MSpace

MSpace is the University of Manitoba’s Institutional Repository. The purpose of MSpace is to acquire, preserve and provide access to the scholarly works of University faculty and students within an open access environment.

 

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Recent Submissions

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Open Access
Optimization of in vitro RNA interference assays to reduce variability in target gene knockdown in the phytopathogen Sclerotinia sclerotiorum
(2025-01-16) Fatema, Reemana; Gerstein, Aleeza (Microbiology); Marcus, Jeffrey (Biological Sciences); Whyard, Steve
Sclerotinia sclerotiorum is a broad host range fungal phytopathogen that is the causal agent of Sclerotinia stem rot in many crop plants, causing devastating yield losses worldwide. Sclerotinia is partially controlled using broad-spectrum chemical fungicides, but new, more species-specific fungicides, such as double-stranded RNAs, are being considered as safer alternatives. This study evaluated the effectiveness of long double-stranded RNAs (dsRNAs) and paperclip RNAs (pcRNAs) to induce gene silencing in two different fungal inoculants of S. sclerotiorum. The three different genes that were targeted in this study, SS1G_01703, SS1G_00005, SS1G_14116, are all involved in different aspects of S. sclerotiorum’s infection process i.e. pathogenicity, encoding 60S ribosomal unit and mycelial growth respectively. Initial dsRNA experiments using small fungal plugs collected from solid media generally resulted in highly variable levels of transcript knockdown, potentially arising from unequal hyphal amounts in treatment tubes. This led to the development of fungal slurries as the inoculum to provide a more uniform distribution of hyphae across all the samples. The slurry method resulted in significant transcript knockdowns using all three long dsRNAs at different doses, although some variability persisted. The variability using the slurry method was linked to potential differences in hyphal developmental stages, age of hyphae and differences in expression levels of each target gene. In contrast, structurally different and shorter in length pcRNAs failed to induce consistent transcript knockdown in both plug and slurry assays, despite earlier reports of their efficacy in reducing lesion sizes on canola leaves. This inefficacy may stem from pcRNAs' reliance on a single siRNA for gene targeting, potentially insufficient to overcome the structural complexity of target mRNAs. These findings highlighted the potential of long dsRNAs for gene knockdown in S. sclerotiorum using optimized slurry-based assays, while suggesting that pcRNAs require further modifications. Future work can explore alternative siRNA designs, increased pcRNA doses, and identifying accessible mRNA regions to improve knockdown efficiency. Additionally, optimizing the methods to assess hyphal developmental stages and minimizing variability during sample preparation are crucial for reproducibility and accuracy in RNAi-based studies.
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Multiplex immunofluorescence studies to characterize early cellular dynamics in laser-ablated non-tumor mouse brain
(2025-02-13) Shanmugam Anandhan, Santhosh; Logue, Susan (Human Anatomy and Cell Science); Kong, Jiming (Human Anatomy and Cell Science); Zeiler, Frederick (Surgery); Klonisch, Thomas; Klonisch, Sabine-Hombach
Introduction: Laser Interstitial Thermal Therapy (LITT) is a minimally invasive, hyperthermia-based laser cytoreductive method used in the treatment of several severe conditions, including inoperable brain tumors, treatment-resistant epilepsy, and radiation necrosis. Laser-induced heat causes damage to the tissue and alters the brain-tissue microenvironment (TME) at the local treatment site and beyond. The regional effects include activation of resident brain cells, release of mediators of inflammation, and immune cell infiltration, which may activate immune responses. Immunomodulatory responses to LITT in the brain microenvironment are largely unknown. One hurdle is the lack of affordable and efficient imaging protocols that enable the simultaneous spatial visualization of multiple immunoreactive cellular targets in the brain. Here, we describe a flexible and affordable multiplex immunofluorescence (mIF) staining method in our established LITT mouse model. In the post-LITT mouse brain, mIF revealed laser ablation-induced complex spatial responses by the resident brain and innate immune cells. Methods: We developed a customizable and affordable mIF detection protocol that uses standard histological and microscopy equipment to assess TME changes on single sections of formalin-fixed paraffin-embedded (FFPE) mouse brain tissues at day 6 post-LITT. This mIF imaging procedure provided a comprehensive spatial analysis of multiple cellular and molecular immunotargets in resident brain cell responses and immune cell profiles in individual tissue sections, as exemplified with a mouse brain microenvironment on day 6 post-LITT. Results: We optimized mIF imaging methodology on LITT-treated mouse brain FFPE serial sections to visualize the spatial distribution of resident brain cell cellular activation states and the emergence and distribution of immune cell populations at the LITT site. Our mIF imaging protocol of mouse brain regions surrounding the LITT site revealed the activation of resident astrocytes and microglial cells and the presence of a diverse innate immune cell population in this region. Conclusion: The combined use of LITT ablation treatment with multiplexed versus regular immunofluorescence offers a significant advancement and aids in our understanding of post-LITT repair and immune responses in the brain. We characterized the thermal ablation-induced modulation of resident brain cells and innate immune cell infiltrations.
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Open Access
Submission to the Standing Committee on Human Resources, Skills and Social Development and the Status of Persons with Disabilities (HUMA)
(2017-10-12) Coalition for Healthy Aging in Manitoba
The Coalition for Healthy Aging in Manitoba (CHAM) would like to congratulate the Standing Committee on Human Resources, Skills and Social Development and the Status of Persons with Disabilities (HUMA) on undertaking the study “Advancing Inclusion and Quality of Life for Canadian Seniors”. The three themes of the report (housing, income security and quality of life), well reflect areas of work that need to be undertaken in Canada and around the world.
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Submission to the Standing Committee on the Status of Women
(2019-03-29) Coalition for Healthy Aging in Manitoba
The World Health Organization’s Global Strategy and Action Plan sets out objectives related to aging that maximize functional ability, and hence, have effects on both intrinsic capacity as well as the external physical and social environments that older adults live in. Thus, there is a strong link with the study that the Status of Women Committee is undertaking. For example, affordable housing and transportation are both key social determinants of health, and are highly linked to quality of life.
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Open Access
2018 Provincial consultations on healthy aging
(2019) Coalition for Healthy Aging in Manitoba
In 2018, the Coalition for Healthy Aging in Manitoba (CHAM) facilitated a series of workshops in various locations in Manitoba. Participants were asked to discuss what Manitoba is doing well and what could be improved with each of the following three strategic areas from the Global Strategy and Action Plan: age-friendly environments, health systems, and long-term care. This report summarizes the comments received across all sites for each of the strategic areas. In general, very similar items were discussed at all locations, even though the communities are quite different in their locations, sizes, demographics, cultural backgrounds, and histories.
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Perspectives on aging
(2016-09-30) Centre on Aging
The University of Manitoba’s Centre on Aging, in collaboration with the Manitoba Association of Senior Centres, hosted a day long symposium in Brandon on September 30, 2016. Following presentations from researchers, participants were invited to take part in an afternoon workshop to discuss the strategic objectives identified in the World Health Organization’s (WHO) Global Strategy and Action Plan: Aligning health systems to the needs of the older populations they now serve; Developing systems for providing long term care; Creating age-friendly environments; Improving measuring, monitoring, and understanding. For each objective discussed, three priority actions were identified by the WHO in relation to meeting each strategic objective, which helped to inform the discussions. A fifth strategic objective, committing to foster healthy ageing in every country, was omitted from discussion since the researcher presentations were focused on this specific objective. Over 50 people participated in the group discussions, where they discussed two of the objectives and identified how well Manitoba was doing in relation to addressing the WHO’s objectives. Participants selected a topic that interested them and each group was assigned a facilitator/note taker.
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WHO Global Strategy & Action Plan on Aging
(2017) Centre on Aging
In February 2017, the Canadian Association on Gerontology and the Centre on Aging at the University of Manitoba applied for a Planning and Dissemination Grant from the Institute of Aging (CIHR). Several partners supported the application. These partners included: Public Health Agency of Canada (PHAC); Canadian Longitudinal Study on Aging (CLSA); AGE-WELL; Canadian Geriatrics Society; Canadian Gerontological Nursing Association; Transportation Option Network for Seniors; Active Aging Canada; Manitoba Association of Senior Centres; Active Living Coalition for Older Adults in Manitoba. In May of 2017, we were informed that we were successful in receiving funding for the Pre-conference Workshop to be held at the Canadian Association on Gerontology Annual Scientific and Educational Meeting. The theme of the Meeting being “Evidence for Action in an Aging World”. Planning for the Workshop included a meeting at the 2017 International Association of Gerontology and Geriatrics World Congress, with several of the partners mentioned above.
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Ageism in Manitoba workshop summary
(2019-01-23) Centre on Aging
As part of the Centre on Aging’s outreach to bring awareness of issues faced by older people in the Province, the Centre hosted a half day workshop on January 23, 2019 that focused on ageism in Manitoba. Over 70 participants took part in this event where ageist stereotypes in society were examined, and participants were asked to reflect on their own internal ageist ideas. A short presentation given by Centre Director Dr. Michelle Porter, outlined the existing definitions in the academic literature and from international organizations (e.g., World Health Organization) on ageism, discussed the effects, and looked at ways to reduce ageism in society. Several group and individual activities required participants to think of and identify examples of ageism; what actions can be taken; and discuss what specific types of strategies or activities/programs could be studied in Manitoba. This document provides a brief synopsis of the individual and group activity reporting.
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Communication Workshops to Re-frame Aging
(2019-11) Centre on Aging
In 2018, the Centre on Aging received funding from the New Horizons for Seniors Program of the Government of Canada for Communication Workshops to Re-frame Aging. The foundational materials that were used to develop the workshop came from the “Gaining Momentum – A Frameworks Communications Toolkit”. The workshops consisted of a presentation about the toolkit, and moved to group activities which helped participants to recognize issues in written materials, and then practice writing short passages incorporating lessons learned from the presentation and resources from the toolkit. All participants received a link to the online toolkit so that they could read through the materials in advance of the workshop, and have the information for future reference.
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Healthy Aging in Manitoba
(2020-01) Centre on Aging
In 2015, the World Health Organization (WHO) launched their first World Report on Ageing and Health (World Report). The World Report “outlines a framework for action to foster Healthy Ageing built around the new concept of functional ability”. In the fall of the same year, the WHO also started consultations on the accompanying draft Global Strategy and Action Plan on Ageing and Health (GSAP). This document, which was ultimately approved by the World Health Assembly in May 2016, focuses on five strategic objectives: commitment to action on Healthy Ageing in every country; developing age-friendly environments; aligning health systems to the needs of older populations; developing sustainable and equitable systems for providing long-term care (home, communities, institutions); and improving measurement, monitoring and research on Healthy Ageing.