Evaluation of Components of Volunteerism in Master Horseman Program Graduates

Providing volunteers with training opportunities affects the overall success of programs with which they are involved. This article describes a study of 98 Louisiana Master Horseman Program graduates. The study addressed their perceived self-efficacy related to equine technical and management skills, their changes in confidence regarding teaching relevant skills to others, and their willingness to teach those skills to others. Results indicated that the training positively affected graduates\u27 horsemanship skills and their confidence and willingness related to teaching such skills. Findings suggest that the Louisiana Master Horseman Program is useful for creating a highly trained volunteer workforce and is a master volunteer program option that could be replicated successfully in other states

A Naturally Occurring Mutation in ropB Suppresses SpeB Expression and Reduces M1T1 Group A Streptococcal Systemic Virulence

Epidemiological studies of group A streptococcus (GAS) have noted an inverse relationship between SpeB expression and invasive disease. However, the role of SpeB in the course of infection is still unclear. In this study we utilize a SpeB-negative M1T1 clinical isolate, 5628, with a naturally occurring mutation in the gene encoding the regulator RopB, to elucidate the role of RopB and SpeB in systemic virulence. Allelic exchange mutagenesis was used to replace the mutated ropB allele in 5628 with the intact allele from the well characterized isolate 5448. The inverse allelic exchange was also performed to replace the intact ropB in 5448 with the mutated allele from 5628. An intact ropB was found to be essential for SpeB expression. While the ropB mutation was shown to have no effect on hemolysis of RBC's, extracellular DNase activity or survival in the presence of neutrophils, strains with the mutated ropB allele were less virulent in murine systemic models of infection. An isogenic SpeB knockout strain containing an intact RopB showed similarly reduced virulence. Microarray analysis found genes of the SpeB operon to be the primary target of RopB regulation. These data show that an intact RopB and efficient SpeB production are necessary for systemic infection with GAS

Determinants of the voltage dependence of G protein modulation within calcium channel β subunits

CaVβ subunits of voltage-gated calcium channels contain two conserved domains, a src-homology-3 (SH3) domain and a guanylate kinase-like (GK) domain with an intervening HOOK domain. We have shown in a previous study that, although Gβγ-mediated inhibitory modulation of CaV2.2 channels did not require the interaction of a CaVβ subunit with the CaVα1 subunit, when such interaction was prevented by a mutation in the α1 subunit, G protein modulation could not be removed by a large depolarization and showed voltage-independent properties (Leroy et al., J Neurosci 25:6984–6996, 2005). In this study, we have investigated the ability of mutant and truncated CaVβ subunits to support voltage-dependent G protein modulation in order to determine the minimal domain of the CaVβ subunit that is required for this process. We have coexpressed the CaVβ subunit constructs with CaV2.2 and α2δ-2, studied modulation by the activation of the dopamine D2 receptor, and also examined basal tonic modulation. Our main finding is that the CaVβ subunit GK domains, from either β1b or β2, are sufficient to restore voltage dependence to G protein modulation. We also found that the removal of the variable HOOK region from β2a promotes tonic voltage-dependent G protein modulation. We propose that the absence of the HOOK region enhances Gβγ binding affinity, leading to greater tonic modulation by basal levels of Gβγ. This tonic modulation requires the presence of an SH3 domain, as tonic modulation is not supported by any of the CaVβ subunit GK domains alone

Letter processing and font information during reading: beyond distinctiveness, where vision meets design

Letter identification is a critical front end of the reading process. In general, conceptualizations of the identification process have emphasized arbitrary sets of distinctive features. However, a richer view of letter processing incorporates principles from the field of type design, including an emphasis on uniformities across letters within a font. The importance of uniformities is supported by a small body of research indicating that consistency of font increases letter identification efficiency. We review design concepts and the relevant literature, with the goal of stimulating further thinking about letter processing during reading

Use of green fluorescent protein for the analysis of protein-protein and protein-DNA interactions

Restriction modification (RM) systems play a crucial role in preventing the entry of foreign DNA into the bacterial cell. The best studied Type I RM system is EcoKI from Escherichia coli K12. Both bacteriophage and conjugative plasmids have developed a variety of strategies to circumvent the host RM system. One such strategy involves the production of antirestriction proteins that mimic a short segment of DNA and efficiently inhibit the RM system. The main aim of this project was to analyse the interaction of EcoKI and its cognate methylase (MTase) with the T7 antirestriction protein, known as overcome classical restriction (Ocr), and various ArdA antirestriction proteins. Currently, there is a paucity of structural data on the complex formed between the Type I system and the antirestriction proteins. The aim of this work was twofold; (i) compare the interaction of MTase with DNA and Ocr and (ii) quantify the strength of interaction between MTase and various ArdA proteins. The MTase was fused to the Green Fluorescent Protein (GFP) to facilitate determination of the orientation of interaction with DNA and Ocr. Time resolved fluorescence measurements were carried out using the GFP-MTase fusion to determine the fluorescence lifetime and anisotropy decay. These experiments were conducted using a time resolved fluorescence instrument fabricated in-house. The values determined in these experiments were then used to perform fluorescence resonance energy transfer (FRET) measurements with fluorescently labelled DNA or Ocr. These measurements gave information concerning the relative orientation of the MTase with either DNA or Ocr. The GFP-MTase fusion was also used to quantify the strength of interaction with various ArdA proteins. Previous attempts to determine the strength of interaction between MTase and ArdA proteins by employing conventional techniques have been unsuccessful. Therefore, a novel method was developed that exploits the interaction of MTase with a cation exchange medium, which can subsequently be displaced upon binding to ArdA. This method facilitated the determination, for the first time, of a set of binding affinities for the MTase and ArdA interaction

The NCAS mobile dual-polarisation Doppler X-band weather radar (NXPol)

In recent years, dual-polarisation Doppler X-band radars have become a widely used part of the atmospheric scientist's toolkit for examining cloud dynamics and microphysics and making quantitative precipitation estimates. This is especially true for research questions that require mobile radars. Here we describe the National Centre for Atmospheric Science (NCAS) mobile X-band dual-polarisation Doppler weather radar (NXPol) and the infrastructure used to deploy the radar and provide an overview of the technical specifications. It is the first radar of its kind in the UK. The NXPol is a Meteor 50DX manufactured by Selex- Gematronik (Selex ES GmbH), modified to operate with a larger 2.4m diameter antenna that produces a 0.98 halfpower beam width and without a radome. We provide an overview of the technical specifications of the NXPol with emphasis given to the description of the aspects of the infrastructure developed to deploy the radar as an autonomous observing facility in remote locations. To demonstrate the radar's capabilities, we also present examples of its use in three recent field campaigns and its ongoing observations at the NERC Facility for Atmospheric Radio Research (NFARR)

Nucleotide flipping by restriction enzymes analyzed by 2-aminopurine steady-state fluorescence

Many DNA modification and repair enzymes require access to DNA bases and therefore flip nucleotides. Restriction endonucleases (REases) hydrolyze the phosphodiester backbone within or in the vicinity of the target recognition site and do not require base extrusion for the sequence readout and catalysis. Therefore, the observation of extrahelical nucleotides in a co-crystal of REase Ecl18kI with the cognate sequence, CCNGG, was unexpected. It turned out that Ecl18kI reads directly only the CCGG sequence and skips the unspecified N nucleotides, flipping them out from the helix. Sequence and structure conservation predict nucleotide flipping also for the complexes of PspGI and EcoRII with their target DNAs (/CCWGG), but data in solution are limited and indirect. Here, we demonstrate that Ecl18kI, the C-terminal domain of EcoRII (EcoRII-C) and PspGI enhance the fluorescence of 2-aminopurines (2-AP) placed at the centers of their recognition sequences. The fluorescence increase is largest for PspGI, intermediate for EcoRII-C and smallest for Ecl18kI, probably reflecting the differences in the hydrophobicity of the binding pockets within the protein. Omitting divalent metal cations and mutation of the binding pocket tryptophan to alanine strongly increase the 2-AP signal in the Ecl18kI–DNA complex. Together, our data provide the first direct evidence that Ecl18kI, EcoRII-C and PspGI flip nucleotides in solution

Stroop effects from newly learned color words : effects of memory consolidation and episodic context

The Stroop task is an excellent tool to test whether reading a word automatically activates its associated meaning, and it has been widely used in mono- and bilingual contexts. Despite of its ubiquity, the task has not yet been employed to test the automaticity of recently established word-concept links in novel-word-learning studies, under strict experimental control of learning and testing conditions. In three experiments, we thus paired novel words with native language (German) color words via lexical association and subsequently tested these words in a manual version of the Stroop task. Two crucial findings emerged: When novel word Stroop trials appeared intermixed among native-word trials, the novel-word Stroop effect was observed immediately after the learning phase. If no native color words were present in a Stroop block, the novel-word Stroop effect only emerged 24 h later. These results suggest that the automatic availability of a novel word's meaning depends either on supportive context from the learning episode and/or on sufficient time for memory consolidation. We discuss how these results can be reconciled with the complementary learning systems account of word learning

Supply market uncertainty:exploring consequences and responses within sustainability transitions

Often it is commercial, not technological, factors which hinder the adoption of potentially valuable innovations. In energy policy, much attention is given to analysing and incentivising consumer demand for renewable energy, but new technologies may also need new supply markets, to provide products and services to build, operate and maintain the innovative technology. This paper addresses the impact of supply constraints on the long-term viability of sustainability related innovations, using the case of bioenergy from organic waste. Uncertainties in the pricing and availability of feedstock (i.e. waste) may generate market deadlock and deter potential investors. We draw on prior research to conceptualise the problem, and identify what steps might be taken to address it. We propose a research agenda aimed at purchasing and supply scholars and centred on the need to understand better the interplay between market evolution and supply uncertainty and 'market shaping' - how stakeholders can legitimately influence supply market evolution - to support the adoption of sustainability related innovation