Propuesta de actividades de autoaprendizaje para el nivel A2 de lengua inglesa en la plataforma Moodle

Los objetivos de este proyecto son proporcionar la teoría, los ejercicios y otros recursos necesarios para que los alumnos de la EUIT de Telecomunicación con un nivel A1 en el Marco Común Europeo de Referencia para las Lenguas (MCERL) puedan obtener el nivel A2 en inglés sin necesidad de asistir a clases ni matricularse en cursos presenciales. La plataforma utilizada para conseguir este fin es Moodle, siendo utilizada en la página web de ILLLab. Este curso online sirve para alcanzar los conocimientos requeridos en la asignatura optativa Introduction to English for Professional and Academic Communication I que parte del nivel B1. Se realiza una propuesta de la gramática con sus correspondientes ejemplos y ejercicios basados todos ellos en adaptaciones de actividades publicadas en un corpus de libros de texto. Se añaden recursos (pequeñas lecturas, videos, enlaces) que se consideran apropiados para el tema tratado. Por otro lado, también se persigue solucionar el problema de los cursos de idiomas basados en e-learning ya que no proporcionan las herramientas necesarias para poner en práctica la expresión oral. Para ello, se aporta una aplicación basada en técnicas de reconocimiento de voz, con tres actividades en las que los resultados han de darse de forma hablada y con la correcta pronunciación. Así, se busca dar una base de conocimientos y experiencias prácticas para futuros proyectos basados en herramientas de síntesis y reconocimiento de voz, además de buscar un nuevo enfoque en el estudio de idiomas. Abstract: The objectives of this project are to provide the theory, exercises and other resources for students at the EUIT Telecommunications with A1 level in the Common European Framework of Reference for Languages (MCERL) in order to get A2 level in English without attending face-to-face courses. The platform used to achieve this aim is Moodle, which is currently being used in ILLLab website. This online course is due to attain the knowledge required in the optional subject Introduction to English for Professional and Academic Communication I which is based on the B1 level. It is a proposal of grammar with corresponding examples and exercises all based on adaptations of activities posted on a corpus of textbooks. It also adds resources (short readings, videos or links) that are appropriate for the subject. On the other hand, this project aims to solve the problem of language courses based on e-learning because these do not usually provide the student with the necessary tools to practice speaking. For this, we develop an application based on speech recognition techniques and propose three activities to practice speaking, and pronunciation. The proposal seeks to provide knowledge and practical experience for future projects based on synthesis tools and voice recognition, and means a new approach to e-learning courses for the study of languages.

Actividades online de vocabulario y pronunciación para el nivel de la lengua inglesa A2

El objetivo del presente proyecto es proporcionar una actividad de la pronunciación y repaso de vocabulario en lengua inglesa para la plataforma Moodle alojada en la página web de Integrated Language Learning Lab (ILLLab). La página web ILLLab tiene el objetivo de que los alumnos de la EUIT de Telecomunicación de la UPM con un nivel de inglés A2 según el Marco Común Europeo de Referencia para las Lenguas (MCERL), puedan trabajar de manera autónoma para avanzar hacia el nivel B2 en inglés. La UPM exige estos conocimientos de nivel de inglés para cursar la asignatura English for Professional and Academic Communication (EPAC) de carácter obligatorio e impartida en el séptimo semestre del Grado en Ingeniería de Telecomunicaciones. Asimismo, se persigue abordar el problema de las escasas actividades de expresión oral de las plataformas de autoaprendizaje se dedican a la formación en idiomas y, más concretamente, al inglés. Con ese fin, se proporciona una herramienta basada en sistemas de reconocimiento de voz para que el usuario practique la pronunciación de las palabras inglesas. En el primer capítulo del trabajo se introduce la aplicación Traffic Lights, explicando sus orígenes y en qué consiste. En el segundo capítulo se abordan aspectos teóricos relacionados con el reconocimiento de voz y se comenta sus funciones principales y las aplicaciones actuales para las que se usa. El tercer capítulo ofrece una explicación detallada de los diferentes lenguajes utilizados para la realización del proyecto, así como de su código desarrollado. En el cuarto capítulo se plantea un manual de usuario de la aplicación, exponiendo al usuario cómo funciona la aplicación y un ejemplo de uso. Además, se añade varias secciones para el administrador de la aplicación, en las que se especifica cómo agregar nuevas palabras en la base de datos y hacer cambios en el tiempo estimado que el usuario tiene para acabar una partida del juego. ABSTRACT: The objective of the present project is to provide an activity of pronunciation and vocabulary review in English language within the platform Moodle hosted at the Integrated Language Learning Lab (ILLLab) website. The ILLLab website has the aim to provide students at the EUIT of Telecommunication in the UPM with activities to develop their A2 level according to the Common European Framework of Reference for Languages (CEFR). In the platform, students can work independently to advance towards a B2 level in English. The UPM requires this level of English proficiency for enrolling in the compulsory subject English for Professional and Academic Communication (EPAC) taught in the seventh semester of the Degree in Telecommunications Engineering. Likewise, this project tries to provide alternatives to solve the problem of scarce speaking activities included in the learning platforms that offer language courses, and specifically, English language courses. For this purpose, it provides a tool based on speech recognition systems so that the user can practice the pronunciation of English words. The first chapter of the project introduces the application Traffic Lights, explaining its origins and what it is. The second chapter deals with theoretical aspects related with speech recognition and comments their main features and current applications for which it is generally used. The third chapter provides a detailed explanation of the different programming languages used for the implementation of the project and reviews its code development. The fourth chapter presents an application user manual, exposing to the user how the application works and an example of use. Also, several sections are added addressed to the application administrator, which specify how to add new words to the database and how to make changes in the original stings as could be the estimated time that the user has to finish the game.

Phosphatidylcholine-specific phospholipase C regulates glutamate-induced nerve cell death

Phosphatidylcholine-specific phospholipase C (PC-PLC) is a necessary intermediate in transducing apoptotic signals for tumor necrosis factor and Fas/Apo-1 ligands in nonneuronal cells. The data presented here show that PC-PLC also is required in oxidative glutamate-induced programmed cell death of both immature cortical neurons and a hippocampal nerve cell line, HT22. In oxidative glutamate toxicity, which is distinct from excitotoxicity, glutamate interferes with cystine uptake by blocking the cystine/glutamate antiporter, indirectly causing a depletion of intracellular glutathione. A PC-PLC inhibitor blocks oxidative glutamate toxicity, and exogenous PC-PLC potentiates glutamate toxicity. The inhibition of PC-PLC uncouples the cystine uptake from glutamate inhibition, allowing the maintenance of glutathione synthesis and cell viability. These data suggest that PC-PLC modulates neuronal cell death through a mechanism that is distinct from that involved in nonneuronal apoptosis.

Bradykinin inhibits M current via phospholipase C and Ca2+ release from IP3-sensitive Ca2+ stores in rat sympathetic neurons

A variety of intracellular signaling pathways can modulate the properties of voltage-gated ion channels. Some of them are well characterized. However, the diffusible second messenger mediating suppression of M current via G protein-coupled receptors has not been identified. In superior cervical ganglion neurons, we find that the signaling pathways underlying M current inhibition by B2 bradykinin and M1 muscarinic receptors respond very differently to inhibitors. The bradykinin pathway was suppressed by the phospholipase C inhibitor U-73122, by blocking the IP3 receptor with pentosan polysulfate or heparin, and by buffering intracellular calcium, and it was occluded by allowing IP3 to diffuse into the cytoplasm via a patch pipette. By contrast, the muscarinic pathway was not disrupted by any of these treatments. The addition of bradykinin was accompanied by a [Ca2+]i rise with a similar onset and time to peak as the inhibition of M current. The M current inhibition and the rise of [Ca2+]i were blocked by depletion of Ca2+ internal stores by thapsigargin. We conclude that bradykinin receptors inhibit M current of sympathetic neurons by activating phospholipase C and releasing Ca2+ from IP3-sensitive Ca2+ stores, whereas muscarinic receptors do not use the phospholipase C pathway to inhibit M current channels.

Requirement of GM2 ganglioside activator for phospholipase D activation

Sequence analysis of a heat-stable protein necessary for the activation of ADP ribosylation factor-dependent phospholipase D (PLD) reveals that this protein has a structure highly homologous to the previously known GM2 ganglioside activator whose deficiency results in the AB-variant of GM2 gangliosidosis. The heat-stable activator protein indeed has the capacity to enhance enzymatic conversion of GM2 to GM3 ganglioside that is catalyzed by β-hexosaminidase A. Inversely, GM2 ganglioside activator purified separately from tissues as described earlier [Conzelmann, E. & Sandhoff, K. (1987) Methods Enzymol. 138, 792–815] stimulates ADP ribosylation factor-dependent PLD in a dose-dependent manner. At higher concentrations of ammonium sulfate, the PLD activator protein apparently substitutes for protein kinase C and phosphatidylinositol 4,5-bisphosphate, both of which are known as effective stimulators of the PLD reaction. The mechanism of action of the heat-stable PLD activator protein remains unknown.

Phospholipase D activity is required for suppression of yeast phosphatidylinositol transfer protein defects

Yeast phosphatidylinositol transfer protein (Sec14p) function is essential for production of Golgi-derived secretory vesicles, and this requirement is bypassed by mutations in at least seven genes. Analyses of such ‘bypass Sec14p’ mutants suggest that Sec14p acts to maintain an essential Golgi membrane diacylglycerol (DAG) pool that somehow acts to promote Golgi secretory function. SPO14 encodes the sole yeast phosphatidylinositol-4,5-bisphosphate-activated phospholipase D (PLD). PLD function, while essential for meiosis, is dispensable for vegetative growth. Herein, we report specific physiological circumstances under which an unanticipated requirement for PLD activity in yeast vegetative Golgi secretory function is revealed. This PLD involvement is essential in ‘bypass Sec14p’ mutants where normally Sec14p-dependent Golgi secretory reactions are occurring in a Sec14p-independent manner. PLD catalytic activity is necessary but not sufficient for ‘bypass Sec14p’, and yeast operating under ‘bypass Sec14p’ conditions are ethanol-sensitive. These data suggest that PLD supports ‘bypass Sec14p’ by generating a phosphatidic acid pool that is somehow utilized in supporting yeast Golgi secretory function.

Loss of virulence in Leishmania donovani deficient in an amastigote-specific protein, A2

Leishmania donovani is the etiologic agent of fatal visceral leishmaniasis in man. During their life cycle, Leishmania exist as flagellated promastigotes within the sandfly vector and as nonflagellated amastigotes in the macrophage phagolysosomal compartment of the mammalian host. The transformation from promastigotes to amastigotes is a critical step for the establishment of infection, and the molecular basis for this transformation is poorly understood. To define the molecular basis for amastigote survival in the mammalian host, we previously identified an amastigote stage-specific gene family termed “A2.” In the present study, we have inhibited the expression of A2 mRNA and A2 protein in amastigotes using antisense RNA and show that the resulting A2-deficient amastigotes are severely compromised with respect to virulence in mice. Amastigotes that did survive in the mice had restored A2 protein expression. These data demonstrate that A2 protein is required for L. donovani survival in a mammalian host, and this represents the first identified amastigote-specific virulence factor identified in Leishmania. This study also reveals that it is possible to study gene function in Leishmania through the expression of antisense RNA.

Altered peptide ligands act as partial agonists by inhibiting phospholipase C activity induced by myasthenogenic T cell epitopes

Myasthenia gravis (MG) is a T cell-regulated, antibody-mediated autoimmune disease. Two peptides representing sequences of the human acetylcholine receptor α-subunit, p195–212 and p259–271, previously were shown to stimulate the proliferation of peripheral blood lymphocytes of patients with MG and were found to be immunodominant T cell epitopes in SJL and BALB/c mice, respectively. Single amino acid-substituted analogs of p195–212 and p259–271, as well as a dual analog composed of the tandemly arranged two single analogs, were shown to inhibit, in vitro and in vivo, MG-associated autoimmune responses. Stimulation of T cells through the antigen-specific T cell receptor activates tyrosine kinases and phospholipase C (PLC). Therefore, in attempts to understand the mechanism of action of the analogs, we first examined whether the myasthenogenic peptides trigger tyrosine phosphorylation and activation of phospholipase C. For that purpose, we measured generation of inositol phosphates and tyrosine phosphorylation of PLC after stimulation of the p195–212- and p259–271-specific T cell lines with these myasthenogenic peptides. Both myasthenogenic peptides stimulated generation of inositol phosphates as well as tyrosine phosphorylation of PLC. However, the single and dual analogs, although inducing tyrosine phosphorylation of PLC, could not induce PLC activity. Furthermore, the single and dual analogs inhibited the induced PLC activity whereas they could not inhibit tyrosine phosphorylation of PLC that was caused by the myasthenogenic peptides. Thus, the altered peptides and the dual analog act as partial agonists. The down-regulation of PLC activity by the analogs may account for their capacity to inhibit in vitro MG-associated T cell responses.

Phospholipase C-linked receptors regulate the ATP-sensitive potassium channel by means of phosphatidylinositol 4,5-bisphosphate metabolism

In the COS7 cells transfected with cDNAs of the Kir6.2, SUR2A, and M1 muscarinic receptors, we activated the ATP-sensitive potassium (KATP) channel with a K+ channel opener and recorded the whole-cell KATP current. The KATP current was reversibly inhibited by the stimulation of the M1 receptor, which is linked to phospholipase C (PLC) by the Gq protein. The receptor-mediated inhibition was observed even when protein kinase C (PKC) was inhibited by H-7 or by chelating intracellular Ca2+ with 10 mM 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetate (BAPTA) included in the pipette solution. However, the receptor-mediated inhibition was blocked by U-73122, a PLC inhibitor. M1-receptor stimulation failed to inhibit the KATP current activated by the injection of exogenous phosphatidylinositol 4,5-bisphosphate (PIP2) through the whole-cell patch pipette. The receptor-mediated inhibition became irreversible when the replenishment of PIP2 was blocked by wortmannin (an inhibitor of phosphatidylinositol kinases), or by including adenosine 5′-[β,γ–imido]triphosphate (AMPPNP, a nonhydrolyzable ATP analogue) in the pipette solution. In inside-out patch experiments, the ATP sensitivity of the KATP channel was significantly higher when the M1 receptor in the patch membrane was stimulated by acetylcholine. The stimulatory effect of pinacidil was also attenuated under this condition. We postulate that stimulation of PLC-linked receptors inhibited the KATP channel by increasing the ATP sensitivity, not through PKC activation, but most probably through changing PIP2 levels.

Inhibition of phospholipase D by lysophosphatidylethanolamine, a lipid-derived senescence retardant

Phospholipid signaling mediated by lipid-derived second messengers or biologically active lipids is still new and is not well established in plants. We recently have found that lysophosphatidylethanolamine (LPE), a naturally occurring lipid, retards senescence of leaves, flowers, and postharvest fruits. Phospholipase D (PLD) has been suggested as a key enzyme in mediating the degradation of membrane phospholipids during the early stages of plant senescence. Here we report that LPE inhibited the activity of partially purified cabbage PLD in a cell-free system in a highly specific manner. Inhibition of PLD by LPE was dose-dependent and increased with the length and unsaturation of the LPE acyl chain whereas individual molecular components of LPE such as ethanolamine and free fatty acid had no effect on PLD activity. Enzyme-kinetic analysis suggested noncompetitive inhibition of PLD by LPE. In comparison, the related lysophospholipids such as lysophosphatidylcholine, lysophosphatidylglycerol, and lysophosphotidylserine had no significant effect on PLD activity whereas PLD was stimulated by lysophosphatidic acid and inhibited by lysophosphatidylinositol. Membrane-associated and soluble PLD, extracted from cabbage and castor bean leaf tissues, also was inhibited by LPE. Consistent with acyl-specific inhibition of PLD by LPE, senescence of cranberry fruits as measured by ethylene production was more effectively inhibited according to the increasing acyl chain length and unsaturation of LPE. There are no known specific inhibitors of PLD in plants and animals. We demonstrate specific inhibitory regulation of PLD by a lysophospholipid.

The Small Mr Ras-like GTPase Rap1 and the Phospholipase C Pathway Act to Regulate Phagocytosis in Dictyostelium discoideum

The function of the small-Mr Ras-like GTPase Rap1 remains largely unknown, but this protein has been demonstrated to regulate cortical actin-based morphologic changes in Dictyostelium and the oxidative burst in mammalian neutrophils. To test whether Rap1 regulates phagocytosis, we biochemically analyzed cell lines that conditionally and modestly overexpressed wild-type [Rap1 WT(+)], constitutively active [Rap1 G12T(+)], and dominant negative [Rap1 S17N(+)] forms of D. discoideum Rap1. The rates of phagocytosis of bacteria and latex beads were significantly higher in Rap1 WT(+) and Rap1 G12T(+) cells and were reduced in Rap1 S17N(+) cells. The addition of inhibitors of protein kinase A, protein kinase G, protein tyrosine kinase, or phosphatidylinositide 3-kinase did not affect phagocytosis rates in wild-type cells. In contrast, the addition of U73122 (a phospholipase C inhibitor), calphostin C (a protein kinase C inhibitor), and BAPTA-AM (an intracellular Ca2+ chelator) reduced phagocytosis rates by 90, 50, and 65%, respectively, suggesting both arms of the phospholipase C signaling pathways played a role in this process. Other protein kinase C–specific inhibitors, such as chelerythrine and bisindolylmaleimide I, did not reduce phagocytosis rates in control cells, suggesting calphostin C was affecting phagocytosis by interfering with a protein containing a diacylglycerol-binding domain. The addition of calphostin C did not reduce phagocytosis rates in Rap1 G12T(+) cells, suggesting that the putative diacylglycerol-binding protein acted upstream in a signaling pathway with Rap1. Surprisingly, macropinocytosis was significantly reduced in Rap1 WT(+) and Rap1 G12T(+) cells compared with control cells. Together our results suggest that Rap1 and Ca2+ may act together to coordinate important early events regulating phagocytosis.