A study on the VEGFR2-ligand multi-physics interactions in Angiogenesis.

Tumorgrowthissustainedbyangiogenesis,i.e. theformationofnewbloodvesselsfrompre-existing ones. Angiogenesis is modulated by the interaction between tyrosine kinase receptors (TKRs), expressed by endothelial cells (ECs), and extracellular ligands, produced by tumor cells. This interaction triggers the activation of intracellular signaling cascades and kinetic processes, including cell deformationandadhesion,whicheventuallycausecelldivisionandproliferation. VascularEndothelial Growth Factor Receptor-2 (VEGFR2) is a pro-angiogenic receptor expressed on ECs. Ligand stimulation induces the polarization of ECs and the relocation of VEGFR2 in cell protrusion or in the basal aspect in cells plated on ligand enriched extracellular matrix (ECM) [1]. EC response to angiogenic growth factors is regulated by distinct sets of inputs conveyed by TRKs and different co-receptors including integrins, membrane proteins that are responsible of stress fibers formation and cell contractility [2]. Although biochemical pathways following VEGFR2 activation are well established, knowledge about the receptor dynamics on the plasma membrane remains limited. A multi-physics model has been developed [3] to describe: i) the diffusion of VEGFR2 on the cellularmembrane;ii)thechemicalkineticsoftheligand-receptorbindingreaction;iii)themechanical adhesion and spreading of the cell onto a ligand-rich extracellular substrate, in finite strain. The identification of the multi-physics interactions that regulate receptor polarization could open new perspectives to develop innovative anti-angiogenic strategies through the modulation of EC activation

A Logistic Model Tree Solution

Beretta, S., Castelli, M., Gonçalves, I., Kel, I., Giansanti, V., & Merelli, I. (2018). Improving eQTL Analysis Using a Machine Learning Approach for Data Integration: A Logistic Model Tree Solution. Journal of Computational Biology, 25(10), 1091-1105. DOI: 10.1089/cmb.2017.0167Expression quantitative trait loci (eQTL) analysis is an emerging method for establishing the impact of genetic variations (such as single nucleotide polymorphisms) on the expression levels of genes. Although different methods for evaluating the impact of these variations are proposed in the literature, the results obtained are mostly in disagreement, entailing a considerable number of false-positive predictions. For this reason, we propose an approach based on Logistic Model Trees that integrates the predictions of different eQTL mapping tools to produce more reliable results. More precisely, we employ a machine learning-based method using logistic functions to perform a linear regression able to classify the predictions of three eQTL analysis tools (namely, R/qtl, MatrixEQTL, and mRMR). Given the lack of a reference dataset and that computational predictions are not so easy to test experimentally, the performance of our approach is assessed using data from the DREAM5 challenge. The results show the quality of the aggregated prediction is better than that obtained by each single tool in terms of both precision and recall. We also performed a test on real data, employing genotypes and microRNA expression profiles from Caenorhabditis elegans, which proved that we were able to correctly classify all the experimentally validated eQTLs. These good results come both from the integration of the different predictions, and from the ability of this machine learning algorithm to find the best cutoff thresholds for each tool. This combination makes our integration approach suitable for improving eQTL predictions for testing in a laboratory, reducing the number of false-positive results.authorsversionpublishe

Hip joint centre position estimation using a dual unscented Kalman filter for computer-assisted orthopaedic surgery

In computer-assisted knee surgery, the accuracy of the localization of the femur centre of rotation relative to the hip-bone (hip joint centre) is affected by the unavoidable and untracked pelvic movements because only the femoral pose is acquired during passive pivoting manoeuvres. We present a dual unscented Kalman filter algorithm that allows the estimation of the hip joint centre also using as input the position of a pelvic reference point that can be acquired with a skin marker placed on the hip, without increasing the invasiveness of the surgical procedure. A comparative assessment of the algorithm was carried out using data provided by in vitro experiments mimicking in vivo surgical conditions. Soft tissue artefacts were simulated and superimposed onto the position of a pelvic landmark. Femoral pivoting made of a sequence of star-like quasi-planar movements followed by a circumduction was performed. The dual unscented Kalman filter method proved to be less sensitive to pelvic displacements, which were shown to be larger during the manoeuvres in which the femur was more adducted. Comparable accuracy between all the analysed methods resulted for hip joint centre displacements smaller than 1 mm (error: 2.2 ± [0.2; 0.3] mm, median ± [inter-quartile range 25%; inter-quartile range 75%]) and between 1 and 6 mm (error: 4.8 ± [0.5; 0.8] mm) during planar movements. When the hip joint centre displacement exceeded 6 mm, the dual unscented Kalman filter proved to be more accurate than the other methods by 30% during multi-planar movements (error: 5.2 ± [1.2; 1] mm)

Multi-physics interactions drive VEGFR2 relocation on endothelial cells.

Vascular Endothelial Growth Factor Receptor-2 (VEGFR2) is a pro-angiogenic receptor, expressed on endothelial cells (ECs). Although biochemical pathways that follow the VEGFR2 activation are well established, knowledge about the dynamics of receptors on the plasma membrane remains limited. Ligand stimulation induces the polarization of ECs and the relocation of VEGFR2, either in cell protrusions or in the basal aspect in cells plated on ligand-enriched extracellular matrix (ECM). We develop a mathematical model in order to simulate the relocation of VEGFR2 on the cell membrane during the mechanical adhesion of cells onto a ligand-enriched substrate. Co-designing the in vitro experiments with the simulations allows identifying three phases of the receptor dynamics, which are controlled respectively by the high chemical reaction rate, by the mechanical deformation rate, and by the diffusion of free receptors on the membrane. The identification of the laws that regulate receptor polarization opens new perspectives toward developing innovative anti-angiogenic strategies through the modulation of EC activatio

Targeting of RET oncogene by naphthalene diimide-mediated gene promoter G-quadruplex stabilization exerts anti-tumor activity in oncogene-addicted human medullary thyroid cancer

Medullary thyroid cancer (MTC) relies on the aberrant activation of RET proto-oncogene. Though targeted approaches (i.e., tyrosine kinase inhibitors) are available, the absence of complete responses and the onset of resistance mechanisms indicate the need for novel therapeutic interventions. Due to their role in regulation of gene expression, G-quadruplexes (G4) represent attractive targets amenable to be recognized or stabilized by small molecules. Here, we report that exposure of MTC cells to a tri-substituted naphthalene diimide (NDI) resulted in a significant antiproliferative activity paralleled by inhibition of RET expression. Biophysical analysis and gene reporter assays showed that impairment of RET expression was consequent to the NDI-mediated stabilization of the G4 forming within the gene promoter. We also showed for the first time that systemic administration of the NDI in mice xenotransplanted with MTC cells resulted in a remarkable inhibition of tumor growth in vivo. Overall, our findings indicate that NDI-dependent RET G4 stabilization represents a suitable approach to control RET transcription and delineate the rationale for the development of G4 stabilizing-based treatments for MTC as well as for other tumors in which RET may have functional and therapeutic implications

Arte, técnica y oficio en metal. Herrería artística en el patrimonio arquitectónico de Montevideo, 1780-1950

This article summarizes some aspects of the project “Iron and bronze. Criteria for the valuation and conservation of ornamental metalworks in the architectural heritage of Uruguay”, whose objective is to establish the heritage attributes of the metallic ornamental elements associated with facades of the national architectural heritage and contribute to their dissemination, enhancement, and conservation. Throughout the research, socio-cultural, artistic, formal, and technical aspects are addressed, and finally, those related to the deterioration of these elements and the guidelines for their preservation and restoration. The methodology is based on the study of primary sources, published and unpublished, on the revision of specialized bibliography and on the site work on a representative sample of metallic ornamental elements applied to the facades of Montevideo architecture. A total of 250 buildings were surveyed, corresponding to the different periods that we distinguished in the evolution of the local ornamental metalwork between the end of the 18th century and the middle of the 20th century. It is an initial contribution that delves into a field that is scarcely studied in our environment about the knowledge and dissemination of metalwork in the context of the Uruguayan architectural and artistic heritage, providing useful information to the academy, the institutions linked to the valuation and heritage conservation and the general public.Este artículo sintetiza algunos aspectos del proyecto “Hierro y bronce. Criterios para la valoración y conservación de la herrería artística en el patrimonio arquitectónico del Uruguay”[i], cuyo objetivo es establecer los atributos patrimoniales de los elementos ornamentales metálicos asociados a fachadas del patrimonio arquitectónico nacional y contribuir a su difusión, puesta en valor y conservación. A lo largo de la investigación se abordan los aspectos socioculturales, artísticos, formales, técnicos y, por último, los relativos al deterioro de estos elementos y las pautas para su preservación y restauración. La metodología se basa en el estudio de fuentes primarias, éditas e inéditas, en la revisión de bibliografía especializada y en el trabajo de campo sobre una muestra representativa de elementos ornamentales metálicos aplicados a las fachadas de la arquitectura montevideana. Fue relevado un total de 250 edificios, correspondientes a los distintos períodos que distinguimos en la evolución de la herrería local entre finales del siglo XVIII y mediados del siglo XX. Se trata de un aporte inicial que profundiza en un campo escasamente abordado en nuestro medio acerca del conocimiento y difusión de los trabajos de herrería en el marco del patrimonio arquitectónico y artístico uruguayo, proporcionando información útil a la academia, las instituciones vinculadas a la valoración y conservación patrimonial y el público general.   Notas [i] Proyecto concursado y financiado en la modalidad Investigación y Desarrollo (I+D), Comisión Sectorial de Investigación científica (CSIC), Universidad de la República, a realizarse entre marzo de 2018 y marzo de 2020, por un equipo interdisciplinario integrado por docentes e investigadores de las facultades de Arquitectura, Diseño y Urbanismo: Miriam Hojman, Valentina Marchese y Tatiana Rimbaud (Instituto de Historia), Gianella Mussio, Leticia Olivera, Carola Romay y Verónica Ulfe (Instituto de Tecnologías); de la Facultad de Humanidades y Ciencias de la Educación: Ernesto Beretta (Instituto de Ciencias Históricas-Historia del Arte); y de la Facultad de Ingeniería: Sofía Aguiar (Instituto de Ensayo de Materiales). El presente artículo fue elaborado por todos los integrantes del equipo

Sulfonates-PMMA nanoparticles conjugates: A versatile system for multimodal application

a b s t r a c t We report herein the viability of a novel nanoparticles (NPs) conjugated system, namely the attachment, based on ionic and hydrophobic interactions, of different sulfonated organic salts to positively charged poly(methylmethacrylate) (PMMA)-based core-shell nanoparticles (EA0) having an high density of ammonium groups on their shells. In this context three different applications of the sulfonates@EA0 systems have been described. In detail, their ability as cytotoxic drugs and pro-drugs carriers was evaluated in vitro on NCI-H460 cell line and in vivo against human ovarian carcinoma IGROV-1 cells. Besides, 8-hydroxypyrene-1,3,6-trisulfonic acid, trisodium salt (HPTS) was chosen for NPs loading, and its internalization as bioimaging probe was evaluated on Hep G2 cells. Overall, the available data support the interest for these PMMA NPs@sulfonates systems as a promising formulation for theranostic applications. In vivo biological data strongly support the potential value of these core-shell NPs as delivery system for negatively charged drugs or biologically active molecules. Additionally, we have demonstrated the ability of these PMMA core-shell nanoparticles to act as efficient carriers of fluorophores. In principle, thanks to the high PMMA NPs external charge density, sequential and very easy post-loading of different sulfonates is achievable, thus allowing the preparation of nanocarriers either with bi-modal drug delivery behaviour or as theranostic systems

Stability and expression levels of HLA-C on the cell membrane modulate HIV-1 infectivity

HLA-C expression is associated with a differential ability to control HIV-1 infection. Higher HLA-C levels may lead to a better control of HIV-1 infection through both a higher efficiency of antigen presentation to cytotoxic T lymphocytes (CTLs), as well as the triggering of activating Killer Immunoglobulin like receptors (KIR) on NK-cells, whereas lower levels may provide a poor HIV-1 control and a rapid progression toward AIDS.We characterized the relative amount of HLA-C heterotrimers (heavy chain/\u3b22m/peptide) and HLA-C free heavy chains on PBMC from healthy blood donors harboring both alleles with stable or unstable binding to \u3b22m/peptide. We analyzed the stability of HLA-C heterotrimers of different allotypes and the infectivity of HIV-1 virions produced by PBMC with various allotypes.We observed significant differences in HLA-C heterotrimers stability and in expression levels. We found that R5 HIV-1 virions produced by PBMC harboring unstable HLA-C alleles were more infectious than those produced by PBMC carrying the stable variants.We propose that HIV-1 infectivity might depend both on the amounts of HLA-C molecules and on their stability as trimeric complex. According to this model, individuals with low expressed HLA-C alleles and unstable binding to \u3b22m/peptide might have a worse control of HIV-1 infection and an intrinsically higher capacity to support viral replication.IMPORTANCE Following HIV-1 infection, some people advance rapidly toward AIDS while others have a slow disease progression. HLA-C, a molecule involved in immunity, is a key determinant of HIV-1 control.Here we reveal how HLA-C variants contribute to modulate viral infectivity. HLA-C is present on the cell surface in two different conformations: the immunologically active conformation is part of a complex that includes \u3b22-microglobulin/peptide; the other conformation is not bound to \u3b22-microglobulin/peptide and can associate with HIV-1, increasing its infectivity. Individuals with HLA-C variants with a predominance of immunologically active conformations would display a stronger immunity against HIV-1, a reduced viral infectivity and an effective control of HIV-1 infection, while subjects with HLA-C variants that easily dissociate from \u3b22-microglobulin/peptide would have a reduced immunological response to HIV-1 and produce more infectious virions.This study provides new information that could be useful to design novel vaccine strategies and therapeutic approaches against HIV-1