Evaluating the pharmacological response in fluorescence microscopy images: The Δm algorithm

Current drug discovery procedures require fast and effective quantification of the pharmacological response evoked in living cells by agonist compounds. In the case of G-protein coupled receptors (GPCRs), the efficacy of a particular drug to initiate the endocytosis process is related to the formation of endocytic vesicles or endosomes and their subsequent internalisation within intracellular compartments that can be observed with high spatial and temporal resolution by fluorescence microscopy techniques. Recently, an algorithm has been proposed to evaluate the pharmacological response by estimating the number of endosomes per cell on time series of images. However, the algorithm was limited by the dependence on some manually set parameters and in some cases the quality of the image does not allow a reliable detection of the endosomes. Here we propose a simple, fast and automated image analysis method?the ?m algorithm- to quantify a pharmacological response with data obtained from fluorescence microscopy experiments. This algorithm does not require individual object detection and computes the relative increment of the third order moment in fluorescence microscopy images after filtering with the Laplacian of Gaussian function. It was tested on simulations demonstrating its ability to discriminate different experimental situations according to the number and the fluorescence signal intensity of the simulated endosomes. Finally and in order to validate this methodology with real data, the algorithm was applied to several time-course experiments based on the endocytosis of the mu opioid receptor (MOP) initiated by different agonist compounds. Each drug displayed a different ?m sigmoid time-response curve and statistically significant differences were observed among drugs in terms of efficacy and kinetic parameters

His452Tyr polymorphism in the human 5-HT2A receptor affects clozapine-induced signaling networks revealed by quantitative phosphoproteomics

Antipsychotic drugs remain the current standard for schizophrenia treatment. Although they directly recognize the orthosteric binding site of numerous monoaminergic G protein-coupled receptors (GPCRs), these drugs, and particularly second-generation antipsychotics such as clozapine, all have in common a very high affinity for the serotonin 5-HT receptor (5-HTR). Using classical pharmacology and targeted signaling pathway assays, previous findings suggest that clozapine and other atypical antipsychotics behave principally as 5-HTR neutral antagonists and/or inverse agonists. However, more recent findings showed that antipsychotics may also behave as pathway-specific agonists. Reversible phosphorylation is a common element in multiple signaling networks. Combining a quantitative phosphoproteomic method with signaling network analysis, we tested the effect of clozapine treatment on the overall level of protein phosphorylation and signal transduction cascades in vitro in mammalian cell lines induced to express either the human 5-HTR or the H452Y variant of the gene encoding the 5-HTR receptor. This naturally occurring variation within the 5-HTR gene was selected because it has been repeatedly associated with schizophrenia patients who do not respond to clozapine treatment. Our data show that short time exposure (5 or 10 min) to clozapine (10 M) led to phosphorylation of numerous signaling components of pathways involved in processes such as endocytosis, ErbB signaling, insulin signaling or estrogen signaling. Cells induced to express the H452Y variant showed a different basal phosphoproteome, with increases in the phosphorylation of mTOR signaling components as a translationally relevant example. However, the effect of clozapine on the functional landscape of the phosphoproteome was significantly reduced in cells expressing the 5-HTR-H452Y construct. Together, these findings suggest that clozapine behaves as an agonist inducing phosphorylation of numerous pathways downstream of the 5-HTR, and that the single nucleotide polymorphism encoding 5-HTR-H452Y affects these clozapine-induced phosphorylation-dependent signaling networks

CHALLENGE 6: Exposing the roots of mental disorders

Mental disorders have devastating and increasing impact in our societies. CSIC researchers face the challenge of determining the biological and social causes and consequences of these disorders, and of finding efficient therapies. To these aims, the collaborative effort of neuroscientists, neurologists, psychiatrists, psychologists and human and social scientists, the use and development of state-of-the-art technologies and the contact with patient associations and pharma industry are required.Peer reviewe

Broken symmetry and the variation of critical properties in the phase behaviour of supramolecular rhombus tilings

The degree of randomness, or partial order, present in two-dimensional supramolecular arrays of isophthalate tetracarboxylic acids is shown to vary due to subtle chemical changes such as the choice of solvent or small differences in molecular dimensions. This variation may be quantified using an order parameter and reveals a novel phase behaviour including random tiling with varying critical properties as well as ordered phases dominated by either parallel or non-parallel alignment of neighbouring molecules, consistent with long-standing theoretical studies. The balance between order and randomness is driven by small differences in the intermolecular interaction energies, which we show, using numerical simulations, can be related to the measured order parameter. Significant variations occur even when the energy difference is much less than the thermal energy highlighting the delicate balance between entropic and energetic effects in complex self-assembly processes

Dopaminergic control of ADAMTS2 expression through cAMP/CREB and ERK: molecular effects of antipsychotics

© The Author(s) 2019.A better understanding of the molecular mechanisms that participate in the development and clinical manifestations of schizophrenia can lead to improve our ability to diagnose and treat this disease. Previous data strongly associated the levels of deregulated ADAMTS2 expression in peripheral blood mononuclear cells (PBMCs) from patients at first episode of psychosis (up) as well as in clinical responders to treatment with antipsychotic drugs (down). In this current work, we performed an independent validation of such data and studied the mechanisms implicated in the control of ADAMTS2 gene expression. Using a new cohort of drug-naïve schizophrenia patients with clinical follow-up, we confirmed that the expression of ADAMTS2 was highly upregulated in PBMCs at the onset (drug-naïve patients) and downregulated, in clinical responders, after treatment with antipsychotics. Mechanistically, ADAMTS2 expression was activated by dopaminergic signalling (D1-class receptors) and downstream by cAMP/CREB and mitogen-activated protein kinase (MAPK)/ERK signalling. Incubation with antipsychotic drugs and selective PKA and MEK inhibitors abrogated D1-mediated activation of ADAMTS2 in neuronal-like cells. Thus, D1 receptors signalling towards CREB activation might participate in the onset and clinical responses to therapy in schizophrenia patients, by controlling ADAMTS2 expression and activity. The unbiased investigation of molecular mechanisms triggered by antipsychotic drugs may provide a new landscape of novel targets potentially associated with clinical efficacy.This work was supported by: SAF2016-76046-R and SAF2013-46292-R (MINECO and FEDER) to B.C.F., PI16/00156 (isciii and FEDER) to J.P.V., LUCHAMOS POR LA VIDA project to F.R.J. and J.P.V., SAF2017-83702-R (MINECO and FEDER), Red TERCEL RD12/0019/0024 (ISCIII) and GVA-PROMETEO 2018/041 (Generalitat Valenciana) to S.M. J.P.V. is supported by the RyC research programme (RYC-2013-14097) and F.R.J. by the predoctoral research programme (BES-2014-070615), from MINECO and FEDER

New discoveries of mud volcanoes on the Moroccan Atlantic continental margin (Gulf of Cádiz): morpho-structural characterization

10th International Conference on Gas in Marine Sediments, 6-11 September 2010, Listvyanka, RussiaDuring the MVSEIS-08 cruise of 2008, ten new mud volcanoes (MVs) were discovered on the offshore Moroccan continental margin (Gulf of Cádiz) at water depths between 750 and 1,600 m, using multibeam bathymetry, backscatter imagery, high-resolution seismic and gravity core data. Mud breccias were recovered in all cases, attesting to the nature of extrusion of these cones. The mud volcanoes are located in two fields: the MVSEIS, Moundforce, Pixie, Las Negras, Madrid, Guadix, Almanzor and El Cid MVs in the western Moroccan field, where mud volcanoes have long been suspected but to date not identified, and the Boabdil and Al Gacel MVs in the middle Moroccan field. Three main morphologies were observed: asymmetric, sub-circular and flat-topped cone-shaped types, this being the first report of asymmetric morphologies in the Gulf of Cádiz. Based on morpho-structural analysis, the features are interpreted to result from (1) repeated constructive (expulsion of fluid mud mixtures) and destructive (gravity-induced collapse and submarine landsliding) episodes and (2) interaction with bottom currentsDivisión de Geología Marina, Instituto Geológico y Minero de España, EspañaCentro Oceanográfico de Málaga, Instituto Español de Oceanografía, EspañaFacultad de Ciencias del Mar, Universidad de Cádiz, EspañaFacultad de Ciencias Geológicas, Universidad Complutense de Madrid, Españ

Innovación y estrategias de intervención en acoso escolar

Los problemas de convivencia escolar, violencia escolar, bullying y cyberbullying, en los centros españoles son una realidad y, por tanto, es necesario que los futuros docentes cuenten con estrategias específicas para el abordaje de estas situaciones cuando se enfrenten a la realidad de las aulas. En este manual, profesionales del área de la convivencia escolar presentan las actuaciones, programas y herramientas disponibles que en estos momentos se están poniendo en práctica en los centros escolares con resultados positivos. Así, se pretende ampliar el conocimiento teórico-práctico en el ámbito de la convivencia, gestión de conflictos y violencia escolar.EducaciónPsicologí

Use of life cycle assessment methodology in the analysis of ecological footprint assessment results to evaluate the environmental performance of universities

The assessment of the environmental performance of an organization is an essential part of the decision making process of an Environmental Management System. Having robust indicators enables a reliable assessment. The Ecological Footprint Assessment is used in different types of organizations, including universities. Its ability to clearly communicate over-consumption by using a land-base unit is an advantage when involving the university community in achieving better environmental performance. However, its lack of standardization makes it difficult to use as an indicator. It is believed that Life Cycle Assessment offers a framework with which to standardize the Ecological Footprint Assessment. In this paper, an Ecological Footprint Assessment considering Life Cycle Assessment methodology is developed as a case study for Universitat Politecnica de Valencia. Findings regarding the critical decisions of the methodology are compared with 23 Ecological Footprint Assessments of universities using a Life Cycle Assessment framework. Only 26% of the studies analyzed reference the Life Cycle Assessment methodology. Critical decisions such as defining a Functional Unit were relevant but not standardized, while the definition of the product system was the most standardized and homogeneous decision. The difficulty of gathering information when Environmental Management Systems are not available makes the Ecological Footprint Assessment a weak indicator. Nevertheless, results show that Life Cycle Assessment can guide an Ecological Footprint Assessment methodology where comparability and reliability is possible. (C) 2016 Elsevier Ltd. All rights reserved.Lo-Iacono-Ferreira, VG.; Torregrosa López, JI.; Capuz-Rizo, SF. (2016). Use of life cycle assessment methodology in the analysis of ecological footprint assessment results to evaluate the environmental performance of universities. Journal of Cleaner Production. 133:45-53. doi:10.1016/j.jclepro.2016.05.046S455313

Interrelación de laboratorios de control y laboratorios de investigación en España para la armonización de metodologías de determinación de toxinas paralizantes

XII Congreso Nacional de Agricultura, Madrid 24-26 de noviembre de 2009Marketing of cultured and harvested shellfish is linked to monitoring programs for granting food safety. Its complexity requires constant cooperation between research and monitoring laboratories in order to improve sampling and analysing performances, achieve legal requirements, etc. for increasing consumer’s health protection but not reducing producer’s benefits. The JACUMAR project «Comparison of methodologies for the evaluation of Paralytic Shellfish Poisoning (PSP) toxins in bivalves. Application for aquaculture in Spain» groups research and monitoring laboratories from Galicia, Andalucía and Cataluña. Efforts are focused on detection and quantification of PSP toxins, searching an analytical method able to fulfil technical and management requirementsEste proyecto está financiado por la Junta Asesora de Cultivos Marinos (JACUMAR), y los programas de control por los gobiernos autónomos de Galicia, Andalucía y CataluñaN

Innocampus Explora: una aproximación multidisciplinar a la problemática ambiental

[ES] Presentamos las actividades del proyecto de innovación Innocampus Explora desarrollado en el campus de Burjassot-Paterna de la Universitat de València y cuyo objetivo principal es mostrar la interrelación existente entre los diferentes grados científicos y técnicos del campus. En la presente anualidad, el equipo de trabajo integrado por estudiantes y profesores de todos las facultades y escuelas del campus de Burjassot-Paterna, ha desarrollado actividades en torno a la problemática medioambiental. Una visión transversal e interdisciplinar de los problemas de los usos del plástico y de la energía nuclear que enlaza con varios de los Objetivos de Desarrollo Sostenible (ODS) dictados por Naciones Unidas. Con el desarrollo de este proyecto contribuimos a una formación transversal de calidad para todos los estudiantes participantes.[EN] We present the activities of the Innocampus Explora innovation project developed on the Burjassot-Paterna campus of the Universitat de València and whose main objective is to show the interrelation between the different scientific and technical degrees on campus. In this year, the work team made up of students and professors from all the faculties and schools of the Burjassot-Paterna campus, have carried out activities around environmental issues. A cross-sectional and interdisciplinary vision of the problems of the uses of plastic and nuclear energy that link with several of the Sustainable Development Goals (SDGs) dictated by the United Nations. With the development of this project we contribute to quality transversal training for all participating students.Moros Gregorio, J.; Quílez Asensio, A.; Jimenez Romero, D.; Blas Medina, A.; Giménez Escamilla, I.; Amorós Hernández, L.; Giner, L.... (2021). Innocampus Explora: una aproximación multidisciplinar a la problemática ambiental. En IN-RED 2020: VI Congreso de Innovación Educativa y Docencia en Red. Editorial Universitat Politècnica de València. 1003-1014. https://doi.org/10.4995/INRED2020.2020.11996OCS1003101