The effect of TFF1 and copper on <i>H. pylori</i> colonization of AGS-AC1 cells.

<p>AGS-AC1 cells, induced and uninduced, were infected with wild type strain (P12), or mutant strain P12ΔHP1191. The infections were carried out in the presence of Cu 100 µM or BCS 500 µM. Adherence was evaluated in comparison to control untreated AGS-AC1 cells. <b>a:</b> Effects of TFF1 and copper on <i>H. pylori</i> P12 cell adhesion. <b>b:</b> Effects of copper on <i>H. pylori</i> P12 cell adhesion. <b>c:</b> Effects of TFF1 and/or BCS on <i>H. pylori</i> P12 cell adhesion. <b>d:</b> Effects of TFF1 and/or copper on <i>H. pylori</i> P12ΔHP1191 cell adhesion.</p

SPR analysis of <i>H. pylori</i> RF-LPS binding to TFF1 C-terminus.

<p>The biotinylated synthetic peptides are composed of the last 16 aa of native TFF1. Panel A, B, C show the analysis of peptide monomers, dimers and scrambled monomers respectively challenged with <i>H. pylori</i> RF-LPS. Panels D, E, F show the analysis of peptide monomers, dimers and scrambled monomers respectively challenged with <i>E. coli</i> RF-LPS.</p

The effect of copper on mucus thickness in HT29-E12 cells.

<p><b>a</b>: Alcian Blue/Neutral Red staining of HT29-E12 cells grown on trans-well filters. Cells were treated or not treated (control) with CuCl₂ 10 µM, 48 hours. Staining shows the mucous secretions (blue) and nuclei of the cell monolayer (red). <b>b</b>: The graph shows the ratio between the areas of mucous secretions and nuclei, in control and treated samples. Values represent means +/− SD. <b>c</b>: Immunofluorescence analysis of TFF1 and MUC5AC expression in HT29-E12 cells cultured on trans-well filters. Staining of filter cross sections are shown. a, b: anti-TFF1 antibody. c,d: anti-MUC5AC antibody.</p

The interaction of <i>H. pylori</i> with AGS-AC1 cells.

<p><b>a</b>: Immunofluorescent staining of <i>H. pylori</i> colonized AGS-AC1 cells. Induced and not induced cells were infected with <i>H. pylori</i>. Blue: DAPI staining for nuclei; Red: <i>H. pylori</i>; Green: TFF1. <b>b</b>: CagA phosphorylation following <i>H. pylori</i> colonization of AGS-AC1 cells. Western immunoblot was probed with anti-CagA and anti-phosphotyrosine antibodies. Lane 1: <i>H. pylori</i> P12 bacterial cell lysate probed with anti-CagA. Lanes 2–5 were probed with anti-phosphotyrosine antibodies. Lane 2: <i>H. pylori</i> P12 bacterial cell lysate; Lane 3: AGS-AC1 cells colonised with <i>H. pylori</i> P12; Lane 4: whole cell lysate of induced AGS-AC1 cells colonized with <i>H. pylori</i>; Lane 5: AGS-AC1 whole cell lysate. <b>c:</b> IL-8 production by uninduced and induced AGS-AC1 cells upon colonization by <i>H. pylori</i>.</p

The effect of copper on <i>H. pylori</i> colonization of HT29-E12 cells.

<p>Colonization of HT29-E12 cells by <i>H. pylori</i> P12 in the presence of excess copper (Cu 100 µM; 2, 4, and 24 hr) (<b>a</b>) and in copper depleted conditions (BCS 500 µM; 24 hr) (<b>b</b>).</p

Structural Evidence of <i>N</i>6‑Isopentenyladenosine As a New Ligand of Farnesyl Pyrophosphate Synthase

<i>N</i>6-isopentenyladenosine (i6A), a modified nucleoside belonging to the cytokinin family, has shown in humans many biological actions, including antitumoral effects through the modulation of the farnesyl pyrophosphate synthase (FPPS) activity. To investigate the relationship between i6A and FPPS, we undertook an inverse virtual screening computational target searching, testing i6A on a large panel of 3D protein structures involved in cancer processes. Experimentally, we performed an NMR investigation of i6A in the presence of FPPS protein. Both inverse virtual screening and saturation transfer difference (STD) NMR outcomes provided evidence of the structural interaction between i6A and FPPS, pointing to i6A as a valuable lead compound in the search of new ligands endowed with antitumoral potential and targeting FPPS protein

Identification of Small-Molecule Enhancers of Arginine Methylation Catalyzed by Coactivator-Associated Arginine Methyltransferase 1

Arginine methylation is a common post-translational modification that is crucial in modulating gene expression at multiple critical levels. The arginine methyltransferases (PRMTs) are envisaged as promising druggable targets, but their role in physiological and pathological pathways is far from being clear due to the limited number of modulators reported to date. In this effort, enzyme activators can be invaluable tools useful as gain-of-function reagents to interrogate the biological roles in cells and in vivo of PRMTs. Yet the identification of such molecules is rarely pursued. Herein we describe a series of aryl ureido acetamido indole carboxylates (dubbed “uracandolates”), able to increase the methylation of histone (H3) or nonhistone (polyadenylate-binding protein 1, PABP1) substrates induced by coactivator-associated arginine methyltransferase 1 (CARM1), both in in vitro and cellular settings. To the best of our knowledge, this is the first report of compounds acting as CARM1 activators