Project description:Taking the advantage that B16F10 cells retain the wild-type p53, we introduced the p53 partner into these cells, the p19Arf. Also, in order to induce a immune stimulation in experiments *in vivo*, we introduced the interferon-beta cDNA. This combination induced several benefits when compared to the use of these factors alone, as seen by propidium iodide staining, tunel staining, several gene expression analysis, tumor growth, mice survival, among others. Because of this, we had the interest to study the effects of the combination of p19Arf plus interferon-beta upon gene regulation of B16F10 cells, compared to the effects of these transgenes alone. Previous works from our group that show several benefits of p19Arf plus interferon-beta combination upon B16F10 cells are described in Merkel et al (2010) doi: 10.1186/1471-2407-10-316, Merkel et al (2013) doi:10.1038/cgt.2013.23 and Medrano et al (2016) doi: 10.1007/s00262-016-1807-8 .
Project description:Fungal effectors play important roles in inciting disease development on host plants. We identified an effector (Secreted in Xylem4, SIX4) in an Arabidopsis infecting isolate (Fo5176) of the root-infecting fungal pathogen Fusarium oxysporum and demonstrated this effector is required for full virulence. To explore the role of Fo5176_SIX4 we use whole transcriptome profiling of root tissues from plants overexpressing this effector (35sSIX4) versus wild-type (Col-0) plants after F. oxysporum infection. Published in DOI:10.1007/978-3-319-42319-7_4. Belowground Defence Strategies in Plants.
Project description:A functional genetic screen to identify genes causing tamoxifen resistance in an estrogen-dependent human breast cancer cell model was performed. By insertion of defective retrovirus into the genome, individual genetic changes were introduced at random in ZR-75-1 cells. Subsequently, infected cells were selected for their ability to proliferate while being exposed to tamoxifen, and from these cultures stable cell lines were established. The retrovirus insertion sites were mapped enabling the identification of several candidate breast cancer anti-estrogen resistance (BCAR) genes. By cDNA transfection of estrogen-dependent cells resulting in their transformation into a tamoxifen-resistant phenotype, proof was provided for the causative role of BCAR genes.A functional genetic screen to identify genes causing tamoxifen resistance in an estrogen-dependent human breast cancer cell model was performed. By insertion of defective retrovirus into the genome, individual genetic changes were introduced at random in ZR-75-1 cells. Subsequently, infected cells were selected for their ability to proliferate while being exposed to tamoxifen, and from these cultures stable cell lines were established. The retrovirus insertion sites were mapped enabling the identification of several candidate breast cancer anti-estrogen resistance (BCAR) genes identified. By cDNA transfection of estrogen-dependent cells resulting in their transformation into a tamoxifen-resistant phenotype, proof was provided for the causative role of BCAR genes (Van Agthoven et al, Breast Cancer Res Treat DOI:10.1007/s10549-008-9969-5, 2008). To elucidate the mechanisms how these individual BCAR genes induce cell proliferation in growth-arrested ZR-75-1 cells, we assessed the gene expression patterns between the different groups of cell lines. Keywords: Expression profiling, reference design RNA was isolated from two independent cultures of 69 tamoxifen-resistant cell lines. Detailed information regarding these cell lines with respect to the viral integration sites, was described previously (Van Agthoven et al, Breast Cancer Res Treat DOI:10.1007/s10549-008-9969-5, 2008). Each sample was hybridized in duplicate/triplicate and once in a dye-swap. The mixture of cell lines used as a reference was detailed previously (Jansen et al, J Clin Oncol 23, 732, 2005).
Project description:Ex vivo colorectal adenocarcinoma samples were analysed by desorption electrospray ionisation using an Orbitrap mass spectrometer. The samples were analysed in negative mode over the m/z range 600-1000. Some of the samples presented in this dataset were analysed as part of [1]. The study makes available imzML files of profile and centroid mode data, as well as low- and high- resolution optical image files of H&E-stained sections. These files can be found in the zip file named after each tissue section. </p> Ref:</br> [1] Gerbig S, Golf O, Balog J, Denes J, Baranyai Z, Zarand A, Raso E, Timar J, Takats Z. Analysis of colorectal adenocarcinoma tissue by desorption electrospray ionization mass spectrometric imaging. Anal Bioanal Chem. 2012 Jun;403(8):2315-25. doi:10.1007/s00216-012-5841-x. PMID:22447214</br>
Project description:Established that 11 human anti-Plasmodium vivax Duffy Binding protein II monoclonal antibodies are not cross reactive with other plasmodium antigens as represented by PfPv500.1 array.
Project description:Aguilar-Toala 2019: Investigators under Andrea Liceaga at Department of Food Science at Purdue University examined the proteomes of chia seeds for their anti-microbial properties and enzyme inhibition [doi:10.1007/s00726-020-02879-4 and doi:10.1007/s12602-020-09653-8]. They followed activity for different size exclusion chromatography fractions of peptides generated through alcalase and flavourzyme digestion. Tandem mass spectra were generated on a Q Exactive Plus by Emma Doud in the Proteomics Core at the Indiana University School of Medicine. The LC-MS/MS experiment representing proteins below 3 kDa was emphasized because it represented the biological activity they sought; the instrument produced 44,208 tandem mass spectra at a rate peaking near 11 Hz. When sequence availability for chia proved to be problematic, Doud made use of PEAKS Studio to infer sequences directly from the fragment ions.
Project description:Insulin analogues are designed to improve the pharmacokinetic parameters compared to regular human insulin. This provides a sustained control of blood glucose levels in diabetic patients. All novel insulin analogues are tested for their mitogenic side effects, however these assays do not take into account the molecular mode-of-action of different insulin analogues. Insulin analogues can bind the insulin receptor (INSR) and the insulin-like growth factor-1 receptor (IGF1R) with different affinities and consequently will activate different downstream signaling pathways. Here we used a panel of MCF7 human breast cancer cell lines that selectively express either one of the isoforms of the INSR (IRA or IRB) or the IGF1R. We sought to study the role of the different receptors (IRA, IRB and IGF1R) in the mitogenic signaling of insulin-like molecules (including insulin, glargine, X10 (or AspB10) and IGF1). MCF7 IRA, MCF7 IRB or MCF7 IGF1R cells (as described in Arch Toxicol. 2014 Apr;88(4):953-66. doi: 10.1007/s00204-014-1201-2. Epub 2014 Jan 25.) were cultured in RPMI supplemented with 5% (v/v) CDFBS (Hyclone) and used for experiments. Cells have been exposed for 1 or 6 hours to 10 nM of the indicated insulin-like molecule. As a control sample a vehicle stimulation was performed that contained everything except the active compound.