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Recent evidence suggests that the prognostic impact of gene mutations in patients with chronic lymphocytic leukemia (CLL) may differ depending on the immunoglobulin heavy variable (IGHV) gene somatic hypermutation (SHM) status. In this study, we assessed the impact of nine recurrently mutated genes (BIRC3, EGR2, MYD88, NFKBIE, NOTCH1, POT1, SF3B1, TP53, and XPO1) in pre-treatment samples from 4580 patients with CLL, using time-to-first-treatment (TTFT) as the primary end-point in relation to IGHV gene SHM status. Mutations were detected in 1588 (34.7%) patients at frequencies ranging from 2.3–9.8% with mutations in NOTCH1 being the most frequent. In both univariate and multivariate analyses, mutations in all genes except MYD88 were associated with a significantly shorter TTFT. In multivariate analysis of Binet stage A patients, performed separately for IGHV-mutated (M-CLL) and unmutated CLL (U-CLL), a different spectrum of gene alterations independently predicted short TTFT within the two subgroups. While SF3B1 and XPO1 mutations were independent prognostic variables in both U-CLL and M-CLL, TP53, BIRC3 and EGR2 aberrations were significant predictors only in U-CLL, and NOTCH1 and NFKBIE only in M-CLL. Our findings underscore the need for a compartmentalized approach to identify high-risk patients, particularly among M-CLL patients, with potential implications for stratified management., Depto. de Bioquímica y Biología Molecular, Fac. de Farmacia, TRUE, pub

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[Background]: Rare inherited thrombocytopenias are caused by alterations in genes involved in megakaryopoiesis, thrombopoiesis and/or platelet release. Diagnosis is challenging due to poor specificity of platelet laboratory assays, large numbers of culprit genes, and difficult assessment of the pathogenicity of novel variants. [Objectives]: To characterize the clinical and laboratory phenotype, and identifying the underlying molecular alteration, in a pedigree with thrombocytopenia of uncertain etiology. [Patients/Methods]: Index case was enrolled in our Spanish multicentric project of inherited platelet disorders due to lifelong thrombocytopenia and bleeding. Bleeding score was recorded by ISTH‐BAT. Laboratory phenotyping consisted of blood cells count, blood film, platelet aggregation and flow cytometric analysis. Genotyping was made by whole‐exome sequencing (WES). Cytoskeleton proteins were analyzed in resting/spreading platelets by immunofluorescence and immunoblotting. [Results]: Five family members displayed lifelong mild thrombocytopenia with a high number of enlarged platelets in blood film, and mild bleeding tendency. Patient's platelets showed normal aggregation and granule secretion response to several agonists. WES revealed a novel nonsense variant (c.322C>T; p.Gln108*) in TPM4 (NM_003290.3), the gene encoding for tropomyosin‐4 (TPM4). This variant led to impairment of platelet spreading capacity after stimulation with TRAP‐6 and CRP, delocalization of TPM4 in activated platelets, and significantly reduced TPM4 levels in platelet lysates. Moreover, the index case displayed up‐regulation of TPM2 and TPM3 mRNA levels. [Conclusions]: This study identifies a novel TPM4 nonsense variant segregating with macrothrombocytopenia and impaired platelet cytoskeletal remodeling and spreading. These findings support the relevant role of TPM4 in thrombopoiesis and further expand our knowledge of TPM4‐related thrombocytopenia.

This article uses Qualitative Comparative Analysis (QCA) to study knowledge-transfer processes that are difficult to observe due to the absence of standardized data sources. It examines university–industry relationships and focuses on firms’ chances of developing R&D projects and services within a local innovation system. The article makes methodological and substantive contributions. First, micro- and meso-level qualitative approaches move towards the tradition of quantitative studies on science through a process that illustrates the construction of quantifiable dimensions and their possibilities of analysis. Second, analysis reveals the conditioning factors that influence non-R&D-intensive small and medium-sized enterprises (SMEs) into using university knowledge. The conclusions highlight the possibilities of this methodology’s cross-fertilization with other approaches in the empirical study of science and innovation., Este artículo utiliza el Análisis Cualitativo Comparado (QCA) para estudiar procesos de transferencia de conocimiento difíciles de observar debido a la ausencia de fuentes de datos estandarizados. Estudia las relaciones entre universidad y empresa focalizando en las posibilidades de las empresas de contratar proyectos y servicios de I+D en un sistema local de innovación. El artículo realiza contribuciones de carácter metodológico y sustantivo. En primer lugar, las aproximaciones cualitativas de carácter micro y meso se acercan a la tradición de los estudios cuantitativos sobre la ciencia a través de un proceso que ilustra la construcción de dimensiones cuantificables y sus posibilidades de análisis. En segundo lugar, el análisis muestra los condicionantes que influyen en que las pequeñas y medianas empresas no intensivas en I+D utilicen conocimiento de la universidad. Las conclusiones resaltan las posibilidades de fertilización cruzada de esta metodología con otros enfoques en el estudio empírico de la ciencia y la innovación.

Interstitial 14q32 deletions involving IGH gene are infrequent events in chronic lymphocytic leukemia (CLL), affecting less than 5% of patients. To date, little is known about their clinical impact and molecular underpinnings, and its mutational landscape is currently unknown. In this work, a total of 871 CLLs were tested for the IGH break-apart probe, and 54 (6.2%) had a 300 kb deletion of 3′IGH (del-3′IGH CLLs), which contributed to a shorter time to first treatment (TFT). The mutational analysis by next-generation sequencing of 317 untreated CLLs (54 del-3′IGH and 263 as the control group) showed high mutational frequencies of NOTCH1 (30%), ATM (20%), genes involved in the RAS signaling pathway (BRAF, KRAS, NRAS, and MAP2K1) (15%), and TRAF3 (13%) within del-3′IGH CLLs. Notably, the incidence of TRAF3 mutations was significantly higher in del-3′IGH CLLs than in the control group (p < .001). Copy number analysis also revealed that TRAF3 loss was highly enriched in CLLs with 14q deletion (p < .001), indicating a complete biallelic inactivation of this gene through deletion and mutation. Interestingly, the presence of mutations in the aforementioned genes negatively refined the prognosis of del-3′IGH CLLs in terms of overall survival (NOTCH1, ATM, and RAS signaling pathway genes) and TFT (TRAF3). Furthermore, TRAF3 biallelic inactivation constituted an independent risk factor for TFT in the entire CLL cohort. Altogether, our work demonstrates the distinct genetic landscape of del-3′IGH CLL with multiple molecular pathways affected, characterized by a TRAF3 biallelic inactivation that contributes to a marked poor outcome in this subgroup of patients.

Inherited antithrombin deficiency, the most severe form of thrombophilia, is predominantly caused by variants in SERPINC1. Few causal structural variants have been described, usually detected by multiplex ligation-dependent probe amplification or cytogenetic arrays, which only define the gain or loss and the approximate size and location. This study has done a complete dissection of the structural variants affecting SERPINC1 of 39 unrelated patients with antithrombin deficiency using multiplex ligation-dependent probe amplification, comparative genome hybridization array, long-range PCR, and whole genome nanopore sequencing. Structural variants, in all cases only affecting one allele, were deleterious and caused a severe type I deficiency. Most defects were deletions affecting exons of SERPINC1 (82.1%), but the whole cohort was heterogeneous, as tandem duplications, deletion of introns, or retrotransposon insertions were also detected. Their size was also variable, ranging from 193 bp to 8 Mb, and in 54% of the cases involved neighboring genes. All but two structural variants had repetitive elements and/or microhomologies in their breakpoints, suggesting a common mechanism of formation. This study also suggested regions recurrently involved in structural variants causing antithrombin deficiency and found three structural variants with a founder effect: the insertion of a retrotransposon, duplication of exon 6, and a 20-gene deletion. Finally, nanopore sequencing was determined to be the most appropriate method to identify and characterize all structural variants at nucleotide level, independently of their size or type.

A través de este trabajo se quiere mostrar la importancia de la enseñanza de una segunda lengua en un aula de Primaria, siempre teniendo en cuenta las características de nuestro alumnado, adaptándonos a sus necesidades con una práctica docente apropiada. Para ello, se ha realizado una planificación y su posterior puesta en práctica de un proyecto en un aula bilingüe fundamentado en las ocho inteligencias múltiples de Howard Gardner. La propuesta, empieza con el planteamiento de un tema novedoso para el alumnado que servirá de hilo conductor de todo el proyecto, a través del cual se pretende que alcancen los objetivos y contenidos propuestos con anterioridad, para la etapa con la que se ha trabajado, Grado en Educación Primaria

Teaching in the 21st century is a huge challenge, and every school must deal with all the changes happening within society and be, at the same time, an enjoyable place for the students to develop their skills to live and function in the current society. Adapted to the times we are living, the United Nations developed and put into practice the 2030 Agenda, but for it to be useful, they need every single action we can do. That is why this proposal gets the 2030 Agenda to the classroom, where the frequently called “citizens of the future” are preparing to learn how to live in this society, La enseñanza en el siglo 21 es un gran desafío, y en cada una de las escuelas se debe lidiar con los constantes cambios que están sucediendo en la sociedad y al mismo tiempo deben ser un lugar donde sus estudiantes disfruten desarrollando las habilidades que necesitan para vivir en la sociedad actual. Adaptado a la época que vivimos, las Naciones Unidas han desarrollado y puesto en práctica la Agenda 2030, pero para que sea útil, necesitan que todos nosotros contribuyamos. Es por ello que se presenta esta propuesta, donde se lleva la Agenda 2030 al aula, donde los llamados “ciudadanos/as del futuro” se están preparando para aprender cómo vivir en sociedad, Grado en Educación Primaria

This Final Degree Work is based on the importance of Storytelling in Early Years. This teaching technique is motivating, challenging and enjoyable, and can help develop positive attitudes towards the foreign language, culture and language learning. Children exercises the imagination; they can get involved in a story as they identify with the characters and try to interpret the narrative and the illustrations. On the other hand, it is really challenging how to implement a storytelling proposal in an infant class in the subject of English as a second language which means that English turns into the vehicular language for learning. A story can be used to develop other subjects such as Knowledge and Understanding of the World, Arts and Crafts, Maths, Music, Drama and Physical Education, RESUMEN Este Trabajo Fin de Grado se centra principalmente en la importancia del Storytelling en el segundo ciclo de Educación Infantil. Esta técnica de enseñanza es motivadora, estimulante y divertida, y puede ayudar a desarrollar actitudes positivas hacia la lengua extranjera, la cultura y el aprendizaje de idiomas. El alumnado ejercita la imaginación, puede involucrarse en un cuento al identificarse con los personajes y tratar de interpretar la narración y las ilustraciones. Por otro lado, es todo un reto implementar una propuesta de Storytelling en una clase de infantil en la asignatura de inglés, como segunda lengua, lo que significa que el inglés se convierte en la lengua vehicular del aprendizaje. Asimismo, un cuento puede servir para desarrollar otras asignaturas como Conocimiento del Entorno, Plástica, Matemáticas, Música, Teatro y Educación Física, Grado en Educación Infantil

For some years now, literature has been a powerful and suggestive tool in Infant Education classrooms all over the world. However, in many occasions, this field only shows the “male” sphere of the society, silencing the female perspective. For this reason, and taking into account the ODS number 5 about gender equality, the main aim of this investigation will be to make a first approach to female relevant figures in the English lessons in Infant Education though literature and art. Moreover, another goal will be to emphasize the current presence of female protagonists who share traits like braveness, effort and persistence. Regarding the structure of my investigation, after the theoretical framework based on the lack of female references in early ages and the use of picture books to introduce them, there will be a didactic proposal using a global methodology based on the Amara Berri’s premises. In this line of argument, it is already known that Infant Education is a crucial educative stage and every single children’s environment element must be taken into consideration. Therefore, an initial feminist approach is essential in order to create “meaning-makers” kids who are respectful and empathic to the Others and contribute from their childhood to promote social and moral values in their daily lives, not only at school but also at home., Desde hace algunos años, la literatura es una herramienta poderosa y sugerente en las aulas de Educación Infantil de todo el mundo. Sin embargo, en muchas ocasiones este campo solo muestra la esfera “masculina” de la sociedad, silenciando la perspectiva femenina. Por ello y, teniendo en cuenta el ODS número 5 sobre igualdad de género, el principal objetivo de esta investigación será realizar un primer acercamiento a figuras femeninas relevantes en las clases de inglés en Educación Infantil a través de la literatura y el arte. Además, otro objetivo será enfatizar la presencial actual de protagonistas femeninas que comparten rasgos como la valentía, el esfuerzo y la perseverancia. En cuanto a la estructura de la investigación, después del marco teórico basado en la falta de referencias femeninas en edades tempranas y el uso de libros ilustrados para presentar éstas mismas, se presentará una propuesta didáctica siguiendo una metodología global basada en las premisas de Amara Berri. En esta línea de argumentación, se sabe que la Educación Infantil es una etapa educativa crucial y debe tenerse en cuenta cada elemento del entorno de los niños y las niñas. Por lo tanto, un enfoque feminista inicial es fundamental para crear niños y niñas “constructores de significado”, respetuosos y empáticos con los otros y contribuyendo desde la infancia a promover los valores sociales y morales en su vida diaria, no solo en la escuela sino también en casa, Grado en Educación Infantil

[Introduction]: Intestinal microbiota is arising as a new element in the physiopathology of cardiovascular diseases. A healthy microbiota includes a balanced representation of bacteria with health promotion functions (symbiotes). The aim of this study is to analyse the relationship between intestinal microbiota composition and arterial stiffness. [Methods and analysis]: An observational case—control study will be developed. Cases will be defined by the presence of at least one of the following: carotid-femoral pulse wave velocity (cf-PWV), Cardio-Ankle Vascular Index (CAVI), brachial ankle pulse wave velocity (ba or ba-PWV) above the 90th percentile, for age and sex, of the reference population. Controls will be selected from the same population as cases. The study will be developed in Primary Healthcare Centres. We will select 500 subjects (250 cases and 250 controls), between 45 and 74 years of age. Cases will be selected from a database that combines data from EVA study (Spain) and Guimarães/Vizela study (Portugal). Measurements: cf-PWV will be measured using the SphygmoCor system, CAVI, ba-PWV and Ankle-Brachial Index will be determined using VaSera device. Gut microbiome composition in faecal samples will be determined by 16S ribosomal RNA sequencing. Lifestyle will be assessed by food frequency questionnaire, adherence to the Mediterranean diet and IPAQ (International Physical Activity Questionnaire). Body composition will be evaluated by bioimpedance. [Ethics and dissemination]: The study has been approved by ‘Committee of ethics of research with medicines of the health area of Salamanca’ on 14 December 2018 (cod. 2018-11-136) and the ’Ethics committee for health of Guimaraes’ (Portugal) on 15 October 2019 (ref: 67/2019).All study participants will sign an informed consent form agreeing to participate in the study, in compliance with the Declaration of Helsinki and the WHO standards for observational studies. The results of this study will allow a better desc

NGS Establishment in Multidisciplinary Healthcare (NEMHESYS) is an Erasmus+ programme with the purpose of providing qualified staff with the essential technical and bioinformatic knowledge and skills on next-generation sequencing (NGS) to be able to carry out NGS studies and perform some of the most common types of analyses. The clinical application of NGS has become easier with advancements in technologies.1 However, the investment needed to bring NGS into medical practice remains significant, with the scale of knowledge required being unprecedented at most hospitals. In addition, these novel technologies bring new challenges in translating NGS to clinical practice, at both technical and regulatory level, in terms of data management, interpretation of the results, and genetic counseling.2,3 All these aspects justify the consideration of what will be the precise role of NGS in diagnosis, risk assessment, response prediction, and treatment monitoring, today and tomorrow. Thus, to evaluate the implementation of NGS in European healthcare/research centers, a mapping survey was carried out, based on previous NGS mapping studies.

[Background]: Several genetic alterations have been identified as driver events in chronic lymphocytic leukemia (CLL) pathogenesis and oncogenic evolution. Concurrent driver alterations usually coexist within the same tumoral clone, but how the cooperation of multiple genomic abnormalities contributes to disease progression remains poorly understood. Specifically, the biological and clinical consequences of concurrent high-risk alterations such as del(11q)/ATM-mutations and del(17p)/TP53-mutations have not been established., [Methods]: We integrated next-generation sequencing (NGS) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 techniques to characterize the in vitro and in vivo effects of concurrent monoallelic or biallelic ATM and/or TP53 alterations in CLL prognosis, clonal evolution, and therapy response., [Results]: Targeted sequencing analysis of the co-occurrence of high-risk alterations in 271 CLLs revealed that biallelic inactivation of both ATM and TP53 was mutually exclusive, whereas monoallelic del(11q) and TP53 alterations significantly co-occurred in a subset of CLL patients with a highly adverse clinical outcome. We determined the biological effects of combined del(11q), ATM and/or TP53 mutations in CRISPR/Cas9-edited CLL cell lines. Our results showed that the combination of monoallelic del(11q) and TP53 mutations in CLL cells led to a clonal advantage in vitro and in in vivo clonal competition experiments, whereas CLL cells harboring biallelic ATM and TP53 loss failed to compete in in vivo xenotransplants. Furthermore, we demonstrated that CLL cell lines harboring del(11q) and TP53 mutations show only partial responses to B cell receptor signaling inhibitors, but may potentially benefit from ATR inhibition., [Conclusions]: Our work highlights that combined monoallelic del(11q) and TP53 alterations coordinately contribute to clonal advantage and shorter overall survival in CLL.

Myelodysplastic syndromes (MDS) are hematological disorders at high risk of progression to secondary acute myeloid leukemia (sAML). However, the mutational dynamics and clonal evolution underlying disease progression are poorly understood at present. To elucidate the mutational dynamics of pathways and genes occurring during the evolution to sAML, next generation sequencing was performed on 84 serially paired samples of MDS patients who developed sAML (discovery cohort) and 14 paired samples from MDS patients who did not progress to sAML during follow-up (control cohort). Results were validated in an independent series of 388 MDS patients (validation cohort). We used an integrative analysis to identify how mutations, alone or in combination, contribute to leukemic transformation. The study showed that MDS progression to sAML is characterized by greater genomic instability and the presence of several types of mutational dynamics, highlighting increasing (STAG2) and newly-acquired (NRAS and FLT3) mutations. Moreover, we observed cooperation between genes involved in the cohesin and Ras pathways in 15-20% of MDS patients who evolved to sAML, as well as a high proportion of newly acquired or increasing mutations in the chromatin-modifier genes in MDS patients receiving a disease-modifying therapy before their progression to sAML.

During platelet activation, arachidonic acid (AA) is released from membrane phospholipids and metabolized to thromboxane A2 (TXA2) through the actions of cyclooxygenase-1 (COX-1) and TXA2 synthase. Note, TXA2 binds to the platelet TXA2 receptor, causing shape change, secretion and platelet aggregation.1 Also, COX-1 (599aa; 70 kDa) has cyclooxygenase and peroxidase activities and it is functionally active as a homodimer, with each COX-1 monomer consisting of four highly conserved domains: an N-terminal signal peptide, a dimerization domain, a membrane-binding domain (MBD) and a large C-terminal catalytic domain2 (Figure 1A). Irreversible COX-1 inhibition by aspirin is a widely established anti-platelet therapy in cardiovascular disease.