Evaluación de la toxicidad de nanovesículas en células de retina

[ES] El objetivo de este trabajo es evaluar la toxicidad in vitro e in vivo de un tipo de nanovesículas denominadas quatsomes, con la finalidad de poder utilizarlas como nanocarriers de drogas que retrasen la degeneración retiniana que ocurre en la patología denominada retinosis pigmentaria. Los quatsomes son unas estructuras vesiculares termodinámicamente estables, constituidas por surfactantes amónicos cuaternarios y esteroles. En este trabajo se evalúa la toxicidad de tres tipos de quatsomes que difieren en los surfactantes por los que están formados (CTAB: Cetyl trimethylammonium bromide; CPC: Cetylpyridinium chloride y MKC: Myristalkonium Chloride ), con el fin de conocer cuál de ellos es el que tiene menor toxicidad. En primer lugar se evalúa la toxicidad in vitro en una línea celular inmortal de fotorreceptores (conos) murinos, la línea 661W. Se realiza una curva de concentración para determinar la concentración de quatsomes óptima que no sea tóxica para las células. Una vez determinada dicha concentración se elegirán los quatsomes con el surfactante menos tóxico. Para llevar a cabo este objetivo se cultivan las células 661W en presencia de los distintos quatsomes durante 24 horas y 48 horas en un rango de concentraciones que varía en función de cada surfactante. A continuación, se realiza un ensayo de viabilidad celular denominado ensayo MTT (metil tiazol tetrazolio) que consiste en un ensayo colorimétrico basado en la reducción del Bromuro de 3-(4,5-dimetiltiazol-2-ilo)-2,5-difeniltetrazol (MTT). Este método permite medir supervivencia y proliferación celular, así como también, determinar la citotoxicidad de potenciales agentes terapéuticos. Una vez determinada la toxicidad in vitro de los quatsomes, se selecciona la nanovesícula menos tóxica y, se administra intravítreamente a una concentración óptima a ratones control para evaluar la toxicidad in vivo. Para ello se evalúan marcadores de muerte celular y de inflamación en retina de ratón. En primer lugar se realiza la técnica de TUNEL (TdT-mediated dUTP-biotin nick-end-labeling) para detectar la fragmentación de ADN en células apoptóticas principalmente y de este modo conocer si las nanovesículas están causando procesos de muerte celular programada en la retina de los ratones tratados. En segundo lugar, se realiza una inmunofluorescencia con el fin de evaluar la presencia de gliosis reactiva como marcadores de inflamación. En concreto, se emplean los anticuerpos contra GFAP (Glial fibrillary acidic protein) e IBA1 (Ionized calcium binding adaptor molecule 1) como marcadores de la glía de Müller y de microglía, respectivamente.[EN] The aim of this study is to assess the in vitro and in vivo toxicity of a type of nanovesicle named quatsomes, in the pursuit of using them as drug nanocarriers to delay the retinal degeneration that happen in the pathology called retinitis pigmentosa. The quatsomes are thermodynamically stable nanovesicle structures, make by quaternary ammonium surfactants and sterols. In this study the toxicity of three types of quatsomes differing in surfactants for which they are made (CTAB: Cetyl trimethylammonium bromide; CPC: Cetylpyridinium chloride y MKC: Myristalkonium Chloride) is evaluated in order to know which of them is the less toxic. Firstly, the in vitro toxicity is assessed in an immortal murine photoreceptors cell line, the 661-W line. A concentration curve is made to establish optimum quatsome concentration non toxic to the cells. Secondly, the quatsomes made by the less toxic surfactant are chosen. To accomplish this goal the 661-W cells are cultured with the different quatsomes for 24 hours and 48 hours in a concentrations range that varies according to each surfactant. Then a cell viability assay called MTT assay (metil tiazol tetrazolio) is made, it is a colorimetric assay based on the Bromure de 3-(4,5-dimetiltiazol-2-ilo)-2,5-difeniltetrazol (MTT) reduction. This method allows to measure cell survival and proliferation, as well as determine the potential therapeutic agents citotoxicity. After determining in vitro toxicity of quatsomes, the least toxic nanovesicle is selected and is administered intravitreally at an optimal concentration to control mice to assess the in vivo toxicity. For this, cell death and inflammation markers of mice retina are evaluated. Firstly, TUNEL (TdT-mediated dUTP-biotin nick end-labeling) is performed to detect DNA fragmentation in apoptotic cells and thus know whether the nanovesicles are causing process of programmed cell death in the mice retina treated. Secondly, immunofluorescence is performed in order to evaluate the presence of reactive gliosis as inflammation markers. Specifically, antibodies against GFAP (Glial fibrillary acidic protein) and IBA1 (Ionized calcium binding adapter molecule 1) as markers of Muller glia and microglia, respectively are used.González Gil, C. (2016). Evaluación de la toxicidad de nanovesículas en células de retina. http://hdl.handle.net/10251/68683.TFG

Does students that receive additional support needs with higher risk for learning disabilities in writing?

Actualmente la escritura es considerada como un proceso cognitivo complejo en el que intervienen diversos factores que conducen a un correcto uso de la escritura en la edad adulta. Este proceso está formado por distintos subprocesos. Entre ellos cabe destacar la transcripción. La capacidad de transcripción adquiere un papel importante en la adquisición de la escritura, siendo de especial importancia en el inicio de la escolaridad. Si los niños son lentos e inexactos en el proceso de la transcripción conlleva a que su capacidad y calidad compositiva global en la producción de textos se vea gravemente perjudicada. La transcripción tiene un efecto significativo sobre la escritura global y el rendimiento académico de los niños en edad escolar, tanto es así que es considerada como un potente predictor para identificar a niños en riesgo de desarrollar déficits más graves en la escritura. Un hallazgo que refuerza la importancia de identificar las dificultades de escritura tan pronto como sea posible. A esto, hay que añadir que actualmente la enseñanza de la escritura en los primeros años de escolaridad se centra casi única y exclusivamente en los aspectos más formales del texto como la caligrafía o la legibilidad, excluyendo la instrucción en la fluidez. La presente investigación analiza las diferencias entre niños que reciben apoyo educativo por presentar retraso en la adquisición de las competencias del currículo escolar con niños que no lo reciben, considerando la fluidez y la exactitud en la escritura. Para ello se seleccionó una muestra de 181 alumnos que reciben apoyo educativo y 181 alumnos que no lo reciben, pertenecientes a los tres primeros cursos de Educación Primaria y cuyas edades oscilan entre 6 y 9 años. Los resultados obtenidos muestran que los niños que reciben apoyo educativo, aún recibiendo este apoyo, presentan mayores carencias en la fluidez y exactitud en la escritura, carencias que sin una apropiada instrucción podrían desembocar en futuras dificultades de aprendizaje en escritura.Nowadays, writing is considered as a complex cognitive process in which several factors are involved that lead to a right writing use in adult age. This process is set up by different sub-processes, between which we can highlight transcription. The transcription skill takes a relevant place in writing acquisition, being of a special importance in the school beginning. If children are imprecise and slow in the transcription process it can lead to a serious setback in to their composition ability and quality. Transcription has a significant effect into the global writing and the academic efficiency in school age even that is considered as a powerful predictor to identify children that are more likely to show severe deficit in the acquisition of writing skills. A discovery that strengthen the importance of identifying writing difficulties as soon as possible. It should be added that nowadays writing teaching at the beginning is focused almost, solely and exclusively, in the most formal text aspects, just like handwriting or legibility, excluding fluency instruction. The current study analyzes the differences in writing fluency and accuracy between children that receive educational support because of their backwardness in the education program and normally achieving children. For this purpose, we selected a sample of 181 normally achieving children and 181 children receiving educational support, belonging to Primary Education and whose ages are in 6 – 9 years old range. Obtained results show that children that receive educational support present a lack of writing fluency and accuracy, problems that without an appropriate instruction could lead in future writing difficulties.peerReviewe

Biosensors Platform Based on Chitosan/AuNPs/Phthalocyanine Composite Films for the Electrochemical Detection of Catechol. The Role of the Surface Structure

Producción CientíficaBiosensor platforms consisting of layer by layer films combining materials with different functionalities have been developed and used to obtain improved catechol biosensors. Tyrosinase (Tyr) or laccase (Lac) were deposited onto LbL films formed by layers of a cationic linker (chitosan, CHI) alternating with layers of anionic electrocatalytic materials (sulfonated copper phthalocyanine, CuPcS or gold nanoparticles, AuNP). Films with different layer structures were successfully formed. Characterization of surface roughness and porosity was carried out using AFM. Electrochemical responses towards catechol showed that the LbL composites efficiently improved the electron transfer path between Tyr or Lac and the electrode surface, producing an increase in the intensity over the response in the absence of the LbL platform. LbL structures with higher roughness and pore size facilitated the diffusion of catechol, resulting in lower LODs. The [(CHI)-(AuNP)-(CHI)-(CuPcS)]2-Tyr showed an LOD of 8.55∙10−4 μM, which was one order of magnitude lower than the 9.55·10−3 µM obtained with [(CHI)-(CuPcS)-(CHI)-(AuNP)]2-Tyr, and two orders of magnitude lower than the obtained with other nanostructured platforms. It can be concluded that the combination of adequate materials with complementary activity and the control of the structure of the platform is an excellent strategy to obtain biosensors with improved performances.Ministerio de Ciencia, Innovación y Universidades - Fondo Europeo de Desarrollo Regional (project RTI2018-097990-B-100)Junta de Castilla y Leon - Fondo Europeo de Desarrollo Regional (project VA275P18)Infraestructuras Red de Castilla y León (grant UVA01

The yin and yang-like clinical implications of the cdkn2a/arf/cdkn2b gene cluster in acute lymphoblastic leukemia

Altres ajuts: This project was supported by the Asociación Española Contra el Cáncer (AECC) (Project reference: GC16173697BIGA), Instituto de Salud Carlos III, CERCA Program/Generalitat de Catalunya.Acute lymphoblastic leukemia (ALL) is a malignant clonal expansion of lymphoid hematopoietic precursors that exhibit developmental arrest at varying stages of differentiation. Similar to what occurs in solid cancers, transformation of normal hematopoietic precursors is governed by a multistep oncogenic process that drives initiation, clonal expansion and metastasis. In this process, alterations in genes encoding proteins that govern processes such as cell proliferation, differentiation, and growth provide us with some of the clearest mechanistic insights into how and why cancer arises. In such a scenario, deletions in the 9p21.3 cluster involving CDKN2A/ARF/CDKN2B genes arise as one of the oncogenic hallmarks of ALL. Deletions in this region are the most frequent structural alteration in T-cell acute lymphoblastic leukemia (T-ALL) and account for roughly 30% of copy number alterations found in B-cell-precursor acute lymphoblastic leukemia (BCP-ALL). Here, we review the literature concerning the involvement of the CDKN2A/B genes as a prognosis marker of good or bad response in the two ALL subtypes (BCP-ALL and T-ALL). We compare frequencies observed in studies performed on several ALL cohorts (adult and child), which mainly consider genetic data produced by genomic techniques. We also summarize what we have learned from mouse models designed to evaluate the functional involvement of the gene cluster in ALL development and in relapse/resistance to treatment. Finally, we examine the range of possibilities for targeting the abnormal function of the protein-coding genes of this cluster and their potential to act as anti-leukemic agents in patients

Adverse prognostic impact of complex karyotype (≥3 cytogenetic alterations) in adult T-cell acute lymphoblastic leukemia (T-ALL)

Cytogenetics; Prognosis; TherapyCitogenética; Pronóstico; TerapiaCitogenètica; Pronòstic; TeràpiaThe potential prognostic value of conventional karyotyping in adult T-cell acute lymphoblastic leukemia (T-ALL) remains an open question. We hypothesized that a modified cytogenetic classification, based on the number and type of cytogenetic abnormalities, would allow the identification of high-risk adult T-ALL patients. Complex karyotype defined by the presence of ≥3 cytogenetic alterations identified T-ALL patients with poor prognosis in this study. Karyotypes with ≥3 abnormalities accounted for 16 % (22/139) of all evaluable karyotypes, corresponding to the largest poor prognosis cytogenetic subgroup of T-ALL identified so far. Patients carrying karyotypes with ≥3 cytogenetic alterations showed a significantly inferior response to therapy, and a poor outcome in terms of event-free survival (EFS), overall survival (OS) and cumulative incidence of relapse (CIR), independently of other baseline characteristics and the end-induction minimal residual disease (MRD) level. Additional molecular analyses of patients carrying ≥3 cytogenetic alterations showed a unique molecular profile that could contribute to understand the underlying molecular mechanisms of resistance and to evaluate novel targeted therapies (e.g. IL7R directed) with potential impact on outcome of adult T-ALL patients.This project was supported by the AECC (GC16173697BIGA); ISCIII (PI19/01828) co-funded by ERDF/ESF "A way to make Europe"/"Investing in your future", CERCA/Generalitat de Catalunya SGR 2017 288 (GRC)/ “La Caixa” P. Barba was supported by the Instituto de Salud Carlos III FIS16/01433 and PERIS 2018-2020 from Generalitat de Catalunya (BDNS357800)

¿Se encuentran los escolares que reciben apoyo educativo en situación de mayor riesgo de presentar dificultades de aprendizaje en la escritura?

Nowadays, writing is considered as a complex cognitive process in which several factors are involved that lead to a right writing use in adult age. This process is set up by different sub-processes, between which we can highlight transcription. The transcription skill takes a relevant place in writing acquisition, being of a special importance in the school beginning. If children are imprecise and slow in the transcription process it can lead to a serious setback in to their composition ability and quality. Transcription has a significant effect into the global writing and the academic efficiency in school age even that is considered as a powerful predictor to identify children that are more likely to show severe deficit in the acquisition of writing skills. A discovery that strengthen the importance of identifying writing difficulties as soon as possible. It should be added that nowadays writing teaching at the beginning is focused almost, solely and exclusively, in the most formal text aspects, just like handwriting or legibility, excluding fluency instruction. The current study analyzes the differences in writing fluency and accuracy between children that receive educational support because of their backwardness in the education program and normally achieving children. For this purpose , we selected a sample of 181 normally achieving children and 181 children receiving educational support, belonging to Primary Education and whose ages are in 6 – 9 years old range. Obtained results show that children that receive educational support present a lack of writing fluency and accuracy, problems that without an appropriate instruction could lead in future writing difficulties.Actualmente la escritura es considerada como un proceso cognitivo complejo en el que intervienen diversos factores que conducen a un correcto uso de la escritura en la edad adulta. Este proceso está formado por distintos subprocesos. Entre ellos cabe destacar la transcripción. La capacidad de transcripción adquiere un papel importante en la adquisición de la escritura, siendo de especial importancia en el inicio de la escolaridad. Si los niños son lentos e inexactos en el proceso de la transcripción conlleva a que su capacidad y calidad compositiva global en la producción de textos se vea gravemente perjudicada. La transcripción tiene un efecto significativo sobre la escritura global y el rendimiento académico de los niños en edad escolar, tanto es así que es considerada como un potente predictor para identificar a niños en riesgo de desarrollar déficits más graves en la escritura. Un hallazgo que refuerza la importancia de identificar las dificultades de escritura tan pronto como sea posible. A esto, hay que añadir que actualmente la enseñanza de la escritura en los primeros años de escolaridad se centra casi única y exclusivamente en los aspectos más formales del texto como la caligrafía o la legibilidad, excluyendo la instrucción en la fluidez. La presente investigación analiza las diferencias entre niños que reciben apoyo educativo por presentar retraso en la adquisición de las competencias del currículo escolar con niños que no lo reciben, considerando la fluidez y la exactitud en la escritura. Para ello se seleccionó una muestra de 181 alumnos que reciben apoyo educativo y 181 alumnos que no lo reciben, pertenecientes a los tres primeros cursos de Educación Primaria y cuyas edades oscilan entre 6 y 9 años. Los resultados obtenidos muestran que los niños que reciben apoyo educativo, aún recibiendo este apoyo, presentan mayores carencias en la fluidez y exactitud en la escritura, carencias que sin una apropiada instrucción podrían desembocar en futuras dificultades de aprendizaje en escritura

Fluidez y exactitud en la copia de letras del alfabeto (manuscrita vs. cursiva): un estudio transversal

Handwriting evolves with the pass of time. The type of script which children begin to learn depends on curriculum in their countries and the educational policy. There are two main types of script: manuscript and cursive. There is a controversial issue about which type of script would be best to use to begin the teaching of handwriting, but has not been a consensus yet. This research analyzes manuscript and cursive script modalities. Our objective was to determine whether there are differences in accuracy and fluency when students are copying the alphabet letters using different types of script (manuscript vs. cursive), and also whether these differences are mediated by the grade (1st, 2nd and 3rd ). A subtest from the test called Early Grade Writing Assessment (EGWA) (Jiménez, 2012) was administered to a sample of children from 1st, 2nd and 3rd grade.La escritura evoluciona con el paso del tiempo. El tipo de letra que los niños comienzan a aprender depende del currículo de su país y la política educativa. Hay dos tipos principales de letra: manuscrita y cursiva. Existe un debate sobre qué tipo de letra sería mejor utilizar para comenzar la enseñanza de la escritura, pero no se ha llegado a un consenso. Esta investigación analiza la escritura con ambos tipos de letra. Nuestro objetivo ha sido averiguar si existen diferencias en la exactitud y fluidez cuando los niños copian el alfabeto utilizando diferentes tipos de letras (manuscrita vs. cursiva), y si estas diferencias están mediatizadas por el curso (1º, 2º y 3º). Para ello se administraron algunos subtests de la prueba denominada Early Grade Writing Assessment (EGWA) (Jiménez, 2012) a una muestra de niños de 1º, 2º y 3º de Educación Primaria.

Genomics improves risk stratification of adults with T-cell acute lymphoblastic leukemia enrolled in measurable residual disease-oriented trials

Genomics; T-cell acute lymphoblastic leukemiaGenòmica; leucèmia limfoblàstica aguda de cèl·lules TGenómica; Leucemia linfoblástica aguda de células TGenetic information has been crucial to understand the pathogenesis of T-cell acute lymphoblastic leukemia (T-ALL) at diagnosis and at relapse, but still nowadays has a limited value in a clinical context. Few genetic markers are associated with the outcome of T-ALL patients, independently of measurable residual disease (MRD) status after therapy. In addition, the prognostic relevance of genetic features may be modulated by the specific treatment used. We analyzed the genetic profile of 145 T-ALL patients by targeted deep sequencing. Genomic information was integrated with the clinicalbiological and survival data of a subset of 116 adult patients enrolled in two consecutive MRD-oriented trials of the Spanish PETHEMA (Programa Español de Tratamientos en Hematología) group. Genetic analysis revealed a mutational profile defined by DNMT3A/ N/KRAS/ MSH2/ U2AF1 gene mutations that identified refractory/resistant patients. Mutations in the DMNT3A gene were also found in the non-leukemic cell fraction of patients with T-ALL, revealing a possible mutational-driven clonal hematopoiesis event to prime T-ALL in elderly. The prognostic impact of this adverse genetic profile was independent of MRD status on day +35 of induction therapy. The combined worse-outcome genetic signature and MRD on day +35 allowed risk stratification of T-ALL into standard or high-risk groups with significantly different 5- year overall survival (OS) of 52% (95% confidence interval: 37-67) and 17% (95% confidence interval: 1-33), respectively. These results confirm the relevance of the tumor genetic profile in predicting patient outcome in adult T-ALL and highlight the need for novel gene-targeted chemotherapeutic schedules to improve the OS of poor-prognosis T-ALL patients.This project was supported by the AECC (GC16173697BIGA); ISCIII (PI19/01828 and PI19/01183), co-funded by ERDF/ESF, "A way to make Europe"/"Investing in your future", CERCA/Generalitat de Catalunya SGR 2017 288 (GRC)/ “La Caixa”. C Gon-zález-Gil was supported by AGAUR grant (ref: 2020 FI_B2 00210)

Recomendador inteligente de vestimenta

En la actualidad más que nunca, una imagen vale más que mil palabras. Esto coincide con una sociedad que cada vez valora más su tiempo y donde desde hace pocas décadas existe un auge de los grandes gigantes de moda, con millones de personas que quieren estar siempre atentos a las últimas tendencias. Este trabajo ha consistido en el desarrollo de una aplicación web que combina todo esto, ayudándonos a mejorar nuestra imagen, adecuándose siempre a cada evento y circunstancia, pero sin perder la esencia personal. Esta aplicación aprenderá de tus gustos y los ajustará a tu agenda, haciendo de esta aplicación una experiencia personal muy satisfactoria. Para ello, se tendrá en cuenta el fondo de armario del usuario, y analizando diversos factores como la meteorología de ese día, los eventos anotados en su agenda y la formalidad requerida, sus preferencias. . . se le dará un look acorde a todo ello. Así mismo, la app podrá hacer recomendaciones de looks que se asemejen a la tendencia de todos los usuarios de la aplicación en ese momento, utilizando prendas lo más parecidas de su armario. Una vez escogido el look, la aplicación aprenderá acerca de su elección

Imipenem heteroresistance but not tolerance in Haemophilus influenzae during chronic lung infection associated with chronic obstructive pulmonary disease

Antibiotic resistance is a major Public Health challenge worldwide. Mechanisms other than resistance are described as contributors to therapeutic failure. These include heteroresistance and tolerance, which escape the standardized procedures used for antibiotic treatment decision-making as they do not involve changes in minimal inhibitory concentration (MIC). Haemophilus influenzae causes chronic respiratory infection and is associated with exacerbations suffered by chronic obstructive pulmonary disease (COPD) patients. Although resistance to imipenem is rare in this bacterial species, heteroresistance has been reported, and antibiotic tolerance cannot be excluded. Moreover, development of antibiotic heteroresistance or tolerance during within-host H. influenzae pathoadaptive evolution is currently unknown. In this study, we assessed imipenem resistance, heteroresistance and tolerance in a previously sequenced longitudinal collection of H. influenzae COPD respiratory isolates. The use of Etest, disc diffusion, population analysis profiling, tolerance disc (TD)-test methods, and susceptibility breakpoint criteria when available, showed a significant proportion of imipenem heteroresistance with differences in terms of degree among strains, absence of imipenem tolerance, and no specific trends among serial and clonally related strains could be established. Analysis of allelic variation in the ftsI, acrA, acrB, and acrR genes rendered a panel of polymorphisms only found in heteroresistant strains, but gene expression and genome-wide analyses did not show clear genetic traits linked to heteroresistance. In summary, a significant proportion of imipenem heteroresistance was observed among H. influenzae strains isolated from COPD respiratory samples over time. These data should be useful for making more accurate clinical recommendations to COPD patients