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

Caspase-8 inhibition represses initial human monocyte activation in septic shock model

In septic patients, the onset of septic shock occurs due to the over-activation of monocytes. We tested the therapeutic potential of directly targeting innate immune cell activation to limit the cytokine storm and downstream phases. We initially investigated whether caspase-8 could be an appropriate target given it has recently been shown to be involved in microglial activation. We found that LPS caused a mild increase in caspase-8 activity and that the caspase-8 inhibitor IETD-fmk partially decreased monocyte activation. Furthermore, caspase-8 inhibition induced necroptotic cell death of activated monocytes. Despite inducing necroptosis, caspase-8 inhibition reduced LPS-induced expression and release of IL-1β and IL-10. Thus, blocking monocyte activation has positive effects on both the pro and anti-inflammatory phases of septic shock. We also found that in primary mouse monocytes, caspase-8 inhibition did not reduce LPS-induced activation or induce necroptosis. On the other hand, broad caspase inhibitors, which have already been shown to improve survival in mouse models of sepsis, achieved both. Thus, given that monocyte activation can be regulated in humans via the inhibition of a single caspase, we propose that the therapeutic use of caspase-8 inhibitors could represent a more selective alternative that blocks both phases of septic shock at the source.Unión Europea, Ministerio de Economía y Competitividad SAF2012-39029Unión Europea, Ministerio de Economía y Competitividad SAF2015-64171REspaña,Junta de Andalucía P10-CTS-649

The Absence of Caspase-8 in the Dopaminergic System Leads to Mild Autism-like Behavior

In the last decade, new non-apoptotic roles have been ascribed to apoptotic caspases. This family of proteins plays an important role in the sculpting of the brain in the early stages of development by eliminating excessive and nonfunctional synapses and extra cells. Consequently, impairments in this process can underlie many neurological and mental illnesses. This view is particularly relevant to dopamine because it plays a pleiotropic role in motor control, motivation, and reward processing. In this study, we analyze the effects of the elimination of caspase-8 (CASP8) on the development of catecholaminergic neurons using neurochemical, ultrastructural, and behavioral tests. To do this, we selectively delete the CASP8 gene in cells that express tyrosine hydroxylase with the help of recombination through the Cre-loxP system. Our results show that the number of dopaminergic neurons increases in the substantia nigra. In the striatum, the basal extracellular level of dopamine and potassium-evoked dopamine release decreased significantly in mice lacking CASP8, clearly showing the low dopamine functioning in tissues innervated by this neurotransmitter. This view is supported by electron microscopy analysis of striatal synapses. Interestingly, behavioral analysis demonstrates that mice lacking CASP8 show changes reminiscent of autism spectrum disorders (ASD). Our research reactivates the possible role of dopamine transmission in the pathogenesis of ASD and provides a mild model of autism.Ministerio de Economía y Competitividad RTI2018-098645-B-I00, PID2019-109569GB-I00, RTI2018-099778-B-I00Junta de Andalucía P18-RT-1372, US-1264806, PI-0080-2017, PI-0009-2017, PI-0134-2018, PEMP-0008-2020, P20_00958, CTS-510Instituto de Salud Carlos III PI18/01691Instituto de Investigación e Innovación en Ciencias Biomédicas de Cádiz-INiBICA LI19/06IN-CO22, IN-C09European Union 95568

Group and call effect in achieving success in a subject: Analyses of the "Biology and Botany" whole life in the E.U.I.T.A. (University of Seville)

La Biología y la Botánica son dos materias fundamentales en la formación de un ingeniero técnico agrícola. Sus contenidos se han ofrecido a veces en asignaturas independientes y, en otras ocasiones como la que nos ocupa, en una sola asignatura. Por otra parte, siempre se ha mirado con interés, e incluso preocupación, el efecto de la variable grupo, de forma independiente o relacionándola con su distribución en turnos de mañana o tarde, así como la importancia de la convocatoria (en los distintos momentos a lo largo del curso académico) en la que el alumno consigue superar la asignatura. Aquí presentamos los resultados en la consecución de objetivos por parte de los alumnos y, por ende, de los profesores y de la Universidad de Sevilla a lo largo de la totalidad de la vida de la asignatura "Biología y Botánica" dentro del plan de estudios 2003/04 para tres titulaciones simultáneas, desde su comienzo en el curso 2003/04 hasta su extinción en el curso 2009/10, de la E.U.I.T.A. (Escuela Universitaria de Ingeniería Técnica Agrícola) de la Universidad de Sevilla. Además se consideran convocatorias pertenecientes al período de extinción añadido. Los análisis toman en cuenta además el éxito en la superación de la asignatura tanto de forma cualitativa (aprobado o suspenso de la evaluación a la que el alumno se ha sometido) como cuantitativa (nota conseguida). Las conclusiones obtenidas permiten mirar los resultados y la consecución de objetivos, así como una potencial toma de decisiones para el futuro, basándose en un marco temporal amplio y objetivo.Biology and Botany are two critical issues in the formation of a technical agricultural engineer. Their topics are sometimes offered in separate subjects and, at other times as here, within a single one. Besides, it has always been considered with interest, and even concern, the effect of the variable group, independently or in relation to their distribution in the morning or afternoon turns, and the importance of the call (at different times during the academic year) at which the student gets to pass the subject. Here we present the results in the achievement of objectives by students, and therefore teachers and the University of Seville, along the entire life of the subject "Biology and Botany" within the teaching 2003/04 plan for three simultaneous academic programs, from its start in the course 2003/04 to its extinction in the course 2009/10, in the E.U.I.T.A. (University School of Technical Agricultural Engineering) of the University of Seville. Calls belonging to the period of extinction are also considered. Analyses are both qualitative, based on having success in passing the subject or not, and quantitative (mark scored). Obtained conclusions let us see the results and the achievement of objectives, as well as a potential decision-making for the future, with a base on a comprehensive and objective frame

The neolithic necropolis of Feixa del Moro (Juberri, Andorra): review and new data

[EN] The data presented in this paper resume all the available information on the Feixa del Moro site, correcting old mistakes and bias, updating the 1980s archaeological registers and presenting new analyses as well. Our aim is to ensure that Feixa del Moro remains a reference site for the Pyrenean and Western Mediterranean Neolithic. At the same time, we wish to encourage other researchers to undertake new analyses and to embrace new perspectives in order to improve our understanding of Neolithic societies.[FR] Le travail que nous présentons ici rassemble toutes les données disponibles sur la Feixa del Moro jusqu’à aujourd’hui, expose les confusions détectées dans les sources, actualise les registres archéologiques obtenus dans les années 1980 et présente les résultats des nouvelles analyses effectuées. Grâce à cette démarche nous souhaitons que ce site continue d’être une référence pour le Néolithique dans les Pyrénées et la Méditerranée occidentale. Nous souhaitons également attirer l’attention d’autres chercheurs afin qu’ils continuent d’analyser et d’apporter de nouveaux éléments et de nouvelles approches pour mieux comprendre les sociétés néolithiques.Peer Reviewe

The Absence of Caspase-8 in the Dopaminergic System Leads to Mild Autism-like Behavior

In the last decade, new non-apoptotic roles have been ascribed to apoptotic caspases. This family of proteins plays an important role in the sculpting of the brain in the early stages of development by eliminating excessive and nonfunctional synapses and extra cells. Consequently, impairments in this process can underlie many neurological and mental illnesses. This view is particularly relevant to dopamine because it plays a pleiotropic role in motor control, motivation, and reward processing. In this study, we analyze the effects of the elimination of caspase-8 (CASP8) on the development of catecholaminergic neurons using neurochemical, ultrastructural, and behavioral tests. To do this, we selectively delete the CASP8 gene in cells that express tyrosine hydroxylase with the help of recombination through the Cre-loxP system. Our results show that the number of dopaminergic neurons increases in the substantia nigra. In the striatum, the basal extracellular level of dopamine and potassium-evoked dopamine release decreased significantly in mice lacking CASP8, clearly showing the low dopamine functioning in tissues innervated by this neurotransmitter. This view is supported by electron microscopy analysis of striatal synapses. Interestingly, behavioral analysis demonstrates that mice lacking CASP8 show changes reminiscent of autism spectrum disorders (ASD). Our research reactivates the possible role of dopamine transmission in the pathogenesis of ASD and provides a mild model of autism

Dendronized Anionic Gold Nanoparticles: Synthesis, Characterization and Antiviral Activity

Anionic carbosilane dendrons decorated with sulfonate functions and with a thiol moiety at the focal point have been used to synthesize water soluble gold nanoparticles (AuNPs) by direct reaction of dendrons, gold precursor and reducing agent in water and also by place-exchange reaction. These nanoparticles have been characterized by nuclear magnetic resonance (NMR), transmission electron microscopy (TEM), thermogravimetric analysis (TGA), X-ray photoelectron spectroscopy (XPS), UV, elemental analysis, and Z potential. Also, the interacting ability of the anionic sulfonate functions was investigated by electron paramagnetic resonance (EPR) using copper(II) as a probe. It was found that the different structures and conformations of the AuNPs modulate the availability of sulfonate and thiol groups to be complexed by copper(II). Toxicity assays of AuNPs showed that those produced by direct reaction were less toxic than those obtained by ligand exchange. Inhibition of HIV-1 infection was higher for dendronized AuNPs than for dendrons.Ministerio de Economía y EmpresaComunidad de MadridUniversidad de Alcal

Diálogos sobre transdisciplina: los investigadores y su objeto de estudio

A la transdisciplinariedad se le ha definido como “una feliz transgresión de las fronteras entre las disciplinas” y es en este tono en que se presenta esta obra, que recopila las experiencias y reflexiones, las discusiones y propuestas de una veintena de investigadores y académicos que hablan sobre o desde la transdisciplina acerca de los temas de su interés o especialidad. La aproximación se da desde perspectivas académicas diversas y se adereza con expresiones estéticas que van desde la poesía hasta la pintura, a través de las cuales se busca ofrecer un espacio a las rutas posibles y limitaciones connaturales de acceder a la realidad para construir conocimiento “de frontera”, “en las fronteras”. Los abordajes son fruto de la exploración, filiación, encantos y desencantos por parte de los autores con la entidad de su búsqueda, quienes buscan contestar, entre otras, las siguientes cuestiones: ¿Cómo establecer un acercamiento transdisciplinar al objeto de estudio? ¿Qué hace a un objeto de estudio transdisciplinar? ¿Cómo impacta la transdisciplinariedad la identidad del académico? Una obra concebida desde una perspectiva más pedagógica que desde la doxa académica, con el interés de aportar una lectura amena para las reflexiones en torno a la trasgresión de las fronteras disciplinarias.ITESO, A.C

Nuclear translocation of glutaminase GLS2 in human cancer cells associates with proliferation arrest and differentiation

Glutaminase (GA) catalyzes the first step in mitochondrial glutaminolysis playing a key role in cancer metabolic reprogramming. Humans express two types of GA isoforms: GLS and GLS2. GLS isozymes have been consistently related to cell proliferation, but the role of GLS2 in cancer remains poorly understood. GLS2 is repressed in many tumor cells and a better understanding of its function in tumorigenesis may further the development of new therapeutic approaches. We analyzed GLS2 expression in HCC, GBM and neuroblastoma cells, as well as in monkey COS-7 cells. We studied GLS2 expression after induction of differentiation with phorbol ester (PMA) and transduction with the full-length cDNA of GLS2. In parallel, we investigated cell cycle progression and levels of p53, p21 and c-Myc proteins. Using the baculovirus system, human GLS2 protein was overexpressed, purified and analyzed for posttranslational modifications employing a proteomics LC-MS/MS platform. We have demonstrated a dual targeting of GLS2 in human cancer cells. Immunocytochemistry and subcellular fractionation gave consistent results demonstrating nuclear and mitochondrial locations, with the latter being predominant. Nuclear targeting was confirmed in cancer cells overexpressing c-Myc- and GFP-tagged GLS2 proteins. We assessed the subnuclear location finding a widespread distribution of GLS2 in the nucleoplasm without clear overlapping with specific nuclear substructures. GLS2 expression and nuclear accrual notably increased by treatment of SH-SY5Y cells with PMA and it correlated with cell cycle arrest at G2/M, upregulation of tumor suppressor p53 and p21 protein. A similar response was obtained by overexpression of GLS2 in T98G glioma cells, including downregulation of oncogene c-Myc. Furthermore, human GLS2 was identified as being hypusinated by MS analysis, a posttranslational modification which may be relevant for its nuclear targeting and/or function. Our studies provide evidence for a tumor suppressor role of GLS2 in certain types of cancer. The data imply that GLS2 can be regarded as a highly mobile and multilocalizing protein translocated to both mitochondria and nuclei. Upregulation of GLS2 in cancer cells induced an antiproliferative response with cell cycle arrest at the G2/M phase

Use of tocilizumab in kidney transplant recipients with COVID-1

Acute respiratory distress syndrome associated with coronavirus infection is related to a cytokine storm with large interleukin-6 (IL-6) release. The IL-6-receptor blocker tocilizumab may control the aberrant host immune response in patients with coronavirus disease 2019 (COVID-19) . In this pandemic, kidney transplant (KT) recipients are a high-risk population for severe infection and showed poor outcomes. We present a multicenter cohort study of 80 KT patients with severe COVID-19 treated with tocilizumab during hospital admission. High mortality rate was identified (32.5%), related with older age (hazard ratio [HR] 3.12 for those older than 60 years, P = .039). IL-6 and other inflammatory markers, including lactic acid dehydrogenase, ferritin, and D-dimer increased early after tocilizumab administration and their values were higher in nonsurvivors. Instead, C-reactive protein (CRP) levels decreased after tocilizumab, and this decrease positively correlated with survival (mean 12.3 mg/L in survivors vs. 33 mg/L in nonsurvivors). Each mg/L of CRP soon after tocilizumab increased the risk of death by 1% (HR 1.01 [confidence interval 1.004-1.024], P = .003). Although patients who died presented with worse respiratory situation at admission, this was not significantly different at tocilizumab administration and did not have an impact on outcome in the multivariate analysis. Tocilizumab may be effective in controlling cytokine storm in COVID-19 but randomized trials are needed