FGGY Carbohydrate Kinase Domain Containing is Induced During Skeletal Muscle Atrophy and Modulates Map Kinase and AKT Signaling

Skeletal muscle atrophy can result from a range of physiological conditions, including denervation, immobilization, hindlimb unweighting, and aging. To better characterize the molecular genetic events of atrophy, a microarray was performed using skeletal muscle isolated from mice after 3 and 14 days of denervation and compared to control muscle to identify novel atrophy-induced genes. The microarray revealed that FGGY carbohydrate kinase domain containing (Fggy) is expressed in skeletal muscle and is induced in response to denervation. Bioinformatic analysis of the Fggy gene locus revealed two validated alternative isoforms, that we have termed Fggy-L-552 and Fggy-S-387, which have distinct transcription initiation sites. In order to confirm that Fggy is expressed in muscle, the cDNAs of the two validated alternative variants were cloned from myoblast cells. Interestingly, two novel alternative splice variants, designated Fggy-L-482 and Fggy-S-344, were also cloned, suggesting that at least four Fggy splice variants are expressed in skeletal muscle. Quantitative RT-PCR (RT-qPCR) was performed using RNA isolated from muscle cells and primers designed to distinguish the four alternative Fggy transcripts. The RT-qPCR data reveals that the Fggy-L transcripts are more highly expressed during myoblast differentiation, while the Fggy-S transcripts show relatively stable expression in proliferating myoblasts and differentiated myotubes. Confocal fluorescent microscopy revealed that the Fggy-L variants appear to localize evenly throughout the cytoplasm, while the Fggy-S variants produce a more punctuate localization pattern throughout the cytoplasm of proliferating muscle cells. Finally, ectopic expression of Fggy-L-552 and Fggy-S-387 resulted in inhibition of muscle cell differentiation and attenuation of the MAP kinase and AKT signaling pathways. The characterization of novel genes induced during neurogenic atrophy helps improve our understanding of the molecular and cellular events that lead to muscle atrophy and may eventually lead to new therapeutic targets for the treatment of muscle wasting

Between security, law enforcement and harm reduction: drug policing at commercial music festivals in England

In this thesis, I use an ethnographic methodology to explore the implementation of drug policing at commercial music festivals in England. I argue that festival drug policing is primarily concerned with the anticipation and mitigation of drug-related risk, and festivals adopt an array of security, enforcement and harm reduction approaches under the ‘3: Ps’ (Prevent, Pursue and Protect) in pursuit of this. With an lens on the in-situ decision making of policing, security and management actors on the ground, I illustrate how drug policies are negotiated between agencies, in order to satisfy their sometimes competing risk-perceptions and interests in their pursuit of drug security

Methods for Measuring Frost Tolerance of Conifers: A Systematic Map

Frost tolerance is the ability of plants to withstand freezing temperatures without unrecoverable damage. Measuring frost tolerance involves various steps, each of which will vary depending on the objectives of the study. This systematic map takes an overall view of the literature that uses frost tolerance measuring techniques in gymnosperms, focusing mainly on conifers. Many different techniques have been used for testing, and there has been little change in methodology since 2000. The gold standard remains the field observation study, which, due to its cost, is frequently substituted by other techniques. Closed enclosure freezing tests (all non-field freezing tests) are done using various types of equipment for inducing artificial freezing. An examination of the literature indicates that several factors have to be controlled in order to measure frost tolerance in a manner similar to observation in a field study. Equipment that allows controlling the freezing rate, frost exposure time and thawing rate would obtain results closer to field studies. Other important factors in study design are the number of test temperatures used, the range of temperatures selected and the decrements between the temperatures, which should be selected based on expected frost tolerance of the tissue and species

Evaluation and Optimization of Mass Spectrometric Settings during Data-dependent Acquisition Mode: Focus on LTQ-Orbitrap Mass Analyzers

Mass-spectrometry-based proteomics has evolved as the preferred method for the analysis of complex proteomes. Undoubtedly, recent advances in mass spectrometry instrumentation have greatly enhanced proteomic analysis. A popular instrument platform in proteomics research is the LTQ-Orbitrap mass analyzer. In this tutorial, we discuss the significance of evaluating and optimizing mass spectrometric settings on the LTQ-Orbitrap during CID data-dependent acquisition (DDA) mode to improve protein and peptide identification rates. We focus on those MS and MS/MS parameters that have been systematically examined and evaluated by several researchers and are commonly used during DDA. More specifically, we discuss the effect of mass resolving power, preview mode for FTMS scan, monoisotopic precursor selection, signal threshold for triggering MS/MS events, number of microscans per MS/MS scan, number of MS/MS events, automatic gain control target value (ion population) for MS and MS/MS, maximum ion injection time for MS/MS, rapid and normal scan rate, and prediction of ion injection time. We furthermore present data from the latest generation LTQ-Orbitrap system, the Orbitrap Elite, along with recommended MS and MS/MS parameters. The Orbitrap Elite outperforms the Orbitrap Classic in terms of scan speed, sensitivity, dynamic range, and resolving power and results in higher identification rates. Several of the optimized MS parameters determined on the LTQ-Orbitrap Classic and XL were easily transferable to the Orbitrap Elite, whereas others needed to be reevaluated. Finally, the Q Exactive and HCD are briefly discussed, as well as sample preparation, LC-optimization, and bioinformatics analysis. We hope this tutorial will serve as guidance for researchers new to the field of proteomics and assist in achieving optimal results

Word Retrieval Treatments for Aphasia: Connected Speech Outcomes

We examined changes in connected speech in individuals with aphasia following errorless naming treatment and gestural facilitation of naming administered in a single-participant crossover design. In addition to picture naming, participants completed two connected speech tasks during baseline and after each training phase. Positive training effect sizes in picture naming were associated with increased use of Correct Information Units and substantive nouns in connected speech. Greater use of CIUs and substantive nouns were evident for a questions task over a picture description task. Open-ended questions tended to be more effective than picture description for documenting speech changes associated with treatment

Restorative Practices in Baltimore City Schools: Research Updates and Implementation Guide

Baltimore City Public Schools (City Schools) and other school districts across the United States are implementing restorative practices (RP) to improve school climate by building meaningful relationships in school communities, reframing school discipline, and supporting student safety, well-being, and success. This transformational approach centers student voice and agency, and enhances students’ engagement and participation in their own learning. The Center for Dispute Resolution at the University of Maryland Francis King Carey School of Law and Open Society Institute – Baltimore (OSI) collaborated to create The Restorative Practices in Baltimore City Public Schools: Research Updates and Implementation Guide. The purpose of the report is to: Consolidate existing work describing the use of RP in public schools in Baltimore, across Maryland, and in other parts of the country; Evaluate progress made by Baltimore City in early implementation of its district-wide restorative practices initiative; Help other districts across the country learn from Baltimore’s successes and challenges and provide a blueprint to follow as they implement restorative practices in their own unique contexts

Fggy Carbohydrate Kinase Domain Containing is Induced During Skeletal Muscle Atrophy and Regulates MAP Kinase Signaling

Skeletal muscle atrophy can result from a range of physiological conditions, including denervation. To better characterize the molecular genetic events of atrophy, skeletal muscle was isolated from mice following 3 and 14 days of denervation. The gene expression profile of the denervated muscle tissue was analyzed by microarray and compared to control muscle to identify novel atrophy-induced genes. The microarray revealed that Fggy carbohydrate kinase domain containing (Fggy) is expressed in skeletal muscle and is induced in response to denervation. Bioinformatic analysis of the Fggy gene locus revealed two validated alternative isoforms, which we have termed Fggy-001 and Fggy-003. To confirm that Fggy is expressed in muscle cells, the cDNA of the two validated alternative variants was cloned from mouse myoblast cells. Interestingly, a novel alternative splice variant for each of the validated alternative isoforms was also cloned from mouse muscle cells, suggesting at least four Fggy splice variants to be expressed in skeletal muscle. Ectopic expression of Fggy-001 and Fggy-003 resulted in inhibition of muscle cell differentiation and attenuation of the MAP kinase signaling pathway. Finally, confocal fluorescent microscopy analysis revealed that the Fggy-001 transcripts appear to localize to the cytoplasm, while the Fggy-003 transcripts produce a more punctuate localization pattern throughout the cytoplasm of proliferating muscle cells. The characterization of novel genes that are activated during neurogenic atrophy helps improve our understanding of the molecular and cellular events that lead to muscle atrophy and could eventually lead to new therapeutic targets for the treatment of muscle wasting

Rider Assist Technologies: Popular Types, Motivations for Use, and Information Sources Consulted by Users

The ubiquity of advanced in-vehicle technologies for cars highlights the relative scarcity of comparable offerings for motorcycles despite the fact that many can be adapted for this mode of transportation. Aspects of Intelligent Transport Systems (ITS) such as advanced rider assist technologies may increase the safety of motorcyclists. In this study, we surveyed motorcycle riders about the types of advance technologies their current bikes were equipped with, as well as, the primary motivation for purchasing the technology, and how they learned to use the technologies (if applicable). The most frequently reported technologies owned by riders were handle bar controls, engine brake controls, and cruise control. The primary motivation reported for using the technology was increased safety and improved riding capabilities. The riders were more likely to consult online resources (e.g., YouTube) instead of the manufacturer’s materials when learning about the advanced features of their bikes. While the accessibility of the information makes it easy to reference, the lack of standardization and validity of the online content could contribute to the potential misuse of the technology