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)

Human marrow mesenchymal stem cells were cultured in a medium containing glycerophosphate, ascorbic acid, and dexamethasone (Dex) on alumina ceramic discs and on tissue culture polystyrene (TCPS) dishes. Cell proliferation followed by osteogenic differentiation was observed to be equal on both culture substrata. The differentiation resulted in the appearance of bone-forming osteoblasts, which fabricated mineralized matrices on these substrata. Stem cells kept at 4 degrees C for 24 h outside a CO2 incubator maintained a viability level of more than 90%. The regenerative cultured bone outside the incubator also maintained high alkaline phosphatase activity for several hours. These results verified that cultured bone fabricated at a cell processing center can be transported to distant hospitals for use in hard tissue repair. To date, the tissue engineered cultured bone formed on alumina ceramics in this environment have already been used in clinical situations, such as total ceramic ankle replacements.
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PMID:Clinical application of marrow mesenchymal stem cells for hard tissue repair. 1525 53

The aim of this study was to compare the viability of human osteoblasts cryopreserved with Me2SO to that of fresh human iliac cancellous bone using cell culture techniques. Osteoblasts were obtained by spontaneous outgrowth of human iliac cancellous bone specimens in experiment I. In experiment II, human iliac cancellous bone was frozen with 10% Me2SO at -80 degrees C for 2 weeks and osteoblasts grew spontaneously after thawing at 37 degrees C by removing Me2SO with sucrose. The cells were grown in culture flasks containing DMEM as a culture medium, supplemented with 10% fetal calf serum. They were kept at 37 degrees C in a humidified atmosphere of 95% air and 5% CO2. Cells from the second passage were plated at a density of 5 times 10(3) cells/cm2 in 24-well plates. For detection of viability and differentiation, WST-1 assay, determination of alkaline phosphatase activity, concentration of procollagen I peptide, concentration of osteocalcin, and indirect immunofluorescence for osteopontin, collagen type I, integrin beta1, and fibronectin were applied. Experiments were conducted at four stages of confluence (days 4, 7, 14, and 21 after plating the cells). Based on the results of this study, we conclude that osteoblast-like cells survived cryopreservation and synthesized a range of markers that were consistent with this cell type.
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PMID:Osteoblast viability and differentiation with Me2SO as cryoprotectant compared to osteoblasts from fresh human iliac cancellous bone. 1629 58

Bone marrow cells have been used for testing biocompatibility of bone substitute materials that would be applied in maxillofacial and orthopedic surgeries. However, it remains unclear whether cells in serial subcultures retain the ability to differentiate into osteoblasts. The purpose of this study was to compare the development of osteoblast phenotype of serially passaged cells from human bone marrow. Cells from first to third passage were cultured (2 x 10(4) cells/well) in supplemented culture medium. Cells were incubated at 37 degrees C in a humidified atmosphere of 5% CO2 and 95% air. Cell attachment was assessed at 4 and 24 h. At 7, 14 and 21 days, cell proliferation, cell viability, total protein content and alkaline phosphatase (ALP) activity were evaluated. Bone-like formation was evaluated at 14 and 21 days. Data were compared by two-way ANOVA and Duncan's multiple range test. Cell attachment, cell viability and total protein content were not affected by serial subcultures. However, serial subcultures did interfered negatively with osteoblast differentiation as shown by osteoblast parameters observed in second and third subcultures, such as continuous cell proliferation, lower ALP activity and bone-like formation in comparison to first subculture. Therefore, it is important to evaluate cell ability to growth and differentiate before selecting the cell population for studies that investigate the biocompatibility of materials to replace bone tissue.
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PMID:Development of the osteoblast phenotype of serial cell subcultures from human bone marrow. 1642 89

Dexamethasone (Dex) has been shown to induce osteoblast differentiation in several cell culture systems. This study investigated the effect of continuous and discontinuous treatment with Dex on osteoblast differentiation of human bone marrow stromal cells (BMSC). Primary culture and first passage were cultured in media with or without Dex 10(-7) M. During the culture period, cells were incubated at 37 degrees C in humidified atmosphere of 5% CO2 and 95% air. At 7, 14, and 21 days, cell proliferation, cell viability, total protein content, alkaline phosphatase (ALP) activity and bone-like formation were evaluated. Data were compared by two-way analysis of variance. Dex did not affect cell viability and total protein content, but reduced cell number. ALP activity and bone-like formation increased when only first passage or both primary culture and first passage were treated with Dex, in comparison to the groups that did not have contact with Dex after first passage. The results of this study indicate that, for human BMSC, continuous presence of Dex did not appear to be required for development of the osteoblast phenotype, but Dex must be present after first passage to allow osteoblast differentiation expressed by reduced cell proliferation and increased ALP activity and bone-like formation.
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PMID:Osteoblast differentiation of human bone marrow cells under continuous and discontinuous treatment with dexamethasone. 1647 12

This study addresses some microbial inactivation phenomena induced by high pressure CO2 over micro-organisms and enzymes. The activity of four selected enzymes was measured before and after treatment with CO2 under pressure in both buffer solutions and natural cellular environment (E. coli cells and tomato paste). Results are reported for acid phosphatase, alkaline phosphatase, ATPase, and pectinase at different conditions of temperature, CO2 pressure, and treatment time (32-40 degrees C, 85-150 bar, 30-70 min). The results obtained show that the high pressure CO2 treatment induces an inactivation of cellular enzymatic activity higher than the one caused on the same enzymes in solution. However, the measured activity difference is not caused by a damage at the enzymes molecular level but is a consequence of the permeabilization of the cellular envelopes which leads to a release of unmodified enzymes from the cells with simultaneous drop of enzymatic cellular activity. The reported data suggest that the bacterial cell death is probably due not to a selective effect of high pressure CO2 treatment but to simultaneous detrimental action of CO2 on cellular membrane and cell wall.
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PMID:A study on the inactivation of micro-organisms and enzymes by high pressure CO2. 1673 96

Increasing osteoblast activity in an anabolic fashion may offer an ideal therapeutic treatment for various orthopedic complications including osteoporosis. The purpose of this study was to evaluate the effect of mevinolin, a clinical statin drug, on osteoblast function (MG 63 Cell Line) and compare its mode of action with the conventionally utilized parathyroid hormone (PTH). MG63 cells were treated with different concentrations (control, low (100 nM), medium (1 uM), and high (10 uM)) of mevinolin or Parathyroid hormone. The cells were incubated for 24, 48, and 72 hours at 37 degrees C in a 95% air and 5% CO2 environmental chamber. Data obtained in this study revealed that: (I) there were significant decreases in cell number after 24 hours upon the exposure of medium and high doses of mevinolin, (II) cell numbers rebounded back toward control after 48 hours and were similar in number at 72 hours, and (III) there were no significant changes in calcium or alkaline phosphatase activity were observed throughout the study. Morphologically, the cells treated with various doses of Mevinolin expressed similar structural changes to those observed using PTH. These changes included pleomorphic characteristics and an occasional hyperchromatic pattern during the entire duration of the study (72 Hours). Other structural features observed were spindle shapes, cluster arrangements and multinucleation. The majority of cells had multiple nucleoli in all treated groups compared to controls. The overall conclusion of this investigation demonstrated that the concentrations used (100 nM and 10 microM) did not appear to affect the mitotic activities of immature phenotypic MG-63 cells. In addition, the concentrations of mevinolin used did not trigger the differentiation process of the cells throughout the experimental phases. This observation led us to suggest that the reason for such an outcome could be attributed to the lack of a response in calcium production or alkaline phosphatase activity (stimulator to differentiation and mineralization process).
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PMID:Comparison between mevinolin and PTH action on MG63 osteoblast-like cells. 1681 19

The isolation and culturation of SSCs of different stage of Wuzhishan Mini Porcine (WZSP) with different way of enzymatic digestion and culturation were deaded in this study. The results of the experiment described are as the following: The proper time of isolation and culturation of SSCs of WZSP is 1-20 old days. Different old of piglets with different method. Using DMEM medium as a fundmental culture medium add different gradient at 34 degrees C in a water-saturated atmosphere of 95% air, 5% CO2. The mulberry-shaped SSCs clusters appeared as original generation in 7-8 days culture. The SSCs clusters developed half-suspendedly in the culture medium. SSCs alkaline phosphatase (AKP) staining expressed positively. Mouse embryonic fibroblast was used as feeder layer for the SSCs passage cultured, The SSCs show good attached attributes, but the number of SSCs decreased quickly after 4 days culture. By seminiferous cord fragment culturation can also appear SSCs clusters in 5 days, The SSCs clusters developed half-suspendedly in the culture medium. In addition, the testes placed in cold (4 degrees C) PBS banlanced salt solution for 24 h also can be used as a good matierials for preparation of SSCs. These results indicate that the method of solation and culturation of SSCs are very correct and efficient, all these can be utilized as a good reference for future studies.
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PMID:[Studies on spermatogonial stem cells cultured in vitro of Wuzhishan Mini Porcine]. 1689 12

Use of moist snuff is widespread in Sweden. In 2004 approximately 8oo,ooo Swedes were daily users which corresponds to 22% of the male population and 3% of the female population. The aim of the present study was to evaluate the effect of Swedish moist snuff extract on PDLfibroblast growth and hard tissue production and compare with moist snuff extract from USA. Periodontal ligament cells (PDL-cells) were obtained from 3 healthy subjects (1 female 14 years, 2 males 14 and 17 years) from the root surface of premolars extracted for orthodontic reasons. The cells were isolated from explants and grown in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal calf serum (FBS) and cultivated in 37 degrees C with 5% CO2 in air. Snuff extract in concentrations 0.3%, 1% and 3% (in DMEM with 1% FBS) was tested. Cells from each individual were tested three times, each time in triplicate. Photographs were taken at o and 24 hours with a digital camera and analysed in terms of growth and morphology. Then the cell suspension was frozen and later thawed for examination of the production of alkaline phosphatase after exposure to different snuff concentrations. This in vitro study has shown that PDL cells from 3 different subjects demonstrated a reduced number of cells at exposure to 3% of both Swedish and American snuff extract. The production of alkaline phosphatase after 2 hours was similarly reduced from cells exposed to 3% snuff extract. Further studies have to be made to understand the effect of smokeless tobacco on periodontal tissues. However, from this study can be concluded that smokeless tobacco has biological effects in terms of reduced PDL cell growth and production of alkaline phosphatase
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PMID:The effect of Swedish and American smokeless tobacco extract on periodontal ligament fibroblasts in vitro. 1723 25

Florida manatees (Trichechus manatus latirostris) are endangered aquatic mammals living in coastal and riverine waterways of Florida and adjacent states. Serum or plasma biochemical analyses are important tools in evaluating the health of free-ranging and captive manatees. The purpose of this study was to measure diagnostically important analytes in the plasma of healthy manatees and to determine whether there was significant variation with respect to location (free-ranging versus captive), age class (small calves, large calves, subadults, adults), and gender. No significant differences in plasma sodium, potassium, bilirubin, glucose, alanine aminotransferase, or creatine kinase were found among these classes of animals. Compared to free-ranging manatees, captive animals had significantly lower mean concentrations of plasma chloride, phosphate, magnesium, triglycerides, anion gap, and lactate. Captive manatees had significantly higher mean values of total CO2, calcium, urea, creatinine, alkaline phosphatase, gamma-glutamyltransferase, total protein, albumin, and albumin/globulin ratio than did free-ranging animals. Differences in the environments of these two groups, including diet, temperature, salinity, and stress, might account for some of these results. The higher plasma lactate and anion gap concentrations and lower total CO2 concentrations of free-ranging manatees were probably due to greater exertion during capture, but the lack of elevated plasma creatine kinase activity relative to captive animals indicates that there was no serious muscle injury associated with capture. Plasma phosphate decreased and total globulins increased with age. Plasma cholesterol and triglyceride concentrations were highest in small calves. Plasma aspartate aminotransferase was higher in large calves than in adults and subadults, and the albumin/ globulin ratio was higher in subadults than in adults. Plasma total CO2 was higher and chloride was slightly lower in females than in males.
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PMID:Clinical biochemistry in healthy manatees (Trichechus manatus latirostris). 1767 11

Healthy Atlantic bottlenose dolphins (Tursiops truncatus) have a sustained postprandial hyperglycemia, producing a prolonged glucose tolerance curve and a transient, diabetes mellitus-like state during 6 to 72 h of fasting. To further assess dolphins as comparative models for diabetes in humans, we hypothesized that a suite of hematological and clinical biochemistry changes during the fasting state may mimic those reported in humans with diabetes. We conducted a retrospective analysis of covariance to compare fasting and nonfasting hematologic and serum biochemical data, including 1161 routine blood samples from 52 healthy bottlenose dolphins (age, 1 to 49 y; male and female) collected during 1998 through 2005. Most changes found in dolphins during the fasting state--including significantly increased glucose, platelets, gamma-glutamyl transpeptidase, and alkaline phosphatase; significantly decreased serum uric acid; and shifts toward a metabolic acidodic state (significantly increased blood CO2)--have been previously associated with diabetes mellitus in humans. Therefore, healthy bottlenose dolphins may be the first complete and natural comparative animal model for diabetes mellitus in humans. Similarities between dolphins and humans, including metabolic changes associated with high-protein, low-carbohydrate diets; large brain-to-mass ratios; high central nervous system demands for glucose; and similarly unique blood glucose-carrying capacities should be further assessed to better understand the potential evolutionary paths of diabetes mellitus in these 2 species.
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PMID:Big brains and blood glucose: common ground for diabetes mellitus in humans and healthy dolphins. 1780 54


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