Inhibition of SHP1 and SHP2 as a molecular targeted therapy against myeloid leukaemias

[EN]Haematopoiesis is a very relevant differentiation process in adult humans where a multipotent cell, the haematopoietic stem cell (HSC), generates a widely varied, fully differentiated progeny, with immune defence, nutrient exchange and volume homeostasis functions. The regulatory cues governing the biology of HSCs must be tightly regulated in order to ensure their own self-renewal, as well as the proper turnover of differentiated cells. These signals are provided by the surrounding environment, known as niche, integrated by both haematopoietic and non-haematopoietic cells. The disruption of this fine equilibrium by alteration of either the external signals or their intracellular transduction in haematopoietic stem and progenitor cells (HSPCs) leads to the development of haematologic malignancies, including leukaemia. Two important blood disorders affecting the myeloid lineage are acute and chronic myeloid leukaemia (AML and CML, respectively). They are especially recurrent among the elderly, with a median age at diagnosis of 65 years for CML and 70 for AML, a fact to be considered due to the increasing life expectancy in Western countries. AML is a highly heterogeneous and aggressive disease with poor prognosis and, in general, no significant therapeutic improvements beyond chemotherapy over the last four decades. An exception to this scenario is the treatment of acute promyelocytic leukaemia (APL), which is highly responsive to pro-differentiative therapy, consisting of all-trans-retinoic acid (ATRA) and arsenic trioxide (ATO). Unlike AML, CML is highly homogeneous in terms of molecular biology, with the expression of the fusion oncokinase breakpoint cluster region-ABL proto-oncogene 1, non-receptor tyrosine kinase (BCR-ABL) as the main pathogenic driver. In the early 2000s, the clinical use of tyrosine kinase inhibitors (TKI) targeting this protein revolutionised the management of CML due to great improvements in treatment response and survival rates. However, this disease remains challenging in particular cases. Despite its heterogeneous nature, AML displays a differentiation blockage as a hallmark. This feature, together with the example of the differentiation-based APL treatment, has prompted the development of an important line of research focusing on the molecular mechanisms governing cell differentiation during haematopoiesis. This knowledge would lead to a better understanding of the dysregulated processes leading to pathogenesis and their subsequent pharmacological targeting to treat the disease. In line with this, the present work sought to assess in detail the involvement of SRC homology 2 domain containing protein tyrosine phosphatases 1 (SHP1) and 2 (SHP2) in the differentiation of leukaemic cells and the potential of these molecules as pharmacological targets for AML. Herein, it was demonstrated the cooperative function of both phosphatases in phorbol ester-induced cell differentiation, with an enhanced differentiated phenotype of cells subjected to simultaneous downregulation of these proteins. In addition, the kinase SRC was identified as a downstream target of SHP2 in this process, which appeared to influence the extent of the differentiation stimulus triggered by phorbol 12-myristate-13-acetate (PMA). Besides, the role of both phosphatases on cell differentiation showed to be partially overlapping through the regulation of β-catenin protein levels. Based on this evidence, the chemical inhibitor of SHP1 and SHP2 NSC 87877 (NSC) was successfully tested to boost the differentiation-inducing effect of phorbol esters in the AML cell line HL-60. Moreover, this compound synergised with the phorbol ester 13-O-acetyl-12-deoxyphorbol or prostratin (PRS) to prevent proliferation of not only in HL-60 cells, but also additional cell lines used as AML models (NB-4, OCI-AML2 and THP-1). Most importantly, the anti-leukaemic activity of this combination was corroborated in vivo with a xenograft mouse model and in primary cells from AML patients ex vivo. On the other hand, the management of CML still requires further improvement, since the success of TKI relies on long-term administration to patients due to the existence of quiescent BCR-ABL independent leukaemic stem cells (LSCs). This is associated with intolerances in patients and great costs for national health systems. Moreover, selective pressure on leukaemic blasts can lead to the emergence of point mutations in BCR-ABL that confer acquired resistance to TKI. To overcome this issue, novel therapeutic approaches based on co-targeting BCR-ABL and other important contributors to CML pathogenesis are subject of intense research. Based on this idea and the involvement of SHP1 and SHP2 in the disease, NSC was tested in combination with TKI in different models of CML. This compound enhanced the effect of the TKI imatinib (IM) and nilotinib (NL) in different CML cell lines (K-562, KCL-22 and BV-173). Immunoblotting analyses allowed the observation of a strong decrease in β-catenin levels upon NSC treatment and a moderate downregulation of c-MYC induced by both IM and NSC. Further gene expression studies identified CCND1, CCND2, CDKN1C and TLE2 as targets of either one or both drugs that might mediate the anti-leukaemic activity of their combination. Additionally, NSC also displayed anti-proliferative potential in patient-derived induced pluripotent stem cells (iPSCs), intrinsically resistant to IM at moderate doses. Treatment of these cells with NSC exerted a dramatic cell cycle arrest concomitant with the downregulation of CTNNB1, encoding β-catenin, as well as CCND1 and CCND2. Additionally, some members of TLE family were modulated by this drug. In summary, the results described in the present work support a promising therapeutic potential of the chemical inhibitor NSC 87877 in different myeloid malignancies where SHP1 and SHP2 are deeply involved. Furthermore, some mechanistic insight on the molecules connected to these phosphatases in both disease biology and pharmacological mode of action of the inhibitor has been provided

Local Government planning: from data to action

Decentralisation is built on the assu mption that decentralized governme nts are more knowledgeable about and responsive to the needs of the poor. This article ex amines the role of local governments in Kenya and the ways in which they make their decisions about the allocation of resources to deliver water and sanitation services. Two major challenges are identified: i) lack of data that accurately reveal which areas are most in need; and ii) inadequate instruments for planning, monitoring and evaluation. In tackling previous shortcomings, this study i) adopts a new specific appr oach for data collection at community level, and ii) exploits these data through simple composite indicator s as policy tools that assist local government with decision-making. It concludes that accurate and compre hensive data are the basis of effective targeting and prioritization, which are fundamental to sector planning.Peer ReviewedPostprint (published version

Monitoring human right to water in rural areas: a Nicaraguan case study

Much effort has gone into the recognition of the hu man right to water. Without doubt, this milestone influences governance and decision making processes at different scales. Now it is essential to shift discussion from legal and conceptual framework to p ractice. Taking this idea into account, the article proposes a methodology for monitoring the implement ation of the right. The work takes the legal and conceptual framework of the right, agreed by intern ational consensus, as a starting point. Moreover, l essons learnt from other indicators and indexes constructi on are considered. It is based on a water & sanitat ion rural services case study carried out in Nicaragua and pa rticularly in Jinotega and Matagalpa -rural departm ents from the north-central region-. The different categ ories of the right to water were included in survey s and structured interviews design that were conducted in rural households and water comities, respectively. Ingeniería sin Fronteras-ApD, an Spanish NGDO, prom oted the study in cooperation with local organizati ons -La Cuculmeca and Coalición de Organizaciones por e l Derecho al Agua (CODA)-. Results show how the inclusion of right to water dimensions gives pertin ent insights both as regards knowledge about the re al situation of rural communities as for improving gov ernance in rural water supply.Peer ReviewedPostprint (published version

Monitoring sanitation and hygiene in the 2030 agenda for sustainable development: A review through the lens of human rights

International monitoring of drinking water and sanitation has been jointly carried out by WHO and UNICEF through their Joint Monitoring Programme (JMP). With the end of the Millennium Development Goals (MDGs) era in 2015, the JMP has proposed a post-2015 framework for integrated monitoring of water and sanitation targets included in the Sustainable Development Goal no. 6. This article discusses how each element of the proposed sanitation target and corresponding indicators can be understood from a human rights perspective. Building on the MDGs, and although some of the weaknesses and gaps persist, the discussion suggests that the post-2015 proposal is a step forward towards a monitoring framework where human rights elements related to sanitation are effectively promoted. In addition, to support the interpretation and implementation of the normative content of human rights obligations related to sanitation, the study proposes a reduced set of easy-to-assess indicators to measure the normative criteria of this right, which are then grouped in a multidimensional framework to describe increasing levels of sanitation service. To do this, the study combines literature review and specific local experience from three case studies. It is shown that the proposed monitoring tools, namely the indicators and the multidimensional indicator framework, provide guidance on monitoring the human right to sanitation. In doing so, they might ultimately help sector stakeholders in the realization of this right.Peer ReviewedPostprint (published version

Detección del discurso de odio en Twitter

[ES] En este Trabajo de Fin de Grado se presenta el diseño e implementación realizado al crear un sistema de alerta que avise de tweets que contengan discurso de odio en su publicación. Para ello, se emplearán diversos tipos de modelos, utilizando técnicas relacionadas con el Aprendizaje Automático, para hacer una clasificación automática de dichas publicaciones. Estos modelos se explicarán y compararán para escoger el que obtenga mejores resultados sobre tweets reales ya etiquetados.[EN] In this Final Degree Project, the design and implementation of an alert system that warns of tweets containing hate speech in their publication is presented. For this purpose, different types of models will be used, using techniques related to Automatic Learning, to make an automatic classification of these publications. These models will be explained and compared to choose the one that obtains better results on real and tagged tweets.[CA] En aquest Treball de Fi de Grau, es presenta el disseny i implementació realitzat al crear un sistema d’alerta que avise de tweets que continguen discurs d’odi en la seua publicació. Per a això, s’empraran diversos tipus de models, utilitzant tècniques relacionades amb el Aprenentatge Automàtic, per a fer una classificació automàtica d’aquestes publicacions. Aquests models s’explicaran i compararan per a triar el que obtinga millors resultats sobre tweets reals ja etiquetats.Pérez Fernández, A. (2020). Detección del discurso de odio en Twitter. http://hdl.handle.net/10251/150500TFG

Metodologías activas con TIC en el área de Educación Física

El presente trabajo busca analizar la repercusión educativa de las metodologías activas en el contexto de la Educación Física (EF). Por lo que para cada modelo seleccionado (Gamificación, Flipped Classroom, Realidad Aumentada, Realidad Virtual, Aprendizaje-Servicio y Aprendizaje Basado en Proyectos) se estudian diferentes aspectos como la conceptualización, los beneficios e inconvenientes y el impacto educativo en las diferentes etapas formativas. Por ello, el trabajo se define como una revisión teórica de las aportaciones científicas más relevantes. En base a esto, es necesario entender que el proyecto también busca trazar un nexo teórico-práctico entre las Tecnologías de la Información y Comunicación (TIC) y un contexto educativo diferente al que supone un aula ordinaria.This work seeks to analyse the educational impact of active methodologies in the context of Physical Education (PE). Therefore, for each selected model (Gamification, Flipped Classroom, Augmented Reality, Virtual Reality, ServiceLearning and Project-Based Learning), different aspects are studied, such as the conceptualization, advantages and disadvantages and the educational impact in different training stages. For this reason, the work is defined as a theoretical review of the most relevant scientific contributions. Based on this, it is necessary to understand that the project also seeks to draw a theoretical-practical link between Information and Communication technologies (ICT) and an educational context different form that of an ordinary classroom

The combined function of Podosphaera xanthii's mechanisms to suppress immunity triggered by chitin.

Fungal pathogens are the main destructive microorganisms for terrestrial plants and pose increasing challenges for global agricultural production. Chitin is a vital building block for fungal cell walls and a widely effective inducer for plant immunity. Chitin-triggered immunity is a powerful plant defense response against fungi. Therefore, phytopathogenic fungi have developed different virulence factors that allow them to suppress the activation of this defensive response. In this study, we intend to evaluate the molecular mechanisms of suppression the chitin-triggered previously identified in Podosphaera xanthii, the main causal agent of powdery mildew in cucurbits. These mechanisms consist of the modification of chitin immunogenic oligomers (CDA), the binding to these oligomers (CHBE) and their degradation (EWCAs). For this, we used RNA interference (RNAi) technology, which consists of the application of double-stranded RNA (dsRNA) designed to suppress the expression of the PxCDA and PxEWCA genes, which would result in the reduction of the three mechanisms of suppression of chitin signaling mentioned above, since CDA and CHBE proteins are products of the same PxCDA gene. Application of dsRNA was carried out using leaf disc assays and infiltration of melon cotyledons. The preliminary results obtained indicate that the application of dsRNA significantly reduces the development of the fungus and the symptoms of powdery mildew disease in melon, suggesting that chitin signaling suppression mechanisms are essential for the development of P. xanthii.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Coordinated role of chitin-triggered immunity suppression mechanisms of Podosphaera xanthii.

Fungal pathogens are the main destructive microorganisms for terrestrial plants and pose increasing challenges for global agricultural production. Chitin is a vital building block for fungal cell walls and a widely effective inducer for plant immunity that, through chitin-triggered immunity, can defend against fungi attack. That is why the phytopathogenic fungi have developed different virulence factors that allow them to suppress the activation of this defensive response. In this study, the molecular mechanisms of chitin-triggered suppression, previously identified in the cucurbit powdery mildew Podosphaera xanthii, have been evaluated in detail. These mechanisms consist of the modification of chitin immunogenic oligomers (CDA), the binding to these oligomers (CHBE) and their degradation (EWCAs). For this, the RNA interference (RNAi) technology, which consists of the application of double-stranded RNA (dsRNA) designed to suppress the expression of target genes, was used. The preliminary results obtained using this strategy significantly reduces the development of the fungus and the symptoms of powdery mildew disease in melon, suggesting that chitin signaling suppression mechanisms are essential for the development of P. xanthii. This work was supported by AEI (PDC2021-121373-C21).Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Optimización de parámetros de operación en proceso de ultrafiltración para la eliminación de materia orgánica en la regeneración de aguas de proceso de una industria papelera

The scientific-technical objective of the project proposal is to study and develop an optimal operating methodology for an ultrafiltration membrane process for the removal of organic matter in the process water regeneration of a paper industry located in Alcoy -Province of Alicante - Spain. The optimization methodology developed will take into account the effects of fouling on the surface and pores of the membrane caused by colloids and suspended solids.El objetivo científico-técnico de la propuesta del proyecto de fin de máster es estudiar y desarrollar una metodología de operación óptima para un proceso por membranas de ultrafiltración para la eliminación de materia orgánica en la regeneración del aguas de proceso de una industria papelera ubicada en Alcoy -Provincia de Alicante - España. La metodología de optimización desarrollada llevará en consideración los efectos del ensuciamiento (fouling) en la superficie y poros de la membrana causados por coloides y sólidos en suspensión. 1. Revisión bibliográfica para establecer el estado del arte de la tecnología de Ultrafiltración atendiendo especialmente a operación y parámetros óptimos de plantas de ultrafiltración; 2. Selección del material de membrana más apropiado para el tipo de sus aguas residuales. 3. Caracterización del agua residual proveniente del tratamiento secundario (reactor biológico) de una industrial de cartón y papel reciclado. Análisis fisicoquímico, potencial Z y distribución de partículas. 4. Puesta a punto de la planta piloto de laboratorio para realizar los ensayos experimentales de caracterización de las membranas (agua pura) y determinación del flux de permeado sobre las condiciones óptimas de proceso (agua tratar). 5. Diseño experimental factorial de las variables; 6. Definir las variables de restricción, variables manipuladas, condiciones y intervalos experimentales de trabajo relativos a concentraciones, composición del agua a tratar, velocidades de flujo, variación de presión en la membrana y temperatura, y desarrollo de experimental en planta piloto; 7. Ensayo de ensuciamiento de las membranas con el agua residual. 8. Mediciones analíticas (fisicoquímicas ), potencial Z y distribución de partículas, de los flux de permeados obtenidos ; 9. Hipótesis y Ajuste del modelado del proceso; 10 . Protocolo y metodología de optimización del proceso de Ultrafiltración UF a partir de las variables de operación y de datos experimentales con el objetivo de maximizar la producción de flujo de permeado y aplicación de dicho protocolo en condiciones residuales reales industriales. 11. Conclusiones finales.Pérez Fernández, DA. (2017). Optimización de parámetros de operación en proceso de ultrafiltración para la eliminación de materia orgánica en la regeneración de aguas de proceso de una industria papelera. http://hdl.handle.net/10251/89492TFG

La responsabilidad penal de las personas jurídicas

El presente trabajo abordará la responsabilidad penal de las personas jurídicas. Tras una breve introducción, analizaremos las reformas que ha sufrido este modelo, que dan lugar a la responsabilidad penal tal y como la conocemos hoy día. También analizaremos el concepto jurídico-penal de la persona jurídica, las personas jurídicas excluidas, así como los dos hechos de conexión previstos en el Código Penal, que permiten exigir responsabilidad penal a la persona jurídica: delitos cometidos por directivos y empleados. Dada la relevancia que han adquirido, analizaremos los modelos de organización y gestión conocidos como Compliance. Tras un repaso a la culpabilidad, circunstancias atenuantes, tentativa y autoría, concluimos el trabajo con las penas previstas en el Código para las personas jurídicas.This thesis will focus on analyzing the criminal liability of legal persons. After a brief introduction, we will analize how this legal regime has changed over time, to reach the model of criminal liability we have nowadays. Once the foundations have been laid, we will proceed to analyze the concept of legal person, within its exclusions. As well, we will study two ways to demand responsaibility reflected in our Legal System: crimes committed by company manager, or employee. Finally, given the relevance Compliance programs have had in recent years, we will study their key aspects and requirements. After cheking criminal culpabilit, extenuating circumstances, attempted commission and authorship, we will conclude conclude this thesis refering to the particular penalties that our Legal System provides. KeyDepartamento de Derecho Penal e Historia y Teoría del DerechoGrado en Derech