MIMU sensors for joint kinematic estimation: anatomical landmarking and frame correction

The ability to capture human motion allows researchers to evaluate an individual’s gait. Gait can be measured in different ways, from camera-based systems to Magnetic and Inertial Measurement Units (MIMU). The former uses cameras to track positional information of photo-reflective markers, while the latter uses accelerometers, gyroscopes, and magnetometers to measure segment orientation. Both systems can be used to measure joint kinematics, but the results vary because of their differences in anatomical calibrations. The objective of this thesis was to study potential solutions for reducing joint angle discrepancies between MIMU and camera-based systems. The first study worked to correct the anatomical frame differences between MIMU and camera-based systems via the joint angles of both systems. This study looked at full lower body correction versus correcting a single joint. Single joint correction showed slightly better alignment of both systems, but does not take into account that body segments are generally affected by more than one joint. The second study explores the possibility of anatomical landmarking using a single camera and a pointer apparatus. Results showed anatomical landmark position could be determined using a single camera, as the anatomical landmarks found from this study and a camera-based system showed similar results. This thesis worked on providing a novel way for obtaining anatomical landmarks with a single point-and-shoot camera, as well aligning anatomical frames between MIMUs and camera-based systems using joint angles.

Investigating the Association Between Computer-Mediated Communication (CMC) and the Health of Canadian Young People: A Mixed Methods Study

Canadian young people are increasingly more connected through technological devices. This computer-mediated communication (CMC) can result in heightened connection and social support but can also lead to inadequate personal and physical connections. As technology evolves, its influence on health and well-being is important to investigate, especially among youth. This study aims to investigate the potential influences of computer-mediated communication (CMC) on the health of Canadian youth, using both quantitative and qualitative research approaches. This mixed-methods study utilized data from the 2013-2014 Health Behaviour in School-aged Children survey for Canada (n=30,117) and focus group data involving Ontario youth (7 groups involving 40 youth). In the quantitative component, a random-effects multilevel Poisson regression was employed to identify the effects of CMC on loneliness, stratified to explore interaction with family communication quality. A qualitative, inductive content analysis was applied to the focus group transcripts using a grounded theory inspired methodology. Through open line-by-line coding followed by axial coding, main categories and themes were identified. The quality of family communication modified the association between CMC use and loneliness. Among youth experiencing the highest quartile of family communication, daily use of verbal and social media CMC was significantly associated with reports of loneliness. The qualitative analysis revealed two overarching concepts that: (1) the health impacts of CMC are multidimensional and (2) there exists a duality of both positive and negative influences of CMC on health. Four themes were identified within this framework: (1) physical activity, (2) mental and emotional disturbance, (3) mindfulness, and (4) relationships. Overall, there is a high proportion of loneliness among Canadian youth, but this is not uniform for all. The associations between CMC and health are influenced by external and contextual factors, including family communication quality. Further, the technologically rich world in which young people live has a diverse impact on their health. For youth, their relationships with others and the context of CMC use shape overall influences on their health.

BLOCK COPOLYMERS CONTAINING A LOW-SURFACE-ENERGY BLOCK AND THEIR APPLICATIONS

This thesis reports the synthesis and/or applications of three types of block copolymers that each bear a low-surface-energy block. First, poly(dimethylsiloxane)-block-poly(2-cinnamoyloxyethyl acrylate) (PDMS-b-PCEA) was synthesized and characterized. Cotton coating using a micellar solution of this block copolymer yielded superhydrophobic cotton fabrics. X-ray photoelectron spectroscopy (XPS) and surface property analyses indicated that the PDMS block topped the polymer coating. Photocuring the cotton swatches crosslinked the underlying PCEA layer and yielded permanent coatings. More interestingly, hydrophilically patterned superhydrophobic cotton fabrics were produced using photolithography that allowed the crosslinking of the coating around irradiated fibers but the removal, by solvent extraction, of the coating on fibers that were not irradiated. Since water-based ink only permeated the uncoated regions, such patterned fabric was further used to print ink patterns onto substrates such as fabrics, cardboard, paper, wood, and aluminum foil. Then, another PDMS-based diblock copolymer poly(dimethylsiloxane)-block-poly(glycidyl methacrylate) (PDMS-b-PGMA) was prepared. Different from PCEA that photocrosslinked around cotton fibers, PGMA reacted with hydroxyl groups on cotton fiber surfaces to get covalently attached. Further, different PGMA chains crosslinked with each other. PDMS-b-PGMA-coated cotton fabrics have been used for oil-water separations. In addition, polymeric nanoparticles were grafted onto cotton fiber surface before PDMS-b-PGMA was used to cover the surfaces of the grafted spheres and the residual surfaces of the cotton fibers. These two types of fabrics, coated by the block copolymer alone or by the polymer nanospheres and then the copolymer, were characterized by scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FT-IR), and water repellency analyses. A comprehensive comparative study was made of their performances in oil-water separation. Finally, a fluorinated ABC triblock copolymer poly(acrylic acid)-block-poly(2-cinnamoyloxyethyl methacrylate)-block-poly(2-perfluorooctylethyl methacrylate) (PAA-b-PCEMA-b-PFOEMA) was used to iii encapsulate air nanobubbles. The produced air nanobubbles were thermodynamically stable in water and were some 100 times more stable than commercially available perfluorocarbon-filled microbubbles under ultrasound. These nanobubbles, due to their small sizes and thus ability to permeate the capillary networks of organs and to reach tumors, may expand the applications of microbubbles in diagnostic ultrasonography and find new applications in ultrasound-regulated drug delivery.

Adiposity, Sex Hormones, and Repetitive Element DNA Methylation in Healthy Postmenopausal Women

INTRODUCTION: Low levels of methylation within repetitive DNA elements, such as long interspersed nuclear element-1 (LINE-1) and Alu repeats, are believed to epigenetically predispose an individual to cancer and other diseases. The extent to which lifestyle factors affect the degree of DNA methylation within these genomic regions has yet to be fully understood. Adiposity and sex hormones are established risk factors for certain types of cancer and other illnesses, particularly amongst postmenopausal women. The aim of the current investigation is to assess the impact of adiposity and sex hormones on LINE-1 and Alu methylation in healthy postmenopausal women. METHODS: A cross-sectional study was conducted using baseline data from an ancillary study of the Alberta Physical Activity and Breast Cancer Prevention (ALPHA) Trial. Current adiposity was measured using a dual-energy x-ray absorptiometry (DXA) scan, computed tomography (CT) scan, and balance beam scale. Historical weights were self-reported in a questionnaire. Current endogenous sex hormone concentrations were measured in fasting blood serum. Estimated lifetime number of menstrual cycles was used as a proxy for cumulative exposure to ovarian sex hormones. Repetitive element methylation was quantified in white blood cells using a pyrosequencing assay. Linear regression was used to model the relations of interest while adjusting for important confounders. RESULTS: Adiposity and serum estrogen concentrations were positively related to LINE-1 methylation but were not associated with Alu methylation. Cumulative ovarian sex hormone exposure had a “U-shaped” relation with LINE-1 regardless of folate intake and a negative relation with Alu methylation amongst low folate consumers. Androgens were not associated with repetitive element DNA methylation in this population. CONCLUSION: Adiposity and estrogens appear to play a role in maintaining high levels of repetitive element DNA methylation in healthy postmenopausal women. LINE-1 methylation may be a mechanism whereby estrogen exposure protects against cardiovascular and neurodegenerative illnesses. These results add to the growing body of literature showing how the epigenome is shaped by our lifestyle choices. Future prospective studies assessing the relation between levels of repetitive element DNA methylation in healthy individuals and subsequent disease risk are needed to better understand the clinical significance of these results.

Maternal and paternal growth regulatory factor expression in hybrid sunfish.

Unidirectional hybridization between bluegill (Lepomis macrochirus) and pumpkinseed (L. gibbosus) sunfish enables researchers to explore the relative expression of paternal and maternal alleles in hybrids. Past studies have found that the metabolic dysfunction in bluegill-pumpkinseed hybrids may be due to incompatibilities between nuclear and mitochondrial genomes. However, the consequences of hybridization on body size and muscle growth have not been examined. This topic is particularly interesting because hybrids grow larger than parentals despite the fact that they are often sired by smaller, precociously mature bluegills. In order to improve our understanding of growth dynamics in hybrid sunfish, I conducted real-time quantitative PCR using species-specific primers on the white muscle tissue of bluegills, pumpkinseeds, and hybrids collected from Lake Opinicon, ON. Five growth factors that have been linked to muscle growth and body size demonstrated similar expression for maternal and paternal alleles. While about half of the hybrids showed the same pattern with myogenin, about half showed very low levels of mRNA for the paternal (bluegill) gene. While this did not explain the heterosis seen in hybrids, it may explain the small body phenotype of the cuckholding bluegill males. I explored the upstream genetic structure of bluegill myogenin and established that four alleles exist within the population. Furthermore, I uncovered a relationship in hybrids between the proximal promoter/ 5’ UTR of myogenin and its transcript level. I found that the hybrids demonstrating low paternal myogenin expression unfailingly possessed A3 or A4 alleles, but future studies will be needed to reveal the molecular links between the genotype and the growth phenotype. A similar genotype-phenotype association was not obvious in parentals, even those that were homozygous for these alleles. Whether this relationship can provide insight into the genetic determinants of bluegill alternative mating strategies has yet to be determined.

THE ELEMENTARY TEACHERS’ PERCEPTIONS OF MOTIVATION FOR AND THE PRINCIPAL’S INFLUENCE ON THEIR ENGAGEMENT IN SELF-DIRECTED PROFESSIONAL DEVELOPMENT

All teachers participate in self-directed professional development (PD) at some point in their careers; however, the degree to which this participation takes place varies greatly from teacher to teacher and is influenced by the leadership of the school principal. The motivation behind why teachers choose to engage in PD is an important construct. Therefore, there is a need for better understanding of the leader’s role with respect to how and why teachers engage in self-directed professional development. The purpose of the research was to explore the elementary teachers’ motivation for and the school principal’s influence on their engagement in self-directed professional development. Three research questions guided this study: 1. What motivates teachers to engage in self-directed professional development? 2. What are the conditions necessary for promoting teachers’ engagement in self-directed professional development? 3. What are teachers’ perceptions of the principal’s role in supporting, fostering, encouraging, and sustaining the professional development of teachers? A qualitative research approach was adopted for this study. Six elementary teachers from one south-eastern Ontario school board, consisting of three novice and three more experienced teachers, provided their responses to a consistent complement of 14 questions. Their responses were documented via individual interviews, transcribed verbatim, and thematically analysed. The findings suggested that, coupled with the individual motivating influences, the culture of the school was found to be a conditional dynamic that either stimulated or dissuaded participation in self-directed PD. The school principal provided an additional catalyst or deterrence via relational disposition. When teachers felt their needs for competency, relatedness, and autonomy were satisfied, the conditions necessary to motivate teachers to engage in PD were fulfilled. A principal who personified the tenets of transformational leadership served to facilitate teachers’ inclinations to take on PD. A leadership style that was collaborative and trustful and allowed for personal autonomy was a dominant foundational piece that was critical for participant participation in self-directed PD. Finally, the principals were found to positively impact school climate by partaking in PD alongside teachers and ensuring there was a shared vision of the school so that teachers could tailor PD to parallel school interests.

UPTAKE OF THE PERIODIC HEALTH EXAM AMONG ADULTS WITH INTELLECTUAL AND DEVELOPMENTAL DISABILITIES FROM 2003 TO 2015

Background: Over the past decade, annual heath exams have been de-emphasized for the general population but emphasized for adults with intellectual and developmental disabilities (IDD). The purpose of this project was to determine if there has been an increase in the uptake of the health exam among adults with IDD in Ontario, to what extent, and the effect on the quality of preventive care provided. Methods: Using administrative health data, the proportion of adults (18-64 years old) with IDD who received a health exam (long appointment, general assessment, and “true” health exam), a high value on the primary care quality composite score (PCQS), and a health exam or high PCQS each year was compared to the proportion in a propensity score matched sample of the general population. Negative binomial and segmented negative binomial regression controlling for age and sex were used to determine the relative risk of having a health exam/high PCQS/health exam or PCQS over time. Results: Pre joinpoint, the long appointment and general assessment health exam definitions saw a decrease and the “true” health exam saw an increase in the likelihood of adults having a health exam. Post joinpoint, all health exam definitions saw a decrease in the likelihood of adults having a health exam. Pre joinpoint, all PCQS measures (high PCQS, long appointment or high PCQS, “true” health exam or high PCQS) saw an increase in the likelihood for adults to achieve a high PCQS or high PCQS/have a health exam. Post joinpoint, all PCQS measures saw a decrease in the likelihood for adults to achieve a high PCQS or high PCQS/have a health exam. Achieving a high PCQS was strongly associated with having a health exam regardless of health exam definition or IDD status. Conclusions: Despite the publication of guidelines, only a small proportion of adults with IDD are receiving health exams. This indicates that the publication of guidelines alone was not sufficient to change practice. More targeted measures, such as the implementation of an IDD-specific health exam fee code, should be considered to increase the uptake of the health exam among adults with IDD.

Academic Integrity and Senior Nursing Undergraduate Clinical Practice

Background: Academic integrity (AI) has been defined as the commitment to the values of honesty, trust, fairness, respect, and responsibility with courage in all academic endeavours. The senior years of nursing studies provide an intersection for students to transition to professional roles through student clinical practice. It is essential to understand what predicts senior nursing students’ intention to behave with AI so that efforts can be directed to initiatives focused on strengthening their commitment to behaving with AI. Research Questions: To what extent do students differ on Theory of Planned Behaviour (TPB) variables? What predicts intention to behave with academic integrity among senior nursing students in clinical practice across three different Canadian Schools of Nursing? Method: The TPB framework, an elicitation (n=30) and two pilot studies (n=59, n=29) resulted in the development of a 38 question (41-item) self-report survey (Miron Academic Integrity Nursing Survey—MAINS: α>0.70) that was administered to Year 3 and 4 students (N=339). Three predictor variables (attitude, subjective norm, perceived behavioural control) were measured with students’ intention to behave with AI in clinical. Age, sex, year of study, program stream, students’ understanding of AI policies, and locations where students accessed AI information were also measured. Results: Hierarchical multiple regression analyses revealed that background, site, and TPB variables explained 32.6% of the variance in intention to behave with academic integrity. The TPB variables explained 26.8% of the variance in intention after controlling for background and site variables. In the final model, only the TPB predictor variables were statistically significant with Attitude having the highest beta value (beta=0.35, p<0.001), followed by Subjective Norm (beta=0.21, p<0.001) and Perceived Behavioural Control (beta=0.12, p<0.02). Conclusion: Student attitude is the strongest predictor to intention to behave with AI in clinical practice and efforts to positively influence students’ attitudes need to be a focus for schools, curricula, and clinical educators. Opportunities for future research should include replicating the current study with students enrolled in other professional programs and intervention studies that examine the effectiveness of specific endeavours to promote AI in practice.

Beyond Black and White: The grisaille exterior of the Netherlandish triptych

This thesis is a study of fifteenth- to mid sixteenth-century Netherlandish triptych exteriors, focusing on the so-called ‘grisaille’ technique. During this period, altarpieces produced in the Low Countries were typically constructed in a tripartite format with folding wings. This arrangement created the opportunity for pictorial representations on both sides of the hinged panels. Painters emphasized the distinction between the triptych’s two faces by executing the exteriors in a strikingly more subdued palette than the interiors. Particular iconographic subject matter was favoured for grisailles, which often depict the Annunciation or saints that reflect the triptych’s patronage or intended location. Jan van Eyck was notable for his emphasis on imitating stone statuary and created three important grisailles, one of which would influence triptych exteriors for years to come. Hieronymus Bosch, an artist working at the turn of the sixteenth century, also brought innovation to his grisailles, further expanding the potential of these reduced-palette paintings. This thesis examines the creative process involved in the production of grisailles and compares the underdrawings of triptych exteriors to those of the corresponding polychromatic interiors. In this study, grisailles are situated in their context as part of multifaceted artworks as well as within the broader church environment. New infrared reflectograms were generated using Queen’s OSIRIS infrared camera to document works in Belgium and the Netherlands. While some aspects of underdrawings could indicate that the figure was meant to imitate statuary, this distinction was not directly linked to triptych exteriors and was related instead to efforts at a trompe-l’oeil effect. Such attempts at mimicry can also be found on triptych interiors. Through a close examination of the underdrawing stage of these paintings it appears that this part of the creative process was not distinguished in any significant way from the underdrawings of triptych interiors.

Understanding arsenic mobility and speciation in lake sediments impacted by ore roasting near Giant mine, NWT

The roasting of gold-bearing arsenopyrite at Giant mine (Northwest Territories) between 1949 and 1999 released approximately 20,000 tonnes of toxic arsenic-bearing aerosols in the local aerial environment. Detailed examination of lake sediments, sediment porewaters, surface waters and lake hydrology sampled from three lakes of differing limnological characteristics was conducted in summer and winter conditions. Samples were analyzed for solid and dissolved elemental concentrations, speciation and mineralogy. The three lakes are located less than 5km from the mine roaster, and downwind, based on predominant wind direction. The objective of the study was to assess the controls on the mobility and fate of arsenic in these roaster-impacted subarctic lacustrine environments. Results show that the occurrence of arsenic trioxide in lake sediments coincides with the regional onset of industrial activities. The bulk of arsenic in sediments is contained in the form of secondary sulphide precipitates, with iron oxides hosting a minimal amount of arsenic near the surface-water interface. The presence of geogenic arsenic is likely contained as dilute impurities in common rock-forming minerals, and is not believed to be a significant source of arsenic to sediments, porewaters or lake waters. Furthermore, the well correlated depth-profiles of arsenic, antimony and gold in sediments may help reveal roaster impact. The soluble arsenic trioxide particles contained in sediments act as the primary source of arsenic into porewaters. Dissolved arsenic in reducing porewaters both precipitate as secondary sulphides in situ, and diffuse upwards into the overlying lake waters. Arsenic diffusion out of porewaters, combined with watercourse-driven residence time, are estimated to be the predominant mechanisms controlling arsenic concentrations in overlying lake waters. The sequestration of arsenic from porewaters as sulphide precipitates, in the study lakes, is not an effective process in keeping lake-water arsenic concentrations below guidelines for the protection of the freshwater environment and drinking water. Seasonal impacts on lake geochemistry derive from ice covering lake waters, cutting them off from of atmospheric oxygen, along with the exclusion of solutes from the ice. Such effects are limited in deep lakes but are can be an important factor controlling arsenic precipitation and mobility in ponds.

The differential impact of oxytocin receptor (OXTR) genotypes on the risk of autism spectrum disorder (ASD) and resulting social communication deficits

Background: Autism spectrum disorder (ASD) is multifactorial and is likely the result of complex interactions between multiple environmental and genetic factors. Recently, it has been suggested that each symptom cluster of the disorder, such as poor social communication, may be mediated by different genetic influences. Genes in the oxytocin pathway, which mediates social behaviours in humans, have been studied with single nucleotide polymorphisms (SNPs) in the oxytocin receptor gene (OXTR) being implicated in ASD. This thesis examines the presence of different oxytocin receptor genotypes, and their associations with ASD and resulting social communication deficits. Methods: The relationship between four OXTR variants and ASD was evaluated in 607 ASD simplex (SPX) families. Cases were compared to their unaffected siblings using a conditional logistic approach. Odds ratios and associated 95 percent confidence intervals were obtained. A second sample of 235 individuals with a diagnosis of ASD was examined to evaluate whether these four OXTR variants were associated with social communication scores on the Autism Diagnostic Interview – Revised (ADI-R). Parameter estimates and associated 95 percent confidence intervals were generated using a linear regression approach. Multiple testing issues were addressed using false discovery adjustments. Results: The rs53576 AG genotype was significantly associated with a lower risk of ASD (OR = 0.707, 95% CI: 0.512-0.975). A single genotype (AG) provided by the rs2254298 marker was found to be significantly associated with higher social communication scores (Parameter estimate = 1.833, SE = 0.762, p = 0.0171). This association was also seen in a Caucasian only and mothers as the respondent samples. No association was significant following false discovery rate adjustments. Conclusion: The findings from these studies provide limited support for the role of OXTR SNPs in ASD, especially in social communication skills. The clinical significance of these associations remains unknown, however, it is likely that these associations do not play a role in the severity of symptoms associated with ASD. Rather, they may be important in the appearance of social deficits due to the rs2254298 markers association with enlarged amygdalas.

The Causes and Effects of Consumer Competence Perceptions

Consumers are constantly making consumption decisions and engaging in marketplace activities that require some level of competence. In other words, consumers possess and require some knowledge, skills, and abilities to engage in the marketplace and obtain what they want. But what causes consumers to infer they are or are not competent? And, what are the consequences of these competence inferences on consumer behaviour? This dissertation examines the role consumption plays in consumers’ inferences of their own competence to enhance our understanding of these issues. By integrating the literature on competence and attributions of blame, this dissertation develops a theory for when and how consumption influences self-perceptions of competence and how these self-perceptions of competence impact future consumer behaviours. Evidence from five studies suggests that consumers infer their own competence from their consumption outcomes, despite who is actually responsible for causing these outcomes. This means consumers potentially see themselves as incompetent for negative outcomes that are entirely firm-caused. This dissertation argues that people infer their own competence from firm-caused outcomes because they conflate their decisions made prior to an outcome with the cause of that outcome. This dissertation also examines how these variations in self-perceptions of competence can influence future consumer behaviours.

Youth sport coaches’ reflections on leadership behaviours during games and practices

Youth sport coaches shape the developmental sporting experience for their athletes (Camiré, Trudel, & Forneris, 2014). Specifically, coaches who form individualized, supportive relationships with their athletes can increase the development of personal and social skills (Fraser-Thomas, Côté, & Deakin, 2005). In light of the value of these relationships, increasing evidence is prompting the application of leadership theories, such as Transformational Leadership (TFL), in youth sport (Vella et al., 2013). The aim of this study was to explore coach perceptions of how and why leadership behaviours are applied in the youth sport context. Eleven coaches (Mage= 42.3, SD= 15.2) were recruited from competitive youth soccer and volleyball clubs (athletes’ Mage= 15.8, SD= 1.9) in Eastern Ontario and participated in a stimulated recall interview. During the interviews, coaches reflected upon their own coaching behaviours and provided insight into the application of leadership behaviours in youth sport. Responses were prompted by relevant video sequences from recorded practice and game sessions. A thematic content analysis revealed that; i) coaches use a variety of leadership behaviours in youth sport, ii) the use of leadership behaviours vary across sport contexts or settings, and iii) contrasting leadership styles (e.g., transactional vs. transformational) are associated with distinctive coach objectives (e.g., promoting confidence vs. establishing respect). These findings have helped identify gaps within coach education, and provide theoretical insight for applying leadership theories, and more specifically TFL, to help improve the sport experiences of young athletes.

Supporting Metacognitive Development in Early Science Education: Exploring Elementary Teachers' Beliefs and Practices in Metacognition

Metacognition is the understanding and control of cognitive processes. Students with high levels of metacognition achieve greater academic success. The purpose of this mixed-methods study was to examine elementary teachers’ beliefs about metacognition and integration of metacognitive practices in science. Forty-four teachers were recruited through professional networks to complete a questionnaire containing open-ended questions (n = 44) and Likert-type items (n = 41). Five respondents were selected to complete semi-structured interviews informed by the questionnaire. The selected interview participants had a minimum of three years teaching experience and demonstrated a conceptual understanding of metacognition. Statistical tests (Pearson correlation, t-tests, and multiple regression) on quantitative data and thematic analysis of qualitative data indicated that teachers largely understood metacognition but had some gaps in their understanding. Participants’ reported actions (teaching practices) and beliefs differed according to their years of experience but not gender. Hierarchical multiple regression demonstrated that the first block of gender and experience was not a significant predictor of teachers' metacognitive actions, although experience was a significant predictor by itself. Experience was not a significant predictor once teachers' beliefs were added. The majority of participants indicated that metacognition was indeed appropriate for elementary students. Participants consistently reiterated that students’ metacognition developed with practice, but required explicit instruction. A lack of consensus remained around the domain specificity of metacognition. More specifically, the majority of questionnaire respondents indicated that metacognitive strategies could not be used across subject domains, whereas all interviewees indicated that they used strategies across subjects. Metacognition was integrated frequently into Ontario elementary classrooms; however, metacognition was integrated less frequently in science lessons. Lastly, participants used a variety of techniques to integrate metacognition into their classrooms. Implications for practice include the need for more professional development aimed at integrating metacognition into science lessons at both the Primary and Junior levels. Further, teachers could benefit from additional clarification on the three main components of metacognition and the need to integrate all three to successfully develop students’ metacognition.

EXAMINATION OF THE EFFECTS OF VALPROIC ACID EXPOSURE OR P300 PROTEIN INHIBITION ON THE BALANCE OF APOPTOSIS AND PROLIFERATION IN P19 EMBRYONAL CARCINOMA CELLS

Valproic acid (VPA), a commonly-used anticonvulsant drug, is associated with increased risk of fetal malformations, including neural tube defects (NTDs). Previous in vivo studies determined that VPA-exposed embryos with a NTD had altered expression of several proteins regulated by p300, a histone acetyltransferase (HAT) protein. p300 is capable of acetylating histones and non-histone proteins through its HAT activity, allowing it to transcriptionally regulate genes as well as modulate the stability and activity of specific proteins. NFκB, Stat3 and Egr1, all of which function as transcription factors, are regulated by p300 through its HAT activity. Together, these proteins all play an important role in maintaining the balance of apoptosis, proliferation and differentiation, the regulation of which is extremely important for proper embryonic development. The studies in this thesis utilized P19 embryonal carcinoma (EC) cells in order to determine the effects of VPA exposure on the expression of p300 and the aforementioned transcription factors, as well as apoptosis and proliferation, in vitro. P19 EC cells were exposed to C646, a selective p300 inhibitor, in order to assess whether the effects observed as a result of VPA exposure were due to p300 protein degradation. It was found that VPA exposure for 24 hours in P19 EC cells in vitro resulted in a significant decrease in p300 protein expression. VPA exposure also significantly decreased NFκB protein expression, while resulting in increased Stat3 protein expression. However, Stat3 acetylation and phosphorylation, which both contribute to Stat3 activation, were significantly decreased as a result of VPA exposure. p300 inhibition resulted in a significant decrease in NFκB, similar to what was observed as a result of VPA exposure, which suggests that VPA-mediated degradation of p300 may play a role in reduced NFκB protein expression following VPA exposure. Conversely, Stat3 protein expression, acetylation and phosphorylation were not significantly changed as a result of p300 inhibition, suggesting that p300 degradation does not play a role in VPA’s effects on Stat3 protein expression and activation. VPA exposure also resulted in a significant increase in apoptosis, while p300 inhibition did not significantly increase apoptosis. These data suggest that p300 degradation plays a role in VPA-mediated teratogenicity, and that VPA may target other cellular mechanisms in order to exert its teratogenic effects.