76 research outputs found
Смешанное обучение английскому языку как форма самостоятельной работы студентов в техническом вузе
Настоящее исследование рассматривает вопросы организации самостоятельной работы студентов по развитию иноязычных монологических умений в условиях смешанного обучения в техническом вузе. В работе проведен анализ трудов зарубежных и отечественных исследователей, занимающихся проблемами смешанного обучения по иностранному языку, что позволило уточнить определение "смешанного обучения", а так же было сформулировано понятие "самостоятельной работы в условиях смешанного обучения".
Для развития умений иноязычной монологической речи студентов технического вуза определено содержание обучения, подходы и принципы, разработан комплекс упражнений, размещенный в электронной среде, обеспечивающие реализацию процесса обучения в условиях смешанного обучения.This research deals with the organization of independent work of students, developing their foreign monologue skills in terms of blended learning in a technical university.
The analysis of the works of foreign and Russian researchers on blended learning in a foreign language, made it possible to clarify the definition of "blended learning", as well as the formulation of the concept - "independent work in a blended learning".
The work describes a pilot training, which proves the efficiency of the developed learning technique of improving students’ foreign monologue skills in a technical university in terms of blended learning
Extracellular Stimuli Specifically Regulate Localized Levels of Individual Neuronal mRNAs
Subcellular regulation of protein synthesis requires the correct localization of messenger RNAs (mRNAs) within the cell. In this study, we investigate whether the axonal localization of neuronal mRNAs is regulated by extracellular stimuli. By profiling axonal levels of 50 mRNAs detected in regenerating adult sensory axons, we show that neurotrophins can increase and decrease levels of axonal mRNAs. Neurotrophins (nerve growth factor, brainderived neurotrophic factor, and neurotrophin-3) regulate axonal mRNA levels and use distinct downstream signals to localize individual mRNAs. However, myelin-associated glycoprotein and semaphorin 3A regulate axonal levels of different mRNAs and elicit the opposite effect on axonal mRNA levels from those observed with neurotrophins. The axonal mRNAs accumulate at or are depleted from points of ligand stimulation along the axons. The translation product of a chimeric green fluorescent protein–β-actin mRNA showed similar accumulation or depletion adjacent to stimuli that increase or decrease axonal levels of endogenous β-actin mRNA. Thus, extracellular ligands can regulate protein generation within subcellular regions by specifically altering the localized levels of particular mRNAs
Localized IRES-Dependent Translation of ER Chaperone Protein mRNA in Sensory Axons
<div><p>Transport of neuronal mRNAs into distal nerve terminals and growth cones allows axonal processes to generate proteins autonomous from the cell body. While the mechanisms for targeting mRNAs for transport into axons has received much attention, how specificity is provided to the localized translational apparatus remains largely unknown. In other cellular systems, protein synthesis can be regulated by both cap-dependent and cap-independent mechanisms. The possibility that these mechanisms are used by axons has not been tested. Here, we have used expression constructs encoding axonally targeted bicistronic reporter mRNAs to determine if sensory axons can translate mRNAs through cap-independent mechanisms. Our data show that the well-defined IRES element of encephalomyocarditis virus (EMCV) can drive internal translational initiation of a bicistronic reporter mRNA in distal DRG axons. To test the potential for cap-independent translation of cellular mRNAs, we asked if calreticulin or grp78/BiP mRNA 5′UTRs might have IRES activity in axons. Only grp78/BiP mRNA 5′UTR showed clear IRES activity in axons when placed between the open reading frames of diffusion limited fluorescent reporters. Indeed, calreticulin’s 5′UTR provided an excellent control for potential read through by ribosomes, since there was no evidence of internal initiation when this UTR was placed between reporter ORFs in a bicistronic mRNA. This study shows that axons have the capacity to translate through internal ribosome entry sites, but a simple binary choice between cap-dependent and cap-independent translation cannot explain the specificity for translation of individual mRNAs in distal axons.</p> </div
EMCV IRES supports cap-independent translation in axons. A-B
<p>, Representative time-lapse sequences from FRAP analyses of DRG neurons transfected with pmCherry<sup>myr</sup>EMCV-eGFP<sup>myr</sup>3′Cal as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040788#pone-0040788-g003" target="_blank">Fig. 3</a> are shown. mCherry and eGFP signals are displayed as a spectrum with white being the brightest signal as indicated. The white boxed regions represent the regions subjected to photobleaching and the arrows indicate the regions of the terminal axon where recovery was quantified. Sequences for cap-dependent translation of mCherry are shown in <b>A</b> and for IRES-dependent translation of eGFP are shown in <b>B</b>. The upper rows for A and B show cultures standard medium and lower rows show cultures pretreated with 150 µM anisomycin [scale bar = 50 µm]. <b>C-D</b>, Quantifications of axonal mCherry and eGFP signals from multiple FRAP sequences from DRG neurons transfected with mCherry<sup>myr</sup>EMCV-eGFP<sup>myr</sup>-3′Cal are shown. Signals in each individual series are normalized to pre-bleach levels and are expressed as average percent prebleach signals ± SEM (n <i>≥</i>8 over at least 4 independent transfections; *p≤0.05, **p<i>≤</i>0.01, and ***p<i>≤</i>0.001 for control vs. anisomycin time points and †p≤0.05, ††p<i>≤</i>0.01, and †††p≤0.001 for control vs. cycloheximide time points by two-way ANOVA compared to t = 0 min post-bleach). Both mCherry and eGFP fluorescence in distal axons shows recovery after photobleaching that is attenuated by protein synthesis inhibitors. These data indicate that the EMCV IRES can drive internal translational initiation in axons.</p
Bicistronic reporters support IRES-dependent translation in sensory neurons.
<p>Representative static images of DRG neurons transfected with bicistronic pmCherry<sup>myr</sup>5′Cal-eGFP<sup>myr</sup>3′Cal (<b>A</b>), pmCherry<sup>myr</sup>EMCV-eGFP<sup>myr</sup>3′Cal (<b>B</b>), and pmCherry<sup>myr</sup>5′BiP-eGFP<sup>myr</sup>3′Cal (<b>C</b>) reporters are shown at 48 h post-transfection. Both mCherry (red, left panel) and eGFP (green, right panel) fluorescence is seen in the cell bodies and axons of mCherry<sup>myr</sup>EMCV-eGFP<sup>myr</sup>3′Cal and mCherry<sup>myr</sup>5′BiP-eGFP<sup>myr</sup>3′Cal expressing neurons (<b>B,C</b>), only the mCherry signals are seen for the mCherry<sup>myr</sup>5′Cal-eGFP<sup>myr</sup>-3′Cal expressing neurons (<b>A</b>). These data suggest that the 5′UTR of grp78/BiP mRNA but not calreticulin’s 5′UTR can function as an IRES in sensory neurons [scale bars = 50 µm].</p
RNA exodus to Israel: RNA controlling function in the far reaches of the neuron
Workshop on RNA Control of Neuronal Functio
A Need for Consistency in Behavioral Phenotyping for ASD: Analysis of the Valproic Acid Model
Autism spectrum disorder (ASD) is a highly prevalent and impairing neurodevelopmental disorder that affects 1 : 54 persons. Over the last several decades, the reported incidence of ASD in the US has increased potentially due to increased awareness and improved diagnostic measurement. Although ASD prevalence is increasing, the etiology of ASD remains relatively unknown. To better understand the neurological basis of ASD, rodent models of ASD have been developed for research. Currently, there is not a standardized set of behavioral tests to quantify ASD-like behavior in rodents. The goal of this review is to present an overview of the methodologies used to analyze ASD-like behaviors in rodents, focusing on the valproic acid (VPA) model, and illustrate inconsistencies between different approaches. Despite that the in utero VPA rodent model for ASD is widely used and extensively characterized, behaviors vary substantially between different researchers. Moving forward, consistency in behavioral method analytics would benefit progress in evaluating interventions for all models of ASD and help to uncover unique qualities underlying mechanisms causing ASD signs and symptoms
5′UTR of calreticulin mRNA does not show IRES activity in HEK-293 cells. A
<p>, Schematic of monocistronic and bicistronic constructs used for mRNA expression in HEK cells and DRG neurons is shown. 5′CAL and 3′CAL correspond to 5′ and 3′UTRs of rat calreticulin mRNA. 5′BiP corresponds to the 5′UTR of grp78/BiP mRNA. EMCV corresponds to the 5′ leader sequence of the encephalomyocarditis virus RNA. The constructs used in HEK cells contained standard mCherry and eGFP, while those for neuronal transfections contained diffusion limited mCherry<sup>myr</sup> and eGFP<sup>myr</sup> reporters. The neuronal constructs also included an axonal targeting 3′UTR (3′Cal). <b>B-D</b>, Representative exposure matched fluorescent images are shown for HEK-293 cells, 48 h post-transfection, for bicistronic pmCherry-5′CAL-eGFP (<b>B</b>), monocistronic p5′CAL-eGFP (<b>C</b>) and bicistronic pmCherry-EMCV-eGFP (<b>D</b>) [scale bars = 200 µm].</p
5′UTR of calreticulin does not support cap-independent translation in sensory neurons. A
<p>, Representative time-lapse sequences from FRAP analyses of DRG neurons transfected with pmCherry<sup>myr</sup>5′Cal-eGFP<sup>myr</sup>3′Cal are shown. mCherry fluorescence is displayed as a spectrum with white being the brightest signal as indicated. The white boxed regions represent the regions subjected to photobleaching and the arrows indicate the regions of the terminal axon where recovery was quantified. The upper row shows cultures standard medium and lower row shows cultures pretreated with 150 µM anisomycin [scale bar = 50 µm]. <b>B</b>, Quantification of normalized mCherry fluorescence intensity in ROI as indicated from multiple FRAP sequences are shown. Signals are shown as the average percent of pre-bleach levels ± SEM (n ≥12 over at least 4 independent transfections; *** = p≤0.001 by two-way ANOVA comparing conditions to t = 0 min for indicated time points).</p
5′UTR of grp78/BiP mRNA can function as an IRES in axons. A-B
<p>, Representative time-lapse sequences from FRAP analyses of DRG neurons transfected with pmCherry<sup>myr</sup>5′BiP-eGFP<sup>myr</sup>3′Cal as in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040788#pone-0040788-g004" target="_blank">Fig. 4</a> are shown. Sequences for cap-dependent translation of mCherry are shown in <b>A</b> and for IRES-dependent translation of eGFP are shown in <b>B</b>. The upper rows for A and B show cultures standard medium and lower rows show cultures pretreated with 150 µM anisomycin [scale bar = 50 µm]. <b>C-D</b>, Quantifications of axonal mCherry and eGFP signals from multiple FRAP sequences from DRG neurons transfected with mCherry<sup>myr</sup>5′BiP-eGFP<sup>myr</sup>-3′Cal are shown. Signals in each individual series are normalized to pre-bleach levels and are expressed as average percent prebleach signals ± SEM (n <i>≥</i>6 over at least 4 independent transfections; *p≤0.05, **p<i>≤</i>0.01, and ***p<i>≤</i>0.001 for control vs. anisomycin time points and †p≤0.05, ††p<i>≤</i>0.01, and †††p≤0.001 for control vs. cycloheximide time points by two-way ANOVA compared to t = 0 min post-bleach). Both mCherry and eGFP fluorescence in distal axons shows recovery after photobleaching that is attenuated by protein synthesis inhibitors. These data indicate that the 5′UTR of rat grp78/BiP mRNA can drive IRES-dependent translation in axons.</p
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