A Quantitative and Mechanistic Assessment of Activated Thrombin-Activatable Fibrinolysis Inhibitor and its Role in Pathological Bleeding and Thrombosis

The coagulation and fibrinolytic systems are linked by the thrombin-thrombomodulin complex which regulates each system through activation of protein C and TAFI, respectively. We have used novel assays and techniques to study the enzymology and biochemistry of TAFI and TAFIa, to measure TAFI activation in hemophilia A and protein C deficiency and to determine if enhancing TAFI activation can improve hemostasis in hemophilic plasma and whole blood. We show that TAFIa not TAFI attenuates fibrinolysis in vitro and this is supported by a relatively high catalytic efficiency (16.41μM-1s-1) of plasminogen binding site removal from fibrin degradation products (FDPs) by TAFIa. Since the catalytic efficiency of TAFIa in removing these sites is ~60-fold higher than that for inflammatory mediators such as bradykinin it is likely that FDPs are a physiological substrate of TAFIa. The high catalytic efficiency is primarily a result of a low Km which can be explained by a novel mechanism where TAFIa forms a binary complex with plasminogen and is recruited to the surface of FDPs. The low Km also suggests that TAFIa would effectively cleave lysines from FDPs during the early stages of fibrinolysis (i.e. at low concentrations of FDPs). Since individuals with hemophilia suffer from premature fibrinolysis as a result of insufficient TAFI activation we quantified TAFI activation in whole blood from hemophilic subjects. Both the rate of activation and the area under the TAFI activation time course (termed TAFIa potential) was determined to be reduced in hemophilia A and the TAFIa potential was significantly and inversely correlated with the clinical bleeding iii phenotype. Using a novel therapeutic strategy, we used soluble thrombomodulin to increase TAFI activation which improved the clot lysis time in factor VIII deficient human plasma and hemophilic dog plasma as well as hemophilic dog blood. Finally, we briefly show in a biochemical case study that TAFI activation is enhanced in protein C deficiency and when afflicted individuals are placed on Warfarin anticoagulant therapy, TAFI activation is reduced. Since TAFIa stabilizes blood clots, this suggests that reducing TAFI activation or inhibiting TAFIa may help restore blood flow in vessels with pathological thrombosis.

Robotic Outcome Assessment in a Non-Human Primate Model of Middle Cerebral Artery Stroke

Stroke is a prevalent disorder with immense socioeconomic impact. A variety of chronic neurological deficits result from stroke. In particular, sensorimotor deficits are a significant barrier to achieving post-stroke independence. Unfortunately, the majority of pre-clinical studies that show improved outcomes in animal stroke models have failed in clinical trials. Pre-clinical studies using non-human primate (NHP) stroke models prior to initiating human trials are a potential step to improving translation from animal studies to clinical trials. Robotic assessment tools represent a quantitative, reliable, and reproducible means to assess reaching behaviour following stroke in both humans and NHPs. We investigated the use of robotic technology to assess sensorimotor impairments in NHPs following middle cerebral artery occlusion (MCAO). Two cynomolgus macaques underwent transient MCAO for 90 minutes. Approximately 1.5 years following the procedure these NHPs and two non-stroke control monkeys were trained in a reaching task with both arms in the KINARM exoskeleton. This robot permits elbow and shoulder movements in the horizontal plane. The task required NHPs to make reaching movements from a centrally positioned start target to 1 of 8 peripheral targets uniformly distributed around the first target. We analyzed four movement parameters: reaction time, movement time (MT), initial direction error (IDE), and number of speed maxima to characterize sensorimotor deficiencies. We hypothesized reduced performance in these attributes during a neurobehavioural task with the paretic limb of NHPs following MCAO compared to controls. Reaching movements in the non-affected limbs of control and experimental NHPs showed bell-shaped velocity profiles. In contrast, the reaching movements with the affected limbs were highly variable. We found distinctive patterns in MT, IDE, and number of speed peaks between control and experimental monkeys and between limbs of NHPs with MCAO. NHPs with MCAO demonstrated more speed peaks, longer MTs, and greater IDE in their paretic limb compared to controls. These initial results qualitatively match human stroke subjects’ performance, suggesting that robotic neurobehavioural assessment in NHPs with stroke is feasible and could have translational relevance in subsequent human studies. Further studies will be necessary to replicate and expand on these preliminary findings.

Constructing the Vulvar Pain Assessment Questionnaire Inventory

The goals of this program of research were to examine the link between self-reported vulvar pain and clinical diagnoses, and to create a user-friendly assessment tool to aid in that process. These goals were undertaken through a series of four empirical studies (Chapters 2-6): one archival study, two online studies, and one study conducted in a Women’s Health clinic. In Chapter 2, the link between self-report and clinical diagnosis was confirmed by extracting data from multiple studies conducted in the Sexual Health Research Laboratory over the course of several years. We demonstrated the accuracy of diagnosis based on multiple factors, and explored the varied gynecological presentation of different diagnostic groups. Chapter 3 was based on an online study designed to create the Vulvar Pain Assessment Questionnaire (VPAQ) inventory. Following the construct validation approach, a large pool of potential items was created to capture a broad selection of vulvar pain symptoms. Nearly 300 participants completed the entire item pool, and a series of factor analyses were utilized to narrow down the items and create scales/subscales. Relationships were computed among subscales and validated scales to establish convergent and discriminant validity. Chapters 4 and 5 were conducted in the Department of Obstetrics & Gynecology at Oregon Health & Science University. The brief screening version of the VPAQ was employed with patients of the Program in Vulvar Health at the Center for Women’s Health. The accuracy and usefulness of the VPAQscreen was determined from the perspective of patients as well as their health care providers, and the treatment-seeking experiences of patients was explored. Finally, a second online study was conducted to confirm the factor structure, internal consistency, and test-retest reliability of the VPAQ inventory. The results presented in these chapters confirm the link between targeted questions and accurate diagnoses, and provide a guideline that is useful and accessible for providers and patients.

The Psychometric Properties of the Developmental Reading Assessment

ABSTRACT The purpose of this study was to examine the technical adequacy of the Developmental Reading Assessment (Beaver & Carter, 2004). Internal consistency analysis, factor analysis, and linear regression analyses were used to test whether the DRA is a statistically reliable measuring of reading comprehension for Grades 7 and 8 students. Correlational analyses, decision consistency analyses, and a focus group of experienced Intermediate (Grades 7 and 8) teachers examined whether there is evidence that the results from the DRA provide valid interpretations regarding students’ reading skills and comprehension. Results indicated that, as currently scored, internal consistency is low and skewness of distribution is high. Factor analyses did not replicate those cited by the DRA developers to prove construct validity. Two-way contingency analyses determined that decision consistency did not vary greatly between the DRA, EQAO, scores and report card marks. Views expressed during the focus group echoed many of the challenges to validity found in the statistical analysis. The teachers found that the DRA was somewhat useful, as there were limited alternative reading assessments available for the classroom, but did not endorse it strongly. The study found little evidence that the DRA provides valid interpretations regarding Intermediate students’ reading skills. Indicated changes to the structure and administration procedures of the DRA may ameliorate some of these issues.

Applying life cycle assessment to analyze the environmental sustainability of public transit modes for the City of Toronto

One challenge related to transit planning is selecting the appropriate mode: bus, light rail transit (LRT), regional express rail (RER), or subway. This project uses data from life cycle assessment to develop a tool to measure energy requirements for different modes of transit, on a per passenger-kilometer basis. For each of the four transit modes listed, a range of energy requirements associated with different vehicle models and manufacturers was developed. The tool demonstrated that there are distinct ranges where specific transit modes are the best choice. Diesel buses are the clear best choice from 7-51 passengers, LRTs make the most sense from 201-427 passengers, and subways are the best choice above 918 passengers. There are a number of other passenger loading ranges where more than one transit mode makes sense; in particular, LRT and RER represent very energy-efficient options for ridership ranging from 200 to 900 passengers. The tool developed in the thesis was used to analyze the Bloor-Danforth subway line in Toronto using estimated ridership for weekday morning peak hours. It was found that ridership across the line is for the most part actually insufficient to justify subways over LRTs or RER. This suggests that extensions to the existing Bloor-Danforth line should consider LRT options, which could service the passenger loads at the ends of the line with far greater energy efficiency. It was also clear that additional destinations along the entire transit line are necessary to increase the per passenger-kilometer energy efficiency, as the current pattern of commuting to downtown leaves much of the system underutilized. It is hoped that the tool developed in this thesis can be used as an additional resource in the transit mode decision-making process for many developing transportation systems, including the transit systems across the GTHA.

Proving My Competency One Test at a Time: Internationally Educated Nurses and the Canadian English Language Benchmark Assessment for Nurses

Abstract Professional language assessment is a new concept that has great potential to benefit Internationally Educated Professionals and the communities they serve. This thesis reports on a qualitative study that examined the responses of 16 Canadian English Language Benchmark Assessment for Nurses (CELBAN) test-takers on the topic of their perceptions of the CELBAN test-taking experience in Ontario in the winter of 2015. An Ontario organization involved in registering participants distributed an e-mail through their listserv. Thematic analyses of focus group and interview transcripts identified 7 themes from the data. These themes were used to inform conclusions to the following questions: (1) How do IENs characterize their assessment experience? (2) How do IENs describe the testing constructs measured by the CELBAN? (3) What, if any, potential sources of construct irrelevant variance (CIV) do the test-takers describe based on their assessment experience? (4) Do IENs feel that the CELBAN tasks provide a good reflection of the types of communicative tasks required of a nurse? Overall, participants reported positive experiences with the CELBAN as an assessment of their language skills, and noted some instances in which they felt some factors external to the assessment impacted their demonstration of their knowledge and skill. Lastly, some test-takers noted the challenge of completing the CELBAN where the types of communicative nursing tasks included in the assessment differed from nursing tasks typical of an IENs country or origin. The findings are discussed in relation to literature on high-stakes large-scale assessment and IEPs, and a set of recommendations are offered to future CELBAN administration. These recommendations include (1) the provision of a webpage listing all licensure requirements (2) monitoring of CELBAN location and dates in relation to the wider certification timeline for applicants (3) The provision of additional CELBAN preparatory materials (4) Minor changes to the CELBAN administrative protocols. Given that the CELBAN is a relatively new assessment format and its widespread use for high-stakes decisions (a component of nursing certification and licensure), research validating IEN-test-taker responses to construct representation and construct irrelevant variance is critical to our understanding of the role of competency testing for IENs.

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.