Constructing a Sense of Story: One Block at a Time

This photo essay focuses upon the literacy practices of two groups of preschoolers as they built, illustrated, and dictated stories in response to their participation in a “Castle Project.” Data, including literacy artifacts, photodocumentation, sociodramatic play scenarios, and conversations are qualitatively analyzed, coded, and evaluated over a three month period. We use a narrative approach to describe the three- and four-year-olds’ talk, actions, and ideas, and the ways block play facilitated their sense of story and motivation to write. We suggest a reciprocity of thinking between the building and composing processes. Preschoolers’ story ideas, we deduce, were conceptualized and rehearsed in the block corner. We illustrate the power of emergent literacy learners to construct understandings of the world with the support and guidance of their teacher, using inquiry based and arts infused approaches to teaching and learning

Constructing a Sense of Story: One Block at a Time

This photo essay focuses upon the literacy practices of two groups of preschoolers as they built, illustrated, and dictated stories in response to their participation in a “Castle Project.” Data, including literacy artifacts, photodocumentation, sociodramatic play scenarios, and conversations are qualitatively analyzed, coded, and evaluated over a three month period. We use a narrative approach to describe the three- and four-year-olds’ talk, actions, and ideas, and the ways block play facilitated their sense of story and motivation to write. We suggest a reciprocity of thinking between the building and composing processes. Preschoolers’ story ideas, we deduce, were conceptualized and rehearsed in the block corner. We illustrate the power of emergent literacy learners to construct understandings of the world with the support and guidance of their teacher, using inquiry based and arts infused approaches to teaching and learning

Effects of Hst3p inhibition in Candida albicans: a genome-wide H3K56 acetylation analysis

Candida spp. represent the third most frequent worldwide cause of infection in Intensive Care Units with a mortality rate of almost 40%. The classes of antifungals currently available include azoles, polyenes, echinocandins, pyrimidine derivatives, and allylamines. However, the therapeutical options for the treatment of candidiasis are drastically reduced by the increasing antifungal resistance. The growing need for a more targeted antifungal therapy is limited by the concern of finding molecules that specifically recognize the microbial cell without damaging the host. Epigenetic writers and erasers have emerged as promising targets in different contexts, including the treatment of fungal infections. In C. albicans, Hst3p, a sirtuin that deacetylates H3K56ac, represents an attractive antifungal target as it is essential for the fungus viability and virulence. Although the relevance of such epigenetic regulator is documented for the development of new antifungal therapies, the molecular mechanism behind Hst3p-mediated epigenetic regulation remains unrevealed. Here, we provide the first genome-wide profiling of H3K56ac in C. albicans resulting in H3K56ac enriched regions associated with Candida sp. pathogenicity. Upon Hst3p inhibition, 447 regions gain H3K56ac. Importantly, these genomic areas contain genes encoding for adhesin proteins, degradative enzymes, and white-opaque switching. Moreover, our RNA-seq analysis revealed 1330 upregulated and 1081 downregulated transcripts upon Hst3p inhibition, and among them, we identified 87 genes whose transcriptional increase well correlates with the enrichment of H3K56 acetylation on their promoters, including some well-known regulators of phenotypic switching and virulence. Based on our evidence, Hst3p is an appealing target for the development of new potential antifungal drugs

BRD9 is a druggable component of interferon-stimulated gene expression and antiviral activity

Interferon (IFN) induction of IFN-stimulated genes (ISGs) creates a formidable protective antiviral state. However, loss of appropriate control mechanisms can result in constitutive pathogenic ISG upregulation. Here, we used genome-scale loss-of-function screening to establish genes critical for IFN-induced transcription, identifying all expected members of the JAK-STAT signaling pathway and a previously unappreciated epigenetic reader, bromodomain-containing protein 9 (BRD9), the defining subunit of non-canonical BAF (ncBAF) chromatin-remodeling complexes. Genetic knockout or small-molecule-mediated degradation of BRD9 limits IFN-induced expression of a subset of ISGs in multiple cell types and prevents IFN from exerting full antiviral activity against several RNA and DNA viruses, including influenza virus, human immunodeficiency virus (HIV1), and herpes simplex virus (HSV1). Mechanistically, BRD9 acts at the level of transcription, and its IFN-triggered proximal association with the ISG transcriptional activator, STAT2, suggests a functional localization at selected ISG promoters. Furthermore, BRD9 relies on its intact acetyl-binding bromodomain and unique ncBAF scaffolding interaction with GLTSCR1/1L to promote IFN action. Given its druggability, BRD9 is an attractive target for dampening ISG expression under certain autoinflammatory conditions

Biallelic JAK1 mutations in immunodeficient patient with mycobacterial infection

Mutations in genes encoding components of the immune system cause primary immunodeficiencies. Here, we study a patient with recurrent atypical mycobacterial infection and early-onset metastatic bladder carcinoma. Exome sequencing identified two homozygous missense germline mutations, P733L and P832S, in the JAK1 protein that mediates signalling from multiple cytokine receptors. Cells from this patient exhibit reduced JAK1 and STAT phosphorylation following cytokine stimulations, reduced induction of expression of interferon-regulated genes and dysregulated cytokine production; which are indicative of signalling defects in multiple immune response pathways including Interferon-γ production. Reconstitution experiments in the JAK1-deficient cells demonstrate that the impaired JAK1 function is mainly attributable to the effect of the P733L mutation. Further analyses of the mutant protein reveal a phosphorylation-independent role of JAK1 in signal transduction. These findings clarify JAK1 signalling mechanisms and demonstrate a critical function of JAK1 in protection against mycobacterial infection and possibly the immunological surveillance of cancer.Wellcome Trust (095198/Z/10/Z and 090233/Z/09/Z); Higher Education Funding Council for England; Great Ormond Street Hospital Children’s Charity; National Institute for Health Research (NIHR); Great Ormond Street Hospital Biomedical Research Centre; NIHR Cambridge Biomedical Research Centre; Alfonso Martin Escudero Foundatio

Development of a Grp94 inhibitor

This document is the Accepted Manuscript version of a Published Work that appeared in final form in the Journal of the American Chemical Society, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://doi.org/10.1021/ja303477g.Heat shock protein 90 (Hsp90) represents a promising therapeutic target for the treatment of cancer and other diseases. Unfortunately, results from clinical trials have been disappointing as off-target effects and toxicities have been observed. These detriments may be a consequence of pan-Hsp90 inhibition, as all clinically evaluated Hsp90 inhibitors simultaneously disrupt all four human Hsp90 isoforms. Using a structure-based approach, we designed an inhibitor of Grp94, the ER-resident Hsp90. The effect manifested by compound 2 on several Grp94 and Hsp90α/β (cytosolic isoforms) clients were investigated. Compound 2 prevented intracellular trafficking of the Toll receptor, inhibited the secretion of IGF-II, affected the conformation of Grp94, and suppressed Drosophila larval growth, all Grp94-dependent processes. In contrast, compound 2 had no effect on cell viability or cytosolic Hsp90α/β client proteins at similar concentrations. The design, synthesis, and evaluation of 2 are described herein

Biallelic RIPK1 mutations in humans cause severe immunodeficiency, arthritis, and intestinal inflammation.

RIPK1 (receptor-interacting serine/threonine kinase 1) is a master regulator of signaling pathways leading to inflammation and cell death and is of medical interest as a drug target. We report four patients from three unrelated families with complete RIPK1 deficiency caused by rare homozygous mutations. The patients suffered from recurrent infections, early-onset inflammatory bowel disease, and progressive polyarthritis. They had immunodeficiency with lymphopenia and altered production of various cytokines revealed by whole-blood assays. In vitro, RIPK1-deficient cells showed impaired mitogen-activated protein kinase activation and cytokine secretion and were prone to necroptosis. Hematopoietic stem cell transplantation reversed cytokine production defects and resolved clinical symptoms in one patient. Thus, RIPK1 plays a critical role in the human immune system

Experimental Nonalcoholic Steatohepatitis and Liver Fibrosis Are Ameliorated by Pharmacologic Activation of Nrf2 (NF-E2 p45-Related Factor 2)

Funding: Supported by grants MR/J001465/1 (JDH, MLJA, JFD and RJMcC) and MR/N009851/1 (JDH, MLJA, ATD-K and DJH) from the Medical Research Council of the UK and grants from Stony Brook Foundation and Reata Pharmaceuticals, Inc. (TH)Background & Aims:  Nonalcoholic steatohepatitis (NASH) is associated with oxidative stress. We surmised that pharmacological activation of NF-E2 p45-related factor 2 (Nrf2) using the acetylenic tricyclic bis(cyano-enone) TBE-31 would suppress NASH because Nrf2 is a transcriptional master regulator of intracellular redox homeostasis. Methods:  Nrf2+/+ and Nrf2-/- C57BL/6 mice were fed a high-fat plus fructose (HFFr) or regular chow (RC) diet for 16 weeks or 30 weeks, and then treated for the final 6 weeks, whilst still being fed the same HFFr or RC diets, with either TBE-31 or DMSO vehicle control. Measures of whole body glucose homeostasis, histological assessment of liver, and biochemical and molecular measurements of steatosis, endoplasmic reticulum (ER) stress, inflammation, apoptosis, fibrosis and oxidative stress were performed in livers from these animals. Results:  TBE-31−treatment reversed insulin resistance in HFFr-fed wild-type mice, but not in HFFr-fed Nrf2-null mice. TBE-31−treatment of HFFr-fed wild-type mice substantially decreased liver steatosis and expression of lipid synthesis genes, whilst increasing hepatic expression of fatty acid oxidation and lipoprotein assembly genes. Also, TBE-31−treatment decreased ER stress, expression of inflammation genes, and markers of apoptosis, fibrosis and oxidative stress in the livers of HFFr-fed wild-type mice. By comparison, TBE-31 did not decrease steatosis, ER stress, lipogenesis, inflammation, fibrosis or oxidative stress in livers of HFFr-fed Nrf2-null mice. Conclusions:  Pharmacological activation of Nrf2 in mice that had already been rendered obese and insulin resistant reversed insulin resistance, suppressed hepatic steatosis, and mitigated against NASH and liver fibrosis, effects that we principally attribute to inhibition of ER, inflammatory and oxidative stress.Publisher PDFPeer reviewe

Fortalecimiento de gestiones a través del Centro de Información de Actividades Porcinas (CIAP) para el desarrollo sustentable de pequeños y medianos productores porcinos familiares de la zona de influencia de la Facultad de Ciencias Agrarias de la Universidad Nacional de Rosario

Fortalecimiento de gestiones a través del Centro de Información de Actividades Porcinas (CIAP) para el desarrollo sustentable de pequeños y medianos productores porcinos familiares de la zona de influencia de la Facultad de Ciencias Agrarias de la Universidad Nacional de RosarioFil: Silva, Patricia. Universidad Nacional de Rosario. Facultad de Ciencias Agrarias; Argentin