A Novel Functional Domain of Tab2 Involved in the Interaction with Estrogen Receptor Alpha in Breast Cancer Cells

Tab2, originally described as a component of the inflammatory pathway, has been implicated in phenomena of gene de-repression in several contexts, due to its ability to interact with the NCoR corepressor. Tab2 interacts also with steroid receptors and dismisses NCoR from antagonist-bound Estrogen and Androgen Receptors on gene regulatory regions, thus modifying their transcriptional activity and leading to pharmacological resistance in breast and prostate cancer cells. We demonstrated previously that either Tab2 knock-down, or a peptide mimicking the Estrogen Receptor alpha domain interacting with Tab2, restore the antiproliferative response to Tamoxifen in Tamoxifen-resistant breast cancer cells. In this work, we map the domain of Tab2 responsible of Estrogen Receptor alpha interaction. First, using both co-immunoprecipitation and pull-down with recombinant proteins, we found that the central part of Tab2 is primarily responsible for this interaction, and that this region also interacts with Androgen Receptor. Then, we narrowed down the essential interaction region by means of competition assays using recombinant protein pull-down. The interaction motif was finally identified as a small region adjacent to, but not overlapping, the Tab2 MEKK1 phosphorylation sites. A synthetic peptide mimicking this motif efficiently displaced Tab2 from interacting with recombinant Estrogen Receptor alpha in vitro, prompting us to test its efficacy using derivatives of the MCF7 breast carcinoma cell lines that are spontaneously resistant to Tamoxifen. Indeed, we observed that this mimic peptide, made cell-permeable by addition of the TAT minimal carrier domain, reduced the growth of Tamoxifen-resistant MCF7 cells in the presence of Tamoxifen. These data indicate a novel functional domain of the Tab2 protein with potential application in drug design

Anomalous venoatrial connections – CT and MRI assessment

Abnormal venous atrial (VA) connections present a congenital heart disease (CHD) challenge for pediatric cardiologists. Fully anatomical evaluation is very difficult in prenatal and perinatal follow-up, but it has a profound impact on surgical correction and outcome. The echocardiogram is first-line imaging and represents the gold standard tool for simple abnormal VA connection. CT and MRI are mandatory for more complex heart disease and “nightmare cases”. 3D post-processing of volumetric CT and MRI acquisition helps to clarify anatomical relationships and allows for the creation of 3D printing models that can become crucial in customizing surgical strategy. Our article describes a ten-year (2013–2022) tertiary referral CHD center of abnormal AV connections investigated with CT and MRI, illustrating most of these complex diseases with the help of volume rendering (VR) or multiplanar reconstructions (MPR). The nightmarish cases will also be addressed due to the complex cardiovascular arrangement that requires a challenging surgical solution for correction along with the post-surgical complications.</p

Differential diagnosis of neurodegenerative dementias with the explainable MRI based machine learning algorithm MUQUBIA

Biomarker-based differential diagnosis of the most common forms of dementia is becoming increasingly important. Machine learning (ML) may be able to address this challenge. The aim of this study was to develop and interpret a ML algorithm capable of differentiating Alzheimer's dementia, frontotemporal dementia, dementia with Lewy bodies and cognitively normal control subjects based on sociodemographic, clinical, and magnetic resonance imaging (MRI) variables. 506 subjects from 5 databases were included. MRI images were processed with FreeSurfer, LPA, and TRACULA to obtain brain volumes and thicknesses, white matter lesions and diffusion metrics. MRI metrics were used in conjunction with clinical and demographic data to perform differential diagnosis based on a Support Vector Machine model called MUQUBIA (Multimodal Quantification of Brain whIte matter biomArkers). Age, gender, Clinical Dementia Rating (CDR) Dementia Staging Instrument, and 19 imaging features formed the best set of discriminative features. The predictive model performed with an overall Area Under the Curve of 98%, high overall precision (88%), recall (88%), and F1 scores (88%) in the test group, and good Label Ranking Average Precision score (0.95) in a subset of neuropathologically assessed patients. The results of MUQUBIA were explained by the SHapley Additive exPlanations (SHAP) method. The MUQUBIA algorithm successfully classified various dementias with good performance using cost-effective clinical and MRI information, and with independent validation, has the potential to assist physicians in their clinical diagnosis

Evaluation of the Mean Corpuscular Volume of Peripheral Blood Mononuclear Cells of HIV Patients by a Coulter Counter To Determine Intracellular Drug Concentrations▿

The mean corpuscular volume (MCV) of peripheral blood mononuclear cells (PBMCs) was determined by Coulter Counter, and data were used to calculate the intracellular drug concentrations. A total of 574 PBMC samples were collected from 190 patients. The MCV was 282.9 fl (minimum, 207.0; maximum, 354.6), with a standard deviation of 8.8%. Previous reports have often used a fixed value of 400 fl for the MCV, which may result in artificially low estimates of the intracellular concentrations of antivirals

A Tab2 mimic peptide displaces the Tab2/ERα interaction.

<p><b>a</b>. Aminoacid sequences of the three synthetic partially overlapping 17-aa each peptides, corresponding to different portions of the Fragment 4 described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0168639#pone.0168639.s002" target="_blank">S2 Fig</a>. (here called Tab2-pept1, Tab2-pept2 and Tab2-pept3). <b>b</b>. MBP pull-down assays, using MBP-Tab2_(406–531), recombinant hERα (1 nM) and the three different synthetic Tab2 peptides (at a concentration of 10 μM each). <b>c</b>. Dose-response curve on pull-down assays, using MBP-Tab2_(406–531), recombinant hERα (1 nM) and the Tab2-pept3 at concentrations ranging from 0 to 10 μM.</p

The central domain of Tab2 contains the major determinants of Tab2/ERα interaction.

<p>a. Scheme of Tab2 fragments of different length expressed in bacteria as MBP fusion proteins. The functional domains are indicated as follows: the CUE domain, the CC (coiled coil) domain including the nuclear export sequence, the NZF (novel zinc finger) domain and the sequence (II) containing two phosphorylation sites (S419 and S423). <b>b</b>. MBP pull-down assays, using ERα-overexpressing HEK293T cell lysate and MBP-Tab2 fusion proteins described in a). The loading control for all Tab2 fragments is provided in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0168639#pone.0168639.s001" target="_blank">S1 Fig</a>. <b>c</b>. MBP pull-down assays using the MBP fusion proteins Tab2 full-length, ΔsinoSTab2 and Tab2_(406–531) as a control, and ERα-overexpressing HEK293T cell lysates in the presence or not of the <i>in vitro</i> transcribed and translated Tab2_(406–531) fragment. The lower bands represent the total amount of MBP-Tab2 fusions present in the assay.</p

The central domain of Tab2 interacts with the conserved N-terminal domain of ERα and AR.

<p><b>a</b>. GST pull-down assay using the ERα fragment encompassing aa 1–45 expressed as GST fusion protein and the <i>in vitro</i> transcribed and translated Tab2_(406–531). <b>b,c,d</b>. Co-immunoprecipitation of T7-tagged Tab2_(406–531) and (<b>b</b>) p65-tagged ERα_(1–60); (<b>c</b>) Flag-tagged full-length ERα; (<b>d</b>) Flag-tagged full-length AR. The plasmids were transiently overexpressed in HEK293T cells, as follows: p65-ERα_(1–60) plus T7-Tab2_(406–531); Flag-full-length ERα (f.l.) plus T7-Tab2_(406–531); Flag-full-length AR (f.l.) plus T7-Tab2_(406–531). Mock transfected cells are untransfected cells. Anti-p65 and anti-T7 immunoprecipitates were carried out from total cell lysates and were analyzed by western blot with anti-ERα, anti-Flag and anti-T7 antibodies.</p

A Novel Functional Domain of Tab2 Involved in the Interaction with Estrogen Receptor Alpha in Breast Cancer Cells - Table 1

<p>A Novel Functional Domain of Tab2 Involved in the Interaction with Estrogen Receptor Alpha in Breast Cancer Cells</p> - Table

Urban Forms of Life. Per una critica delle forme di vita urbana

Che forme assume la vita nei contesti urbani? Da che deriva la loro presunta, attuale «illeggibilità»? Che cosa impedisce il fiorire di forme di vita appaganti, degne, vitali? È possibile rendere visibile e articolabile l’«elemento» urbano in cui siamo immersi? Quali esperimenti tentare per risvegliare un nuovo senso (comune) di vita urbana? Che ruolo pos- sono assumere le forme di vita urbana nei confronti del contesto più ampio, umano e non-umano, da cui dipendono sotto molteplici aspetti? Quali opportunità e quali insidie si prospettano con l’inarrestabile penetrazione delle nuove tecnologie digitali e di intelligenza artificiale nel tessuto urbano? A partire da queste e altre domande è nato nel 2017 un progetto di ricerca in filosofia, presentato all’Ateneo della Sapienza – Università di Roma, che lo ha finanziato e di cui questo libro è uno dei risultati, insieme a una serie di incontri e seminari, a partire da quello tenutosi presso il Centre Marc Bloch di Berlino nel settembre del 2018What forms does life take in urban contexts? Where does their sup- posed, current “unreadability” come from? What prevents the flour- ishing of fulfilling, worthy, vital forms of life? Is it possible to make visible and articulable the urban “element” in which we are immersed? What experiments should be attempted to awaken a new (common) sense of urban life? What role can urban forms of life assume vis-à- vis the larger human and non-human context on which they depend in multiple respects? What opportunities and pitfalls lie ahead with the unstoppable penetration of new digital and artificial intelligence technologies into the urban fabric? Starting from these and other ques- tions, a research project in philosophy was presented to the Sapienza - University of Rome in 2017, of which this book is one of the results, along with a series of meetings and seminars, starting with the one held at the Marc Bloch Centre in Berlin in September 201