Multimodality and deaf children's participation in informal literature socialization contexts

En este artículo realizamos un análisis del discurso y la interacción multimodal de una sesión de cuentacuentos realizada en una biblioteca en la que el narrador está acompañado por una intérprete de lengua de signos española (LSE). El caso forma parte de una investigación más amplia que examina sesiones de cuentacuentos en bibliotecas, librerías infantiles y parques como espacios de socialización literaria en contextos de aprendizaje informal. Analizamos, por una parte, la relación cambiante entre los medios/modos desplegados en el relato y las dificultades que esto genera para los espectadores sordos y, por otra parte, la propia participación de los niños sordos en esta sesión. El análisis hace explícita la complejidad del acto de interpretación en LSE y la situación asimétrica en la que se encuentra la infancia sorda en estas sesiones de cuentacuentos. Igualmente, el episodio analizado invita a plantear el desarrollo de una conciencia multimodal como parte de las competencias literarias y discursivas que potencialmente favorecen la clase de eventos narrativos analizadosIn this article we present an analysis of discourse and multimodal interaction in a storytelling session that took place in a library in which the main narrator is accompanied by a Spanish Sign Language (LSE) interpreter. This case is part of a larger project that examines storytelling sessions in libraries, bookstores and parks as informal literature socialization contexts. On one hand, we examine the changing relationship between media/modes displayed in the story and the difficulties that this generates for deaf children. On the other hand, we explore deaf children's participation in the event. Our analysis makes explicit the complexities involved in LSE interpretation and the asymmetrical standing of deaf children in these types of events. Also, with the analysis of this episode we consider multimodal awareness as a possible discursive and literary competence that is fostered in these narratives event

RNA-Seq transcriptomic profiling of primary murine microglia treated with LPS or LPS+IFNγ

Microglia, the main resident immune cells in the CNS, are thought to participate in the pathogenesis of various neurological disorders. LPS and LPS + IFNγ are stimuli that are widely used to activate microglia. However, the transcriptomic profiles of microglia treated with LPS and LPS + IFNγ have not been properly compared. Here, we treated murine primary microglial cultures with LPS or LPS + IFNγ for 6 hours and then performed RNA-Sequencing. Gene expression patterns induced by the treatments were obtained by WGCNA and 11 different expression profiles were found, showing differential responses to LPS and LPS + IFNγ in many genes. Interestingly, a subset of genes involved in Parkinson's, Alzheimer's and Huntington's disease were downregulated by both treatments. By DESeq analysis we found differentially upregulated and downregulated genes that confirmed LPS and LPS + IFNγ as inducers of microglial pro-inflammatory responses, but also highlighted their involvement in specific cell functions. In response to LPS, microglia tended to be more proliferative, pro-inflammatory and phagocytic; whereas LPS + IFNγ inhibited genes were involved in pain, cell division and, unexpectedly, production of some inflammatory mediators. In summary, this study provides a detailed description of the transcriptome of LPS- and LPS + IFNγ treated primary microglial cultures. It may be useful to determine whether these in vitro phenotypes resemble microglia in in vivo pathological conditions

Myeloid C/EBPβ deficiency reshapes microglial gene expression and is protective in experimental autoimmune encephalomyelitis

Background CCAAT/enhancer binding protein β (C/EBPβ) is a transcription factor that regulates the expression of important pro-inflammatory genes in microglia. Mice deficient for C/EBPβ show protection against excitotoxic and ischemic CNS damage, but the involvement in this neuroprotective effect of the various C/EBPβ-expressing cell types is not solved. Since C/EBPβ-deficient microglia show attenuated neurotoxicity in culture, we hypothesized that specific C/EBPβ deficiency in microglia could be neuroprotective in vivo. In this study, we have tested this hypothesis by generating mice with myeloid C/EBPβ deficiency. Methods Mice with myeloid C/EBPβ deficiency were generated by crossing LysMCre and C/EBPβfl/fl mice. Primary microglial cultures from C/EBPβfl/fl and LysMCre-C/EBPβfl/fl mice were treated with lipopolysaccharide ± interferon γ (IFNγ) for 6 h, and gene expression was analyzed by RNA sequencing. Gene expression and C/EBPβ deletion were analyzed in vivo in microglia isolated from the brains of C/EBPβfl/fl and LysMCre-C/EBPβfl/fl mice treated systemically with lipolysaccharide or vehicle. Mice of LysMCre-C/EBPβfl/fl or control genotypes were subjected to experimental autoimmune encephalitis and analyzed for clinical signs for 52 days. One- or two-way ANOVA or Kruskal-Wallis with their appropriate post hoc tests were used. Results LysMCre-C/EBPβfl/fl mice showed an efficiency of C/EBPβ deletion in microglia of 100 and 90% in vitro and in vivo, respectively. These mice were devoid of female infertility, perinatal mortality and reduced lifespan that are associated to full C/EBPβ deficiency. Transcriptomic analysis of C/EBPβ-deficient primary microglia revealed C/EBPβ-dependent expression of 1068 genes, significantly enriched in inflammatory and innate immune responses GO terms. In vivo, microglial expression of the pro-inflammatory genes Cybb, Ptges, Il23a, Tnf and Csf3 induced by systemic lipopolysaccharide injection was also blunted by C/EBPβ deletion. CNS expression of C/EBPβ was upregulated in experimental autoimmune encephalitis and in multiple sclerosis samples. Finally, LysMCre-C/EBPβfl/fl mice showed robust attenuation of clinical signs in experimental autoimmune encephalitis. Conclusion This study provides new data that support a central role for C/EBPβ in the biology of activated microglia, and it offers proof of concept for the therapeutic potential of microglial C/EBPβ inhibition in multiple sclerosis

Retrospective evaluation of whole exome and genome mutation calls in 746 cancer samples

Funder: NCI U24CA211006Abstract: The Cancer Genome Atlas (TCGA) and International Cancer Genome Consortium (ICGC) curated consensus somatic mutation calls using whole exome sequencing (WES) and whole genome sequencing (WGS), respectively. Here, as part of the ICGC/TCGA Pan-Cancer Analysis of Whole Genomes (PCAWG) Consortium, which aggregated whole genome sequencing data from 2,658 cancers across 38 tumour types, we compare WES and WGS side-by-side from 746 TCGA samples, finding that ~80% of mutations overlap in covered exonic regions. We estimate that low variant allele fraction (VAF < 15%) and clonal heterogeneity contribute up to 68% of private WGS mutations and 71% of private WES mutations. We observe that ~30% of private WGS mutations trace to mutations identified by a single variant caller in WES consensus efforts. WGS captures both ~50% more variation in exonic regions and un-observed mutations in loci with variable GC-content. Together, our analysis highlights technological divergences between two reproducible somatic variant detection efforts

Generation of mice deficient in C/EBPbeta in microglial cells using the Cre/LoxP system

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Deleció condicional de C/EBPß a les cèl·lules de la micròglia. Nova eina per a l'estudi i la modulació de l'activació glial en models de neurodegeneració

[cat] Treballs publicats recentment demostren que el factor de transcripció CCAAT/enhancer binding protein beta (C/ EBPbeta) regula l'activació microglial. Aquests resultats suggereixen que la inhibició de EBPbeta a les cèl.lules de la micròglia podria resultar neuroprotectora en trastorns del SNC on la inflamació crònica resulta ésser patològica. Amb l'objectiu de corroborar aquesta hipòtesi en models animals de neurodegeneració, durant el desenvolupament d'aquesta tesi doctoral s'ha generat la colònia LysMCre-C/ EBPbeta (fl/fl) amb deficiència selectiva de C/EBPbeta a les cèl.lules de la micròglia. Aquests animals són fèrtils, viables i l'estudi histològic no mostra cap alteració als òrgans i teixits, contràriament a allò observat als animals on el factor de transcripció ha estat eliminat a tots els tipus cel.lulars. Cultius primaris de micròglia mostren que la deleció de C/EBPbeta s'ha produït al 100% de les cèl.lules, mentre que l'expressió del factor de transcripció als astròcits no es veu alterada. La deleció de C/EBP8 a les cèl.lules microglials inhibeix la producció de NO alhora que augmenta la capacitat d'aquestes cèl.lules per a fagocitar i eliminar bacteris. A més, la seqüenciació de l'ARN de la micròglia primària mostra una gran afectació del transcriptoma d'aquest tipus cel.lular en resposta als tractaments amb LPS i LPS+IFNy. En aquestes mostres, la deleció de C/EBPbeta afecta l'expressió de 1068 gens involucrats principalment en les respostes inflamatòria i immune, fet que suggereix la implicació del factor de transcripció en aquests processos biològics. Per tal de corroborar aquests resultats en ratolins adults, s'ha aïllat la micròglia dels SNC d'aquests animals. Les dades obtingudes mostren que el 90% de les cèl.lules micròglials no expressen el factor de transcripció, alhora que l'expressió de gens proinflamatoris es veu clarament redu•la. Finalment, l'expressió de C/EBPbeta augmenta marcadament al model animal d'esclerosi múltiple, EAE, per aquesta raó s'ha induït la patologia als animals LysMCre- EBPbeta (fl/fl) ". Aquests ratolins mostren un retard en l'aparició dels primers símptomes, així com una marcada atenuació de la simptomatologia patològica. Així doncs, els resultats obtinguts en aquest treball demostren que C/EBPbeta és un factor clau en la regulació de l'activació microglial i que la seva inhibició resulta ésser una estratègia terapèutica.[eng] Recently published reports demonstrate that the transcription factor CCAAT/enhancer binding protein beta (C/EBPbeta) regulates microglia activation. This suggests that microglial C/EBPbeta inhibition could have therapeutical potential in CNS disorders with a pathogenic neuroinflammatory component. To test this hypothesis in animal models of neurodegenerative diseases, we have therefore generated mice with specific microglial C/EBPbeta deficiency, crossing mice with Cre expression under the microglial/macrophage promoter LysM with C/EBPbetaflm mice. Double mutants LysMCre-C/EBPbetaflm were selected. These animals showed normal fertility, survival and histology in contrast to C/EBPbeta deficient mice. In primary microglial cultures, lack of C/EBPbeta was observed in virtually 100% of microglial cells, whereas astrocytes showed normal C/EBPbeta expression. Microglial C/EBPbeta absence resulted in the almost blockade of NO production induced by LPS+IFNy and in an altered bacterial phagocytic function. Furthermore, RNA sequencing of cultured microglia shows dramatic changes in the microglia transcriptome in response to LPS and LPS+IFNy. In these samples C/EBPbeta deletion alters the expression of 1068 mainly involved in immune and inflammatory responses, suggesting and important role of this transcription factor in this biological processes. To corroborate these findings in adult mice, microglia was acutely isolated from the CNS and analyzed. Data revealed a very high (90%) proportion of C/EBPbeta negative microglial cells also in vivo, as well as, a remarkably reduction of proinflammatory genes expression. Finally, because C/EBPbeta was markedly upregulated by experimental autoimmune encephalomyelitis (EAE) in wild-type mice, control and LysMCre-C/EBPbetaflm mice were subjected to EAE. LysMCre-C/EBPbetaflm mice presented a delayed EAE onset and EAE severity was markedly attenuated. Alltogether, these findings support the hypothesis that C/EBPbeta is a key regulator of proinflammatory gene expression in microglial cells and that its inhibition has therapeutical potential

Multimodalidad y participación de la infancia sorda en contextos de socialización literaria informales.

In this article we present an analysis of discourse and multimodal interaction in a storytelling session that took place in a library in which the main narrator is accompanied by a Spanish Sign Language (LSE) interpreter. This case is part of a larger project that examines storytelling sessions in libraries, bookstores and parks as informal literature socialization contexts. On one hand, we examine the changing relationship between media/modes displayed in the story and the difficulties that this generates for deaf children. On the other hand, we explore deaf children's participation in the event. Our analysis makes explicit the complexities involved in LSE interpretation and the asymmetrical standing of deaf children in these types of events. Also, with the analysis of this episode we consider multimodal awareness as a possible discursive and literary competence that is fostered in these narratives events.En este artículo realizamos un análisis del discurso y la interacción multimodal de una sesión de cuentacuentos realizada en una biblioteca en la que el narrador está acompañado por una intérprete de lengua de signos española (LSE). El caso forma parte de una investigación más amplia que examina sesiones de cuentacuentos en bibliotecas, librerías infantiles y parques como espacios de socialización literaria en contextos de aprendizaje informal. Analizamos, por una parte, la relación cambiante entre los medios/modos desplegados en el relato y las dificultades que esto genera para los espectadores sordos y, por otra parte, la propia participación de los niños sordos en esta sesión. El análisis hace explícita la complejidad del acto de interpretación en LSE y la situación asimétrica en la que se encuentra la infancia sorda en estas sesiones de cuentacuentos. Igualmente, el episodio analizado invita a plantear el desarrollo de una conciencia multimodal como parte de las competencias literarias y discursivas que potencialmente favorecen la clase de eventos narrativos analizados. Palabra

Gene expression profiling of LPS- and LPS+IFN-activated primary murine microglia by RNAseq

Trabajo presentado en el IX Simposi de Neurobiologia Experimental, celebrado en Barcelona, España, el 22 y 23 de octubre de 2014Peer Reviewe

Generation of LyzM-Cre x C/EBPβfl/fl mice to obtain selective depletion of the transcription factor C/EBPβ in microglia

Trabajo presentado al I Congrés Internacional de Biologia de Catalunya: Global questions on advanced biology, celebrado en Barcelona del 9 al 12 de julio de 2012.Complete deficiency of the transcription factor CCAAT/enhancer binding protein β (C/EBPβ) protects from excitotoxic and ischemic insults suggesting the involvement of C/EBPβ in neurotoxicity in these models. C/EBPβ can be expressed by numerous cell types and the cell types involved in the potential detrimental effects of C/EBPβ have not been determined. We have recently shown that C/EBPβ absence results in the attenuated expression of pro-inflammatory genes in primary mixed glial cultures and in the abrogation of the neurotoxicity elicited by activated microglia in neuron-microglia co-cultures. In order to determine the neuroprotective potential of C/EBPβ inhibition in microglia we have generated a mouse transgenic line in which C/EBPβ gene is flanked by LoxP sequences (C/EBPβfl/fl) and Cre-recombinase is expressed under the control of the microglia/macrophage promoter Lysozyme M. In primary mixed glial cultures from wild-type or from the two parental lines, C/EBPβ was expressed both in astrocytes and microglia. Primary mixed glial cultures prepared from the cerebral cortex of neonatal LyzM-Cre x C/EBPβfl/fl mice revealed that virtually all microglial cells, identified by CD11b or Iba1 immunostaining, lacked C/EBPβ expression whereas C/EBPβ expression in astrocytes, identified by GFAP immunostaining, was similar to control lines. Interestingly, LPS or LPS+IFNγ-induced NO production in mixed glial cultures, which is of microglial origin, was markedly attenuated in LyzM-Cre x C/EBPβfl/fl cultures when compared to control lines and the same effect was observed in microglial-enriched cultures. We are in the process of obtaining samples to analyze gene expression profile of control and activated microglial cultures from LyzM-Cre x C/EBPβfl/fl and wild-type mice. In parallel, we are interested to determine whether the cell specific absence of C/EBPβ in microglia in LyzM-Cre x C/EBPβfl/fl mice is also observed in vivo. In this case, LyzM-Cre x C/EBPβfl/fl mice would be a useful tool to analyze the involvement of microglial C/EBPβ in neurodegeneration in vivo and to test the hypothesis that inhibition of microglial C/EBPβ could have potential for the treatment of neurological disorders in which neuroinflammation plays a pathogenic role.Supported by ISCIII PI08/1396 and PI10/0378.Peer reviewe

Is microglial C/EBPß deficiency neuroprotective in EAE? A new mouse model to study its implications in vitro and in vivo

Trabajo presentado en la XII European Meeting on Glial Cells in Health and Disease, celebrada en Bilbao, España, del 15 al 18 de julio de 2105Peer Reviewe