Fluorescent Calcium Imaging and Subsequent In Situ Hybridization for Neuronal Precursor Characterization in Xenopus laevis

Spontaneous intracellular calcium activity can be observed in a variety of cell types and is proposed to play critical roles in a variety of physiological processes. In particular, appropriate regulation of calcium activity patterns during embryogenesis is necessary for many aspects of vertebrate neural development, including proper neural tube closure, synaptogenesis, and neurotransmitter phenotype specification. While the observation that calcium activity patterns can differ in both frequency and amplitude suggests a compelling mechanism by which these fluxes might transmit encoded signals to downstream effectors and regulate gene expression, existing population-level approaches have lacked the precision necessary to further explore this possibility. Furthermore, these approaches limit studies of the role of cell-cell interactions by precluding the ability to assay the state of neuronal determination in the absence of cell-cell contact. Therefore, we have established an experimental workflow that pairs time-lapse calcium imaging of dissociated neuronal explants with a fluorescence in situ hybridization assay, allowing the unambiguous correlation of calcium activity pattern with molecular phenotype on a single-cell level. We were successfully able to use this approach to distinguish and characterize specific calcium activity patterns associated with differentiating neural cells and neural progenitor cells, respectively; beyond this, however, the experimental framework described in this article could be readily adapted to investigate correlations between any time-series activity profile and expression of a gene or genes of interest

Movement analysis to indicate discomfort in vehicle seats

Long distance travel is associated with discomfort and fatigue. It is a significant challenge to design a seat that remains comfortable for the occupant over the several hours required for many long-distance journeys. When designing seats, an indication of the perception of comfort/discomfort can be useful either for research and development purposes or potentially for automated systems to take actions that might mitigate discomfort. This paper considered a system that uses measurements of body movement in a seat to provide an objective measure of perceptions of discomfort. The system uses cameras and image processing to recognize when a seat occupant makes a movement in the seat which could be associated with relief of discomfort. The system was validated using a laboratory driving simulator. 10 participants volunteered to complete a study in which they drove for 90 minutes and gave subjective ratings of discomfort every 10 minutes, whilst also being observed using the camera system. It was shown that using a simple algorithm an association could be made between the movements of the driver and subjective ratings of discomfort. However, there remain challenges to improve reliability, optimize movement detection thresholds, and to make it more robust to naturalistic driving scenarios

Characterization of tweety gene (ttyh1-3) expression in Xenopus laevis during embryonic development

The tweety family of genes encodes large-conductance chloride channels and has been implicated in a wide array of cellular processes including cell division, cell adhesion, regulation of calcium activity, and tumorigenesis, particularly in neuronal cells. However, their expression patterns during early development remain largely unknown. Here, we describe the spatial and temporal patterning of ttyh1, ttyh2, and ttyh3 in Xenopus laevis during early embryonic development. Ttyh1 and ttyh3 are initially expressed at the late neurula stage are and primarily localized to the developing nervous system; however ttyh1 and ttyh3 both show transient expression in the somites. By swimming tadpole stages, all three genes are expressed in the brain, spinal cord, eye, and cranial ganglia. While ttyh1 is restricted to proliferative, ventricular zones, ttyh3 is primarily localized to postmitotic regions of the developing nervous system. Ttyh2, however, is strongly expressed in cranial ganglia V, VII, IX and X. The differing temporal and spatial expression patterns of ttyh1, ttyh2, and ttyh3 suggest that they may play distinct roles throughout embryonic development. (C) 2014 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Fluorescent Calcium Imaging and Subsequent In Situ Hybridization for Neuronal Precursor Characterization in Xenopus laevis

Spontaneous intracellular calcium activity can be observed in a variety of cell types and is proposed to play critical roles in a variety of physiological processes. In particular, appropriate regulation of calcium activity patterns during embryogenesis is necessary for many aspects of vertebrate neural development, including proper neural tube closure, synaptogenesis, and neurotransmitter phenotype specification. While the observation that calcium activity patterns can differ in both frequency and amplitude suggests a compelling mechanism by which these fluxes might transmit encoded signals to downstream effectors and regulate gene expression, existing population-level approaches have lacked the precision necessary to further explore this possibility. Furthermore, these approaches limit studies of the role of cell-cell interactions by precluding the ability to assay the state of neuronal determination in the absence of cell-cell contact. Therefore, we have established an experimental workflow that pairs time-lapse calcium imaging of dissociated neuronal explants with a fluorescence in situ hybridization assay, allowing the unambiguous correlation of calcium activity pattern with molecular phenotype on a single-cell level. We were successfully able to use this approach to distinguish and characterize specific calcium activity patterns associated with differentiating neural cells and neural progenitor cells, respectively; beyond this, however, the experimental framework described in this article could be readily adapted to investigate correlations between any time-series activity profile and expression of a gene or genes of interest

Using medical education as a tool to train doctors as social innovators

IntroductionCurrent medical education prepares doctors to diagnose, assess and treat individual patients yet lacks the expectation to be responsible for the care of the wider community. Learning the skills to recognise and redress the social determinants of health are increasingly being recognised as an essential part of medical education.ObjectivesThe goal of this research was (1) to investigate how medical education can be leveraged to reduce health inequalities through the role and practice of doctors and (2) to elucidate how key innovations in medical education are a necessity that can support doctors as ‘change agents.’MethodsTwo international multidisciplinary roundtable focus groups with 23 healthcare leaders from various backgrounds were facilitated. The discussions were audiorecorded, transcribed and then thematically analysed with the qualitative analysis software QDA Miner.ResultsEight themes emerged: (1) Social innovation training in medical education; (2) Linking community working with social innovation; (3) Future curricula development; (4) Settings, context, environment and leaving the classroom; (5) Developing links with third sector organisations and community, including low-income and middle-income countries; (6) Including learners’ perspectives and lived experience; (7) Medical roles are political and need political support and (8) The need to address power imbalances and impact of discrimination.ConclusionsMedical education needs to fundamentally widen its focus from the individual doctor–patient relationship to the doctor–community relationship. Doctors’ training needs to help them become social innovators who can balance interventions with prevention, promote good health on a community and societal scale and tailor their treatments to the individuals’ contexts.</jats:sec

Use of virtual reality in the education of orthopaedic procedures : a randomised control study in early validation of a novel virtual reality simulator

Background Virtual reality (VR) simulation is a potential solution to the barriers surgical trainees are facing. There needs to be validation for its implementation within current training. We aimed to compare VR simulation to traditional methods in acquiring surgical skills for a TFN-ADVANCED™ Proximal Femoral Nailing System (TFNA; DePuy Synthes, Auckland, New Zealand) femoral nailing system. Methods Thirty-one surgical trainees were randomised to two groups: traditional-training group (control group) and a VR-training group (intervention group) for insertion of a short cephalomedullary TFNA nail. Both groups then inserted the same TFNA system into saw-bone femurs. Surveys evaluated validity of the relevant activities, perception of simulation, confidence, stress and anxiety. The primary outcomes were tip-apex distance (TAD) and user anxiety/confidence levels. Secondary outcomes included number of screw- and nail-guidewire insertion attempts, the time taken to complete and user validity of the VR system. Results There was no statistical difference in TAD between the intervention and control groups (9mm vs 15mm, p=0.0734). The only TAD at risk of cut-out was in the control group (25mm). There was no statistical difference in time taken (2547.5ss vs 2395ss, p=0.668), nail guide-wire attempts (two for both groups, p=0.355) and screw guide-wire attempts (one for both groups, p=0.702). The control group versus intervention had higher anxiety levels (50% vs 33%) and had lower confidence (61% vs 84%). Interpretation There was no objective difference in performance on a saw-bone model between groups. However, this VR simulator resulted in more confidence and lower anxiety levels whilst performing a simulated TFNA. Whilst further studies with larger sample sizes and exploration of transfer validity to the operating theatre are required, this study does indicate potential benefits of VR within surgical training