Bloodstream form trypanosoma brucei depend upon multiple metacaspases associated with RAB11-positive endosomes

Trypanosoma brucei possesses five metacaspase genes. Of these, MCA2 and MCA3 are expressed only in the mammalian bloodstream form of the parasite, whereas MCA5 is expressed also in the insect procyclic form. Triple RNAi analysis showed MCA2, MCA3 and MCA5 to be essential in the bloodstream form, with parasites accumulating pre-cytokinesis. Nevertheless, triple null mutants (Δmca2/3Δmca5) could be isolated after sequential gene deletion. Thereafter, Δmca2/3Δmca5 mutants were found to grow well both in vitro in culture and in vivo in mice. We hypothesise that metacaspases are essential for bloodstream form parasites, but they have overlapping functions and their progressive loss can be compensated for by activation of alternative biochemical pathways. Analysis of Δmca2/3Δmca5 revealed no greater or lesser susceptibility to stresses reported to initiate programmed cell death, such as treatment with prostaglandin D2. The metacaspases were found to colocalise with RAB11, a marker for recycling endosomes. However, variant surface glycoprotein (VSG) recycling processes and the degradation of internalised anti-VSG antibody were found to occur similarly in wild type, Δmca2/3Δmca5 and triple RNAi induced parasites. Thus, the data provide no support for the direct involvement of T. brucei metacaspases in programmed cell death and suggest that the proteins have a function associated with RAB11 vesicles that is independent of known recycling processes of RAB11-positive endosomes

Characterisation of Leishmania major metacaspase

Metacaspases are distant orthologues of mammalian caspases and have been proposed to play a role in programmed cell death in yeast and plants, but little is known about their function in parasitic protozoa. In this study, I found that the single MCA gene of Leishmania major (LmajMCA or MCA) is expressed in actively replicating amastigotes and promastigotes, whereas expression level is significantly lower in non-dividing stationary phase promastigotes. In order to determine the role of the metacaspase during promastigote division, an analysis of the events of the L. major cell cycle was performed. Based on these findings, immunofluorescence experiments revealed the presence of LmajMCA in punctate structures throughout the cytoplasm of interphase cells and its accumulation in the kinetoplast (mitochondrial DNA) at the time of the organelle's segregation, LmajMCA also translocates to the nucleus during mitosis, where it associates with the mitotic spindle. LmajMCA null mutants could not be generated using standard gene deletion techniques. When LmajMCA was expressed from an episome, the only mutants that were viable were those expressing LmajMCA at physiological levels. Over-expression of LmajMCA in promastigotes leads to changes in ploidy and a severe growth retardation, associated with defects in kinetoplast segregation and nuclear division and an impairment of cytokinesis. Several LmajMCA variants lacking their N-terminal and/or C-terminal domains or with active site mutation(s) (C201G, C201-C202G, C202G, H147A) were expressed, with or without tags, in L. major promastigotes, in Escherichia coli, in the yeast Pichia pastoris, and using an in vitro translation kit. In all the systems used, the expression of most LmajMCA variants was barely detectable and was apparently toxic for the host cells. Some encouraging results were obtained using P. pastoris, with the expression and secretion in the culture medium of N- and C-terminally truncated LmajMCA. Thus, this system may facilitate production of sufficient enzyme for the characterisation of the biochemical properties of LmajMCA. Insights into the putative function of the different domains of LmajMCA were gained when it was observed, that the first 101 N-terminal amino acids of LmajMCA worked, in L.major, as a mitochondrion targeting signal. A potential role of the LmajMCA proline-, glutamine- and tyrosine-rich C-terminal domain in protein- protein interactions was also hypothesized. Together these data suggest that LmajMCA is essential for the correct segregation of the nucleus and kinetoplast and cytokinesis and that the tight regulation of the amount of LmajMCA within the cell is crucial for the development of Leishmania promastigotes. Trypanosoma brucei possesses five metacaspase genes. Two of the encoded proteins, TbMCA1 and TbMCA4, possess substitutions at their putative active site residues and thus might lack peptidase activity. Therefore functional studies were performed only on the three other metacaspases. Triple RNAi analysis showed TbMCA2, TbMCA3 and TbMCA5 to be essential in the bloodstream form, with induced parasites showing a delay in kinetoplast segregation and being blocked in pre-cytokinesis. Triple null mutants (DeltaTbmca2/3DeltaTbmca5) were found to be initially affected in their development, but recovered after several weeks of in vitro adaptation. The initial slow growth rate of the DeltaTbmca2/3DeltaTbmca5 mutant was associated with a delayed entry in cytokinesis. This phenotype was similar, although less extreme, to that produced upon triple RNAi of TbMCA2, TbMCA3 and TbMCA5. These data suggest that these metacaspases are essential for T. brucei bloodstream forms, but they have overlapping functions and their progressive loss can be compensated for by activation of alternative biochemical pathways. Analysis of DeltaTbmca2/3DeltaTbmca5 revealed no greater or lesser susceptibility to stresses reported to initiate programmed cell death, such as treatment with prostaglandin D2. These metacaspases co-localise with RAB11, a marker for recycling endosomes but variant surface glycoprotein (VSG) recycling processes and the degradation of internalised anti-VSG antibody were found to occur similarly in wild type, DeltaTbmca2/3DeltaTbmca5 and triple RNAi-induced parasites. Thus the data provide no support for the direct involvement of T. brucei metacaspases in programmed cell death and suggest that the proteins have a function associated with RAB11 vesicles that is independent of known recycling processes of RAB11-positive endosomes. As both the L major and the T. brucei metacaspases appear to play crucial roles in cell cycle progression and because metacaspases are absent from the mammalian genome, these metacaspases have potential as novel drug targets

Personal Assistants in the Promotion of Independent Living for Persons with Intellectual Disability: A Basic Applied Investigation

Background: The personal assistant is a support figure aimed at improving the autonomy and independence of persons with intellectual disability, and is coherent with the new models of intervention with such groups. Method: Two sub-studies were conducted. The first one, comprising 120 persons with intellectual disability and 147 relatives, was aimed at identifying perceptions on support needs for independent living. The second was a case study with 4 participants who received support from personal assistants. Results: In the first sub-study, persons with intellectual disability showed a clear preference for a greater degree of autonomy and self-determination in their life projects. The second one highlights the effect of personal assistance on the individuals’ empowerment and the improvement of their autonomy. Discussion: The results are used to suggest a competency profile of the personal assistant, including ten key competencies, and a critical analysis of the most relevant findings is made.Planteamiento del problema: El asistente personal es una figura de apoyo a la mejora de la autonomía y la vida independiente, coherente con los nuevos modelos de intervención con personas con discapacidad intelectual. Método: Se desarrollan dos subestudios. El primero cuantitativo, con 120 personas con discapacidad intelectual y 147 familiares, tiene el objetivo de conocer la percepción de las necesidades de apoyo para la vida independiente. La segunda fase es un estudio de casos con 4 participantes, en los que se aplica la figura del asistente personal. Resultados: Destaca del primer subestudio la clara disposición de las personas con discapacidad intelectual a tener más autonomía y autodeterminación en sus proyectos de vida. En el segundo subestudio resalta el impacto de la asistencia personal en el empoderamiento y en la mejora de la autonomía. Conclusiones: Con base en los resultados, se propone un perfil competencial del asistente personal con diez competencias clave y se realiza un análisis crítico de los hallazgos más relevantes

Diagnósticos obtenidos mediante biopsia pulmonar guiada por tomografía computerizada

Treball Final de Grau en Medicina. Codi: MD1158. Curs acadèmic: 2020/2021Objetivo: Estudiar la precisión diagnóstica de la punción aspiración por aguja fina (PAAF), la biopsia con aguja gruesa (BAG) y la combinación de ambas técnicas para obtener el diagnóstico certero de nódulos pulmonares sin contacto pleural, usando como guía la tomografía computerizada (TC), así como comprobar cuál de ellas es más precisa según la distancia a la que se encuentre el nódulo de la piel (40mm). Materiales y métodos: Se llevaron a cabo un total de 202 biopsias de pulmón clasificadas en tres grupos en función de la técnica utilizada: Grupo 1 (PAAF, n= 86), Grupo 2 (BAG, n=16) y Grupo 3 (PAAF y BAG, n= 100). Se calculó la precisión diagnóstica de cada grupo, sus aciertos respecto a la distancia a la que se encontraba el nódulo pulmonar y si existía una asociación estadísticamente significativa entre estos. Para terminar, efectuamos un análisis descriptivo de las características principales de las variables, incluyendo el análisis molecular. Resultados: El método con el que se logró una mayor cantidad de diagnósticos certeros fue la combinación de PAAF y BAG, alcanzando un 95 % de aciertos. La BAG obtuvo un 87.5% de aciertos, mientras que el método menos certero fue la PAAF, con un 80.2% de aciertos. No se ha encontrado una asociación estadísticamente significativa entre la realización de una técnica (PAAF, BAG o PAAF más BAG) y la obtención de un diagnóstico certero, entre los diagnósticos correctos de cada técnica y las franjas interna (40mm), pero sí existe esta relación con la franja media (20-40mm). Conclusiones: No se ha encontrado una asociación estadísticamente significativa (p<0,05) entre pertenecer a un grupo de técnica de biopsia con la obtención de un diagnóstico certero, entre los diagnósticos correctos de cada técnica y las franjas interna (<20mm) y externa (>40mm), pero sí existe esta relación con la franja media (20-40mm).Objective: To study the diagnostic precision of fine needle aspiration (FNAB), core needle biopsy (CNB) and the combination of both techniques to obtain an accurate diagnosis of pulmonary nodules without pleural contact, using computed tomography (CT) as a guide, as well as to see which of them is more accurate according to the distance where the nodule is from the skin ( 40mm). Materials and methods: A total of 202 lung biopsies were carried out, classified into 3 groups according to the technique used: Group 1 (FNAB, n = 86), Group 2 (CNB, n = 16) and Group 3 (FNAB and CNB, n = 100). The diagnostic precision of each group was calculated, and their correct answers regarding the distance where the pulmonary nodule was located and whether there was a statistically significant association between them. Finally, a descriptive analysis was carried out of the main characteristics of the variables, including molecular analysis. Results: The method with which a greater amount of accurate diagnoses was achieved was the combination of FNAB and CNB, reaching 95% correct diagnosis. The CNB obtained 87.5% of correct answers, while the least certain method was the FNA, with 80.2% of correct answers. There is no statistically significant association between performing a technique (FNAB, CNB or FNAB and CNB) and obtaining a correct diagnosis, between the correct diagnoses of each technique and the internal ( 40mm) stripes, but this relationship does exist with the middle stripe (20-40mm). Conclusions: No significant statistically association (p <0.05) was found between belonging to a group of biopsy technique with obtaining an accurate diagnosis, between the correct diagnoses of each technique and the internal stripes ( 40mm), but this relationship does exist with the middle stripe (20-40mm)

Análisis de usabilidad y aplicación de los resultados en el desarrollo de plataformas web

Este trabajo trata cómo se pueden aplicar las técnicas de análisis de usabilidad al desarrollo de plataformas web. Actualmente es común que los servicios sean ofrecidos mediante plataformas web para un grupo muy heterogéneo de personas. Por otra parte, los análisis de usabilidad son una herramienta muy útil para conocer cómo interactúan las personas con los ordenadores y mejorar el diseño de las aplicaciones. Realizar un buen diseño permite mejorar la experiencia de usuario, factor fundamental para el éxito de cualquier producto que requiera interacción con el usuario. A continuación se describen las diferentes fases de los test de usabilidad y se detalla cómo han sido aplicadas durante el desarrollo del proyecto. Finalmente, se presentarán los resultados obtenidos durante la evaluación de la plataforma y el análisis de los mismos indicando cómo han afectado al diseño de la plataforma. ---ABSTRACT---This document discusses how to apply usability test techniques over web platform development. Nowadays, it is common that services are offered through web platforms for a large group of heterogeneous people. Moreover, usability tests are a very useful tool to understand human-computer interaction and improve the design of the applications. A good design can improve user experience, which is essential for the success of any product that requires user interaction. The following pages describes the different phases of usability testing and detail how these have been applied during the development of the project. Finally, the results obtained during the platform evaluation are presented and analysed, explaining how they have affected the design of the platform

Análisis y propuesta de mejora de la administración electrónica en el Ayuntamiento de Beniparrell

[ES] Tras la publicación de la Ley 11/2007 de 22 de junio de acceso electrónico de los ciudadanos a los servicios públicos, analizaré la situación en la que se encuentra el Ayuntamiento de Beniparrell en su administración electrónica, teniendo en cuenta que es un municipio inferior a 2500 habitantes con un alto porcentaje de gente longeva. Analizaré y propondré mejoras para tener acceso al mayor número de trámites posibles.[EN] After the publication of Law 11/2007 of June 22 of electronic access of citizens to public services, I will analyze the situation in which the municipality of Beniparrell in its electronic administration, taking into account that it is a municipality of less than 2500 inhabitants with a high percentage of long-lived people. Analyze and propose improvements to have access to as many procedures as possible.Ambit Jiménez, J. (2018). Análisis y propuesta de mejora de la administración electrónica en el Ayuntamiento de Beniparrell. http://hdl.handle.net/10251/110545TFG

La figura del asistente personal en la promoción de la autonomía y la vida independiente de las personas con discapacidad intelectual

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Formación de Profesorado y Educación, Departamento de Didáctica y Teoría de la Educación. Fecha de lectura : 28 de abril de 2016Uno de los retos de los modelos de intervención con personas con discapacidad intelectual es la promoción de su autonomía y vida independiente. Esta finalidad responde a un derecho básico de cualquier persona: ser el agente protagonista de su propio proyecto de vida. En esta Tesis Doctoral se realizan dos investigaciones que tratan de validar la figura del Asistente Personal como profesional que puede favorecer la autonomía y la formación para la vida independiente de las personas con discapacidad intelectual. En un primer estudio, se investigan las necesidades percibidas de vida independiente por 120 jóvenes con discapacidad intelectual y 147 familiares. Los resultados de este estudio demuestran que tanto las personas con discapacidad intelectual participantes como las familias, son conscientes de la importancia de contar con apoyos profesionales en este ámbito. Estos resultados contribuyen a crear un modelo de Asistente Personal que se aplica en el segundo estudio a través del análisis de 10 casos con distintas necesidades de apoyo. La evaluación de estas experiencias confirma que la figura del Asistente Personal puede favorecer la autonomía y la vida independiente. La tesis doctoral incluye una Guía Orientativa para Asistentes Personales en formato digital.One of the challenges of the models of intervention with people with intellectual disabilities is to promote their autonomy and independent living. This aim reflects a basic right of any person: being the protagonist agents of their own life project. In this Doctoral Thesis two studies that try to validate the figure of the Personal Assistant which can promote autonomy and training for independent living for people with intellectual disabilities are made. In a first study, the perceived needs of supporting independent life are investigated by both 120 young people with intellectual disabilities and 147 families. The results of this study demonstrate that individuals with intellectual disabilities and families, are aware of the importance of professional support in this area. These results contribute to create a model of Personal Assistant that applies in the second study through the analysis of 10 cases with different support needs. The evaluation of these experiences confirms that the figure of the Personal Assistant can promote autonomy and independent living. The doctoral thesis includes a digital Orientation Guide for Personal Assistants

Utilidad de los sistemas optoelectrónicos de seguimiento de trayectorias para determinar movilidad articular y curvas fuerza/elongación de materiales elásticos

Esta tesis tuvo como objetivos principales los siguientes : 1. Determinar la validez y fiabilidad de las medidas de movilidad articular activa de las articulaciones del miembro superior establecidas con Veloflex en sujetos sin deficiencias locales, y también su fiabilidad en sujetos con ellas. 2. Evaluar la validez y fiabilidad de las medidas de movilidad articular activa cervical establecidas con Veloflex en sujetos sin deficiencias locales, y también su fiabilidad en sujetos con ellas. 3. Identificar las curvas de fuerza/elongación de las bandas CLX para los 7 grados de resistencia que utiliza Therband®, así como, determinar el valor de resistencia máximo de elongación para cada uno de ellas. 4. Analizar si las curvas de fuerza/elongación de las bandas elásticas CLX son estables ante la modificación de la velocidad de elongación, la longitud inicial de la banda en reposo o las fases de acortamiento/estiramiento. Metodología: Para abordar los dos primeros objetivos, se realizaron dos estudios secuenciales. Participaron sujetos sin deficiencias y sujetos con deficiencias en la movilidad articular. Los participantes fueron evaluados en dos sesiones. Se utilizó Veloflex para medir los rangos de movimiento, y se compararon los resultados con los instrumentos "Gold estándar" (goniómetro universal y C-ROM). Para el tercer y cuarto objetivo, se llevó a cabo un estudio experimental secuencial en el que se evaluaron las bandas CLX de Theraband® en diferentes condiciones. Se utilizaron distintos diseños experimentales para determinar las curvas de fuerza/elongación de las bandas y analizar su estabilidad ante cambios en la velocidad de elongación, longitud inicial y fases de acortamiento/estiramiento. Se utilizo Velowin para medir la velocidad y longitud de elongación de las bandas, y se registró la fuerza ejercida mediante un dinamómetro. Para llevar a cabo los dos primeros objetivos se realizaron análisis estadísticos utilizando medidas descriptivas, coeficiente de correlación de Pearson y gráficos de Bland-Altmann. En el tercer estudio, se analizaron curvas de fuerza/elongación y se ajustaron líneas de regresión logarítmica lineal. Se comparó la tensión en diferentes experimentos. Todos los análisis se realizaron con SPSS 24.0. Resultados: En los primeros dos estudios participaron 68 sujetos asintomáticos y 40 sujetos con deficiencias. Se obtuvieron datos sobre la movilidad articular activa en el miembro superior y la columna cervical utilizando Veloflex. Los resultados mostraron una buena validez y fiabilidad de las medidas en sujetos sin deficiencias y también en aquellos con deficiencias locales. En el tercer estudio, se analizaron las curvas de fuerza/elongación de las bandas CLX de Theraband®. Se establecieron las curvas para cada una de las graduaciones de resistencia y se determinó el valor máximo de elongación para cada una de ellas. Además, se encontró que las curvas de fuerza/elongación de las bandas eran estables ante cambios en la velocidad de elongación, longitud inicial y fases de acortamiento/estiramiento. Conclusiones de la tesis: 1. El Veloflex proporciona medidas de la movilidad articular activa de las articulaciones del miembro superior que son válidas y fiables en sujetos sin y con deficiencias locales. 2. El presente estudio determino que Veloflex aporta medidas de la movilidad articular activa cervical que son válidas y fiables en sujetos sin y con deficiencias locales. 3. Cada una de las bandas CLX de Theraband posee una curva fuerza/elongación diferente, y este estudio ofrece una ecuación asociada a cada una el cual permite predecir sus valores en cualquier posición del rango de elongación 0 a 100%. 4. Factores habituales de la práctica con resistencia elástica, como son las fases de alargamiento/acortamiento, la velocidad de alargamiento de las bandas CLX y la longitud inicial de reposo, no influyen en el comportamiento fuerza/elongación de las bandas elásticas.This thesis had the following main objectives: 1. Determine the validity and reliability of active joint mobility measurements in the upper limb established with Veloflex in subjects without local deficiencies, as well as their reliability in subjects with deficiencies. 2. Evaluate the validity and reliability of active cervical joint mobility measurements established with Veloflex in subjects without local deficiencies, as well as their reliability in subjects with deficiencies. 3. Identify the force/elongation curves of CLX bands for the 7 resistance levels used by Theraband®, and determine the maximum elongation resistance value for each of them. 4. Analyze if the force/elongation curves of CLX elastic bands remain stable when modifying elongation speed, initial resting band length, or stretching/shortening phases. Methodology: To address the first two objectives, two sequential studies were conducted. Subjects without deficiencies and subjects with joint mobility deficiencies participated in the studies. The participants were evaluated in two sessions. Veloflex was used to measure the ranges of motion, and the results were compared with "Gold standard" instruments (universal goniometer and C-ROM). For the third and fourth objectives, a sequential experimental study was conducted, evaluating Theraband® CLX bands under different conditions. Various experimental designs were used to determine the force/elongation curves of the bands and analyze their stability when changes were made to elongation speed, initial length, and stretching/shortening phases. Velowin was used to measure the speed and length of elongation of the bands, and the force exerted was recorded using a dynamometer. Statistical analysis was performed using descriptive measures, Pearson's correlation coefficient, and Bland-Altmann plots to address the first two objectives. In the third study, force/elongation curves were analyzed and logarithmic linear regression lines were fitted. Tension was compared across different experiments. All analyses were performed using SPSS 24.0. Results: The first two studies involved 68 asymptomatic subjects and 40 subjects with deficiencies. Data on active joint mobility in the upper limb and cervical spine were obtained using Veloflex. The results showed good validity and reliability of the measurements in subjects without deficiencies and also in those with local deficiencies. In the third study, force/elongation curves of Theraband® CLX bands were analyzed. Curves were established for each resistance level, and the maximum elongation value for each band was determined. Additionally, it was found that the force/elongation curves of the bands remained stable when modifying elongation speed, initial length, and stretching/shortening phases. Thesis conclusions: 1. Veloflex provides valid and reliable measures of active joint mobility in the upper limb for subjects with or without local deficiencies. 2. This study determined that Veloflex provides valid and reliable measures of active cervical joint mobility for subjects with or without local deficiencies. 3. Each Theraband CLX band has a different force/elongation curve, and this study provides an equation associated with each band that allows predicting its values at any position within the 0-100% elongation range. 4. Common factors in elastic resistance practice, such as stretching/shortening phases, elongation speed of CLX bands, and initial resting length, do not influence the force/elongation behavior of the elastic bands

Técnicas complementarias de relajación y analgesia no farmacológicas durante el parto: revisión sistemática

Introduction:There is a need to provide analgesia strategies that encourage and promote women's participation in decision-making at the time of delivery and relaxation techniques could be a complementary and/or alternative non-pharmacological analgesic method to the widely used epidural anaesthesia in standard labour care. Objective: The objective of this study is to analyze the obstetric effects of relaxation techniques on pain management during labour. Method: A systematic review is performed with critical reading of included studies. The search for studies was carried out in the main databases MEDLINE, Cochrane Library, Cuiden, LILACS and SciELO. Studies published in English or Spanish between 2015 and February 2021 were included. Eleven studies were included, six of which are systematic reviews and five are randomised clinical trials. The interventions analysed were relaxation techniques such as hypnosis, intradermal injection of sterile water, warm water immersion, massage, acupuncture, music therapy, aromatherapy, continuous support and mind-body practices like relaxing breathing, yoga and meditation, among others. Conclusion: The main conclusion of this study is that relaxation techniques may decrease the level of pain during labour, although the current scientific evidence is limited and the methodological quality varies from low to moderate. More randomised controlled trials are needed to support this research.Introducción: Existe la necesidad de proporcionar estrategias de analgesia que alienten y promuevan la participación de la mujer en la toma de decisiones en el momento del parto y las técnicas de relajación podrían ser un método analgésico no far-macológico complementario y/o alternativo a la anestesia epidural ampliamente utilizado. en la estándar atención del trabajo de parto. Objetivo: El objetivo de este estudio es analizar los efectos obstétricos de las técnicas de relajación en el manejo del dolor durante el parto. Método: Se realiza una revisión sistemática con lectura crítica de los estudios incluidos. La búsqueda de estudios se realizó en las principales bases de datos MEDLINE, Cochrane Library, Cuiden, LILACS y SciELO. Se incluyen estudios publicados en inglés o español entre 2015 y febrero de 2021. Se incluyen una vez estudios, seis de los cuales son revisados sistemáticamente y cinco son ensayos clínicos aleatorios. Las intervenciones analizadas fueron técnicas de relajación como hipnosis, inyección intradérmica de agua estéril, inmersión en agua tibia, masaje, acu-puntura, musicoterapia, aromaterapia, apoyo continuo y prácticas mente-cuerpo como respiración relajante, yoga y meditación, entre otras.Conclusión: La principal conclusión de este estudio es que las técnicas de relajación pueden disminuir el nivel de dolor durante el trabajo, aunque la evidencia científica actual es limitada y la calidad metodológica varía de baja a moderada. Se necesitan más ensayos controlados aleatorios para apoyar esta investigación