Illustration of Skeletal Muscle Calsequestrin Complex Formation by Cell Overlay and Two-Dimensional Blot Overlay Methodology

Based on the recent finding that a peroxidase-conjugated calsequestrin probe retains its biological binding properties, we have used it in two-dimensional blot and cell overlay procedures. Labeling of nitrocellulose replicas of electrophoretically separated microsomal proteins from predominantly fast versus slow fibres revealed self-aggregation of fast calsequestrin with molecular species of differing isolectric points. Incubation of transverse cryosections showed restricted calsequestrin interactions with elements in the fibre interior, mostly representing calsequestrin itself. This confirms that fast calsequestrin exists in large aggregates under native conditions and demonstrates the usefulness of blot and cell overlay techniques in identifying and locating supramolecular protein assemblies

A qualitative exploration of individual and couple’s experiences of movement disorders

The thesis titled ‘A qualitative exploration of individual and couple’s experiences of movement disorders’ explores experiences relevant to two common movement disorders; Parkinson's and cervical dystonia (CD). Section one presents a meta-ethnography, synthesising 19 qualitative studies relevant to the couple’s experience of Parkinson’s. The analysis of which produced three themes, which were: 1) Disruption of the couple’s roles and responsibilities and attempting to maintain the relationship; 2) Challenges to communication and closeness which can lead to increased resilience; and 3) Increased cognitive and behavioural symptoms leading to grief, burden and isolation. The findings highlight the challenges that couples experience and the individual and relational resources that support them to cope. The relevant strengths, weaknesses, and implications for clinical practice and future research are discussed. Section two presents a qualitative interpretative phenomenological analysis (IPA) study exploring the experience of diagnosis in CD. Six participants were interviewed, and their data analysed. The analysis produced three themes: 1) Losing control over one’s body and attempts to regain control: ‘You don't know what's going on and your whole life is kinda falling apart’; 2)Feeling powerless in response to the health system: ‘I had this change and I couldn't do anything about it, and nobody seemed to have any kind of solution’; 3) Mixed feelings upon diagnosis: ‘It's kinda like a relief, but scary at the same time’. The findings provide further insight and interpretation of the experience of diagnosis and highlight how this can be a significant and distressing experience. Clinical implications and suggestions for further study are discussed. Section three presents a critical appraisal where the author has discussed broader research issues and reflections on the reports contained in sections one and two. Considered, are the implications for the findings presented, their strengths, limitations, and suggestions for future research

Increased sensitivity of the ryanodine receptor to halothane-induced oligomerization in malignant hyperthermia-susceptible human skeletal muscle

Mutations in the skeletal muscle RyR1 isoform of the ryanodine receptor (RyR) Ca2+-release channel confer susceptibility to malignant hyperthermia, which may be triggered by inhalational anesthetics such as halothane. Using immunoblotting, we show here that the ryanodine receptor, calmodulin, junctin, calsequestrin, sarcalumenin, calreticulin, annexin-VI, sarco(endo)plasmic reticulum Ca2+-ATPase, and the dihydropyridine receptor exhibit no major changes in their expression level between normal human skeletal muscle and biopsies from individuals susceptible to malignant hyperthermia. In contrast, protein gel-shift studies with halothane-treated sarcoplasmic reticulum vesicles from normal and susceptible specimens showed a clear difference. Although the alpha2-dihydropyridine receptor and calsequestrin were not affected, clustering of the Ca2+-ATPase was induced at comparable halothane concentrations. In the concentration range of 0.014-0.35 mM halothane, anesthetic-induced oligomerization of the RyR1 complex was observed at a lower threshold concentration in the sarcoplasmic reticulum from patients with malignant hyperthermia. Thus the previously described decreased Ca2+-loading ability of the sarcoplasmic reticulum from susceptible muscle fibers is probably not due to a modified expression of Ca2+-handling elements, but more likely a feature of altered quaternary receptor structure or modified functional dynamics within the Ca2+-regulatory apparatus. Possibly increased RyR1 complex formation, in conjunction with decreased Ca2+ uptake, is of central importance to the development of a metabolic crisis in malignant hyperthermia

Brain dystrophin-glycoprotein complex: Persistent expression of beta-dystroglycan, impaired oligomerization of Dp71 and up-regulation of utrophins in animal models of muscular dystrophy

BACKGROUND: Aside from muscle, brain is also a major expression site for dystrophin, the protein whose abnormal expression is responsible for Duchenne muscular dystrophy. Cognitive impairments are frequently associated with this genetic disease, we therefore studied the fate of brain and skeletal muscle dystrophins and dystroglycans in dystrophic animal models. RESULTS: All dystrophin-associated glycoproteins investigated were reduced in dystrophic muscle fibres. In Dp427-deficient mdx brain and Dp71-deficient mdx-3cv brain, the expression of α-dystroglycan and laminin was reduced, utrophin isoforms were up-regulated and β-dystroglycan was not affected. Immunofluorescence localization of β-dystroglycan in comparison with glial, endothelial and neuronal cell markers revealed co-localization of von Willebrand factor with β-dystroglycan. Its expression at the endothelial-glial interface was preserved in dystrophin isoform-deficient brain from mdx and mdx-3cv mice. In addition, chemical crosslinking revealed that the Dp71 isoform exists in mdx brain predominantly as a monomer. CONCLUSIONS: This suggests an association of β-dystroglycan with membranes at the vascular-glial interface in the forebrain. In contrast to dystrophic skeletal muscle fibres, dystrophin deficiency does not trigger a reduction of all dystroglycans in the brain, and utrophins may partially compensate for the lack of brain dystrophins. Abnormal oligomerization of the dystrophin isoform Dp71 might be involved in the pathophysiological mechanisms underlying abnormal brain functions

Exploring the experience of residents during the first six months of family medicine residency training

Background: The shift from undergraduate to postgraduate education signals a new phase in a doctor’s training. This study explored the resident’s perspective of how the transition from undergraduate to postgraduate (PGME) training is experienced in a Family Medicine program as they first meet the reality of feeling and having the responsibility as a doctor.Methods: Qualitative methods explored resident experiences using interpretative inquiry through monthly, individual in-depth interviews with five incoming residents during the first six months of training.  Focus groups were also held with residents at various stages of training to gather their reflection about their experience of the first six months. Residents were asked to describe their initial concerns, changes that occurred and the influences they attributed to those changes.Results: Residents do not begin a Family Medicine PGME program knowing what it means to be a Family Physician, but learn what it means to fulfill this role. This process involves adjusting to significant shifts in responsibility in the areas of Knowledge, Practice Management, and Relationships as they become more responsible for care outcomes.Conclusion: This study illuminated the resident perspective of how the transition is experienced. This will assist medical educators to better understand the early training experiences of residents, how these experiences contribute to consolidating their new professional identity, and how to better align teaching strategies with resident learning needs

Genetic dissection of MHC-associated susceptibility to Lepeophtheirus salmonis in Atlantic salmon

Background: Genetic variation has been shown to play a significant role in determining susceptibility to the salmon louse, Lepeophtheirus salmonis. However, the mechanisms involved in differential response to infection remain poorly understood. Recent findings in Atlantic salmon (Salmo salar) have provided evidence for a potential link between marker variation at the major histocompatibility complex (MHC) and differences in lice abundance among infected siblings, suggesting that MHC genes can modulate susceptibility to the parasite. In this study, we used quantitative trait locus (QTL) analysis to test the effect of genomic regions linked to MHC class I and II on linkage groups (LG) 15 and 6, respectively. Results: Significant QTL effects were detected on both LG 6 and LG 15 in sire-based analysis but the QTL regions remained unresolved due to a lack of recombination between markers. In dam-based analysis, a significant QTL was identified on LG 6, which accounted for 12.9% of within-family variance in lice abundance. However, the QTL was located at the opposite end of DAA, with no significant overlap with the MHC class II region. Interestingly, QTL modelling also revealed evidence of sex-linked differences in lice abundance, indicating that males and females may have different susceptibility to infection. Conclusion: Overall, QTL analysis provided relatively weak support for a proximal effect of classical MHC regions on lice abundance, which can partly be explained by linkage to other genes controlling susceptibility to L. salmonis on the same chromosom

An experimental assessment of detection dog ability to locate great crested newts ( Triturus cristatus ) at distance and through soil

Detection dogs are increasingly used to locate cryptic wildlife species, but their use for amphibians is still rather underexplored. In the present paper we focus on the great crested newt (Triturus cristatus), a European species which is experiencing high conservation concerns across its range, and assess the ability of a trained detection dog to locate individuals during their terrestrial phase. More specifically, we used a series of experiments to document whether a range of distances between target newts and the detection dog (odour channelled through pipes 68 mm in diameter) affects the localisation, and to assess the ability and efficiency of target newt detection in simulated subterranean refugia through 200 mm of two common soil types (clay and sandy soil, both with and without air vents to mimic mammal burrows, a common refuge used by T. cristatus). The detection dog accurately located all individual T. cristatus across the entire range of tested distances (0.25 m– 2.0 m). The substrate trials revealed that the detection dog could locate individuals also through soil. Contrary to existing studies with detection dogs in human forensic contexts, however, detection was generally slower for T. cristatus under sandy soil compared to clay soil, particularly when a vent was absent. Our study provides a general baseline for the use of detection dogs in locating T. cristatus and similar amphibian species during their terrestrial phase

The complex multiscale structure in simulated and observed emission maps of the proto-cluster cloud G0.253+0.016 (\u27the Brick\u27)

The Central Molecular Zone (the central ∼500 pc of the Milky Way) hosts molecular clouds in an extreme environment of strong shear, high gas pressure and density, and complex chemistry. G0.253+0.016, also known as \u27the Brick\u27, is the densest, most compact, and quiescent of these clouds. High-resolution observations with the Atacama Large Millimetre/submillimetre Array (ALMA) have revealed its complex, hierarchical structure. In this paper we compare the properties of recent hydrodynamical simulations of the Brick to those of the ALMA observations. To facilitate the comparison, we post-process the simulations and create synthetic ALMA maps of molecular line emission from eight molecules. We correlate the line emission maps to each other and to the mass column density and find that HNCO is the best mass tracer of the eight emission lines within the simulations. Additionally, we characterize the spatial structure of the observed and simulated cloud using the density probability distribution function (PDF), spatial power spectrum, fractal dimension, and moments of inertia. While we find good agreement between the observed and simulated data in terms of power spectra and fractal dimensions, there are key differences in the density PDFs and moments of inertia, which we attribute to the omission of magnetic fields in the simulations. This demonstrates that the presence of the Galactic potential can reproduce many cloud properties, but additional physical processes are needed to fully explain the gas structure

An experimental assessment of detection dog ability to locate great crested newts (Triturus cristatus) at distance and through soil

Detection dogs are increasingly used to locate cryptic wildlife species, but their use for amphibians is still rather underexplored. In the present paper we focus on the great crested newt (Triturus cristatus), a European species which is experiencing high conservation concerns across its range, and assess the ability of a trained detection dog to locate individuals during their terrestrial phase. More specifically, we used a series of experiments to document whether a range of distances between target newts and the detection dog (odour channelled through pipes 68 mm in diameter) affects the localisation, and to assess the ability and efficiency of target newt detection in simulated subterranean refugia through 200 mm of two common soil types (clay and sandy soil, both with and without air vents to mimic mammal burrows, a common refuge used by T. cristatus). The detection dog accurately located all individual T. cristatus across the entire range of tested distances (0.25 m-2.0 m). The substrate trials revealed that the detection dog could locate individuals also through soil. Contrary to existing studies with detection dogs in human forensic contexts, however, detection was generally slower for T. cristatus under sandy soil compared to clay soil, particularly when a vent was absent. Our study provides a general baseline for the use of detection dogs in locating T. cristatus and similar amphibian species during their terrestrial phase