A Comprehensive Tyrosine Phosphoproteomic Analysis Reveals Novel Components of the Platelet CLEC-2 Signaling Cascade

This is an Accepted Manuscript of an article published by Thieme Publishing Group in Thrombosis and Haemostasis on 04 January 2020, available online at https://www.thieme-connect.de/products/ejournals/abstract/10.1055/s-0039-3400295C-type lectin-like receptor 2 (CLEC-2) plays a crucial role in different platelet-related physiological and pathological processes. It signals through a tyrosine kinase-mediated pathway that is highly dependent on the positive feedback exerted by the platelet-derived secondary mediators, adenosine diphosphate (ADP) and thromboxane A2 (TXA2). Here, we aimed to analyze the tyrosine phosphoproteome of platelets activated with the CLEC-2 agonist rhodocytin to identify relevant phosphorylated tyrosine residues (p-Tyr) and proteins involved in platelet activation downstream of this receptor. We identified 363 differentially p-Tyr residues, corresponding to the majority of proteins previously known to participate in CLEC-2 signaling and also novel ones, including adaptors (e.g., DAPP1, Dok1/3, CASS4, Nck1/2), kinases/phosphatases (e.g., FAK1, FES, FGR, JAK2, SHIP2), and membrane proteins (e.g., G6F, JAM-A, PECAM-1, TLT-1). To elucidate the contribution of ADP and TXA2 at different points of the CLEC-2 signaling cascade, we evaluated p-Tyr levels of residues identified in the analysis and known to be essential for the catalytic activity of kinases Syk(p-Tyr525+526) and Src(p-Tyr419), and for PLCγ2 activity (p-Tyr759). We demonstrated that Syk phosphorylation at Tyr525+526 also happens in the presence of ADP and TXA2 inhibitors, which is not the case for Src-pTyr419 and PLCγ2-pTyr759. Kinetics studies for the three phosphoproteins show some differences in the phosphorylation profile. Ca2+ mobilization assays confirmed the relevance of ADP and TXA2 for full CLEC-2-mediated platelet activation. The present study provides significant insights into the intracellular events that take place following CLEC-2 activation in platelets, contributing to elucidate in detail the CLEC-2 signalosomeThis study was supported by the Spanish Ministry of Economy and Competitiveness (MINECO) [grant No. SAF2016-79662-R], co-funded by the European Regional Development Fund (ERDF); and the Consellería de Cultura, Educación e Ordenación Universitaria, Xunta de Galicia [ED431C 2018/21; predoctoral grant Plan I2C 2014; and Centro Singular de investigación de Galicia accreditation 2016-2019, ED431G/05], co-funded by the European Regional Development Fund (ERDF). The study also received funding from the European Union's Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement No 766118. J.A.E. is supported by Deutsche Forschungsgemeinschaft [DFG grant: EB177/13-1]S

A Dimer of the Toll-Like Receptor 4 Cytoplasmic Domain Provides a Specific Scaffold for the Recruitment of Signalling Adaptor Proteins

The Toll-like receptor 4 (TLR4) is a class I transmembrane receptor expressed on the surface of immune system cells. TLR4 is activated by exposure to lipopolysaccharides derived from the outer membrane of Gram negative bacteria and forms part of the innate immune response in mammals. Like other class 1 receptors, TLR4 is activated by ligand induced dimerization, and recent studies suggest that this causes concerted conformational changes in the receptor leading to self association of the cytoplasmic Toll/Interleukin 1 receptor (TIR) signalling domain. This homodimerization event is proposed to provide a new scaffold that is able to bind downstream signalling adaptor proteins. TLR4 uses two different sets of adaptors; TRAM and TRIF, and Mal and MyD88. These adaptor pairs couple two distinct signalling pathways leading to the activation of interferon response factor 3 (IRF-3) and nuclear factor κB (NFκB) respectively. In this paper we have generated a structural model of the TLR4 TIR dimer and used molecular docking to probe for potential sites of interaction between the receptor homodimer and the adaptor molecules. Remarkably, both the Mal and TRAM adaptors are strongly predicted to bind at two symmetry-related sites at the homodimer interface. This model of TLR4 activation is supported by extensive functional studies involving site directed mutagenesis, inhibition by cell permeable peptides and stable protein phosphorylation of receptor and adaptor TIR domains. Our results also suggest a molecular mechanism for two recent findings, the caspase 1 dependence of Mal signalling and the protective effects conferred by the Mal polymorphism Ser180Leu

Responding to the need of postgraduate education for Planetary Health: Development of an online Master's Degree

Data availability statement: The original contributions presented in the study are included in the article/Supplementary material, further inquiries can be directed to the corresponding author/s.Copyright © 2022 O'Callaghan-Gordo, Moreno, Bosque-Prous, Castro-Sanchez, Dadvand, Guzmán, García-Juanatey, Gascon, Grau, Jordana, Lowe, March, Medina, Mélon, Navas, Núñez Casal, Ruiz-Mallén, Sánchez-Valdivia, Tonne, Triguero-Mas, Zografos and Antó. . Planetary Health has emerged as a new approach to respond to the existential risks that the clime and global environmental crises pose to human societies. As stated by various stakeholders, the challenges involved in Planetary Health are of such magnitude that education must be at the forefront to obtain a meaningful response. Universities and higher education institutions have been specifically called to embed the concept of planetary stewardship in all curricula and train the next generation of researchers and change makers as a matter of urgency. As a response to this call, the Universitat Oberta de Catalunya (UOC), the Universitat Pompeu Fabra (UPF), and the Barcelona Institute for Global Health (ISGlobal) developed the first online and asynchronous Master in Science (MSc) in Planetary Health. The aim of the programme is to train a new generation of academics and professionals who understand the challenges of Planetary Health and have tools to tackle them. This article describes the development of the curriculum of this MSc, presents the main characteristics of the programme and discusses some of the challenges encountered in the development of the programme and its implementation. The design of this MSc was based on: the alignment of the programme with the principles for Planetary Health education with a focus on human health; a multi-, inter-, and trans-disciplinary approach; the urgency to respond to the Anthropocene challenges; and the commitment to the 2030 Agenda. The MSc was recognized as an official degree by the Agency for Quality of the Catalan University System, included in the European Quality Assurance Register for Higher Education, and the Spanish National Academic Coordination body in April 2021 and launched in October 2021. There are currently more than 50 students enrolled in the program coming from a broad range of disciplines and geographic locations. The information presented in this article and the discussion on challenges encountered in developing and implementing the programme can be useful for those working in the development of similar programs.Spanish Ministry of Science and Innovation and State Research Agency through the Centro de Excelencia Severo Ochoa 2019–2023 Program (CEX2018-000806-S); Generalitat de Catalunya through the CERCA Program

Responding to the need of postgraduate education for Planetary Health: development of an online Master's Degree

The Universitat Oberta de Catalunya (UOC), the Universitat Pompeu Fabra (UPF), and the Barcelona Institute for Global Health (ISGlobal) have developed an online and asynchronous Master in Science (MSc) in Planetary Health. The aim of the programme is to train a new generation of academics and professionals who understand the challenges of Planetary Health and have tools to tackle them. The design of this MSc was based on: the alignment of the programme with the principles for Planetary Health education with a focus on human health; a multi-, inter-, and trans-disciplinary approach; the urgency to respond to the Anthropocene challenges; and the commitment to the 2030 Agenda. The MSc was recognized as an official degree by the Spanish academic system on April 2021 and launched in October 2021. There are currently more than 50 students enrolled in the program coming from a broad range of disciplines and geographic locations. This article describes the development of the curriculum of this MSc, presents the main characteristics of the programme and discusses some of the challenges encountered in the development of the programme and its implementation

Perforación tras colonoscopia: experiencia en 16 años Perforation after colonoscopy: our 16 year experience

Objetivo: el propósito de este estudio retrospectivo fue conocer la incidencia de perforaciones de colon por colonoscopia diagnóstica y terapéutica y valorar el manejo de las mismas en nuestro medio. Pacientes y método: se revisaron las perforaciones por colonoscopias realizadas en nuestro hospital entre enero de 1991 y diciembre de 2006. Se analizaron la finalidad del procedimiento, el mecanismo de la lesión, su presentación clínica y radiológica, el retraso diagnóstico, el estado general de los pacientes, el manejo terapéutico y sus resultados. Resultados: en el periodo de estudio hubo 15 perforaciones (0,09%) de un total de 16.285 colonoscopias realizadas (9 varones, 6 mujeres). Nueve ocurrieron tras colonoscopias diagnósticas y 6 terapéuticas. El 60% de las perforaciones fueron advertidas por el endoscopista durante el procedimiento (en el 88,6% de las diagnósticas y en el 16,6% de las terapéuticas). En el 73,7% de los casos el dolor con distensión abdominal fue el síntoma más frecuente y en 11 pacientes con pruebas de imagen se apreció gas extraluminal en el 100% de los casos. El retraso diagnóstico (> 24 horas) ocurrió en el 40% de los pacientes (rango: 1-6 días). Fueron intervenidos 12 pacientes (80%) de los que 4 eran ASA II y 8 ASA III/IV. La morbilidad postoperatoria fue del 44,44% y la mortalidad del 25%. La evolución de los pacientes tratados conservadoramente fue satisfactoria. Conclusión: la perforación de colon por colonoscopia es una complicación rara de consecuencias graves e incluso letales. El tratamiento conservador puede realizarse en casos seleccionados y bajo un control clínico estricto. El tipo de cirugía va a depender de la localización, tamaño de la lesión, patología colónica concomitante y grado de contaminación fecal. La mortalidad parece estar más relacionada con el estado general del paciente que con la propia técnica quirúrgica realizada.<br>Objective: the aim of this retrospective study was to evaluate the incidence of colon perforations from diagnostic and therapeutic colonoscopies, and to assess their management in our hospital. Patients and method: perforations resulting from colonoscopy in our hospital were reviewed for the period January, 1991 to December, 2006. The study analyzed: purpose of procedure, lesion mechanisms, clinical and radiological presentations, delays in diagnosis, patient status, therapeutic handling, and outcome. Result: fifteen perforations (0.09%) (9 males and 6 females) out of a total of 16,285 colonoscopies carried out were seen to have taken place during the study period. Nine of these occurred after diagnostic colonoscopies, and 6 occurred after therapeutic endoscopies. Around 60% of perforations were detected by the endoscopist while carrying out the procedure (88.6% during diagnostic endoscopy, and 16.6% during therapeutic endoscopy). In 73.7% of cases abdominal pain and distension were the most frequent symptoms; extraluminal gas was seen in 100% of cases with imaging techniques. Delayed diagnosis (> 24 hours) occurred in 40% of patients (range: 1-6 days). Twelve patients were operated upon (80%), of whom 4 were ASA II and 8 were ASA III/IV. Postoperative morbidity was 44.44%, and mortality was 25%. The outcome of patients receiving conservative treatment was found to be satisfactory. Conclusion: perforation of the colon during colonoscopy is a rare complication with serious -even lethal- consequences. Conservative treatment can be provided for selected cases under strict clinical control. Type of surgery will depend on the lesion location and size, concomitant colon pathology, and degree of fecal contamination. Mortality seems to be related to general status rather than the surgical technique used

Neumopericardio por fístula gastropericárdica Pneumopericardium caused by gastropericardial fistula

Presentamos el caso de un varón de 56 años que acude a Urgencias por un cuadro de dolor epigástrico irradiado a hombro izquierdo y a cuello. El diagnóstico de pneumopericardio fue realizado tras una radiografía simple de tórax confirmándose una fístula gastropericárdica con la realización de un tránsito con contraste hidrosoluble. Fue tratado quirúrgicamente practicándose gastrectomía total con reconstrucción esofagoyeyunal y drenaje pericárdico, siendo su evolución favorable.This is the case of a 56-year-old male who presented at our Emergency Room complaining of severe epigastric pain that radiated towards the left shoulder and neck. Pneumopericardium was diagnosed with plain chest X-rays, which indicated a gastropericardial fistula. A contrast X-ray film was taken using a water-soluble contrast. A full gastrectomy was performed with esophagojejunostomy reconstruction and pericardial drainage, which resulted in a favourable patient outcome

Calidad de vida y síntomas persistentes tras hospitalización por COVID-19. Estudio observacional prospectivo comparando pacientes con o sin ingreso en UCI

[Abstract] Background: Hospitalized COVID-19 patients are prone to develop persistent symptoms and to show reduced quality of life following hospital admission. Methods: Prospective cohort study of COVID-19 patients admitted to a hospital from March 1 to April 30, 2020. The primary outcome was to compare health related quality of life and persistent symptoms six months after hospital admission, of COVID-19 patients who required ICU admission with those who did not. Results: Among the 242 patients hospitalized during the defined period of time, 44 (18.2%) needed ICU admission. Forty (16.5%) patients died during hospital admission. Two hundred and two (83.5%) patients were discharged alive from the hospital. At six months, 183 (75.6%) patients completed the questionnaires (32 ICU patients and 151 non ICU patients). Ninety-six (52.4%) reported decreased quality of life and 143 (78.1%) described persistent symptoms. More ICU patients showed worsening of their quality of life (71.9 vs. 43.7%, P = 0.004). There were no differences in the proportion of patients with persistent symptoms between ICU and non ICU patients (87.5 vs. 76.2%, P = 0.159). ICU patients showed more frequently dyspnea on exertion (78.1 vs. 47.7%, P = 0.02), dyspnea on light exertion (37.5 vs. 4.6%, P < 0.001), and asthenia (56.3 vs. 29.1, P = 0.003). Conclusions: Survivors of COVID-19 needing hospitalization had persistent symptoms and a decline in the quality of life. ICU patients referred a large decrease of their quality of life compared with non ICU patients.[Resumen] Antecedentes/Contexto. Existe una tendencia en los pacientes hospitalizados por COVID-19 a desarrollar síntomas persistentes y a presentar una disminución en su calidad de vida tras el ingreso hospitalario. Métodos. Estudio de cohorte prospectivo de pacientes COVID-19 con ingreso hospitalario entre el 1 de marzo al 30 de abril de 2020. El objetivo primario fue comparar la calidad de vida relacionada con la salud y la presencia de síntomas persistentes seis meses después del ingreso, comparando los pacientes que requirieron ingreso en UCI con los que no lo precisaron. Resultados. De los 242 pacientes hospitalizados durante el período de estudio, 44 (18,2%) necesitaron ingreso en UCI. 40 (16,5%) pacientes fallecieron durante el ingreso hospitalario. 202 (83,5%) pacientes fueron dados de alta del hospital. A los seis meses, 183 (75,6%) pacientes completaron los cuestionarios (32 pacientes UCI y 151 pacientes no UCI). 96 (52,4%) refirieron disminución de la calidad de vida y 143 (78,1%) describieron síntomas persistentes. Un número mayor de pacientes de UCI mostraron un empeoramiento de su calidad de vida (71,9% vs 43,7%, P = 0,004). No hubo diferencias en la proporción de pacientes con síntomas persistentes entre los pacientes con UCI y sin UCI (87,5% vs 76,2%, P = 0,159). Los pacientes de UCI mostraron con mayor frecuencia disnea de esfuerzo (78,1% vs 47,7%, P = 0,02), disnea de pequeños esfuerzos (37,5% vs 4,6%, P < 0,001) y astenia (56,3 vs 29,1, P = 0,003). Conclusiones. Los supervivientes de COVID-19 que necesitaron hospitalización presentaron síntomas persistentes y un deterioro de su calidad de vida. Los pacientes de UCI refirieron una mayor disminución de su calidad de vida en comparación con los pacientes que no precisaron UCI