The Organisation of Ebola Virus Reveals a Capacity for Extensive, Modular Polyploidy

BACKGROUND: Filoviruses, including Ebola virus, are unusual in being filamentous animal viruses. Structural data on the arrangement, stoichiometry and organisation of the component molecules of filoviruses has until now been lacking, partially due to the need to work under level 4 biological containment. The present study provides unique insights into the structure of this deadly pathogen. METHODOLOGY AND PRINCIPAL FINDINGS: We have investigated the structure of Ebola virus using a combination of cryo-electron microscopy, cryo-electron tomography, sub-tomogram averaging, and single particle image processing. Here we report the three-dimensional structure and architecture of Ebola virus and establish that multiple copies of the RNA genome can be packaged to produce polyploid virus particles, through an extreme degree of length polymorphism. We show that the helical Ebola virus inner nucleocapsid containing RNA and nucleoprotein is stabilized by an outer layer of VP24-VP35 bridges. Elucidation of the structure of the membrane-associated glycoprotein in its native state indicates that the putative receptor-binding site is occluded within the molecule, while a major neutralizing epitope is exposed on its surface proximal to the viral envelope. The matrix protein VP40 forms a regular lattice within the envelope, although its contacts with the nucleocapsid are irregular. CONCLUSIONS: The results of this study demonstrate a modular organization in Ebola virus that accommodates a well-ordered, symmetrical nucleocapsid within a flexible, tubular membrane envelope

MECP2 Isoform-Specific Vectors with Regulated Expression for Rett Syndrome Gene Therapy

BACKGROUND:Rett Syndrome (RTT) is an Autism Spectrum Disorder and the leading cause of mental retardation in females. RTT is caused by mutations in the Methyl CpG-Binding Protein-2 (MECP2) gene and has no treatment. Our objective is to develop viral vectors for MECP2 gene transfer into Neural Stem Cells (NSC) and neurons suitable for gene therapy of Rett Syndrome. METHODOLOGY/PRINCIPAL FINDINGS:We generated self-inactivating (SIN) retroviral vectors with the ubiquitous EF1alpha promoter avoiding known silencer elements to escape stem-cell-specific viral silencing. High efficiency NSC infection resulted in long-term EGFP expression in transduced NSC and after differentiation into neurons. Infection with Myc-tagged MECP2-isoform-specific (E1 and E2) vectors directed MeCP2 to heterochromatin of transduced NSC and neurons. In contrast, vectors with an internal mouse Mecp2 promoter (MeP) directed restricted expression only in neurons and glia and not NSC, recapitulating the endogenous expression pattern required to avoid detrimental consequences of MECP2 ectopic expression. In differentiated NSC from adult heterozygous Mecp2(tm1.1Bird)+/- female mice, 48% of neurons expressed endogenous MeCP2 due to random inactivation of the X-linked Mecp2 gene. Retroviral MECP2 transduction with EF1alpha and MeP vectors rescued expression in 95-100% of neurons resulting in increased dendrite branching function in vitro. Insulated MECP2 isoform-specific lentiviral vectors show long-term expression in NSC and their differentiated neuronal progeny, and directly infect dissociated murine cortical neurons with high efficiency. CONCLUSIONS/SIGNIFICANCE:MeP vectors recapitulate the endogenous expression pattern of MeCP2 in neurons and glia. They have utility to study MeCP2 isoform-specific functions in vitro, and are effective gene therapy vectors for rescuing dendritic maturation of neurons in an ex vivo model of RTT

Investigating the Marketization of the Nonprofit Sector: A Comparative Case Study of Two Nonprofit Organizations

It has been well documented in recent years that nonprofits are becoming increasingly marketized. What is less well understood from this body of research is the variation of marketization in the nonprofit sector and how it is made manifest in nonprofit organizations. This article aims to fill this gap in knowledge by recounting the results of a comparative case study of two nonprofits. We examine their marketization through a multifaceted theoretical framework that allows us to document the marketization comprehensively and to posit some preliminary explanations as to why this variation is occurring. Our findings indicate that one organization is adopting a strong entrepreneurial orientation while the other is integrating its traditional community orientation with more professionalization. The differentiation witnessed in the case organizations suggests that marketization is best understood as a situated process that may in part be explained by varying strategic responses to institutional pressures

Effectiveness of Physical Activity Interventions for Preschoolers: A Meta-Analysis

Purpose: The purpose of the meta-analysis was to examine the effectiveness of physical activity interventions on physical activity participation among preschoolers. A secondary purpose was to investigate the influence of several possible moderator variables (e.g., intervention length, location, leadership, type) on moderate-to-vigorous physical activity (MVPA). Method: Nine databases were systematically searched for physical activity interventions. Studies were included if they contained statistics necessary to compute an effect size (ES), were written in or translated into English, examined physical activity in preschoolers, incorporated a physical activity intervention, and targeted preschool-aged children. Fifteen studies satisfied these criteria. ESs were calculated using a random-effects model. Results: Results indicated that overall, interventions had a small-to-moderate effect on general physical activity (Hedges g = 0.44, p \u3c .05, n = 73 ESs) and a moderate effect on MVPA (Hedges g = 0.51, p \u3c .05, n = 39 ESs). The greatest effects for MVPA were identified for interventions that were less than 4 weeks in duration, were offered in an early-learning environment, were led by teachers, involved outdoor activity, and incorporated unstructured activity. Conclusions: This meta-analysis provides an overview and synthesis of physical activity interventions and highlights effective strategies for future interventions aimed at increasing physical activity levels among preschoolers

Teaching adults to read braille using phonological methods: Single-case studies

Four women with visual impairments were taught 13 braille letters as phonemes and another 13 braille letters as graphemes and then were taught 10 braille words as onset-rime and another 10 braille words as whole words. Phoneme and onset-rime instruction resulted in faster and more accurate performance