Quantitative phosphoproteome analysis for the detection of early alterations of signal transduction by dioxin (TCDD) Melanie Schulz, Ulrich Andrae, Martin R. Larsen2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is an extremely toxic pollutant and regarded the most potent chemical carcinogen in experimental animals. Most of its biological effects are mediated by binding of TCDD to the cytosolic Ah receptor (AHR). In addition, TCDD cause AHR-dependent alterations in signal transduction that are independent of changes in gene expression. In this study, we have therefore conducted a quantitative phospho proteomic study on TCDD-induced alterations. |  | |
Sialylation and Metastasis: from tumor-associated antigens discovery to therapeutic development Giuseppe Palmisano, Rikke Leth-Larsen and Martin R. LarsenAberrant sialylation affects the metastatic behaviour of cancer cells.We have quantified the sialylated cell surface glycoproteins from cells with different metastatic behaviour.The membrane proteins from isogenic human cell lines were analyzed by a method based on TiO2-HILIC-MS.1200 sialylated glycosites, 116 enzymes involved in the glycoconjugate metabolism and for the first time 4 glycosylation sites of Her2 receptor were identified, showing the importance of this tool for discovery and thera |  | |
Proteomic non-small cell lung carcinoma biomarker screening in bronchoalveolar lavage fluid Tonio Oumeraci 1, Bernd Schmidt 3, Thomas Wolf 1,4, Marc Zapatka 4, Andreas Pich 2, Benedikt Brors 4, Roland Eils 4,5, Brigitte Schlegelberger 1, Nils von Neuhoff 1Using a standardized method to acquire MALDI-TOF proteome profile spectra of bronchoalveolar lavage fluid (BALF), we have shown the upregulation of histatin 3 and calgranulin C in a small pilot cohort of NSCLC patients. This pilot study serves to demonstrate that it is feasible to screen a larger NSCLC patient cohort for BALF proteome level biomarkers in a clinical setting. |  | |
Some thiazole derivatives as potent antiproliferative agents with CDK2/Cycin E/A inhibitory activities Mahesh Chhabria1, Shailesh Patel*1, Maulik Suthar2We report synthesis and biological evaluation of novel thiazole derivatives. High throughput screening of various compounds for their anti-proliferative activity led to the identification of thiazolo-thiones as potential candidate for Lung cancer (NCl-H23, NClH510A, NCI-H522) and breast cancer (MDA-MB-453/231/468, MCF-7). The most potent compounds were also screened for their inhibitory activity against CDK2/ Cyclin E/A which are considered as promising targets for lung and breast cancer. |  | |
Predicting hepatotoxicity: Reactive metabolite trapping using glutathione and freshly isolated hepatocytes Birks, V., Webber, G., Geoffroy, S., Cole, R., and Wood, S.This poster presents our results to date using clozapine (a compound known to be associated with GSH-adduct formation) as substrate and using stable isotope GSH (GSH13C2,15N) to enhance specificity. In addition, all analyses have been conducted using an Waters Acquity UPLC-MS/MS. Results we have obtained in hepatocytes are compared against findings using human liver microsomes (HLM). |  | |
A simple, fast and quantitative single-step dead-cell indicator for flow cytometry Jixiang Liu, Jolene Bradford, Chris LangsdorfWe have evaluated a series of new compounds for dead cell stain and identified a new product, SYTOX® AADvancedTM dead cell stain, which demonstrates improved properties over 7-AAD. These properties make the SYTOX® AADvancedTM dead cell stain a simple, fast and quantitative single-step no-wash dead-cell indicator as well as ideal for use in multicolor application requiring DNA content. |  | |
Protein array-based screening of autoantibody signatures Zingaretti C., Arigò M., Colombatto P., Brunetto M., Muratori L., Bonino F., Bianchi F., Pagani M., De Francesco R., Abrignani S., Bombaci M.The evidence for an association between autoimmune diseases and chronic HCV infection has been clearly established. To this aim, a protein array was employed to analyze serum samples of HCV patients w/wo autoimmune complications, of patients with autoimmune hepatitis but not infected with HCV and of healthy donors as controls. A panel of autoantigens able to discriminate among the three groups of patients was identified for potential use as biomarkers. |  | |
PROTEOME WIDE PLASMA PROFILING USING ANTIBODY SUSPENSION BEAD ARRAYS Maja Neiman, Ulrika Igel, Burcu Ayoglu, Kimi Drobin, Mathias Uhlén, Peter Nilsson and Jochen M. SchwenkA newly developed antibody suspension bead array assay allows for a systematic and high-throughput plasma profiling. This microtiter based assay uses antibody-coupled beads for a multiplexed analysis of minute amounts of directly labelled samples. The key requirement of a?nity reagents towards all human proteins is met by the Human Protein Atlas project. |  | |
Five noncovalent peptidic ligands show different affinity rankings in solution and gas phase Andrey Dyachenko , Michael Goldflam , Marta Vilaseca , Ernest Giralt Stability of noncovalent complexes of VEGF protein with 5 peptidic ligands is studied. Experiments were conducted in solution (NMR CSP, ITC) and in gas phase (CID TOF MS). Each ligand differs from others in chirality of one amino acid. It was shown, that trend of stability of the studied noncovalent complexes is reversed in the gas phase relatively to the solution. An explanation of this behavior is presented. |  | |
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