Isotope-Edited Variable Temperature Infrared Spectroscopy for Measuring Transition Temperatures of Single A-T Watson-Crick Base Pairs in DNA Duplexes.

Experimental methods to determine transition temperatures for individual base pair melting events in DNA duplexes are lacking despite intense interest in these thermodynamic parameters. Here, we determine the dimensions of the thymine (T) C2═O stretching vibration when it is within the DNA duplex via isotopic substitutions at other atomic positions in the structure. First, we determined that this stretching state was localized enough to specific atoms in the molecule to make submolecular scale measurements of local structure and stability in high molecular weight complexes. Next, we develop a new isotope-edited variable temperature infrared method to measure melting transitions at various locations in a DNA structure. As an initial test of this "sub-molecular scale thermometer", we applied our T13C2 difference infrared signal to measure location-dependent melting temperatures (TmL) in a DNA duplex via variable temperature attenuated total reflectance Fourier transform infrared (VT-ATR-FTIR) spectroscopy. We report that the TmL of a single Watson-Crick A-T base pair near the end of an A-T rich sequence (poly T) is ∼34.9 ± 0.7°C. This is slightly lower than the TmL of a single base pair near the middle position of the poly T sequence (TmL ∼35.6±0.2°C). In addition, we also report that the TmL of a single Watson-Crick A-T base pair near the end of a 50% G-C sequence (12-mer) is ∼52.5 ± 0.3°C, which is slightly lower than the global melting Tm of the 12-mer sequence (TmL ∼54.0±0.9°C). Our results provide direct physical evidence for end fraying in DNA sequences with our novel spectroscopic methods.

A community-driven and evidence-based approach to developing mental wellness strategies in First Nations: a program protocol.

BACKGROUND: Mental health, substance use/addiction and violence (MSV) are important issues affecting the well-being of Indigenous People in Canada. This paper outlines the protocol for a research-to-action program called the Mental Wellness Program (MWP). The MWP aims to increase community capacity, promote relationship-building among communities, and close gaps in services through processes that place value on and supports Indigenous communities' rights to self-determination and control. The MWP involves collecting and using local data to develop and implement community-specific mental wellness strategies in five First Nations in Ontario. METHODS: The MWP has four key phases. Phase 1 (data collection) includes a community-wide survey to understand MSV issues, service needs and community strengths; in-depth interviews with individuals with lived experiences with MSV issues to understand, health system strengths, service gaps and challenges, as well as individual and community resilience factors; and focus groups with service providers to improve understanding of system weaknesses and strengths in addressing MSV. Phase 2 (review and synthesis) involves analysis of results from these local data sources and knowledge-sharing events to identify a priority area for strategic development based on local strengths and need. Phase 3 (participatory action research approach) involves community members, including persons with lived experience, working with the community and local service providers to develop, implement, and evaluate the MWP to address the selected priority area. Phase 4 (share) is focused on developing and implementing effective knowledge-sharing initiatives. Guidelines and models for building the MWP are shared regionally and provincially through forums, webinars, and social media, as well as cross-community mentoring. DISCUSSION: MWP uses local community data to address MSV challenges by building on community supports and resilience factors. Drawing on local data and each community's system of formal and informal supports, the program includes sharing exemplary knowledge-to-action models and wellness strategies developed by and for First Nations people that can be used by other First Nations to identify shared wellness priorities in each community, and determine and execute next steps in addressing areas of main concern.

Research governance in NunatuKavut: engagement, expectations and evolution.

Some of the world's most southern Inuit populations live along central and the southeastern coast of Labrador in the territory of NunatuKavut and are represented by the NunatuKavut Community Council (NCC). Southern Inuit and NCC staff have been actively collaborating with researchers and research ethics boards since 2006 on research ethics and the governance of research in NunatuKavut. As self-determining peoples, Southern Inuit, like many Indigenous communities, are reclaiming control of research through a number of highly effective community consent contracts and ethical review processes and protocols. These community-driven research agreements have both shaped, and been shaped by, academic writings on the issue of collective consent to research. This case report describes the evolution of NCC research governance from 2006 to 2018, emphasising the ethics and engagement that is required to conduct research with Southern Inuit or within the territory of NunatuKavut.

Research with Aboriginal peoples: authentic relationships as a precursor to ethical research.

Recent ethics guidelines and policies are changing the way health research is understood, governed, and practiced among Aboriginal communities in Canada. This provides a unique opportunity to examine the meanings and uses of such guidelines by Aboriginal communities themselves. This qualitative study, conducted in Labrador, Canada, with the Innu, Inuit, and Inuit-Metis, examined how communities and researchers collaborate in a co-learning environment whereby mutual interests and agendas are discussed and enacted throughout the entire research process-a process referred to an authentic research relationship. The purpose of this study was to answer the following questions: (1) Why are authentic research relationships important? (2) What is authenticity in research? (3) How do we achieve authenticity in research with Aboriginal peoples? This shift to more wholistic methodologies can be used in various contexts in Canada and internationally. This is the first study by an Aboriginal person to examine the perspectives of Aboriginal people, in an Aboriginal context, using Aboriginal methodologies.

Children having spinal surgery to correct scoliosis: a qualitative study of parents' experiences.

In order to assist parents to cope with the stress of their child undergoing major spinal surgery, health professionals need an understanding of what the key stressors are. In this study, 13 interviews and questionnaires from parents in the UK and USA were carried out and analysed using Interpretative Phenomenological Analysis (IPA). Five themes emerged: 'Information'; 'Parenting role'; 'Confidence in professionals'; 'Pain'; and 'Effect on life'. Findings suggested that parents need appropriate information and support from health professionals throughout their experience to help minimize uncertainty and distress and that pain management is a major source of stress to parents.