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)

The effects of cryopreservation on periodontal regeneration of transplanted rat molars were investigated histologically and histochemically in rats. Bilateral first and second maxillary molars of 4-week-old Wistar rats were gently extracted and transplanted into the abdominal subcutaneous connective tissue immediately or after cryopreservation in liquid nitrogen overnight. Donor teeth were slowly frozen by a rate-controlling freezer (program freezer) using 5% dimethylsulfoxide (DMSO) and 6% hydroxyethyl starch (HES) as cryoprotectants. One-four weeks after transplantation, they were carefully excised with the surrounding tissues. Regeneration of acellular cementum, periodontal ligament, and alveolar bone were observed 2 weeks after immediate transplantation. The pulp was repaired by the ingrowth of granulation tissue from the root apex followed by the formation of calcified tissue. The regenerated periodontal ligament was positive for alkaline phosphatase (ALP). Small or mononuclear tartrate resistant acid phosphatase (TRAP) positive cells were scattered on the newly formed alveolar bone and on the hard tissue in the pulp, but there was no external or internal progressive root resorption at 4 weeks. Cryopreserved teeth had acellular cementum with a rough surface at 1 week, but with the increase of cementoblasts and the appearance of periodontal ligament and alveolar bone, the surface became smooth at 3 weeks. Epithelial rests of Malassez (ERM) also revived. After regeneration of the periodontal tissues at 4 weeks, there was no evidence of root resorption. Although the process proceeded slowly, the cryopreserved teeth showed the periodontal regeneration substantially similar to that of the immediately transplanted teeth without progressive root resorption, indicating that they could be applicable for clinical use.
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PMID:Periodontal regeneration of transplanted rat molars after cryopreservation. 1469 98

The present study was performed to investigate the subacute effect of alpha-cypermethrin (alpha-CP) in rats. Alfacypermethrin a synthetic pyrethroid insecticide, dissolved in dimethyl sulfoxide (DMSO) and oral LD50 was investigated after administering orally different doses in rats and was determined as 145 mg/kg. Other groups of rats were given repeated daily oral dose (1/10 LD50) of alpha-CP for 30 days. The animals were sacrificed on 31st day. Activities of various enzymes, cytochrome P450 and b5 contents in liver, hepatic antioxidant status, tissue residue concentration, haemogram and pathological changes were studied. It increased the serum aminotransaminases (AST, ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) activities and blood glucose level significantly. alpha-CP decreased RBC count, PCV and Hb level significantly. It significantly decreased cytochrome P450 in liver. Residues were present in different tissues. It increased malondialdehyde (MDA) level, while decreased the activities of catalase (CAT), superoxide dismutase (SOD) and glycogen level in liver significantly. Mild to moderate histological alterations were observed in lungs, liver, stomach, kidneys, testes and cerebellum. So repeated daily oral doses of alpha-CP at 1/10LD50 altered the biochemical parameters, decreased cytochrome P450 content, antioxidant status, which correlated with histopathological changes of tissues.
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PMID:Repeated dose toxicity of alfa-cypermethrin in rats. 1536 39

Green tea polyphenol (GTP) together with dimethylsulphoxide (DMSO) were added to a freezing solution of osteoblastic cells (rat calvarial osteoblasts and human osteosarcoma cells) exposed to repeated freeze/thaw cycles (FTC) to induce oxidative stress. When cells were subjected to 3 FTCs, freezing medium containing 10% (v/v) DMSO and 500 mug GTP ml(-1) significantly (p<0.05) suppressed cell detachment and growth inhibition by over 63% and protected cell morphology. Furthermore, the alkaline phosphatase activity of osteoblastic cells was appreciably maintained after 2 and 3 FTCs in this mixture. Polyphenols may thus be of use as a cell cryopreservant and be advantageous in such fields as cell transplantation and tissue engineering.
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PMID:Protection of osteoblastic cells from freeze/thaw cycle-induced oxidative stress by green tea polyphenol. 1597 73

Semen samples from 12 bucks Were extended with 10 different extenders containing glycerol, DMSO, glycerol + DMSO, and glycerol + lactose in varying concentrations as cryoprotective agents. The activities of acrosin, hyaluronidase, alkaline phosphatase (AKP), aspartate aminotransferase (AST), alanine amino transferase (ALT) and lactic dehydrogenase (LDH) were assayed in equilibrated (Prefreeze) and frozen thawed (Postfreeze) semen samples. Significantly (P < 0.01) higher intracellular activity of acrosin was recorded in semen samples extended with lactose than with the other extenders, with the maximum being with Tris yolk glycerol lactose (TYGL(180)). Effects of extenders on acrosin activity were significant (P < 0.01) at both of the pre-and postfreeze stages. However, extracellular activities of hyaluronidase, alkaline phosphatase, transaminases (AST and ALT), and lactic dehydrogenase were significantly higher in extenders containing DMSO than lactose. Leakage of these enzymes was found to increase from the prefreeze to the post freeze stage.
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PMID:Effect of cryoprotectants on release of various enzymes from buck spermatozoa during freezing. 1672 4

Polychlorinated biphenyls (PCBs) are persistent environmental pollutants. Two PCB mixtures, Aroclors 1221 and 1254 have been suggested to have estrogenic and anti-estrogenic properties, respectively. We have examined whether these PCB mixtures modulate bone turnover and vertebral histology in intact and ovariectomized (ovx) rat models. Thirty-two adult female rats were divided into four groups subcutaneously receiving 4% DMSO (control), A1221 (10 mg/kg), A1254 (10 mg/kg) oestradiol (E2, 30 microg/kg). These compounds were injected to the animals for a period of 6 weeks at two daily intervals. In the second model, rats (n=32) were ovx and allowed to recover for a period of 3 weeks. Control group received vehicle (4% DMSO) alone. Remaining rats were divided into three groups and injected (s.c.) with A1221, A1254 and E2 for 5 weeks. Urine samples were collected prior to end of the experiments. Then, all animals were decapitated. Serum parathyroid hormone (PTH), calcitonin and osteocalcin levels were determined by immunoradiometric method. Serum concentrations of alkaline phosphatase (ALP), calcium and inorganic phosphate were determined by enzymatic-colorimetric method. Urinary deoxypyridinoline (DPD) was measured by ELISA. Lumbar vertebrae (L2) of all animals were dissected out and processed for light microscopy. Levels of urinary DPD were significantly lowered in E2 -treated intact rats (p<0.001). Ovx significantly increased urinary DPD excretion (p<0.01) compared to intact control values. Administration of A1221 and A1254 had no significant effects in intact rats, however, they significantly reduced (p<0.05) and increased (p<0.001) urinary DPD levels in ovx rats, respectively. Neither of the PCB mixtures significantly changed serum osteocalcin and ALP levels in intact or ovx rats (except A1221 increased ALP in intact model, p<0.01). Both PCB mixtures had differential effects on serum PTH, calcitonin, calcium and inorganic phosphate concentrations. Treatment with A1221 reversed the adverse effects of ovariectomy on L2 histology. However, A1254 produced necrotic areas in vertebral bone, and this effect was expanded in ovx animals. Our findings suggest that both Aroclor compounds interfere with bone turnover mechanisms, particularly in ovx rats.
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PMID:Modulatory effects of Aroclors 1221 and 1254 on bone turnover and vertebral histology in intact and ovariectomized rats. 1697 6

Our group is developing a novel technology, enzyme-mediated cancer imaging and therapy (EMCIT), that aims to entrap radioiodinated compounds within solid tumors for noninvasive tumor detection and therapy. In this approach, a water-soluble, radioiodinated prodrug is hydrolyzed in vivo to a highly water-insoluble compound by an enzyme overexpressed extracellularly by tumor cells. We have synthesized and characterized the water-soluble prodrug, 2-(2'-phosphoryloxyphenyl)-6-[(125)I]iodo-4-(3H)-quinazolinone [(125)I]5, which is readily hydrolyzed by alkaline phosphatase, an enzyme expressed by many tumor cell lines, to a water-insoluble drug, 2-(2'-hydroxyphenyl)-6-[(125)I]iodo-4-(3H)-quinazolinone [(125)I]1. In the course of our study, we discovered that ammonium 2-(2'-phosphoryloxyphenyl)-6-tributylstannyl-4-(3H)-quinazolinone, an intermediate in the radioiodination of the prodrug, exists as two isomers (3 and 4) whose radioiodination leads, respectively, to [(125)I]6 and [(125)I]5. These prodrugs have different in vitro and in vivo biologic activities. Compound 6 is not hydrolyzed by alkaline phosphatase (ALP), whereas 5 is highly soluble (mg/mL) in aqueous solution and is rapidly dephosphorylated in the presence of ALP to 1, a water-insoluble molecule (ng/mL). Mouse biodistribution studies indicate that [(125)I]6 has high uptake in kidney and liver and [(125)I]5 has very low uptake in all normal organs. Compounds 3 and 6 are converted, respectively, to 4 and 5 after incubation in DMSO. The stability of 5 in human serum is high. The minimum ALP concentration needed to hydrolyze 5 is much greater than the ALP level in the blood of patients with cancer, and the latter should not affect the pharmacokinetics of the compound. Incubation of 5 with viable human and mouse tumor-cell lines--but not with normal human cells and mouse tissues--leads to its hydrolysis and the formation of large crystals of 1. We expect that 5 will also be hydrolyzed in vivo by tumor cells that express phosphatase activity extracellularly and anticipate the specific precipitation of radioiodinated 1 within tumor cell clusters. This should lead to high tumor-to-normal-tissue ratios and enable imaging (SPECT/PET) and radionuclide therapy of solid tumors.
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PMID:Evaluation of chemical, physical, and biologic properties of tumor-targeting radioiodinated quinazolinone derivative. 1738 3

Puerarin, a natural isoflavonoid found in Chinese Pueraria lobata (Wild.) Ohwi, has received increasing attention because of its possible role in the prevention of osteoporosis. However, the relationship between puerarin and bone formation remains unknown. In the present study, rat osteoblasts isolated from newborn Wistar rats were used to investigate the effect of puerarin on osteoblasts, and its possible molecular mechanism. Data showed that puerarin caused a significant increase in cell viability, alkaline phosphatase (ALP) activity and mineral nodules formation in osteoblasts, suggesting that puerarin had a stimulatory effect on osteoblastic bone formation. This functional improvement by puerarin was accompanied by activation and nuclear translocation of Akt. Furthermore, puerarin-stimulated osteoblastic growth, Akt activation and redistribution were significantly blocked by the specific PI3K inhibitor, LY294002. These results strongly suggested that puerarin stimulated osteoblastic proliferation and Akt activation in a PI3K-dependent manner. In summary, puerarin derived from Chinese Pueraria lobata (Wild.) Ohwi can promote bone formation in cultured rat osteoblasts, which might be mediated by activation of the PI3K/Akt pathway. DMEM:Dulbecco's modification of Eagel's medium PBS:phosphate buffered saline DMSO:dimethyl sulfoxide EDTA:ethylene diamine tetraacetic acid SDS:sodium dodecyl sulfate SDS-PAGE:sodium dodecylsulfate polyacrylamide gel electrophoresis FITC:fluorescein isothiocyanate HRP:horseradish peroxidase PI3K:phosphatidylinositol 3-kinase.
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PMID:Stimulatory effect of puerarin on bone formation through activation of PI3K/Akt pathway in rat calvaria osteoblasts. 1744 35

This study was aimed to examine effects of genistein on leukocyte adhesion in femur microcirculation in relation to bone-loss induced in ovariectomized female rats. Sixty-four female Wistar rats were divided into 4 groups: sham (daily treated with vehicle; DMSO, sc; 100 microl/day), ovariectomized rat treated with vehicle (OVX(veh)), 17beta-estradiol treated-ovariectomized rat (OVX(E2), 5 microg/kg/day, s.c.) and genistein treated-ovariectomized rat (0.25 mg/kg/day, s.c.; OVX(gen)). One and three weeks after the ovariectomy, blood flow perfusion (BF) in femur tissue was measured using laser Doppler flowmetry. Leukocyte adhesion in femur venules (15-30 microm in diameter) of each group was evaluated by intravital fluorescence videomicroscopy. The bone mineral content (BMC) was measured and expressed in terms of ratio of ash-to-dry matter weight. Serum osteocalcin and alkaline phosphatase levels were determined using chemiluminescence immunoassay. In both one and three week-OVX(veh), leukocyte adhesion increased significantly, compared to their age-matched sham groups, but it decreased significantly in OVX(gen), compared to OVX(veh) (p<0.05). In three week-OVX(gen), both BF and BMC increased significantly, but osteocalcin and alkaline phosphatase levels decreased, compared to those of three week-OVX(veh). In conclusion, genistein supplementation could effectively prevent bone-loss and microvascular endothelial dysfunction induced by estrogen deficiency.
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PMID:Preventive effects of genistein on leukocyte adhesion in femur venules and on bone-loss induced in ovariectomized female rats. 1833 78

Human embryonic stem (hES) cells are a valuable tool for studying human development in addition to their potential applications in regenerative medicine and drug discovery. The role of genetic background and epigenetic influences in development as well as in response to external influences such as drugs and therapies is well recognized. The great ethnic diversity in the Indian subcontinent translates to interindividual variability in drug response and disease susceptibility. For these reasons, new hES cell lines representing Indian genetic diversity will be valuable in studies of tissue-differentiation, cellular-function and for aspects of characterization of responses to drugs. We have derived two new hES cell lines, BJNhem19 and BJNhem20 from the inner cell mass (ICM) of discarded grade III human embryos that were not suitable for in vitro fertility treatment. Human leukocyte antigen (HLA) isotype analysis shows that they are genetically distinct from existing hES cell lines. Short tandem repeat (STR) analysis shows that the two cell lines are derived from sibling embryos. These cell lines show an undifferentiated phenotype in culture for more than 65 passages, show normal karyotype and express pluripotency markers such as TRA-1-60, TRA-1-81, stage-specific embryonic antigen-4 (SSEA-4), alkaline phosphatase, DNMT3B, GABRB3, GDF3, OCT4, NANOG, SOX2, TERF1, TDGF, LEFTA, THY1, and REX1. While both cell lines can differentiate into derivatives of all three germ layers in vitro, only BJNhem20 can form teratomas when transplanted into mice. We observe an increased frequency of cardiomyocyte differentiation from BJNhem20 embryoid bodies in feeder-free cultures upon induction with DMSO. Cardiomyocytes purified from such cultures survive and show rhythmic contractions for several weeks in culture. These hES cell lines have been accepted for deposit in the U.K. Stem Cell Bank and will be a useful resource for the international stem cell community.
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PMID:Derivation and characterization of two sibling human embryonic stem cell lines from discarded grade III embryos. 1861 85

Fish embryonic stem (ES) cells derived from of blastulae (64 cell stage embryo) of Labeo rohita were propagated in culture and retained their ES cell-like properties after cryogenic storage (-196 degrees C, i.e., liquid nitrogen). Toxic effect of DMSO (dimethyl sulphoxide) on stem cells during preservation process has been reported to restrict therapeutic applications. In this study we reduced the concentration of DMSO and added the non-toxic cryoprotective agent (CPA) trehalose. Cryopreservation of ES cell colonies was done at 5, 25 and 52 passages with 0.2 M trehalose and 0.8 M (DMSO). A combination of both the cryoprotective agents (non-toxic and toxic) demonstrated better survival and recovery of ES cells than the DMSO used alone. Use of this CPA combination in the freezing media gave an optimum viability of more than 83 % in a slow freezing protocol. Trehalose showed a definite advantage over DMSO in terms of viability and intactness of ES cell colonies with evenly distributed morphology. There was no significant difference observed in the expression levels of cell surface markers like stage specific embryonic antigen-1 (SSEA-1) and alkaline phosphatase (ALP) between early and late passages after 60 days of post-thawing. More than 90 % of the ES cell colonies showed extensive expression of ALP and positive expression of SSEA-1 from an early stage of ES cells culture up to passage 52 (in our study) in the presence of leukemia inhibitory factor (LIF) and without feeder cells. Further, thawed ES cells showed a normal karyotype and maintained an undifferentiated state through out the study. This study on ES cell cryopreservation and subsequent retention of stem cell properties without feeder cells using a non-toxic cryoprotectant trehalose would be highly useful for future in vitro differentiation, manipulation of fish ES cells and as a model for mammalian ES cell culture.
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PMID:Use of the non-toxic cryoprotectant trehalose enhances recovery and function of fish embryonic stem cells following cryogenic storage. 1907 57


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