Activity-dependent regulation of thalamic interneuron and microglia in the visual thalamus

Los circuitos talámicos están formados por neuronas de proyección excitadoras y neuronas inhibidoras. De manera similar a como ocurre en la corteza, las neuronas excitadoras talámicas nacen de progenitores que se encuentran en la zona proliferativa del tálamo en desarrollo, mientras que las interneuronas inhibidoras locales nacen fuera del tálamo y necesitan migrar hasta él para integrarse en el circuito. En ratón, las interneuronas locales del tálamo se encuentran principalmente en el núcleo dorso-lateral geniculado (dLGN), el núcleo visual primario, encargado de recibir los axones retinales y proyectar a la corteza visual primaria (V1). La integración de estas interneuronas en el circuito comienza durante el desarrollo postnatal temprano en ratón. A pesar del estado inmaduro de las conexiones neuronales a esta edad, los circuitos son activos de manera espontánea, con patrones definidos de disparo. Por lo general, se ha observado que la actividad periférica es relevante para la correcta migración e integración de las interneuronas talámicas. Sin embargo, aún no está claro si estos procesos se ven afectados por patrones específicos de actividad que surgen en paralelo y son mediadas por otras fuentes. En este proyecto, describimos cómo las interneuronas se comportan de manera diferente a distintas etapas del desarrollo, desde estadíos tardíos embrionarios hasta estadíos postnatales tempranos, tras interrumpir la actividad retinal o talámica. El bloqueo de la actividad retinal mediante diferentes procedimientos confirma resultados previos que sugieren que los axones retinales son necesarios por la colocación de las interneuronas en el dLGN. Asimismo, hemos observado que la actividad intrínseca del tálamo es también importante para la velocidad de migración de estas interneuronas locales talámicas. Además, hemos visto que la actividad espontánea talámica durante el desarrollo embrionario es necesaria para el correcto posicionamiento de las interneuronas corticales en V1, principalmente aquellas que expresan SST y PV. Es interesante observar que los resultados obtenidos combinando todos los modelos muestran que el dLGN necesita llegar a un número concreto de interneuronas locales talámicas. Puesto que hay evidencias previas que sugieren que la maduración de la microglia y las interneuronas podría estar conectada, buscamos también estudiar las células microgliales en nuestros modelos. De esta forma, encontramos que la actividad intrínseca del tálamo afecta a la densidad de microglia talámica, así como a la microglia cortical en estadíos postnatales, mientras que la supresión de la actividad retinal no afecta a estas células. Así, la combinación de todos estos resultados sugiere que los patrones tempranos de actividad talámica son un factor novedoso involucrado en la correcta integración de las interneuronas en el sistema visual

Urban ageing, gender and the value of the local environment: The experience of older women in a central neighbourhood of Madrid, Spain

This article belongs to the Special Issue Land, Ageing, Gender and Environment: Problems and Challenges from Different Disciplines.Urban ageing is an emerging domain that mixes two challenges of current societies: the ageing of the population and the increasing urbanisation. While ageing in place has demonstrated numerous benefits, some social sectors question whether the city is the right environment for ageing, since cities are home to many of the social problems that characterise contemporary societies. Urban environments are widely described as rootless in most academic articles, with a focus on the impersonality, transience, and segmentation of links between city dwellers. However, this portrayal coexists with contrasting views of urban life that instead emphasise the importance of the local setting and other experiences of attachment to the place of residence. From the age and gender perspectives, in some urban areas, the neighbourhood plays a fundamental role in the lives of many older women, as a natural setting for interaction and an area conducive to collaborative relationships and practical and emotional support in times of need. This article analyses the role that the local space plays in the lives of older women, the value they attach to it and the meaning they attribute to neighbourhood relations in the local urban environment. In order to analyse this reality, the Universidad (Malasaña) neighbourhood, has been selected as a case study, a central area in Madrid (Spain) exposed to numerous processes of transformation, which shows the highest rate of residential mobility in the area. Despite this reality, in a context marked by new difficulties, the conclusion shows that elderly women have a strong attachment to their neighbourhood. This attachment is not necessarily characterised by deep friendships, but by the existence of a significant social network that responds in case of need

A general protocol to afford enantioenriched linear homoprenylic amines

The reaction of a readily obtained chiral branched homoprenylamonium salt with a range of aldehydes, including aliphatic substrates, affords the corresponding linear isomers in good yields and enantioselectivities.We thank the Spanish Ministerio de Ciencia e Innovación for their financial support (CTQ2011-24165). I. B. acknowledges the Generalitat Valenciana for a predoctoral fellowship (ACIF/2011/159)

Isolation, Culture, and Immunophenotypic Analysis of Bone Marrow HSPCs From Patients With Myelodysplastic Syndromes

Drug testing assays in hematopoietic stem and progenitor cells (HSPCs) are fundamental in biological studies of myelodysplastic syndromes (MDS) but have historically entailed a technical challenge. This protocol allows the efficient isolation of MDS HSPCs from bone marrow mononuclear cell fractions and their culturing with the support of stromal cells for improved maintenance during drug testing. Lastly, specific steps are given to quantify surviving cells and assess changes in the HSPC hierarchies. For complete details on the use and execution of this protocol, please refer to Ganan-Gomez et al. (2022)

A cryopreservation method for bioengineered 3D cell culture models

Technologies to cryogenically preserve (a.k.a. cryopreserve) living tissue, cell lines and primary cells have matured greatly for both clinicians and researchers since their first demonstration in the 1950s and are widely used in storage and transport applications. Currently, however, there remains an absence of viable cryopreservation and thawing methods for bioengineered, three-dimensional (3D) cell models, including patients' samples. As a first step towards addressing this gap, we demonstrate a viable protocol for spheroid cryopreservation and survival based on a 3D carboxymethyl cellulose scaffold and precise conditions for freezing and thawing. The protocol is tested using hepatocytes, for which the scaffold provides both the 3D structure for cells to self-arrange into spheroids and to support cells during freezing for optimal post-thaw viability. Cell viability after thawing is improved compared to conventional pellet models where cells settle under gravity to form a pseudo-tissue before freezing. The technique may advance cryobiology and other applications that demand high-integrity transport of pre-assembled 3D models (from cell lines and in future cells from patients) between facilities, for example between medical practice, research and testing facilities

A Synthesis of the Producer-Consumer Problem Using NowPit

In recent years, much research has been de- voted to the visualization of multicast applications; on the other hand, few have enabled the simulation of IPv6. In our research, authors prove the construction of write-ahead logging. Our focus in this position paper is not on whether the UNIVAC computer and robots are rarely incompatible, but rather on proposing new self-learning archetypes (NowPit)

Aproximación computacional basada en métodos de inteligencia artificial para la predicción de complejos proteína-proteína

De novo protein design has reached a new height in recent years thanks to new computational procedures, such as AlphaFold2, an artificial intelligence (AI) that predicts the three-dimensional structure of proteins from an amino acid sequence. This method allows for the design of proteins with high structural precision and complexity, capable of performing functions of medical and biotechnological relevance. This work aims to explore how the latest artificial intelligence methods, specifically AlphaFold2, can improve the prediction of protein tertiary structures and determine which proteins are more likely to successfully bind to therapeutic targets of interest, such as the RBD domain of SARS-CoV-2. Additionally, it aims to optimize current computational methodologies. Specifically, the prediction of structures of protein-protein complexes and ligand complexes competing for the same receptor has been carried out, analyzing various parameters regarding the quality of the prediction and the physicochemical properties of the proteins. This approach aims to optimize the computational methodology and develop a ranking of the affinities of proteins that could bind to the RBD receptor. The proteins studied have been designed both by other laboratories in previously published studies and by the Protein Design and Modeling group at the Institute of Molecular Biology of Barcelona (IBMB), for which experimental data are available. This provides an ideal framework for working on the optimization of computational methodologies.El disseny computacional de proteïnes de novo ha experimentat un creixement en els últims anys gracies a nous procediments computacionals, com AlphaFold2, una intel·ligència artificial (IA) que prediu l'estructura tridimensional de proteïnes a partir d'una seqüencia d'aminoàcids. Aquest mètode permet el disseny de proteïnes amb alta precisió estructural i complexitat, capaces de desenvolupar funcions de rellevància mèdica i biotecnològica. Aquest treball té com a objectiu explorar com els mètodes més recents d'intel·ligència artificial, concretament AlphaFold2, poden millorar la predicció de l'estructura terciària de proteïnes i determinar quines tenen més probabilitat d'unir-se exitosament a dianes terapèutiques d'interès, com el domini RBD de SARS-CoV-2. A més, es pretén optimitzar les metodologies computacionals actuals. Específicament, s'ha realitzat la predicció d'estructures de complexos proteïna-proteïna i de complexos de lligands competint pel mateix receptor, analitzant diversos paràmetres sobre la qualitat de la predicció i propietats fisicoquímiques de les proteïnes. Aquest enfocament busca optimitzar la metodologia computacional i desenvolupar un rànquing d'afinitat de les proteïnes que podrien unir-se al receptor RBD. Les proteïnes estudiantes han estat dissenyades tant per altres laboratoris en estudis prèviament publicats com pel grup Protein Design and Modeling de l'Institut de Biologia Molecular de Barcelona (IBMB), per a les quals hi ha dades experimentals disponibles. Això proporciona un marc ideal per treballar en l'optimització de les metodologies computacionals.El diseño computacional de proteínas de novo ha experimentado un auge en los últimos años gracias a nuevos procedimientos computacionales, como AlphaFold2, una inteligencia artificial (IA) que predice la estructura tridimensional de proteínas a partir de una secuencia de aminoácidos. Este método permite el diseño de proteínas con alta precisión estructural y complejidad, capaces de desempeñar funciones de relevancia médica y biotecnológica. Este trabajo tiene como objetivo explorar como los métodos más recientes de inteligencia artificial, concretamente AlphaFold2, pueden mejorar la predicción de la estructura terciaria de proteínas y determinar cuales tienen mayor probabilidad de unirse exitosamente a dianas terapéuticas d interés, como el dominio RBD de SARS-CoV-2. Además, se pretende optimizar las metodologías computacionales actuales. Específicamente, se ha realizado la predicción de estructuras de complejos proteína-proteína y de complejos de ligandos compitiendo por el mismo receptor, analizando diversos parámetros sobre la calidad de la predicción y propiedades fisicoquímicas de las proteínas. Este enfoque busca optimizar la metodología computacional y desarrollar un ranking de afinidades de las proteínas que podrían unirse al receptor RBD. Las proteínas estudiadas han sido diseñadas por otros laboratorios en estudios previamente publicados como el grupo Protein Design and Modeling del Instituto de Biología Molecular de Barcelona (IBMB), para las cuales hay datos experimentales disponibles. Esto proporciona un marco ideal para trabajar en la optimización de las metodologías computacionales.Objectius de Desenvolupament Sostenible::3 - Salut i Benesta

FSH and IGF-I signal via AKT to regulate beta-catenin accumulation and estradiol production in granulosa cells

Estradiol is the female sex hormone that profoundly influences the female reproductive system from puberty to fertility. Circulating estradiol in women is synthesized primarily by ovarian granulosa cells in response to the pituitary glycoprotein follicle-stimulating hormone (FSH). In granulosa cells FSH triggers a signaling cascade that subsequently induces expression of aromatase (Cyp19a1), a steroidogenic enzyme that catalyzes the aromatization of testosterone into estradiol. While FSH stimulates estradiol production, estradiol concentrations are regulated by intra-ovarian signaling molecules wingless-type mammary tumor virus integration-site (WNT) family molecules and insulin-like growth factor-I (IGF-I). Intracellular signaling cascades elicited by FSH, WNT, and IGF-I eventually overlap at beta-catenin, a transcription co-factor. In granulosa cells beta-catenin accumulates in response to FSH, and WNT, and is required for Cyp19a1 expression. Although it is evident that granulosa cells require beta-catenin to maintain estradiol production, there is still much to be identified about its role, regulators, and downstream effectors. In this dissertation, evidence is presented that enhances our understanding of the complex intracellular regulation of estradiol biosynthesis. Data demonstrates beta-catenin accumulation in response to FSH and IGF-I occur via phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT) pathway as inhibition of PI3K signaling reduced beta-catenin and estradiol concentrations. Additionally, IGF-I rescued FSH-mediated Cyp19a1 expression and estradiol production from the inhibitory effects of WNT3A. To elucidate the mechanism, beta-catenin accumulation, phosphorylation status of beta-catenin and Forkhead box O protein were analyzed by Western blot and Axin2 mRNA expression by real-time PCR. Data indicates IGF-I did not modulate expression of the above mentioned target markers and therefore was ruled out as potential mechanisms. A noteworthy discovery was in comparing animal models and identifying bovine but not rat granulosa cells have increased beta-catenin accumulation with IGF-I treatment which further adds to the complex nature of estradiol production. In the final study it was confirmed that the crucial mechanism by which beta-catenin regulates Cyp19a1 transcription is through its association with T-cell Factor on the promoter. Together, these data provide a new appreciation and understanding of the complex regulation of beta-catenin in estradiol production