production of Monoclonal antibody against ALKALINE PHOSPHATASE for application in immunohistochemical and immunocytochemical techniques such as ALKALINE PHOSPHATASE anti-ALKALINE PHOSPHATASE (APAAP) method.   Material and Methods: In this investigation female Balb/c mice were immunized by several injections of ALKALINE PHOSPHATASE and the antibody titer in their sera were measured after each injection. The spleen lymphocytes of immunized mice and Sp2/0 myeloma cells were fused using 50% polyethylen glycol as fusing agent and hybridoma cells were selected by HAT medium. Identification and selection of anti-ALKALINE PHOSPHATASE producing clones were done by performing ELISA test on supernatants of all the resulting clones. Limiting dilution was performed twice on antiboby producing clones for their seperation and the resulted subclones were propagated. Since APAAP complex must be enzymatically active for using in immunohistochemical techniques the adhesion of Ab molecule to enzyme molecule must not affect the enzyme activity. For investigation of this effect, an ELISA technique was planned and the supernatants of selected hybridoma clones were studied by this method. For production of concentrated Ab the hybridoma cells were injected to peritoneal cavity of mice and the ascetic fluids were obtained. Finally the antibodies isotypes were determined.   Results: After 6 fusion experiments 104 hybridoma clones were abtained and two clones (A1G9 and A1G8) which were antibody producing and had the highest absorbance in ELISA test were selected. Using the limiting dilution method finally two monoclonal subclones A1G8F7 and A1G9G3 were selected. ELISA experiments showed that antibodies which were produced by selected hybridoma clones did not react with active site of the enzyme and did not interfer with enzymatic activity. Electrophoresis of ascetic fluids of hybridoma injected mice showed a prominent band in γ position. Isotype determination of monoclonal antibodies showed that both hybridoma clones produce antibody from IgG class with k light chain.   Conclusion: Because monoclonal antibodies which are produced by the obtained hybridoma cell lines are from IgG class and do not affect the enzyme activity, it seem's that they are suitable for APAAP complex formation. Other steps of this study are now being performed until APAAP complex formation and it's application in immunohistochemistry.  

ALKALINE PHOSPHATASE (ALP; EC 3.1.3.1) is mainly derived from the liver, bones and in lesser amounts from intestines, placenta, kidneys and leukocytes. A raised level of ALKALINE PHOSPHATASE in the blood frequently indicates a variety of diseases. The examination of the ALP isoenzyme can be performed by electrophoresis. This examination can be helpful in disease classification of those cases with hyperALKALINEPHOSPHATASEmia. Of several ALP isoenzymes, biliary ALP isoenzyme is mentioned for its clinical usefulness in detection of biliary obstruction. Here, the authors performed a study to investigate the clinical usefulness of biliary ALKALINE PHOSPHATASE isoenzyme in biliary obstruction. Of interest, there is no significant difference of serum ALP level between malignant and benign biliary obstruction group. But there is a significant difference of serum biliary - ALP isoenzyme between malignant (range 28 U/L – 365 U/l) and benign biliary obstruction (range 20 – 140 U/L). Nevertheless, the average biliary - ALP level of the cholangiocarcinoma cases (range 105 – 365 U/L) is significant higher than the other malignant biliary obstruction cases (range 28 – 50 U/L). According to our study, the biliary - ALP isoenzyme determination can be use as a marker for malignant biliary obstruction.

ALKALINE PHOSPHATASE, an orthophosphoric monoester phosphohydrolase Ec 3.1. 3.1., is a glycoprotein dimer. It is active in ALKALINE PH and needs mg2+ ion as an activator. This membrane-bound enzyme is widely distributed in nature. In this study, it was purified from human placenta in seven steps. Placenta was homogenated and the enzyme was extracted through following steps: Butanol extraction, filtration on sephadex G200, ammonium sulfate precipitation, aceton precipation and DEAE cellulose chromatography with a gradient of sodium chloride. A peak of the enzyme activity at sodium chloride concentrations of 0.087 to 0.093 mg was observed on DEAE cellulose chromatography. The specific activity of the enzyme and purification factor were observed to be 209.9 U/mg and 611.8 respectively. At the final steps of preparative electrophoresis, the purified enzyme was. subjected to polyacrylamide disc gel electrophoresis. P-nitrophenyl phosphate was used, instead of N-naphthyl phosphate, to stain ALKALINE PHOSPHATASE. The appearance of a yellaw band proved the purification of the enzyme.

Background and Purpose: ALKALINE PHOSPHATASE (EC: 3.1.3.1) is synthesized by kidney, liver, bone, Intestine and placenta. This enzyme is a glycoprotein and dimmer 4 Zn+2 and Mn+2 in each dimmer. It hydrolyzes mono ester phosphate to organic compound and phosphate_in ALKALINE medium. The purpose of this research is to compare this enzyme with placental ALKALINE PHOSPHATASE.Methods and Materials: Human Molehydatiform was purified by folds of precipitation of bybutanol, acetone, Amoniumm sulphate, Sephadex G200, affinity chromatography and preparative electrophoresis.Results: Human Molehydatiform was purified 611.8 times. We obtained specific activity, optimum temperature and optimum Ph equaling to 611.8 U/mg, 40 centigrade degrees and 10.4 respectively.Conclusion: Purified Human Molehydatiform Alkalie PHOSPHATASE is different from Human placental ALKALINE PHOSPHATASE due to optimum pH and optimum temperature.

introduction: Crystal is one the pernicious addictive drugs which has unfortunately been imported to Iran in the recent years. As Crystal is the purified form of Heroin, its adverse effects are much more serious than those of Heroin. Apart from psychological effects, addiction to Crystal may have adverse effects on organs and tissues. So, the evaluation of enzymatic changes could be helpful in identifying the origin of damages. ALKALINE PHOSPHATASE is present in most tissues and the most prevalent cause of its elevation is liver and bone diseases. Since liver damages may be caused by Heroin consumption, assaying ALP levels could be useful for investigating the adverse effects of this drug.Materials and Methods: This study is a case control study which was carried out on 50 control persons and 108 crystal addicts who referred to Salehabad Center of Drug Addiction Treatment (Torbat jam) for the first time during 2008-2010. The activity level of ALKALINE PHOSPHATASE in the serum was measured with Calorimetry-Spectrophotometry Using computer software SPSS-16, t-test was used to analyze the data.Results: The results of this study indicate that there is a significant relationship between Crystal consumption and the serum level of ALP in both groups (P= 0.001). Besides, the relationships between the duration of Crystal consumption with ALKALINE PHOSPHATASE level and cigarette smoking were significant.Conclusion: Our results revealed that Crystal consumption is an important factor for elevating the ALP serum level in Crystal addicts.

ALKALINE PHOSPHATASE is an enzyme with widespread use in research and industry such as protein labeling, dephosphorylation of nucleic acids, and enzyme based biosensors. In the present study, ALKALINE PHOSPHATASE gene was inserted into the pET15b vector. The recombinant DNA was then expressed using IPTG as an inducer. The expression of this enzyme was optimized by changing expression conditions such as IPTG concentration, time, and temperature. The obtained results demonstrate that with the expression system used, large quantities of active ALKALINE PHOSPHATASE can be produced in E. coli.

Background: In tissue engineering, scaffold characteristics play an important role in the biological interactions between cells and the scaffold. Cell adhesion, proliferation, and activation depend on material properties used for the fabrication of scaffolds.Objective: In the present investigation, we used collagen with proper characteristics including mechanically stability, biodegradability and low antigenicity. Optimization of the scaffold was done by immobilization of ALKALINE PHOSPHATASE on the collagen surface via cross-linking method, because this enzyme is one of the most important markers of osteoblast, which increases inorganic phosphate concentration and promote mineralization of bone formation.Methods: ALKALINE PHOSPHATASE was immobilized on a collagen surface by 1-ethyl-3- (dimethylaminopropyl) carbodiimide hydrochloride, as a reagent. Then, rat mesenchymal stem cells were cultured in osteogenic medium in control and treated groups. The osteogenesis-related genes were compared between treatments (differentiated cells with immobilized ALKALINE PHOSPHATASE/collagen scaffold) and control groups (differentiated cells on collagen surface without ALKALINE PHOSPHATASE) on days 3 and 7 by quantitative real-time PCR (QIAGEN software).Results: Several genes, including ALKALINE PHOSPHATASE, collagen type I and osteocalcine associated with calcium binding and mineralization, showed upregulation in expression during the first 3 days, whereas tumor necrosis factor-α, acting as an inhibitor of differentiation, was down-regulated during osteogenesis.Conclusion: Collagen scaffold with immobilized ALKALINE PHOSPHATASE can be utilized as a good candidate for enhancing the differentiation of osteoblasts from mesenchymal stem cells.