tissue biomarkers as predictors of response to therapy

... immunohistochemically for type II collagen as described above Isolation of total RNA Total RNA was isolated from cells cultured in a monolayer (passage 1) and from day pellets with the use of an RNeasy ... determined as the cycle number at which all samples were in the exponential phase of amplification By using the formulas below, a value (Y) was obtained as a measure of the gene expression correlated to ... magnifications of portions of (a), (d) and (g), respectively Another reason that the implant has to withstand mechanical stress is that systematic redifferentiation signalling, as part of the disease condition,...

Ngày tải lên: 09/08/2014, 06:22

... Evaluation of Human SPHK Inhibitors After the amount of DAG was optimized, the DAGK assay was carried out as a counter screening for all of the synthetic compounds Compounds, as well as DMS as a control, ... Evaluation of Human SPHK Inhibitors not specific to SPHK It was found to inhibit not only two isozymes of human SPHKs (SPHK1 & SPHK2), but other kinases such as PKC (Igarashi and Hakomori 1989; Igarashi ... minutes To terminate the reactions, 20μl of HCl (1N) was added To extract the lipids, 0.8ml of chloroform: methanol: HCl (100:200:1, v/v) mixture was added to the samples, mixed and left to stand...

Ngày tải lên: 11/09/2015, 16:06

... calculated based on the knowledge of the average mass of a base pair The calculation method is addressed below: The average mass of a base pair in DNA is about 615 dalton according to their structures ... was reverse transcribed to cDNA using MuLV reverse transcriptase RNase H (Fermentas, #EP0451) according to the manufacturer’s instructions Based on the optimized conditions, initially, RNA was ... below); a dalton (Da), named in honor of the British chemist John Dalton (1766-1844), is equivalent to 1/12 the mass of the 12C which equals 1.66053873E-27 kg, i.e., Da = atomic mass unit Average...

Ngày tải lên: 11/09/2015, 16:06

... MSCs, was measured when exogenous S1P was added into the cells Pertusis toxin (PTX) was used to study the role of S1PRs involvement in S1P triggered cell proliferation, as PTX is able to bind ... Moreover, half amount of calcium release triggered by 10μM S1P was not through its receptors in both types of cells, as 300ng/ml PTX abolished the same amount of calcium release as 100ng/ml PTX did ... when 1μM S1P was added into the culture media to promote the stem cells proliferation, what it functioned as: did it function as an extracellular mediator to bind with its receptors and promote...

Ngày tải lên: 11/09/2015, 16:06

... Conclusion and Future Work being one of the most potent and specific SPHK1 inhibitors (chapter 2), DMS was continuously used as a reference inhibitor It is interesting to note that CP6 and DMS affected ... differentiation of stem cells, for future 154 Chapter Conclusion and Future Work therapeutic applications to balance the abnormal portions of osteoblasts and adipocytes observed in bone diseases such as osteoporosis ... decreased but the adipose tissue increases (Jaiswal et al., 2000) It also suggests the potential for the use of SPHK inhibitors in preconditioning stem cells before transplanting them into the...

Ngày tải lên: 11/09/2015, 16:06

... World J Gastroenterol 10(19): 2818-22 Shu, X., W Wu, et al (2002) "Sphingosine kinase mediates vascular endothelial growth factor-induced activation of ras and mitogen-activated protein kinases." ... (1999) "Analysis of three suggested psoriasis susceptibility loci in a large Swedish set of families: confirmation of linkage to chromosome 6p (HLA region), and to 17q, but not to 4q." Hum Hered ... "Discovery and evaluation of inhibitors of human sphingosine kinase." Cancer Res 63(18): 5962-9 Friedenstein, A J., K V Petrakova, et al (1968) "Heterotopic of bone marrow.Analysis of precursor cells...

Ngày tải lên: 11/09/2015, 16:06

... in cardiovascular system Interestingly, while S1PR1 has been shown to promote cell survival/proliferation, S1PR2 has been shown to activate stress-associated kinases leading to apoptosis (Kluk ... research As the 3-D structure of SPHK remains unknown, it has been difficult to design targetspecific inhibitors, or structurally-based inhibitors In order to develop specific SPHK inhibitor, some ... hyper-proliferative role of SPHK and S1P has been proposed to contribute to the early stages of diabetic nephropathy (Katsuma et al., 2002 and 2003; Geoffroy et al., 2004) In recent reports, streptozotocin-induced...

Ngày tải lên: 11/09/2015, 16:06

... this activation occurs as a response to DNA damage or as a response to alterations to chromatin structure (which activate ATM) is open In vivo studies that measure the impact of ATM mutations on ... are required to suppress the checkpoint responses to damaged DNA This would be an exciting class of mutations to explore, since restoration of checkpoints or targeting specific apoptotic pathways ... to a dramatic loss of proliferative capacity of such cells (Knoepfler et al 2002) As long as questions of redundancy between different members of the myc gene family and compensation by loss of...

Ngày tải lên: 25/10/2013, 21:20

... Proliferation of lung fibroblasts was assayed with a kit from Roche (Indianapolis, IN, USA) that monitors the incorporation of BrdU into newly synthesized DNA The BrdU was detected using anti-BrdU-peroxidase ... incorporation of 5-bromo-2¢-deoxy-uridine (BrdU) into the cells was assayed As shown in Fig 1A, incubation of HFL-1 lung fibroblasts with a-defensin-1 induced dose-dependent increases in BrdU incorporation, ... specific inhibitor of PI3K, for 1–24 h, after which the protein contents of phosphorylated p38 MAP kinase, total p38 MAP kinase, phosphorylated Akt, total Akt, phosphorylated GSK3b, total GSK3b,...

Ngày tải lên: 18/02/2014, 06:20

... consequence of cell rounding, the action of cytochalasin D, an inhibitor of actin polymerization, was studied on GC cultured on LN substratum Addition of cytochalasin D to the culture medium of GC ... specific role of LN-induced cytoskeleton changes on GC functions to be assessed In our experiments, the observed effects of cytochalasin were likely to be the consequence of both cytoskeleton disruption ... 20 0.1 Cytochalasin D ( g/ml) 10 b 60 a c (c) a a a 0.1 a 10 Cytochalasin D ( g/ml) Figure cytochalasin D on morphology of GC cultured on LN substratum Effect of2 Effect of cytochalasin D on...

Ngày tải lên: 05/03/2014, 17:20

... effect of DHEA upon CASKI cells is more cytotoxic than cytostatic Table Percentage of cells in each phase of the cell cycle as evaluated by flow cytometry Percentage of cells in the phases of the ... experiment representative of three independent assays as well as apoptotic degradation, the type of cell death was determined using fluorochrome-labeled inhibitors of caspases (FLICA) and propidium ... Histograms show the percentage of cells in each phase of the cell cycle as evaluated by flow cytometry (see Experimental procedures) The percentage of cells in each phase of the cell cycle was...

Ngày tải lên: 16/03/2014, 00:20

... of the peptide hormone apelin to decrease the activation of caspase 9, as well as caspases and In mouse osteoblasts, this inhibition of caspase activity and the apoptotic activity induced by serum ... members of the Bcl2 family at the mitochondrial membrane, inducing the release of cytochrome c from the mitochondria and triggering caspase-mediated apoptosis Akt phosphorylation of Bad leads to its ... of a number of cell cycle regulatory proteins [11], and its ability to inactivate pro-apoptotic factors, such as Bad, caspase-9 and forkhead (FH) transcription factors [12] In contrast, it is...

Ngày tải lên: 16/03/2014, 05:20

... response to mitogen stimulation (Fig 7E) In contrast with MAP kinase, Rho mediates growth factor-dependent activation of c-fos via the serum response factor (SRF) [30] Protein kinase C-related kinase ... however, the increase in cytosolic activity was only marginal (Fig 3B) Activation of arginine-dependent mART would be expected to lead to an increase in the post-translational modification of select proteins ... effect of a selective inhibitor of poly(ADP-ribosyl)ation, PD128763 [24], on the stimulation of DNA synthesis by mitogen (2% fetal bovine serum) PD128763 was unable to prevent the increase in...

Ngày tải lên: 17/03/2014, 09:20

... an epitope in its cytoplasmic domain Increased expression of E-cadherin and tissue inhibitor of matrix metalloproteinase (TIMP2) by DPIV-transfected cells has been reported [22] b-Catenin associates ... dipeptidyl peptidase IV enhances expression of topoisomerase II alpha and sensitivity to apoptosis induced by topoisomerase II inhibitors Br J Cancer 89, 1366–1374 34 Aldinucci D, Poletto D, Lorenzon ... percentages of GFP-expressing cells by use of flow cytometry [38] In vitro wound-healing assay The wound-healing assay was performed as described [12] 293T cells were plated onto plastic coated...

Ngày tải lên: 23/03/2014, 11:20

... Caspase-dependent effects in neuroblast apoptosis changes and a decrease in the level of prenylation of Ras and RhoA proteins [12] Rho small GTPases are important regulators of the actin cytoskeleton ... cleavage of PKCd during lovastatin-induced neuroblast apoptosis is completely dependent of caspase activity However, our results suggest that activation of other caspases different than caspase-3 ... B Caspase-dependent effects in neuroblast apoptosis apoptosis induced by lovastatin [12] Therefore, to test the ability of mevalonate at preventing the effect of lovastatin on FAK and p130Cas...

Ngày tải lên: 23/03/2014, 11:20

... low-grade dysplasias, and strongly positive in high-grade dysplasias In invasive squamous cervical carcinomas, cytoplasms of basal-type tumour cells were negative, while squamous-type dysplastic cells ... and the effect of the phase of cell cycle, differentiation and apoptosis on expression of this gene was also studied Furthermore, evidence was presented for the association of PP17b to lipid droplets ... adipocytes [22], while a tissue- specific distribution of PP17 variants was also discovered, as PP17b was ubiquitously expressed, while PP17a expression was restricted to steroidogenic tissues only [5];...

Ngày tải lên: 23/03/2014, 21:20

... The thickness of the interphases may be as great as 10 pm, but Voyutskii and Vasenin argue that layers as thin as 1-2 nm should be sufficient to produce order -of- magnitude increases in joint ... between the phases Such an ‘interphase’ has a structure, and possibly a composition, distinct from that of either of the bulk phases Recognizing the ubiquitous presence of interphases, the following ... the extra tasks associated with the completion of their contribution to this volume We would also like to acknowledge the contribution of Mr Theodore Reinhart of the University of Dayton Research...

Ngày tải lên: 26/04/2014, 15:36

... motility and invasion was examined in CRC cell lines that were subjected to CK2a knockdown or to the CK2a activity inhibitor emodin Further analysis was conducted to elucidate the mechanisms of CK2a ... [mean: 50.5]) consisted of serrate adenomas, 22 canalicular adenomas, villous adenomas, and tubulovillous adenomas Immunohistochemistry Immunohistochemical staining was performed using a Dako ... http://www.translational-medicine.com/content/9/1/97 Tumor cell invasion assay Warm serum-free medium was added to the top chamber of the cell invasion chamber (Chemicon, Temecula, CA, USA) to rehydrate the ECM layer for h at...

Ngày tải lên: 18/06/2014, 19:20

... colorimetric assay for measuring cellular growth It has been developed by Mosmann It can also be used to determine cytotoxicity of toxic materials Yellow MTT is reduced to purple formazan in the mitochondria ... opportunity for cytological detection of DNA interchange This technic is used as a sensitive means of monitoring DNA damage It is useful for assessing the cytogenic impact of clastogenic agents ... proliferation) sensitivity to borax toxicity was based on the cytotoxicity index (CI) The percent of cytotoxicity index (%CI = [1-OD570 treated/OD570 control] × 100) was calculated for each borax concentration...

Ngày tải lên: 20/06/2014, 00:20