Investigating the Role of PDE4D1/2 in Vascular Smooth Muscle Cell Desensitization

Vascular smooth muscle cell (VSMC) behaviour and phenotypic modulation is critical to vessel repair following damage, and the progression of various cardiovascular diseases. The second messenger cyclic adenosine monophosphosphate (cAMP) plays a key role in VSMC function under the synthetic/activated phenotype, which is typically associated with unhealthy cell behaviour. Consequently, cAMP signaling is often targeted in attempts to impact several pathological diseases, including atherosclerosis, restenosis, and pulmonary arterial hypertension (PAH). The cyclic nucleotide phosphodiesterases (PDEs) catalyze hydrolysis of cAMP to an inactive form, and therefore directly regulate cAMP signaling. The PDE4D family dominates in synthetic VSMCs, and there is considerable interest in determining how distinct PDE4D isoforms affect cell function. Specifically, we are interested in the potential link between short isoforms of PDE4D and VSMC desensitization to pharmacological agents that impact cardiovascular disease via cAMP signaling. This study extends on previous work that assessed the expression of PDE4D splice variants in rat aortic VSMCs following prolonged challenge with cAMP-elevating agents. It was determined that PDE4D1 and PDE4D2 were uniquely expressed in synthetic VSMCs incubated with these agents, and that this upregulation impacted PDE activity and cAMP accumulation in these cells. Here, we report that PDE4D1 and PDE4D2 are markedly upregulated in synthetic human aortic smooth muscle cells (HASMCs) following prolonged challenge with cAMP-elevating agents. Using a combination of RNAi-based and pharmacological approaches, we establish that this upregulation is reflected in levels of cAMP PDE activity, and restricted to the cytosolic sub-cellular compartment. Our results suggest a role for localized PDE4D1 and PDE4D2 activity in regulating cAMP-mediated desensitization in HASMCs, and highlight their therapeutic potential in treating various cardiovascular diseases.

Queen's University Journal (1910-1911)

The Journal has been Queen's main student newspaper since it was founded in 1873. It appears twice a week on campus with a mix of news, sports, and entertainment stories, editorials, letters to the editor, and photographs. The paper is students' most important source of news and general information and has been a training ground for scores of Canadian journalists.

If People Were Kaleidoscopes: Sharpening the Resolution on Patient Engagement in Safety

Healthcare is unacceptably error prone. The question remains why, with 20 years of evidence, is error and harm reduction not being effective? While precise numbers may be debated, all stakeholders recognize the frequency of healthcare errors is unacceptable, and greater efforts to ensure safety must occur. In recent years, one of these strategies has been the inclusion of the patient and their family as partners in safety, and has been a required organizational practice of Accreditation Canada in many of their standard sets. Existing patient advisories created to encourage engagement, have typically not included patient perspectives in their development or been comprehensively evaluated. There are no existing tools to determine if and how a patient wants to be involved in safety engagement. As such, a multi-phased study was undertaken to advance our knowledge about the client’s and family’s role in promoting safety. Phase 1 of the study was a scoping review to methodically review the existing literature about patients’ and families’ attitudes, beliefs and behaviours about their involvement in healthcare safety. Phase 2 was designed to inductively explore how a group of patients in an Ontario, Canada, community hospital, describe healthcare safety and see their role in preventing error. The study findings, which include the narratives of 30 patients and 4 family members, indicate that although there are shared themes that influence a patient’s engagement in patient safety, every individual has unique, changing beliefs, experiences and reasons for involvement. Five conceptual themes emerged from their narratives: Personal Capacity, Experiential Knowledge, Personal Character, Relationships, and Meaning of Safety. These study results will be used to develop and test a pragmatic, accessible tool to enable providers a way to collaborate with patients for determining their personal level and type of safety involvement. The most ethical and responsible approach to healthcare safety is to consider every facet and potential way for improvement. This exploratory study provides fundamental insights into the complexity of patient engagement in safety, and evidence for future steps.

A Restorative Theory of Criminal Justice

In this project, I defend a restorative theory of criminal justice. I argue that the response to criminal wrongdoing in a just society should take the form of an attempt to heal the damage done to the community resulting from crime. I argue that the moral responsibilities of wrongdoers as wrongdoers ought to provide the framework for how a just society should respond to crime. Following the work of R.A. Duff, I argue that wrongdoers incur second-order duties of moral recognition. Wrongdoers owe it to others to recognize their wrongdoing for what it is, i.e. wrongdoing, and to shoulder certain burdens in order to express their repentant recognition to others via a meaningful apology. In short, wrongdoers owe it to their victims and others in the community to make amends. What I will deny, however, is the now familiar claim in the restorative justice literature that restoring the normative relationships in the community damaged by criminal forms of wrongdoing requires retributive punishment. In my view, how we choose to express the judgement that wrongdoers are blameworthy should flow from an all things considered judgment that is neither reducible to the judgement that the wrongdoer is culpably responsible for wronging others, nor the judgement that the wrongdoer in some basic sense “deserves to suffer” (or “deserves punishment,” etc.).

Paternalism and Competent Patient Choice in the Standards of Care for the Health of Transsexual, Transgender, and Gender-Nonconforming People

In 2016 physicians in Ontario will be granted the authority to refer patients with gender dysphoria for sex reassignment surgery. In order to be granted this authority physicians must be trained in the World Professional Association for Transgender Health’s Standards of Care, which outlines healthcare procedures for the treatment of gender dysphoria and provides background information concerning transgender health. The Standards of Care require that patients undergo a process of 12 months of continuous living in a gender role that is congruent with their gender identity prior to being given access to sex reassignment surgery. While this requirement can sometimes be helpful it can also cause more harm than benefit. This paper argues that the requirement is strongly paternalistic in its current form and should no longer be mandatory in most cases.

Communication-efficient decentralized sequential detection

The problem of decentralized sequential detection is studied in this thesis, where local sensors are memoryless, receive independent observations, and no feedback from the fusion center. In addition to traditional criteria of detection delay and error probability, we introduce a new constraint: the number of communications between local sensors and the fusion center. This metric is able to reflect both the cost of establishing communication links as well as overall energy consumption over time. A new formulation for communication-efficient decentralized sequential detection is proposed where the overall detection delay is minimized with constraints on both error probabilities and the communication cost. Two types of problems are investigated based on the communication-efficient formulation: decentralized hypothesis testing and decentralized change detection. In the former case, an asymptotically person-by-person optimum detection framework is developed, where the fusion center performs a sequential probability ratio test based on dependent observations. The proposed algorithm utilizes not only reported statistics from local sensors, but also the reporting times. The asymptotically relative efficiency of proposed algorithm with respect to the centralized strategy is expressed in closed form. When the probabilities of false alarm and missed detection are close to one another, a reduced-complexity algorithm is proposed based on a Poisson arrival approximation. In addition, decentralized change detection with a communication cost constraint is also investigated. A person-by-person optimum change detection algorithm is proposed, where transmissions of sensing reports are modeled as a Poisson process. The optimum threshold value is obtained through dynamic programming. An alternative method with a simpler fusion rule is also proposed, where the threshold values in the algorithm are determined by a combination of sequential detection analysis and constrained optimization. In both decentralized hypothesis testing and change detection problems, tradeoffs in parameter choices are investigated through Monte Carlo simulations.

The effects of the non-native invertebrate predator Bythotrephes longimanus and declining aqueous calcium on crustacean zooplankton communities in Canadian Shield lakes

Over the last several decades, human activities have resulted in environmental changes that have increased the number of stressors that can act on a single environment. In Canadian Shield lakes, two recent stressors, the invasion of Bythotrephes longimanus and calcium decline, have been documented. Widespread acidification of hundreds of North American lakes has resulted in the precipitous decline of lake water calcium concentration. Crustacean zooplankton with high calcium demands are likely to be vulnerable to calcium decline, especially <1.5 mg Ca/L, where survival and reproduction rates are reduced. These taxa are also vulnerable to predation by Bythotrephes that has been implicated in the loss of pelagic biodiversity in soft water lakes. Despite laboratory and field studies aimed at understanding the independent impact of these stressors, it is unclear how their co-occurrence will influence community response. Using a combination of data from a large regional lake survey and field experiments, I examined the individual and joint effects of Bythotrephes and calcium decline on native zooplankton community structure. Results demonstrated that much is known about Bythotrephes and our findings of reduced total zooplankton and species richness, due to the loss of Cladocera, are consistent with field surveys and other experimental studies. While we did not detect strong evidence for an effect of calcium on zooplankton using the lowest calcium concentration among invaded lakes (1.2 mg Ca/L), there is evidence that, as lake water calcium concentrations fall <1 mg Ca/L, per capita growth rates of a broad variety of taxa are expected to decline. At the regional scale, negative effects of Bythotrephes and calcium on abundances of small cladocerans and Daphnia pulicaria, respectively, were in agreement with my experimental observations. We also observed significant interactions between Bythotrephes and calcium for a broad variety of taxa. As Bythotrephes continues to spread and invade lakes that are also declining in aqueous calcium, both stressors are likely to amplify negative effects on Cladocera that appear the most vulnerable. Loss of these important zooplankton in response to both Bythotrephes and calcium decline, is likely to lower zooplankton productivity, with potential effects on phytoplankton and higher trophic levels.

Polymer Light-Emitting Electrochemical Cells with Embedded Bipolar Electrodes: Visualizing Bipolar Electrochemistry in Solid State

The work presented in this thesis examines the properties of BPEs of various configurations and under different operating conditions in a large planar LEC system. Detailed analysis of time-lapsed fluorescence images allows us to calculate the doping propagation speed from the BPEs. By introducing a linear array of BPEs or dispersed ITO particles, multiple light-emitting junctions or a bulk homojunction have been demonstrated. In conclusion, it has been observed that both applied bias voltages and sizes of BPEs affected the electrochemical doping from the BPE. If the applied bias voltage was initially not sufficiently high enough, a delay in appearance of doping from the BPE would take place. Experiments of parallel BPEs with different sizes (large, medium, small) demonstrate that the potential difference across the BPEs has played a vital role in doping initiation. Also, the p-doping propagation distance from medium-sized BPE has displayed an exponential growth over the time-span of 70 seconds. Experiments with a linear array of BPEs with the same size demonstrate that the doping propagation speed of each floating BPE was the same regardless of its position between the driving electrodes. Probing experiments under high driving voltages further demonstrated the potential of having a much more efficient light emission from an LEC with multiple BPEs.

Alteration mineralogy and pathfinder element inventory of the McArthur River unconformity-related uranium deposit, Canada

The chemical compositions, modal mineralogy, and textural variability of interstitial minerals in sandstones of the Athabasca Group strata in the vicinity of the McArthur River unconformity-related uranium deposit were characterized using a combination of short wave infrared spectroscopy (SWIR), lithogeochemistry, scanning electron microscopy (SEM), electron probe microanalysis (EPMA) and laser ablation mass spectrometry (LA-ICP-MS) to determine the residence sites of pathfinder trace elements. The importance of integrating in-situ mineral chemistry with whole-rock analyses resides in the possibility to establish the mineralogical and paragenetic context of geochemical signatures in defining the footprint of the deposit. Located in the Athabasca Basin, Saskatchewan, Canada, the deposit is situated below ~550 m of quartz arenitic sandstones that are strongly silicified between depths of approximately 200-400 m. The silicified layer exhibits significant control on the distribution of alteration minerals, and appears to have restricted both the primary and secondary dispersion of pathfinder trace elements, which include U, radiogenic Pb isotopes, V, Ni, Co, Cu, Mo, As, Zn, and REEs. Diagenetic background sandstones contain assemblages of illite, dickite, aluminum-phosphate-sulfate (APS) minerals, apatite, and Fe-Ti oxide minerals. Altered sandstones contain assemblages of Al-Mg chlorite (sudoite), alkali-deficient dravite, APS minerals, kaolinite, illite, and oxide minerals. Throughout the sandstones, APS minerals account for the majority of the Sr and LREE concentrations, whereas late pre-ore chlorite, containing up to 0.1 wt.% Ni, accounts for the majority of Ni concentrations. Cobalt, Cu, Mo, and Zn occur predominantly in cryptic sub-micron sulfide and sulfarsenide inclusions in clay mineral aggregates and in association with paragenetically-late Fe-Ti oxides. Uranium occurs predominantly in cryptic micro-inclusions associated with pyrite in late-stage quartz overgrowths, and with paragenetically late Fe-Ti oxide micro-inclusions in kaolinite. Additionally, up to 0.2 wt.% U is cryptically distributed in post-ore Fe-oxide veins. Early diagenetic apatite, monazite and apatite inclusions in detrital quartz, and detrital zircon also contribute significant U and HREE to samples analyzed with an aggressive leach such as Aqua Regia. Detailed LA-ICP-MS chemical mapping of interstitial assemblages, detrital grains, and cements provides new insights into the distribution and inventory of pathfinder elements in the footprint of the McArthur River uranium deposit.

Performance and Assessment of Rehabilitated Steel Culverts

This thesis explores the effects of rehabilitation on the structural performance of corrugated steel culverts. A full-scale laboratory experiment investigated the effects of grouted slip-liners on the performance of two buried circular corrugated steel culverts. One culvert was slip-lined and grouted using low strength grout, while the other was slip-lined and grouted using high strength grout. The performances of the culverts were measured before and after rehabilitation under service loads using single wheel pair loading at 0.45m of cover. Then, the rehabilitated culverts were loaded to their ultimate limit states. Results showed that the low and high strength grouted slip-liners provided strength well beyond requirements, with the low strength specimen failing at a load 2.4 times the fully factored service load, while the high strength specimen did not reach an ultimate limit state before bearing failure of the soil stopped testing. Results also showed that the low strength specimen behaved rigidly under service loads and flexibly under higher loads, while the high strength specimen behaved rigidly under all loads. A second full-scale experiment investigated the effect of a paved invert rehabilitation procedure on the performance of a deteriorated horizontal ellipse culvert. The performance of the culvert before and after rehabilitation was examined under service loads using tandem axle loading at 0.45m of cover. The rehabilitated culvert was then loaded up to its ultimate limit state. The culvert failed due to the formation of a plastic hinge at the West shoulder, while the paved invert cracked at the invert. Results showed that the rehabilitation increased the structural performance of the culvert, increasing the system stiffness and reducing average strains and local bending at critical locations in the culvert under service loads. A sustainability rating tool specifically for the evaluation of deteriorated culvert replacement or rehabilitation projects was also developed. A module for an existing tool, called GoldSET, was created and tested using two case studies, each comparing the replacement of a culvert using a traditional open-cut method with two trenchless rehabilitation techniques. In each case, the analyses showed that the trenchless techniques were the better alternatives in terms of sustainability.

Experience, Education and Subjectivity: A Comparison of John Dewey's and Maurice Merleau-Ponty's Conceptions of Experience and Their Implications for Education

This thesis compares John Dewey’s philosophy of experience and Maurice Merleau-Ponty’s phenomenology, and illustrates how Merleau-Ponty’s phenomenology can strengthen and further Dewey’s philosophy of education. I begin by drawing the connection between Dewey’s philosophy of experience and his philosophy of education, and illustrate how Dewey’s understanding of growth, and thinking in education, is rooted in and informed by his detailed philosophy of experience. From there, I give an interpretation of Merleau-Ponty’s phenomenology with a focus on his descriptions of subjectivity that he presents in the Phenomenology of Perception. Following this, I outline some of the implications Merleau-Ponty’s phenomenology has on our understanding of rationality, expression and existence. In the final chapter, I make the comparison between Dewey’s philosophy of experience and Merleau-Ponty’s phenomenology. After demonstrating how these two philosophies are not only similar but also complementary, I then look to Merleau-Ponty’s phenomenology to provide insight into and to advance Dewey’s philosophy of education. I will illustrate how Merleau-Ponty’s understanding of subjectivity helps to support, and reinforce the rationale behind Dewey’s inquiry-based approach to education. Furthermore, I will show how Merleau-Ponty’s phenomenology and its implications for rationality, expression and existence support Dewey’s democratic ideal and add a hermeneutical element to Dewey’s philosophy of education.

Heavy Minerals in Soils from the Athabasca Basin and the Implications for Exploration Geochemistry of Uranium Deposits at Depth

The Centennial deposit is a high grade (~8% U3O8), deeply buried (~950m), unconformity-related U deposit located in the south-central region of the Athabasca Basin in northern Saskatchewan, Canada. The mineral chemistry of fine fractions (<63 μm) of soils from grids above the Centennial deposit were examined to understand possible controls on the geochemistry and radiogenic 207Pb/206Pb ratios measured in the clay-size (<2 μm) fractions used for exploration. Soil samples distal and proximal to the deposit projection to the surface and geophysically defined structures were selected. Mineral abundances were determined using the scanning electron microscope and Mineral Liberation Analysis. Zircon was the only U-rich mineral identified with modal abundances >0.02% by weight. Monazite, which can be U-rich, was identified, but not in significant abundances. The source of the zircon and other heavy minerals is interpreted to be from sub-cropping sources that are >100 km up-ice from Centennial. Trace element analysis using laser ablation inductively coupled plasma mass spectrometry of hydroseparated zircon grains indicate that zircon abundances and zircon Pb concentrations in surficial samples have minimal effect on the radiogenic 207Pb/206Pb ratios in the clay-fraction of the samples, with the dominant source of radiogenic Pb being clay mineral surfaces that trapped Pb during secondary dispersion from the Centennial uranium deposit through faults and fractures to the surface. The REE patterns indicate HREE enrichment in the clay-fractions of samples that have higher abundances of zircon in the <20 μm fraction. Immobile elements such as HREE that are concentrated in zircon can be used as indicators of radiogenic Pb being sourced from minerals at the surface rather than being sourced from secondary dispersion from deeply buried U deposits.

An Examination of Current Approaches to Integrative Indigenous and Western Knowledge System Implementation in Water Research and Management: A Case Study Encompassing the Colonized Geographies of Canada, Australia, New Zealand, and the United States

The commodification of natural resources and the pursuit of continuous growth has resulted in environmental degradation, depletion, and disparity in access to these life-sustaining resources, including water. Utility-based objectification and exploitation of water in some societies has brought us to the brink of crisis through an apathetic disregard for present and future generations. The ongoing depletion and degradation of the world’s water sources, coupled with a reliance on Western knowledge and the continued omission of Indigenous knowledge to manage our relationship with water has unduly burdened many, but particularly so for Indigenous communities. The goal of my thesis research is to call attention to and advance the value and validity of using both Indigenous and Western knowledge systems (also known as Two-Eyed Seeing) in water research and management to better care for water. To achieve this goal, I used a combined systematic and realist review method to identify and synthesize the peer-reviewed, integrative water literature, followed by semi-structured interviews with first authors of the exemplars from the included literature to identify the challenges and insights that researchers have experienced in conducting integrative water research. Findings suggest that these authors recognize that many previous attempts to integrate Indigenous knowledges have been tokenistic rather than meaningful, and that new methods for knowledge implementation are needed. Community-based participatory research methods, and the associated tenets of balancing power, fostering trust, and community ownership over the research process, emerged as a pathway towards the meaningful implementation of Indigenous and Western knowledge systems. Data also indicate that engagement and collaborative governance structures developed from a position of mutual respect are integral to the realization of a given project. The recommendations generated from these findings offer support for future Indigenous-led research and partnerships through the identification and examination of approaches that facilitate the meaningful implementation of Indigenous and Western knowledge systems in water research and management. Asking Western science questions and seeking Indigenous science solutions does not appear to be working; instead, the co-design of research projects and asking questions directed at the problem rather than the solution better lends itself to the strengths of Indigenous science.

Structural and Functional Characterization of the Human Tribbles Homologue 2 Pseudokinase

The Tribbles Homologues are a family of three eukaryotic pseudokinases (Trb1, Trb2, Trb3) that act as allosteric inhibitors and regulatory scaffold sites in pathways governing adipogenesis, cell proliferation and insulin signaling. The Tribbles Homologues have the same overall tertiary structure of the eukaryotic protein kinase domain, but lack multiple residues necessary to catalysis in the nucleotide-binding P-loop and the Mg2+-coordinating DFG motif. Trb1 has been shown conclusively to be incapable of binding ATP, whereas a recent study presents evidence that Trb2 autophosphorylates independently of Mg2+ in vitro. This finding is surprising given the high degree of sequence similarity between the two proteins (71%), and suggests unique nucleotide binding and phosphotransfer mechanisms. The goal of this project was to investigate whether Trb2 possesses kinase activity or not and determine its structural basis. A method for the high-yield recombinant expression and purification of stable Trb2 was developed. Trb2 nucleotide binding and autophosphorylation could not be detected across multiple experimental approaches, including thermal shift assays, MANT-ATP fluorescence, radiolabeled phosphate incorporation, and nonspecific ATPase activity assays. Further characterization also revealed that Trb2 forms homomultimers with possible functional consequences, and extensive crystallization screening has yielded multiple promising conditions that could produce diffraction-quality crystals with further optimization. This project explores the difficulties in functionally characterizing putatively active pseudokinases, and proposes a structural basis for the conserved pseudokinase features of the Tribbles homologues.

La voz del otro: rompiendo con la tradición del museo

Existe una actitud predominante en el mundo de los museos según la cual las diversas culturas se presentan y se respetan de manera adecuada, dándose un diálogo efectivo entre los museos y las comunidades culturales a las que sirven. Sin embargo, el presente trabajo disiente de dicha creencia, expone sus motivos, y sugiere una trayectoria por la que los museos pueden acercarse con éxito a tales objetivos. Lo que está en juego aquí es el ethos contemporáneo e histórico de los museos y su concepto de la identidad propia, lo que interfiere con su capacidad para incluir a “los otros”, excepto en sus propios términos. El propósito del museo en este sentido es mantener su voz preeminente y su prerrogativa intelectual en sus comunicaciones con el público al que sirve. La idea de que hay múltiples voces que necesitan ser escuchadas no es un concepto fácil de contemplar para los museos, y un paso positivo en aquella dirección representaría un cambio importante con consecuencias de largo alcance.