The Bovine Coronavirus 2\u27-\u3cem\u3eO\u3c/em\u3e-Methyltransferase Binds \u3cem\u3eCis\u3c/em\u3e-Acting Stem-Loop IV In the 5-Prime Untranslated Region Of The Viral Genome

The positive-stranded coronavirus genome, at 32 kilobases in length, is the largest known viral RNA genome, and internal cis-signaling elements directing its replication have been described only within the last ten years. The bovine coronavirus genome encodes 26 proteins in the region between the 5’-terminal 210-nt untranslated region and the 3’-terminal 298-nt untranslated region. Here, genes for 5 of the 26 proteins were cloned into bacterial expression plasmids for the long-term goals of characterizing enzymatic and RNA binding properties. These genes encode enzymes postulated to interact directly with the cis-acting RNA elements and carry out RNA synthesis, namely, the RNA-dependent RNA polymerase, the helicase, the exonuclease, the endonuclease, and the 2’-O-methyltransferase. For a detailed analysis, bacterially-expressed BCoV 2’-O-Methyltransferase was purified and (i) tested for enzymatic activity, which is presumably a 2’-O-methylation of 5’-terminal cap structures, and (ii) tested for its binding to terminal genomic regions known to contain cis-acting replication elements. Methyltransferase activity was not found, suggesting the proper conditions were not met or the proper template was not used, or perhaps, as with many viral enzymes made from a polyprotein precursor, it does not function as a unit-length molecule. Using the electrophoretic mobility shift assay, the 2’-O-Methyltransferase was found to bind cis-acting stem-loop IV in the 5’ untranslated region, but does not bind other cis-acting elements, including the region in gene 1 containing stem-loops V and VI or the 3’-proximal cis-acting bulged stem-loop and pseudoknot. The results of this study suggest that the putative bovine coronavirus 2’-O-Methyltransferase uses stem-loop IV as a binding site to carry out methyltransferase function(s) yet to be discovered

How practice size impacts on the implementation and dissemination of post occupancy evaluation

There has been much research on the subject of environmentally sustainable design (ESD), with emerging techniques and technologies improving rapidly and informing sustainability higher education teaching to architects and prospective architects. By examining the success of sustainable designs using post occupancy evaluations, architectural practices might also increase their knowledge of sustainable building practice. Post occupancy evaluations could be useful for improving the designs of future buildings and the design processes that generated them. This paper aims to evaluate these claims by asking: &quot;Do sustainable design practices use the feedback gained from post occupancy evaluations?,&quot; &quot;How does the feedback refine the design process?,&quot; &quot;How is the information gained in these evaluations absorbed within the firm\u27s design practices?,&quot; and, &quot;Does the size of a practice impact on its implementation anddissemination of POE?&quot; This paper investigates the questions posed above through the questioning of architectural practices that have gained a reputation for environmentally sustainable design by having a strong sustainable design philosophy and/or by being recognised for this by winning a sustainability design award. The interviewed practices will have provided some form of post occupancy evaluation as a service or employed them to add to their own knowledge.<br /

Need Fulfillment in Intercollegiate Student-Athletes’ Dual Roles: A Mixed-Methods, Person-Oriented Investigation

Individuals competing in varsity intercollegiate sport are tasked with managing the dual roles of athletes and students. The purpose of current study was to simultaneously explore student-athletes’ perceived satisfaction of autonomy, competence and relatedness in sport and academics. A mixed-methods, person-oriented design was utilized. Quantitative data was collected with a sample of N = 238 student-athletes from various varsity intercollegiate sports. For sport, four clusters were revealed: ‘Low Need Satisfaction,’ ‘Moderately Low Need Satisfaction,’ ‘Moderate Relatedness,’ and ‘High Need Satisfaction.’ For academics, four clusters were revealed: ‘Low Need Satisfaction,’ ‘Moderate Relatedness,’ ‘Moderate Autonomy and Competence,’ and ‘High Need Satisfaction.’ Reflexive thematic analysis of semi-structured interviews with a sub-sample of n = 12 student-athletes representing all clusters revealed four themes: (a) global factors sensitized student-athletes to the experience of basic psychological needs, (b) contextual factors determined fluctuations in student-athletes’ need fulfillment, (c) student-athletes perceived interaction effects in the satisfaction of the three basic psychological needs within the same domain, and (d) student-athletes’ sport participation had a cross-contextual influence on their need fulfillment in academics. Findings provide an understanding of student-athletes’ perceived basic psychological needs across the achievement domains of academics and sport

Defective myogenesis in the absence of the muscle-specific lysine methyltransferase SMYD1

AbstractThe SMYD (SET and MYND domain) family of lysine methyltransferases harbor a unique structure in which the methyltransferase (SET) domain is intervened by a zinc finger protein–protein interaction MYND domain. SMYD proteins methylate both histone and non-histone substrates and participate in diverse biological processes including transcriptional regulation, DNA repair, proliferation and apoptosis. Smyd1 is unique among the five family members in that it is specifically expressed in striated muscles. Smyd1 is critical for development of the right ventricle in mice. In zebrafish, Smyd1 is necessary for sarcomerogenesis in fast-twitch muscles. Smyd1 is expressed in the skeletal muscle lineage throughout myogenesis and in mature myofibers, shuttling from nucleus to cytosol during myoblast differentiation. Because of this expression pattern, we hypothesized that Smyd1 plays multiple roles at different stages of myogenesis. To determine the role of Smyd1 in mammalian myogenesis, we conditionally eliminated Smyd1 from the skeletal muscle lineage at the myoblast stage using Myf5cre. Deletion of Smyd1 impaired myoblast differentiation, resulted in fewer myofibers and decreased expression of muscle-specific genes. Muscular defects were temporally restricted to the second wave of myogenesis. Thus, in addition to the previously described functions for Smyd1 in heart development and skeletal muscle sarcomerogenesis, these results point to a novel role for Smyd1 in myoblast differentiation

Coaching and/or education intervention for parents with overweight/obesity and their children: Study protocol of a single-centre randomized controlled trial

Background: In Canada, a majority of children and adults are insufficiently active for health gains, and about one in seven children and over 20% of adults are overweight or obese. Overweight and obesity are risk factors for many chronic diseases in both adults and children and can result in lower quality and quantity of life. Children whose parents are overweight or obese are more likely to become overweight themselves. Thus, parent/child interventions are important for reducing obesity and promoting long-term healthy weights among members of the family unit. Programs using Co-Active coaching have resulted in positive behaviour changes among adults with overweight/obesity; however, little research has explored the effects of Co-Active coaching on parents, and the consequent impact on the family unit (i.e. all parents and children in the same household). This protocol paper provides a detailed methodological account of a coaching-based program targeting parent and child dyads, in hopes of enhancing health behaviours within the family unit. Methods: Using a randomized controlled trial design, the researchers aim to identify the impact of coaching plus education (intervention) compared to education only (control) on parents with overweight/obesity and their children (ages 2.5-10, of any weight). A total of 50 dyads are being recruited and randomly assigned using a 1:1 ratio into the control or intervention group. The control group receive 6 webinar-based education sessions focused on physical activity and nutrition. The intervention group receive the same education sessions and nine, 20-min telephone-based sessions with a certified coach. Coaching and health education sessions are conducted with the parent/guardian of the dyad. This paper provides a detailed methodological account of this program. Discussion: The expected findings from this research will advance coaching literature, research, and practice on this topic by determining whether coaching and education are more effective than education alone at producing behaviour changes among a family unit. If proven effective, this approach may be applied more broadly through public health interventionists to parent and child populations in hopes of affecting change with both individuals and their families. Trial registration: ISRCTN ISRCTN69091372. Retrospectively registered 24 September 2018

Perspectives and impact of a parent-child intervention on dietary intake and physical activity behaviours, parental motivation, and parental body composition: A randomized controlled trial

Adults and children in Canada are not meeting physical activity guidelines nor consuming sufficient nutrient-rich foods. High engagement in these unhealthy behaviours can lead to obesity and its associated diseases. Parent-child interventions aimed at obesity prevention/treatment have assisted families with making positive changes to their nutrition and physical activity behaviours. Given that the home environment shapes early health behaviours, it is important to target both parents and children when addressing diet and physical activity. One method that has been shown to improve health outcomes is co-active coaching. The current study explored the impact of a threemonth co-active coaching and/or health education intervention on the dietary intake and physical activity behaviours of parents with overweight/obesity and their children (ages 2.5–10; of any weight). Body composition (i.e., body mass index [BMI] and waist circumference), changes in parental motivation with respect to physical activity and dietary behaviours, and parental perceptions of program improvements were collected. A concurrent mixed methods study comprised of a randomized controlled trial and a descriptive qualitative design was utilized. Fifty parent-child dyads were recruited and randomly assigned to the control (n = 25) or intervention (n = 25) group. Assessments were completed at baseline, mid-intervention (six weeks), postintervention (three months), and six-month follow-up. A linear mixed effects model was utilized for quantitative analysis. Inductive content analysis was used to extract themes from parent interviews. No significant results were observed over time for the dependent measures. Parents in both control and intervention groups reported varied program experiences, including developing changes in perspective, increased awareness of habits, and heightened accountability for making positive changes in themselves, and consequently, their families. Parents also shared barriers they faced when implementing changes (e.g., time, weather, stress). Qualitatively, both groups reported benefitting from this program, with the intervention group describing salient benefits from engaging in coaching. This research expands on the utility of coaching as a method for behaviour change, when compared to education only, in parents with overweight/obesity and their children

Novel PTH gene mutations causing isolated hypoparathyroidism

Context: PTH gene mutations represent a rare cause of familial isolated hypoparathyroidism (FIH). These defects can cause hypoparathyroidism with increased or decreased serum levels of PTH through 1) impaired PTH synthesis; 2) induction of parathyroid cell apoptosis; or 3) secretion of bioinactive PTH molecules. Eight pathogenic mutations of this gene have been described previously. Objective: Through describing two novel mutations of the PTH gene, we aim to extend the molecular basis for FIH and further refine the proposed mechanisms by which PTH mutations cause hypoparathyroidism. Design: Proband case reports with extended family analysis. Patients and Other Participants: The probands in both kindreds presented before 10-days-of-age with hypocalcemia and elevated phosphate levels. Proband A had low PTH levels, while these levels were elevated in Proband B. Proband B was initially diagnosed with pseudohypoparathyroidism. Interventions: Methylation analysis of CpG dinucleotides within three GNAS differentially methylated regions; Whole genome sequencing; and PTH infusion with analysis of nephrogenous cyclic AMP. Results: Proband A had a novel heterozygous sequence change in exon 2 of the PTH gene, c.46_47delinsAA (p.Ala16Lys) and Proband B had a novel homozygous nucleotide transition in PTH exon 3 (c.128G>A; p.G43E) that led to replacement of glycine by glutamic acid at position 12 of PTH 1-84. PTH 1-34 infusion demonstrated that renal responsiveness to PTH was intact and not antagonized by circulating bioinactive PTH. Conclusions: PTH gene mutations are uncommon causes of hypoparathyroidism, but can be misdiagnosed as disorders of gland development or receptor function if PTH levels are decreased or elevated, respectively. Genetic testing should be considered early in the diagnostic approach to these presentations