Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Films of Ti-Ca-P-C-O-(N), Ti-Ca-C-O-(N) and Ti-Zr-C-O-(N) were deposited by DC magnetron sputtering or ion implantation-assisted magnetron sputtering of composite targets TiC0.5 + 10%Ca10(PO4)6(OH)2, TiC0.5 + 20%(CaO + TiO2) and TiC0.5 + 10%ZrO2 in an Ar atmosphere or reactively in a gaseous mixture of Ar + 14%N2. The microstructure, elemental and phase composition of films were studied by means of X-ray diffraction, transmission electron microscopy, scanning force microscopy, X-ray photoelectron spectroscopy and energy-dispersive X-ray spectroscopy. The films were characterized in terms of their hardness, Young's modulus, elastic recovery, adhesion strength, and friction and wear both in air and under physiological solution. Particular attention was paid to the analysis of deformation and fracture for various film/substrate systems during scratch testing. The biocompatibility of the films was evaluated by both in vitro and in vivo experiments. In vitro studies involved the investigation of adhesion, spreading, and proliferation of MC3T3-E1 osteoblasts and IAR-2 epithelial cells, morphometric analysis, actin cytoskeleton, focal contacts staining, alkaline phosphatase activity and von Kossa staining of osteoblastic culture. Cell culture experiments demonstrated an increase of osteoblastic proliferation on Ca- and P-incorporated films. In vivo studies were fulfilled by subcutaneous implantation of Teflon plates coated with the tested films in mice and analysis of the population of adherent cells on their surfaces. The results obtained show that multicomponent nanostructured Ti-(Ca, Zr)-(C, N, O, P) films possess a combination of high hardness, wear resistance and adhesion strength, reduced Young's modulus, low friction coefficient and high biocompatibility.
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PMID:Multifunctional Ti-(Ca,Zr)-(C,N,O,P) films for load-bearing implants. 1653 Aug 25

Immobilized metal affinity chromatography (IMAC) and titanium oxide (TiO2) chromatography are simple, widely used, and cost-effective methods to enrich phosphopeptides, but the sample loading buffer composition, desalting procedure, and control of loading amount are critical to avoid nonspecific interactions and to achieve efficient phosphopeptide enrichment. Although the combination of MS3 analysis and high-resolution mass spectrometry (MS) is helpful to identify phosphopeptides, the quality of many MS/MS spectra having a neutral loss peak of phosphate is still too poor to allow sequence identification, and this results in many false-negative as well as false-positive identifications. Here, we present a novel strategy, which is based on the use of alkaline phosphatase to remove phosphates and analysis of phospho/dephosphopeptide retention times to increase the reliability of identification. The use of phospho/dephosphopeptide retention time ratios allows the identification of phosphopeptides with high confidence with the aid of a focused database of dephosphopeptides. This approach was very effective to identify multiple phophorylations in tryptic peptides. A 'true' phosphorylation data set should contain about 90% phospho-Ser and a few percent phospho-Tyr, and this ratio can be used as a quality criterion for evaluation of data sets. By applying this efficient approach, we were able to identify more than one thousand phosphopeptides.
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PMID:Enhancement of the efficiency of phosphoproteomic identification by removing phosphates after phosphopeptide enrichment. 1733 Sep 47

The surface topography and chemistry of titanium are postulated to be two major factors that affect the osseointegration capacity of titanium implants. However, it is extremely difficult to control one factor without changing the other, which prevents the isolation of the genuine effect of one factor. This study aimed to determine whether surface chemistry of titanium alone affects osteoblastic function. Two different titanium surfaces were prepared by sputter depositioning of titanium (Ti; 99.99% purity) or titanium dioxide (TiO2; 99.99% purity) (50-nm thick for each) onto machined commercially pure titanium disks. Rat bone marrow-derived osteoblastic cells were cultured on each of the two surfaces. TiO2 surface showed 4.4 times higher elemental oxygen concentration and higher water wettability than Ti surface. Scanning electron microscopic and atomic force microscopic examination revealed no differences in surface topography and roughness values between the two surfaces. The cell proliferated more on TiO2 than on Ti by up to 60%. Although the expression of collagen I gene increased more rapidly on TiO2 at early culture stage of day 3, the late stage marker genes for osteoblastic differentiation, including osteopontin and osteocalcin, were not modulated between the two cultures. The alkaline phosphatase positive area and mineralized nodule area were approximately two times larger on TiO2 than on Ti. In conclusion, titanium materials having different superficial chemistry, that is, titanium or titanium dioxide, may exert different biological capacity of osteoblasts; titanium dioxide may induce superior osteoconduction, primarily because of the increased osteoblastic proliferation.
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PMID:The effect of superficial chemistry of titanium on osteoblastic function. 1760 Mar 32

It is widely accepted that implant surface factors affect the quality of the bone-to-implant interface. Recent additional treatments superimposed on moderately rough cpTitanium surface provide further enhancement of bone-to-implant contact. The aim of this study was to compare osteoinductive and bone-specific gene expression in cells adherent to titanium dioxide-grit blasted (TiO2) versus TiO2 grit blasted and HF treated (TiO2/HF) cpTitanium implant surfaces. MC3T3-E1 cells were grown in osteogenic supplements on the titanium disk surfaces for 1-14 days. Real-time PCR was used to measure RUNX-2, Osterix, and bone sialoprotein (BSP) mRNA levels. Implants were placed in rat tibia and, following harvesting at 1-7 days after placement, real-time PCR was used to measure RUNX-2, alkaline phosphatase (ALP), and BSP mRNA levels in implant adherent cells. In cell culture, RUNX-2 and Osterix levels were significantly increased (p<0.05) on the TiO2/HF surfaces as compared to the TiO2 and smooth surfaces through the cultural period, while BSP expression was elevated on both TiO2 and TiO2/HF surfaces when compared to a machined surface control. In cells adherent to implants retrieved from rat tibia, RUNX-2 mRNA levels were 2-fold and 8-fold greater on the TiO2/HF surfaces at 1-3 and 7 days following implantation. This was paralleled by significantly greater levels of ALP at 3 and 7 days and BSP mRNA at 7 days following implantation. As a marker of osteoinduction, the increased levels of RUNX-2 in cells adherent to the TiO2/HF surfaces suggest that the additional HF treatment of the TiO2 grit blasted surface results in surface properties that support adherent cell osteoinduction. In vivo assessments of implant adherent cell phenotypes provide further insight into the mechanisms affecting alloplast-tissue interactions.
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PMID:The effect of hydrofluoric acid treatment of TiO2 grit blasted titanium implants on adherent osteoblast gene expression in vitro and in vivo. 1786 50

A novel detection method for the analysis of multi-phosphopeptides using microcolumn high performance liquid chromatography-electrospray ionization tandem mass spectrometry (microHPLC-ESI-MS/MS) was proposed by the dephosphorylation treatment of alkaline phosphatase (AP). After the selective enrichment by a microcolumn packed with TiO2, phosphopeptides from the tryptic digests of beta-casein were dephosphorylated by AP. Through the removal of phosphate groups, the detection of multi-phosphopeptides according to the non-phosphorylated ones was achieved by ESI-MS/MS. By comparing the chromatograms before and after the AP treatment, mono-phosphopeptides were identified based on the relative molecular mass (Mr) difference of 80. Furthermore, since more peaks appeared after the treatment, the existence of multi-phosphopeptides was proven. By controlling the treatment procedure, the partial dephosphorylation of multi-phosphopeptides was performed, and the multi-phosphorylated stees of the digest of beta-casein were found to be on serine residues at the possible sites of 17, 18 and 19.
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PMID:[Detection of multi-phosphopeptide sites using microcolumn high performance liquid chromatography-electrospray ionization tandem mass spectrometry]. 1797 96

Protein phosphorylation is a central cell signaling event that underlies a broad spectrum of key physiological processes. Advances in affinity chromatography and mass spectrometry are now providing the ability to identify and quantitate thousands of phosphorylation sites simultaneously. Comprehensive phosphoproteome analyses present sizable analytical challenges in view of suppression effects of phosphopeptides and the variable quality of MS/MS spectra. This work presents an integrated enzymatic and data mining approach enabling the comprehensive detection of native and putative phosphopeptides following alkaline phosphatase digestion of titanium dioxide (TiO2)-enriched cell extracts. The correlation of retention times of more than 750 phospho- and dephosphopeptide pairs from J774 macrophage cell extracts indicated that removal of the phosphate groups can impart a gain or a loss in hydrophobicity that is partly explained by the formation of a salt bridge with proximal amino groups. Dephosphorylation also led to an average 2-fold increase in MS sensitivity that facilitated peptide sequencing. More importantly, alkaline phosphatase digestion enhanced the overall population of putative phosphopeptides from TiO2-enriched cell extracts providing a unique approach to profile multiphosphorylated cognates that would have remained otherwise undetected. The application of this approach is demonstrated for differential phosphoproteome analyses of mouse macrophages exposed to interferon-gamma for 5 min. TiO2 enrichment enabled the identification of 1143 phosphopeptides from 432 different proteins of which 125 phosphopeptides showed a 2-fold change upon interferon-gamma exposure. The use of alkaline phosphatase nearly doubled the number of putative phosphopeptides assignments leading to the observation of key interferon-gamma signaling events involved in vesicle trafficking, production of reactive oxygen species, and mRNA translation.
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PMID:Combined enzymatic and data mining approaches for comprehensive phosphoproteome analyses: application to cell signaling events of interferon-gamma-stimulated macrophages. 1800 92

The detection of phosphopeptides, especially multi-phosphopeptides, by tandem electrospray ionization mass spectrometry (ESI-MS/MS) is a great challenge due to their low abundance and the poor ionization efficiency of samples. In our recent study, a strategy was proposed for the analysis of trace multi-phosphopeptides which combined selective enrichment of phosphorylated peptides by TiO2 and dephosphorylation by alkaline phosphatase (AP). After separation by muHPLC, the profiles of enriched peptides before and after AP treatment were compared, and the additional peaks appearing in the latter case hinted at the existence of multi-phosphopeptides. Subsequently, an incomplete dephosphorylation reaction was performed to partially remove the phosphate groups so that the phosphorylation sites of the multi-phosphopeptides might be estimated. Through analysis of the digests of beta-casein and extracted proteins of bovine milk, more information on the multi-phosphopeptides was obtained by muHPLC-ESI-MS/MS than that obtained without AP treatment, which demonstrated that such a strategy might supply some potential information about trace multi-phosphopeptides lost in shotgun analysis.
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PMID:Characterization of multi-phosphopeptides by muHPLC-ESI-MS/MS with alkaline phosphatase treatment. 1821 Mar 78

Ti being bioinert shows poor bone cell adhesion with an intervening fibrous capsule. Ti could be made bioactive by several methods including growing in situ TiO2 layer on Ti-surface. TiO2 nanotubes were grown on Ti surface via anodization process and the bone cell-material interactions were evaluated. Human osteoblast cell attachment and growth behavior were studied using an osteoprecursor cell line for 3, 7, and 11 days. An abundant amount of extracellular matrix (ECM) between the neighboring cells was noticed on anodized nanotube surface with filopodia extensions coming out from cells to grasp the nanoporous surface of the nanotube for anchorage. To better understand and compare cell-materials interactions, anodized nanoporous sample surfaces were etched with different patterns. Preferential cell attachment was noticed on nanotube surface compare to almost no cells in etched Ti surface. Cell adhesion with vinculin adhesive protein showed higher intensity, positive contacts on nanoporous surface and thin focal contacts on the Ti-control. Immunochemistry study with alkaline phosphatase showed enhanced osteoblastic phenotype expressions in nanoporous surface. Osteoblast proliferation was significantly higher on anodized nanotube surface. Surface properties changed with the emergence of nanoscale morphology. Higher nanometer scale roughness, low contact angle and high surface energy in nanoporous surface enhanced the osteoblast-material interactions. Mineralization study was done under simulated body fluid (SBF) with ion concentration nearly equal to human blood plasma to understand biomimetic apatite deposition behavior. Although apatite layer formation was noticed on nanotube surface, but it was nonuniform even after 21 days in SBF.
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PMID:TiO2 nanotubes on Ti: Influence of nanoscale morphology on bone cell-materials interaction. 1849 67

To evaluate the acute lung toxicity of intratracheally instilled nano-TiO2 in Kunming mice, healthy adult male Kunming mice were randomly grouped by their body weight (5 mice in each group). The lungs of mice were intratracheally instilled with 1 or 10 mg/kg x bw of nano-TiO2. The control group was intratracheally instilled with the same volume of physiological saline. After 1 d, 7 d, 14 d and 28 d of exposure, the bronchoalveolar lavage fluid (BALF) and lung tissue were collected. The indices of BALF were examined. Lung tissues were assess histopathologically. The results showed that all indices of 10 mg/kg x bw groups were obviously higher than those of the control group and the group of nano-1 mg/kg x bw, respectively. Activities of lactate dehydrogenase (LDH) on the 1st, 7th, 14th and 28th day post-exposure (pe), the amounts of malodialdehyde (MDA) on the 1st, 7th and, 14th day pe and total protein (TP) on the 1st and 7th day pe as well as the amounts of leukocyte on the 1st and 7th day pe of 10 mg/kg x bw groups were significantly different as compared with controls (P < 0.05). There were no obvious changes observed in the activity of alkaline phosphatase (ALP) within groups (P > 0.05). Histopathological examination revealed that the lungs of 10 mg/kg x bw groups presented marked increase in pulmonary inflammation. Many TiO2 particles were still clearly found in the interstitium at 28 days pe. In contrast, low-dose instillation put forward a low risk potential for producing adverse effects on pulmonary health. We conclude that the inflammatory reaction gradually ceased after 28 days. Under the same experimental condition, the effect of lung injury was severer in high-dose nano-TiO2 than in low-dose nano-TiO2.
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PMID:[Bio-effects of nano-TiO2 on lungs of mice]. 1981 15

The biological response of fetal rat calvarial cells on a TiO2 nanotubular surface (Ti-NT) was evaluated by cell viability assay, alkaline phosphatase (ALP) activity and reverse transcription polymerase chain reaction (RT-PCR) analysis. The cell viability assay showed no significant difference between the Ti-NT and smooth titanium surfaces (Ti-S). Ti-NT had better cellular responses with regard to the ALP activity and bone-associated markers, such as bone sialoprotein and osteocalcin mRNA than Ti-S. These results suggest that Ti-NT stimulate the differentiation into osteoblasts of fetal rat calvarial cells, potentially contributing to rapid osseointegration.
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PMID:The effect of nanotubular titanium surfaces on osteoblast differentiation. 2035 4


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