VictorJuiz/thesis huggingface dataset:
Stubborn Subject: The Persistence of Self in Dramatic Acting

Dramatic acting, as a theatrical event that mysteriously fuses actor with character, is also a process that is deeply invested in the idea of Self. From Diderot’s early paradox through Stanislavski’s highly influential psychotechnique and into the present day, notions of who actors are – and how they precisely feel – when portraying characters have remained at the centre of Euroamerican discussions about dramatic performance. Conversely, for most of the twentieth and into the twenty-first century, the concept of the Self as an organic, reflexive, self-directed and differentiated human being in the world has been not only “deconstructed” within every dominant critical and artistic discourse but has also – until recently – been pretty much left for dead. Yet within the realm of acting in the traditional theatre’s models, the tenacity of the Self’s hold as the centrepiece of both pedagogy and methodology appears to be the hallmark of a concept that, if somewhat bruised and battered, is still very much alive. Is there something inherent – or at least necessary – within the study and practice of dramatic acting that perpetually reconstitutes the stubborn idea of an autonomous and cohesive subjectivity within the actor? Through a freewheeling, and sometimes unorthodox, methodology of interrogation, this dissertation asserts that, for the modern dramatic actor, a demonstrable sense of self is an inescapable vocational reality. This position is argumentatively framed through a valorization of the actor’s phenomenological and psychological experience as fundamental to assessing how the Actor and the Self creatively collide. Though drawing from a broad range of interdisciplinary discourses, the author’s subjective and self-historicizing experience as someone who has worked as a professional actor in the Canadian theatre and other media since childhood, for over forty years, is everywhere implicit in this analysis.

Keywords: acting
character
drama
mirror
self
subjectivity
Advisor: Johnson, Stephen
Investigating the Role of microRNAs in Cell Fate Determination

miRNAs are key regulators of a myriad of cellular processes, including proliferation, differentiation, and signaling. These developmentally regulated and abundantly expressed RNA species elicit their effects by controlling the expression of multiple genes in a spatial and temporal manner. microRNAs are differentially expressed in the extra-embryonic trophoblast and embryonic inner cell mass of the preimplantation embryo and their stem cell models, but while they are required for trophoblast development, it is unknown whether they have the capacity to specify or determine trophoblast fate in the preimplantation embryo. Knockout phenotypes of microRNA processor genes have not identified a role for microRNAs in this early cell fate choice, and expression of microRNAs in embryonic stem cells (ESCs) has not yet induced a trophoblast cell state. In this thesis I have identified miRNAs with trophoblast function and interrogated their role in cell fate specification. First, I employed stem cell models of the early embryo and confirm that overexpression of these miRNAs in ESCs induces a trophoblast phenotype, resulting in the derivation of a novel mural trophectoderm-like cell (mTELC) type. Second, by ectopically expressing these miRNAs in the preimplantation embryo ex vivo, I provide evidence that miRNAs may be involved in trophoblast fate specification by modulating components of the Hippo signaling pathway. Finally, I elaborate on the role of these miRNAs in trophoblast development and reveal that they could be driving a state of dormancy in this lineage by impinging on mTOR signaling. Together, these studied reveal a role for miRNAs in modulating embryo developmental transitions and specification of cell lineages.

Keywords: Embryo
Embryonic Stem Cells
Genetics
microRNA
Trophoblast
Advisor: Cox, Brian J
School Travel for Families Living with Childhood Disability

The rise of automobility during the past century has contributed to children becoming more sedentary and isolated from public life and, correspondingly, exhibiting reduced independent mobility and a troubling range of chronic health problems. Concern over these issues has led to an increase in attention to school transportation among planning scholars and practitioners. School transportation has come to be regarded as an opportunity for creating change since converting sedentary trips to and from school into trips involving active modes could help to mitigate the children’s health issues. Unfortunately, the increased attention to school transportation has come to pass with little regard for childhood disability. This dissertation engages school transportation’s childhood disability gap. Using a critical ableist studies perspective, attention is given to questioning and unsettling the ableist normative ordering of childhood disability as excludable within the planning and design of school transportation and school sites. A systematic review of disability’s treatment in the active school travel and children’s independent mobility literatures demonstrates and offers insight into the childhood disability gap. Following this is a presentation of an ethnographic research design that used photovoice, institutional ethnographic conceptual tools, and numerous inclusive measures to study how families living with childhood disability experience everyday school travel across Ontario, Canada’s Greater Toronto and Hamilton Area. Pilot study findings are then presented and discussed to demonstrate the value of piloting qualitative research designs involving disabled children. Lastly, findings about how families of disabled children experience parking at school are considered. The findings show how the families are expected and relied on to take on various types of inequitable work (e.g., work that is social, physical, and temporal in nature) to overcome normalized, ableist school site designs.

Keywords: childhood disability
critical ableist studies
inclusive design
photovoice
pilot study
school transport
Advisor: Buliung, Ronald
Inpatient Rehabilitation for Patients with Traumatic Brain Injury: Process and Content

The goals of this thesis were to a) compare the content and process of inpatient rehabilitation (IR) between patients with Traumatic Brain Injury (TBI) who were treated in one Canadian and nine US facilities, b) investigate the association of therapeutic factors with short-term functional outcomes, and c) describe the functional recovery from admission to nine months post-discharge at 4 time points as well as explore the predictive value of therapeutic and post-discharge determinants to explain the variation in long-term outcomes in patients with TBI who were admitted to a Canadian facility. This thesis was based on three studies. The results of the first study showed that there are significant differences between components of IR for patients with TBI who were treated in Canadian versus US facilities. These findings provide informative data for practitioners and researchers about variations in service provision for patients with similar profiles. The findings of the second study revealed that level of effort was associated with higher discharge cognitive scores. Additionally, more intensity of time spent in complex occupational and physical therapy activities were significant predictors of better discharge motor scores. These findings provide evidence to consider patient level of effort and intensity of specific activities in the process of goal setting. The results of the third study demonstrated that after three months post-discharge, patients did not show significant variations in functional outcomes. Also, the number of post-discharge health issues was negatively associated with both cognitive and motor scores at nine months post-discharge, and a greater intensity of time spent in complex occupational therapy activities was a predictor of better motor scores at nine months follow-up. These findings inform health care providers about the temporal influence of IR and the necessity of investigating the association of continuous treatments with long-term outcomes after IR discharge. In summary, this thesis provides valuable information for stakeholders to consider the effect of variations of pattern and content of care on short and long-term outcomes following IR and to direct the resources for providing adequate and appropriate level of care.

Keywords: Inpatient Rehabilitation
Outcome
Post-discharge
Therapeutic Activities
Traumatic Brain Injury
Advisor: Colantonio, Angela
Cullen, Nora
Development of a 3 French Dual Frequency IVUS Catheter

There is a need for imaging modalities capable of more fully visualizing the features of atherosclerotic plaque and monitoring treatments. Currently, only invasive modalities are able to precisely assess fibrous cap thickness, plaque size, the extent of positive remodelling, and the correct expansion of stents. No individual modality can visualize all of these features at the present time, and considerable effort has been made to develop hybrid imaging catheters for broader capabilities. In line with that goal, this thesis describes the development of a dual frequency intravascular ultrasound (DF IVUS) catheter incorporating a bidirectional transducer stack with centre frequencies of 30 and 80 MHz. 30 MHz is within the range of currently available clinical IVUS, and 80 MHz was chosen to provide the desired superficial resolution. The prototype was designed to have a clinically-relevant size and form factor to ensure its applicability for this purpose. The design and fabrication process are explained in detail, and the characterization results show the successful performance of the prototype with respect to the design criteria. Imaging of phantoms, in vitro human atherosclerotic coronary arteries and in vivo healthy porcine coronary arteries demonstrate that the prototype is able to provide complementary, coplanar images; with superficial axial resolution as low as 16 μm, lower-resolution depth penetration up to 6 mm, penetration through blood at both 30 and 80 MHz, and improved visualization of a stent relative to the vessel wall. Features of plaque and the vessel wall were well-visualized compared with conventional IVUS (30 MHz). These results suggest that DF IVUS is technologically and clinical feasible as a technique for improved coronary imaging. Other design approaches to DF IVUS may provide additional benefits, including contrast-enhanced imaging, and the procedures described here may be relevant to the development of these and other sub-millimetre ultrasound devices.

Keywords: Atherosclerosis
Dual frequency
High frequency
Intravascular imaging
Intravascular ultrasound
Transducers
Advisor: Foster, F. Stuart
Stratospheric Ballooning With SPIDER and BIT

Stratospheric ballooning is an important platform for astrophysics and cosmology. This work presents the development of Spider, a CMB polarimeter, and BIT, a visible light and near-UV imager, both of which are balloon experiments. Both experiments require gondolas and pointing systems to track or scan on the sky, and their development is an important area of research in the field. Spider aims to observe the relics of inflation by constraining the tensor to scalar ratio, r. It flew an initial flight in 2014-15 and the data analysis is presented here. This work includes the development of the Spider noise model which is an important component of any CMB analysis. Development work on the second Spider gondola is also presented, preparing it for a second Antarctic campaign in the near future. BIT is attempting to constrain Ω_m , the energy density of matter and σ_8 , the scale size of early universe fluctuations, through the use of weak lensing. Although no weak lensing data has been obtained yet, this thesis presents the development of the instrument from its earliest stages through its two successful test flights. Specific emphasis is given to the development of the star camera subsystem, which was integral to the success of the project.

Keywords: B Modes
CMB
Star Cameras
Stratospheric Ballooning
Weak Lensing
Advisor: Netterfield, C Barth
Functional Neural Correlates of Cognitive Impairment and Neural Compensation in Mild to Moderate Cerebral Small Vessel Disease

Rationale: Aging is associated with both structural and functional changes along with decline in certain cognitive domains. Reduced white matter integrity is a common age-related structural change that is accelerated in those cerebral small vessel disease (CSVD), a disease characterized by white matter hyperintensities and subcortical lesions. To what extent these lesions contribute to age-related cognitive decline is unclear as there is considerable heterogeneity in the cognitive abilities of those with similar amounts of vascular lesion burden. Research Aims: The aim of this dissertation is to use a combination of neuropsychological testing, structural brain imaging and multimodal functional neuroimaging to investigate individual differences in cognitive function among older adults with CSVD. Hypothesis: I hypothesize that poor executive function in those with CSVD will relate to deficits in top-down attentional control as indexed by electrophysiological markers of attention as well as changes in functional brain activity. Results: In chapter 3, six trends within the CSVD functional imaging literature are identified and summarized within a proposed “distributed network disruption and interference model”. This model proposes that cognitive impairment in CSVD results from diffuse disruption of fronto-subcortical circuits. This in turn contributes to DMN hyperactivation and compensatory frontal activation. Subsequent chapters build upon this model with chapter 4 confirming the presence of high cognitive heterogeneity in CSVD and identifying distinct behavioural subgroups within the CSVD population. Whereas Chapter 5 presents evidence that poor attentional control in CSVD is related to a specific deficit in suppression of irrelevant stimuli as indexed by an attenuated selection negativity. Chapter 6 relates individual differences in executive dysfunction and the suppression deficit to differences in task-related brain activation. Interpretation: Cumulatively these results point to a mechanism for understanding cognitive impairment and compensation in CSVD which is rooted in the ability to suppress irrelevant stimuli and promote greater involvement of the frontoparietal control network. This may have implications for guiding the development and assessment of cognitive interventions for this growing patient population.

Keywords: Attention
Cerebral Small Vessel Disease
Cognitive Reserve
Executive Function
Vascular Cognitive Impairment
White Matter Hyperintensities
Advisor: Levine, Brian
The Tansmembrane Associations of HIV-1 Vpu and its Human Targets

Many viruses evade the innate immune response through the expression of accessory proteins that modulate the activity of immune system proteins. The HIV-1 protein Vpu functions to downregulate a number of host cell surface proteins within infected cells. For several target proteins, including the NTB-A and PVR receptors and the restriction factor tetherin, downregulation occurs through direct interaction of the proteins’ transmembrane (TM) domains. The mechanism through which a single TM domain can interact with a wide range of targets that share no obvious interaction motifs is poorly understood. In this thesis we characterize the TM domain homo and heterooligomerization of Vpu and its human targets, tetherin, NTB-A and PVR. By using a series of fluorophore labeled TM domain peptides we used Förster resonance energy transfer (FRET) to characterize the interactions of Vpu with itself as well as its human targets. Our data show target TM domains compete for interaction with Vpu, suggesting that all targets bind the same helical face, also that formation of each heterooligomer has a similar free energy of association to the Vpu homooligomer. This leads to a model in which Vpu monomers, Vpu homooligomers and Vpu-target heterooligomers coexist and suggests that the binding surface of the Vpu TM domain has been selected for weak binding to multiple targets. We also document the role of the lipid bilayer, specifically lipid phase and hydrophobic thickness, on TM domain assembly within the Vpu-target system. We show that thicker membranes and gel phase lipids non-specifically promote associations of Vpu and its targets without affecting secondary structure. The properties of the lipid bilayer are an important factor in Vpu-target oligomerization and may explain differences in Vpu-target oligomerization in different subcellular compartments. This work provides new insight into the molecular mechanism of Vpu and furthers our knowledge of membrane protein assembly

Keywords: FRET
HIV-1
transmembrane
Vpu
Advisor: Sharpe, Simon J
Plk4 Regulates Cell Motility through Arhgef1 and Rho GTPase Activation

Cell migration is a fundamental process essential for embryogenesis, immune cell function and maintenance of homeostasis. It is driven primarily by polymerization of the actin cytoskeleton, which is regulated by a family of small Rho GTPases. Rho GTPases Rac1 and Cdc42 are known as the major regulators of actin polymerization and RhoA is mostly thought of as the regulator of actin stress fiber formation and actomyosin contraction. Rho GTPases are activated by GEFs and deactivated by GAPs. Polo-like kinase 4 (Plk4) is well-acknowledged as the master regulator of centriole duplication. Nevertheless, our laboratory has shown that Plk4 activates RhoA during cytokinesis to ensure proper cleavage furrow positioning. We also identified a novel role for Plk4 in regulation of spreading, migration and invasion of normal and cancer cells. However, the mechanisms through which Plk4 regulates these processes are not completely understood. I hypothesized that Plk4 regulates the activation of RhoA and possibly other Rho GTPases to indice cell migration and spreading. In support of this hypothesis, I show here that Plk4 acts as an activator of Rho GTPases RhoA and Rac1 in cycling cells. I also identify 12 GEFs that possess Plk4 consensus phosphorylation motif and show that Plk4 physically interacts with the GEFs Arhgef1 and P-Rex2. Plk4 functionally interacted with Arhgef1 to affect cell migration, showing at least partial dependence of Plk4 migration phenotype on Arhgef1. Further delineation of their interaction showed that Plk4 binds the DH/PH domains and phosphorylates the L-DH/PH fragment of Arhgef1, and that Plk4 enhances Arhgef1 GEF activity. Finally, I show that Plk4 is not likely to activate Arhgef1-RhoA pathway to enhance cell spreading but may act through P-Rex2-Rac1. These results identify a novel mechanism through which Plk4 regulates cell migration. Plk4 has emerged as a potential oncogene, promoter of tumour progression and metastasis formation, prompting the creation of Plk4 inhibitors for therapeutic purposes. The data presented here deepen our understanding of the pathways that may be affected by the Plk4-targeted therapy.

Keywords: Arhgef1
GEFs
migration
motility
Plk4
Rho GTPases
Advisor: Swallow, Carol J
Microfluidic Devices for High-resolution Cellular Profiling

Rapid and accurate detection, quantification and analysis of rare cells in biological samples is of critical importance for a variety of clinical applications. These cells, which can be outnumbered a billion to one by non-target background cells, can have important implications in cancer diagnostics, diagnosis of viral infections and regenerative medicine. Microfluidics allows the processing of large sample volumes quickly while maintaining rigorous control over dynamical and chemical conditions at the microscale. Though many technologies have been developed to identify bulk populations of rare cells, some applications require higher resolution detection methods, increased throughput, or improved purity to advance current research and clinical practices. This thesis reports the design and application of a novel, high-resolution cell sorter incorporating a high-performance ferromagnetic alloy. Incubating biological samples with magnetic nanoparticles specifically bound to a cell surface marker of interest yields cell subpopulations exhibiting differential magnetic loading. These cells are then deflected laterally by rows of cobalt-based ribbons and directed to one of several outlets corresponding to their level of magnetic loading. Using integrated graphene Hall sensors, we demonstrate for the first time concurrent automated profiling of single cells and CTC clusters. We demonstrate the utility of our approach by profiling cancer cells derived from a mouse xenograft model. We then modify our approach with an eye towards maximizing throughput, and develop a modified design capable of sorting 30 million target cells per hour. With multiple devices running in parallel, we sort hundreds of millions of cells in a genome-wide CRISPR-Cas9 loss-of-function screen to identify genetic regulators of CD47, a clinically relevant protein overexpressed on the surface of many tumour cells. Our screen hits were then validated by side-by-side comparative screens run with the two most widespread cell sorting technologies, FACS and MACS. Overlapping hits between the three distinct methods reveal as many as five heretofore unexplored potential targets for cancer immunotherapy.

Keywords: Cancer
Cell sorting
Circulating tumour cells
CRISPR
Microfluidics
Advisor: Kelley, Shana
Receptor Binding Domains and Coronavirus Adaptation and Evolution

Coronaviruses are enveloped RNA viruses that cause respiratory diseases in humans and other animals. The continued circulation of human coronaviruses and the large reservoir of non-human coronaviruses that have the potential to infect humans represent a significant health concern. Similar to other RNA viruses, coronaviruses are characterized by a high mutation rate, and evolution and adaptation has accompanied the diversification of coronaviruses. The coronavirus spike protein is a viral membrane protein involved in host receptor binding and fusion of the viral and host cell membranes. Reported here is work performed on the spike proteins of two human common cold causing coronaviruses: HCoV-229E and HCoV-OC43. Specifically, the X-ray crystal structures of the following were determined: i) the receptor binding domain (RBD) of the S-protein of HCoV-229E in complex with human aminopeptidase N (hAPN) and ii) the carbohydrate binding domain (CBD) of the S-protein of HCoV-OC43 in complex with 9-O-Acetyl-N-acetylneuraminic acid (Neu5,9Ac2). In addition, viral sequence analysis, binding studies, and comparative sequence and structural analysis among closely related coronaviruses were performed. The observation that the greatest variability shown by natural viral isolates of HCoV-229E is located in the receptor binding loops is quite surprising from a receptor binding standpoint. However, this observation suggests that the receptor binding region is changing due to viral selection. Indeed, our results indicate that the optimization of receptor binding affinity and/or immune evasion, two known determinants of viral fitness, are operative in the emergence of new viral isolates. Although less well characterized at this point, natural viral sequence variation at or near the carbohydrate binding site of HCoV-OC43 is also observed, an indication that similar selection pressures are operative. Together, our results provide mechanistic insights into coronavirus adaptation and evolution.

Advisor: Rini, James