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Development of sphingosine kinase (SPHK) inhibitors and the role of sphingolipids in adult stem cell proliferation and differentiation 2
... mixed and left to stand at room temperature for 5-10 minutes The organic and aqueous phases were separated by adding 240μl of chloroform and 240 μl of 2N KCl The mixture was then vortexed, and ... cells were pretreated with 10μM of each of the synthesized compounds, or with 10μM of DMS, for 30 minutes prior to stimulation Cells were then stimulated with 5nM of C5a and warmed to 37°C for ... phosphorylated and non-phosphorylated peptides were separated by standard electrophoresis, corresponding bands can be excised for quantification by spectrophotometry, densitometry or spectrofluorometry...
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Development of sphingosine kinase (SPHK) inhibitors and the role of sphingolipids in adult stem cell proliferation and differentiation 3
... differentiation in the presence of 1μM of DMS cultured for 14 or 28 days, or in the presence of 0.5μM of CP6 for 28 days, or 1μM of CP6 for 14 or 28 days, or with 5μM of CP6 for 14 days, there was no ... differentiation media together with 0.5μM of DMS for 14 or 28 days, with 1μM of DMS for 14 or 28 days, with 0.5μM of CP6 for 14 or 28 days, or with 1μM of CP6 for 14 days, the osteocalcin expression ... the stem cells growth at amounts of 10μM or higher However, for human AD-MSCs (Figure 3.2B), DMS showed to inhibit stem cells growth significantly only when at amounts of 5μM or higher, and CP6...
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Development of sphingosine kinase (SPHK) inhibitors and the role of sphingolipids in adult stem cell proliferation and differentiation 4
... four days and cells were cultured for 12 days After that, cells were counted, and equal amount of cells were seeded in 48-well plates for 24 hours for 141 Chapter Role of S1P in human BM- and AD- ... the stem cells proliferation, what it functioned as: did it function as an extracellular mediator to bind with its receptors and promote cells proliferation? Or it penetrated into the cells and ... receptors and shown to bind/activate Gi subunit (Section 1.1 SPHK and S1P) 144 Chapter Role of S1P in human BM- and AD- MSCs proliferation Cells were pre-treated or not with 100ng/ml and 300ng/ml of...
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Development of sphingosine kinase (SPHK) inhibitors and the role of sphingolipids in adult stem cell proliferation and differentiation 5
... for a more controlled and facilitate differentiation of stem cells, for future 154 Chapter Conclusion and Future Work therapeutic applications to balance the abnormal portions of osteoblasts and ... types of stem cells Also, it should be noticed that both human BMand AD- MSCs are primary cells, and thus they may possess certain variations between different donor cells Therefore, more work ... expanded to study other stem cells and other differentiations pathways, to further validate and confirm the potential usage of SPHK inhibitors in stem cell differentiation, and S1P roles on stem...
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Development of sphingosine kinase (SPHK) inhibitors and the role of sphingolipids in adult stem cell proliferation and differentiation 6
... (2007) "Whole genome analysis of human neural stem cells derived from embryonic stem cells and stem and progenitor cells isolated from fetal tissue." Stem Cells 25(5): 1298-306 Shmelkov, S V., S Meeus, ... 590-2 Safford, K M and H E Rice (2005) "Stem cell therapy for neurologic disorders: therapeutic potential of adipose-derived stem cells. " Curr Drug Targets 6(1): 57-62 Safford, K M., S D Safford, ... growth factor in the maintenance of human embryonic stem cells. " Stem Cells 23(10): 1541-8 Peng, H and J Huard (2003) "Stem cells in the treatment of muscle and connective tissue diseases." Curr...
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Development of sphingosine kinase (SPHK) inhibitors and the role of sphingolipids in adult stem cell proliferation and differentiation
... applications and significance of using SPHK and S1P in stem cells research, derived from the current understanding on human stem cells and the strategies used for stem cell expansion and differentiation, ... mediated proliferation and survival of murine mesoangioblasts A more recent study, by Pébay et al (2005), investigated the roles of S1P and SPHK in human stem cells and showed that for human ... SPHK and/ or S1P in stem cell proliferation and multipotency maintenance It is well known that in stem cells, there is a balance between cell proliferation and differentiation, which directs the cells...
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Effects of high glucose concentrations on the expression of genes involved in proliferation and cell fate specification of mouse embryonic neural stem cells
... essential for the growth of both dorsal and ventral regions of the diencephalon and anterior midbrain regions (Britto et al., 2002; Ishibashi and McMahon, 2002) In addition, removal of notochord in ... roof plate cells of the neural tube (Liem, Jr et al., 1995) The role of BMPs signaling in the dorsal patterning of the neural tube has been widely reported It has been reported that dorsal and ... equivalent of early stage of human anencephaly) (Harris and Juriloff, 1999) In this case, the brain development ceases and the vault of the skull fails to form Failure of the caudal neuropore to...
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Báo cáo khoa học: Hexadecylphosphocholine inhibits phosphatidylcholine biosynthesis and the proliferation of HepG2 cells pot
... biosynthesis of PC by using choline as exogenous precursor Cells were incubated for h with [methyl-14C]choline either in the presence or absence of HePC HePC inhibited the incorporation of choline into ... the tumoral hepatic cell line HepG2, HePC hinders the incorporation of radiolabelled choline into PC, the endproduct of the CDP–choline pathway This reduction in the incorporation of choline was ... chloroform layer was dried by evaporation with a stream of nitrogen whilst the water-methanol phase was collected and used to determine water-soluble metabolites We analysed the incorporation of...
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Báo cáo khoa học: A comparative analysis of the transcriptome and signal pathways in hepatic differentiation of human adipose mesenchymal stem cells doc
... maternal origin from human placenta Stem Cells 22, 1338–1345 Bieback K, Kern S, Kluter H & Eichler H (2004) Critical parameters for the isolation of mesenchymal stem cells from umbilical cord blood Stem ... of AT-MSCs, without the necessity for co-culture with fetal or adult hepatocytes We have shown that our system induced transplantable cells with morphological and functional characteristics of ... ethics, cost and time Considerable emphasis has therefore been placed on the development of in vitro test systems Although primary human hepatocytes are the best source of cells for such systems, their...
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Báo cáo khoa học: "Functional recovery and neural differentiation after transplantation of allogenic adipose-derived stem cells in a canine model of acute spinal cord injury" potx
... assessment of cord swelling and hemorrhage, and T2 weighted images were valuable for assessment of fluid infiltration into the cord, i.e edema [24] Cord swelling occurs due to disruption of vasculature ... Ireland) and saline at a dose of 40 μL/kg body weight for 12 h It took approximately 30 Fig Green fluorescence protein (GFP) labeled canine adipose-derived stem cells (ASCs) (A) Typical morphological ... medians for Olby scores and the means ± SD for SEP values and Luxol fast blue positive areas Statistical analysis used SPSS 12.0 software (SPSS, USA) Kruskal-Wallis analysis for Olby scores and oneway...
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Báo cáo khoa học: "Herpes simplex virus type 2 tegument protein UL56 relocalizes ubiquitin ligase Nedd4 and has a role in transport and/or release of virions" ppt
... UL56 or gG showed little or no localization to late endosome/MVBs Therefore, our finding Figure ons A model 7of maturation, transport and release of HSV-2 viriA model of maturation, transport and ... localization of Nedd4-GFP and mRFP-UL56 was not stable; the pattern of overlap changed rapidly, and even merge or separation of the two proteins was observed (Fig 1E-3 and 1E-4) Nedd4-GFP and a part of ... 5B) If HSV-2 exploited MVBs and/ or late endosomes for assembly or release of virions, and if Nedd4 and UL56 were involved in the process, a greater amount of Nedd4 and UL56 should colocalize...
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Báo cáo y học: " Gene expression profiling of human prostate cancer stem cells reveals a pro-inflammatory phenotype and the importance of extracellular matrix interactions" pps
... data analysis and design SL performed the stem cell isolations from benign samples PB performed stem cell isolations from tumors CH performed culturing and isolation of stem cells, and NF-κB experiments ... the stem cell population TJP1 (ZO-1) and TJP2 (ZO-2) proteins were also expressed by the majority of progenitor and stem cells from tumor cell cultures (Figure 4c,d), whereas only a minority of ... populations before fractionation according to [4] Positive controls for invasive capacity were cell lines MCF7 and PC3M whose invasion score was 18-36%, whereas normal cell lines PNT2 and PNT1a and BPH/primary...
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Generation of porcine induced pluripotent stem cells and their differentiation into cardiac lineages
... differentiation of Human IPS cell lines 27 3.5 Generation of Porcine Induced Pluripotent Stem Cells 29 3.5.1 Factors used for reprogramming of porcine cells 29 3.5.2 Reprogramming techniques of porcine ... recording of IPS derived cardiomyocytes 56 5.5 Culture of porcine fibroblast cells 57 5.6 Reprogramming process of porcine fibroblast cells 58 5.6 Formation of Porcine IPS colonies and ... Morphology of porcine IPS cells 89 6.2 Expression of pluripotent genes in porcine IPS cells 89 6.3 Function of LIF in the culture of porcine IPS cells 90 6.4 Importance of doxycycline in the...
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Báo cáo khoa học: Lipid droplet and milk lipid globule membrane associated placental protein 17b (PP17b) is involved in apoptotic and differentiation processes of human epithelial cervical carcinoma cells pptx
... factors, AP-2, GCF and PAX-2 [49]; (h) factors abundant in placenta, AHR [50], AP-2 and PPARc; (i) proliferation and/ or apoptosis regulators, AP-2, c-MYC [51] and NF-jB; (j) embryo- and organogenic ... apoptosis and differentiation of HeLa cells (Eur J Biochem 270) 1185 Fig PP17 immunostaining of apoptotic and differentiated HeLa cells A, C, E and G show control cells, B, D, and F paclitaxel (10 nM for ... the transcription of PP17 gene include: (a) general activators or repressors GCF, SP-1, YY1 [34] and USF [35]; (b) coactivators, AP-4 [36] and P-300 [37]; (c) cAMP/PKA, PKC or phorbol ester responsive...
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báo cáo hóa học:" Human fallopian tube: a new source of multipotent adult mesenchymal stem cells discarded in surgical procedures" potx
... purposes) Journal of Translational Medicine 2009, 7:46 Immunofluorescence and Western Blot Analysis Immunofluorescence (IF) Immunofluorescence localization of dystrophin was performed on muscle-differentiated ... Therefore, based on the experience of our research group in the identification, and characterization of potential sources of adult stem cells [7,10,1215], the aim of this study was to isolate, expand, ... growth and regeneration, in order to provide the unique environment required for the maintenance of male and female gamete viability, fertilization, and early embryo development as well as transport...
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Báo cáo sinh học: "Autologous Transplantation of Adipose-Derived Mesenchymal Stem Cells Markedly Reduced Acute Ischemia-Reperfusion Lung Injury in a Rodent Model" potx
... for each animal was then determined by summation of all numbers divided by Immunofluorescent (IF) Studies and Crowded Score of Lung Parenchyma IF staining was performed for the examinations of ... YHK, and YCL were responsible for the laboratory assay and troubleshooting SC, THT, MF, and SFK participated in data acquisition, analysis, and interpretation SL and HKY conceived of the study, and ... injury Moreover, immunofluorescent staining identified substantially higher number of infiltrated CD68+ cells (inflammatory cells of macrophages) in injured lung parenchyma in IR group than in normal...
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Báo cáo hóa học: " Impacts of Post-metallisation Processes on the Electrical and Photovoltaic Properties of Si Quantum Dot Solar Cells" potx
... The cells investigated in this work have areas in the range 2–10 mm2 Removal of localised Aluminium shunts Fig Normalised photoluminescence for Si QDs of various sizes in SiO2 matrix [1] solar cells ... existence of localised Al shunts lying across the p-type and n-type regions as the annealing improves the contact of the Al shunts to both p- and n-regions of the cell thus shorting the cell more effectively ... is in the order of MX.cm2 The short circuit current increases by a factor of three due to the decrease of Rs Contact and Sheet Resistances To identify the origin of the large series resistance,...
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báo cáo khoa học: "Bostrycin inhibits proliferation of human lung carcinoma A549 cells via downregulation of the PI3K/Akt pathway" pdf
... of medium (for the control group) and incubated for 24, 48 or 72 hours Culture supernatant from each group was pooled and the cells were fixed for 12 h with ml of 75% ethanol (106 cells/ ml) and ... microRNA-638 and microRNA-923 in A549 cells treated with bostrycin detected by microRNA real time PCR A549 cells were treated with 10 μM Bostrycin for 72 h and total RNA was isolated for microRNA ... expression of microRNAs Preparation of total RNA sample A549 cells were cultured in 6-well plates (1.5 × 105 cells per well) and treated for 72 h with 10 μmol/L bostrycin for the bostrycin group or with...
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báo cáo khoa học: "Effect of anti-CD52 antibody alemtuzumab on ex-vivo culture of umbilical cord blood stem cells" doc
... mature cells and lymphocytes were actively removed in culture with the maintenance of hematopoietic progenitors and stem cells, and a relative expansion of myeloid and megakaryocytic precursors; ... incorporate the expansion of http://www.jhoonline.org/content/1/1/19 one unit of cord blood following the infusion of another unexpanded unit in a double cord blood transplant setting The unexpanded ... Secondary culture for day 35 CD34+ cells To study the long term effects of alemtuzumab on hematopoietic progenitors and stem cells, cells were cultured for up to 35 days and CD34+ cells were isolated...
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