6,730 research outputs found

    The applications of computers in biological research

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    Research in many fields could not be done without computers. There is often a great deal of technical data, even in the biological fields, that need to be analyzed. These data, unfortunately, previously absorbed much of every researcher's time. Now, due to the steady increase in computer technology, biological researchers are able to make incredible advances in their work without the added worries of tedious and difficult tasks such as the many mathematical calculations involved in today's research and health care

    Qualitative Assessment of the PAX Good Behavior Game Implementation

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    This paper reports on a program evaluation of the PAX Good Behavior Game (GBG), an evidence-based practice intervention designed to create a nurturing environment conducive to learning in elementary schools. To evaluate and improve the PAX Good Behavior Game, a focus group was conducted at the end of the 2016-17 academic year. A total of ten teachers and school administrators from schools who implemented the PAX Good Behavior Game (PAX professionals) participated in a focus group session and provided feedback about the program. Focus group questions assessed four program objectives: (1) environmental change, (2) personal well-being and stress levels, (3) engagement with parents, and (4) networking with other PAX professionals. Results indicated that the PAX GBG decreased problematic classroom behaviors, provided more instructional time for teachers, and generated public interest of the program in the home and community

    Carbon Cycle Science Data and Services at the Goddard Earth Sciences Data Information and Services Center (GES DISC)

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    The Goddard Earth Sciences Data Information and Services Center (GES DISC) archives and distributes a number of observational and model carbon cycle science data sets. We also provide services that facilitate data discovery, intercomparison, and visualization of these heterogeneous datasets for both research and applications users, such as subsetting, format conversion, How-To documentation, and the Help Desk

    CRISPR/Cas9-mediated gene manipulation to create single-amino-acid-substituted and floxed mice with a cloning-free method.

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    Clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) technology is a powerful tool to manipulate the genome with extraordinary simplicity and speed. To generate genetically modified animals, CRISPR/Cas9-mediated genome editing is typically accomplished by microinjection of a mixture of Cas9 DNA/mRNA and single-guide RNA (sgRNA) into zygotes. However, sgRNAs used for this approach require manipulation via molecular cloning as well as in vitro transcription. Beyond these complexities, most mutants obtained with this traditional approach are genetically mosaic, yielding several types of cells with different genetic mutations. Recently, a growing body of studies has utilized commercially available Cas9 protein together with sgRNA and a targeting construct to introduce desired mutations. Here, we report a cloning-free method to target the mouse genome by pronuclear injection of a commercial Cas9 protein:crRNA:tracrRNA:single-strand oligodeoxynucleotide (ssODN) complex into mouse zygotes. As illustration of this method, we report the successful generation of global gene-knockout, single-amino-acid-substituted, as well as floxed mice that can be used for conditional gene-targeting. These models were produced with high efficiency to generate non-mosaic mutant mice with a high germline transmission rate

    Role of metal-dependent regulation of ESX-3 secretion in intracellular survival of Mycobacterium tuberculosis

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    More people die every year from Mycobacterium tuberculosis infection than from infection by any other bacterial pathogen. Type VII secretion systems (T7SS) are used by both environmental and pathogenic mycobacteria to secrete proteins across their complex cell envelope. In the nonpathogen Mycobacterium smegmatis, the ESX-1 T7SS plays a role in conjugation, and the ESX-3 T7SS is involved in metal homeostasis. In M. tuberculosis, these secretion systems have taken on roles in virulence, and they also are targets of the host immune response. ESX-3 secretes a heterodimer composed of EsxG (TB9.8) and EsxH (TB10.4), which impairs phagosome maturation in macrophages and is essential for virulence in mice. Given the importance of EsxG and EsxH during infection, we examined their regulation. With M. tuberculosis, the secretion of EsxG and EsxH was regulated in response to iron and zinc, in accordance with the previously described transcriptional response of the esx-3 locus to these metals. While iron regulated the esx-3 expression in both M. tuberculosis and M. smegmatis, there is a significant difference in the dynamics of this regulation. In M. smegmatis, the esx-3 locus behaved like other iron-regulated genes such as mbtB. In M. tuberculosis, both iron and zinc modestly repressed esx-3 expression. Diminished secretion of EsxG and EsxH in response to these metals altered the interaction of M. tuberculosis with macrophages, leading to impaired intracellular M. tuberculosis survival. Our findings detail the regulatory differences of esx-3 in M. tuberculosis and M. smegmatis and demonstrate the importance of metal-dependent regulation of ESX-3 for virulence in M. tuberculosis
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