Characterizing the Role of the Previously Undescribed Protein Caskin2 in Vascular Biology

Maintenance of vascular homeostasis is an active process that is dependent on continuous signaling by the quiescent endothelial cells (ECs) that line mature vessels. Defects in vascular homeostasis contribute to numerous disorders of significant clinical impact including hypertension and atherosclerosis. The signaling pathways that are active in quiescent ECs are distinct from those that regulate angiogenesis but are comparatively poorly understood. Here we demonstrate that the previously uncharacterized scaffolding protein Caskin2 is a novel regulator of EC quiescence and that loss of Caskin2 in mice results in elevated blood pressure at baseline. Caskin2 is highly expressed in ECs from various vascular beds both in vitro and in vivo. When adenovirally expressed in vitro, Caskin2 inhibits EC proliferation and migration but promotes survival during hypoxia and nutrient deprivation. Likewise, loss of Caskin2 in vivo promotes increased vascular branching and permeability in mouse and zebrafish models. Caskin2 knockout mice are born in normal Mendelian ratios and appear grossly normal during early adulthood. However, they have consistently elevated systolic and diastolic blood pressure at baseline and significant context-dependent abnormalities in systemic metabolism (e.g., body weight, fat deposition, and glucose homeostasis). Although the precise molecular mechanisms of these effects remain unclear, we have shown that Caskin2 interacts with several proteins known to have important roles in endothelial biology and cardiovascular disease including the serine/threonine phosphatase PP1, the endothelial receptor Tie1, and eNOS, which is a critical regulator of vascular homeostasis. Ongoing work seeks to further characterize the functions of Caskin2 and its mechanisms of action with a focus on how Caskin2-mediated regulation of endothelial phenotype relates to its systemic effects on cardiovascular and metabolic function.

Epigenetic Basis of Centromere Maintenance and Inheritance

Centromeres are essential chromosomal loci at which kinetochore formation occurs for spindle fiber attachment during mitosis and meiosis, guiding proper segregation of chromosomes. In humans, centromeres are located at large arrays of alpha satellite DNA, contributing to but not defining centromere function. The histone variant CENP-A assembles at alpha satellite DNA, epigenetically defining the centromere. CENP-A containing chromatin exists as an essential domain composed of blocks of CENP-A nucleosomes interspersed with blocks of H3 nucleosomes, and is surrounded by pericentromeric heterochromatin. In order to maintain genomic stability, the CENP-A domain is propagated epigenetically over each cell division; disruption of propagation is associated with chromosome instabilities such as aneuploidy, found in birth defects and in cancer.

The CENP-A chromatin domain occupies 30-45% of the alpha satellite array, varying in genomic distance according to the underlying array size. However, the molecular mechanisms that control assembly and organization of CENP-A chromatin within its genomic context remain unclear. The domain may shift, expand, or contract, as CENP-A is loaded and dispersed each cell cycle. We hypothesized that in order to maintain genome stability, the centromere is inherited as static chromatin domains, maintaining size and position within the pericentric heterochromatin. Utilizing stretched chromatin fibers, I found that CENP-A chromatin is limited to a sub-region of the alpha satellite array that is fixed in size and location through the cell cycle and across populations.

The average amount of CENP-A at human centromeres is largely consistent, implying that the variation in size of CENP-A domains reflects variations in the number of CENP-A subdomains and/or the density of CENP-A nucleosomes. Multi-color nascent protein labeling experiments were utilized to examine the distribution and incorporation of distinct pools of CENP-A over several cell cycles. I found that in each cell cycle there is independent CENP-A distribution, occurring equally between sister centromeres across all chromosomes, in similar quantities. Furthermore, centromere inheritance is achieved through specific placement of CENP-A, following an oscillating pattern that fixes the location and size of the CENP-A domain. These results suggest that spatial and temporal dynamics of CENP-A are important for maintaining centromere and genome stability.

Social-Cognitive Determinants of Success in Online Communities for Weight Management

Background: Online communities may be an effective, convenient, and relatively inexpensive intervention platform for individuals seeking assistance with weight management. Recent research suggests that these communities may be as effective as in-person treatments for weight management; however, very little is known about the characteristics that predict weight loss amongst those using an online community. Methods: Within a social-cognitive framework, we sought to identify the psychosocial characteristics that are associated with successful weight management for users of MyFitnessPal, a popular online community for weight management. We recruited participants who were new to the online community and asked them to complete 2 surveys (one at baseline and one 3 months later) that assessed various psychosocial constructs as well as self-reported height and weight. Results: Participants in our sample reported losing, on average, 4.55 kg during the 3-month time period. We found that engaging in weight control behaviors (e.g., monitoring food intake, weighing oneself, etc.) fully mediated the relationship between several of our variables of interest (i.e., baseline self-efficacy and perceived social support within the community) and weight loss. We also found that participants who expected to lose more weight at baseline were significantly more likely to have lost more weight at follow-up. Conclusions: On average, participants in our study lost a clinically meaningful amount of weight. Predictors of weight loss within this community included perceived support within the community (mediated by weight control behaviors), baseline self-efficacy (mediated by weight control behaviors), and baseline outcome expectations. Results of this study can ultimately serve to inform the design of future eHealth interventions for weight management.

Connectivity Mapping (ssCMap) To Predict A20 Inducing Drugs: Anti-inflammatory Action In Cystic Fibrosis

Cystic Fibrosis (CF) lung disease is characterised by a chronic and exaggerated inflammation in the airways. Despite recent developments to therapeutically overcome the underlying functional defect in CFTR (cystic fibrosis transmembrane conductance regulator), there is still an unmet need to also normalise the inflammatory response. The prolonged and heightened inflammatory response in CF is in part mediated by a lack of intrinsic downregulation of the pro-inflammatory NF-kB pathway. We have previously identified reduced expression of the NF-kB down-regulator A20 in CF as a key target to normalise the inflammatory response. Here we have used publically available gene array expression data together with sscMap (statistically significant connections’map)to successfully predict drugs already licensed for the use in humans to induce A20 mRNA and protein expression and thereby reduce inflammation. The effect of the predicted drugs on A20 and NFkB (p65) expression (mRNA) as well as pro-inflammatory cytokine release (IL-8) in the presence and absence of bacterial LPS was shown in bronchial epithelial cells lines (16HBE14o-, CFBE41o-) and in primary nasal epithelial cells (PNECs) from patients with CF (Phe508del homozygous) and non-CF controls. Additionally, the specificity of the drug action on A20 was confirmed using cell lines with TNFAIP3 (A20) knockdown (siRNA). We also show that the A20 inducing effect of ikarugamycin and quercetin is lower in CF derived airway epithelial cells than in non-CF cells.

A Review of the Dodo and its Ecosystem: Insights from a Vertebrate Concentration Lagerstätte in Mauritius

The dodo Raphus cucullatus Linnaeus, 1758, an extinct and flightless, giant pigeon endemic to Mauritius, has fascinated people since its discovery, yet has remained surprisingly poorly known. Until the mid-19th century, almost all that was known about the dodo was based on illustrations and written accounts by 17th century mariners, often of questionable accuracy. Furthermore, only a few fragmentary remains of dodos collected prior to the bird’s extinction exist. Our understanding of the dodo’s anatomy was substantially enhanced by the discovery in 1865 of subfossil bones in a marsh called the Mare aux Songes, situated in southeastern Mauritius. However, no contextual information was recorded during early excavation efforts, and the majority of excavated material comprised larger dodo bones, almost all of which were unassociated. Here we present a modern interdisciplinary analysis of the Mare aux Songes, a 4200-year-old multitaxic vertebrate concentration Lagerst€atte. Our analysis of the deposits at this site provides the first detailed overview of the ecosystem inhabited by the dodo. The interplay of climatic and geological conditions led to the exceptional preservation of the animal and associated plant remains at the Mare aux Songes and provides a window into the past ecosystem of Mauritius. This interdisciplinary research approach provides an ecological framework for the dodo, complementing insights on its anatomy derived from the only associated dodo skeletons known, both of which were collected by Etienne Thirioux and are the primary subject of this memoir.

Through Class Darkly: Class in the British TV Noir

Class has always been at the heart of the television crime drama. Whether it is the post-war paternalism of Dixon of Dock Green (1955 – 1976), the harsh social realism of The Sweeney (1975-1978), or the almost mythical evocations of Britain in Heartbeat (1992 – 2010) and Midsomer Murders (1997- present), class and crime have always been seen as being inextricably linked. Since the 1990s, the British crime drama has been influenced by successive waves of cultural imports from, firstly, the US and then from Scandinavia. There is now a recognisable ‘genre’ for what we might think of as British TV Noir. Beginning with shows such as Cracker (1993 – 2006), Prime Suspect (1991 – 2006) and Messiah (2001) and continuing with dramas like Red Riding (2008), Southcliffe (2013) and Hinterland (2013 – present), the British TV Noir employs narratives and stylistic tropes that might usually be associated with the cinema of the 1940s. Although drawing influence from high profile shows such as Twin Peaks (1990 – 1991), Millennium (1996) and (latterly) The Wire (2002 – 2008), CSI (2000 – present) and The Killing (2007) these British Noir shows also articulate the nation’s shifting class system. As Susan Sydney-Smith has ably demonstrated, the crime drama is “historically contingent” (Sydney-Smith, 2002, p. 5) and shaped by the surrounding socio-political, as well aesthetic, context. To this end, this chapter traces the depiction of class in three key crime series – Prime Suspect, Red Riding and Southcliffe - and explores how social class, and more importantly, its changing face provides a constant background to the narratives and characterisations. These three texts were each produced at pivotal moments in Britain’s relationship to class – Prime Suspect was shown 6 months after Margaret Thatcher vacated office; Red Riding was produced in the midst of the global recession in 2008 and Southcliffe was made in the shadows of stringing welfare and immigration reforms. These texts span three successive political administrations and over two decades of social and political change. Understanding the relationship between criminal activity and class in these dramas however is far more complicated than simply reading the historical context through the text. Commensurate with its cinematic incarnation, TV Noir is both reflective and productive, employing visual and narrative tropes to manipulate, as well reflect, its audience’s moral and social positioning. The picture that emerges from an examination of class and the British TV Noir is one of suspicion and discontent. As Andrew Spicer suggests (with reference to British cinema) the Noir sensibility both depicts and critiques a society that it sees as being “class-ridden, racist and misogynist” (Spicer, 2002, p.202). This is certainly the case with the texts that are being examined here, as social positions and taxonomies are constantly being redefined and renegotiated.

Adaptive Crowd Algorithms for Open-Ended Problems

Thesis (Ph.D.)--University of Washington, 2016-06

"It's Equal but It's not Fair": The experiences of Teacher Candidates of Color in a Predominantly White Institution

Thesis (Ph.D.)--University of Washington, 2016-06

A social-ecological impact assessment for public lands management: application of a conceptual and methodological framework

According to the U.S. National Environmental Policy Act of 1969 (NEPA), federal action to manipulate habitat for species conservation requires an environmental impact statement, which should integrate natural, physical, economic, and social sciences in planning and decision making. Nonetheless, most impact assessments focus disproportionately on physical or ecological impacts rather than integrating ecological and socioeconomic components. We developed a participatory social-ecological impact assessment (SEIA) that addresses the requirements of NEPA and integrates social and ecological concepts for impact assessments. We cooperated with the Bureau of Land Management in Idaho, USA on a project designed to restore habitat for the Greater Sage-Grouse (Centrocercus urophasianus). We employed questionnaires, workshop dialogue, and participatory mapping exercises with stakeholders to identify potential environmental changes and subsequent impacts expected to result from the removal of western juniper (Juniperus occidentalis). Via questionnaires and workshop dialogue, stakeholders identified 46 environmental changes and associated positive or negative impacts to people and communities in Owyhee County, Idaho. Results of the participatory mapping exercises showed that the spatial distribution of social, economic, and ecological values throughout Owyhee County are highly associated with the two main watersheds, wilderness areas, and the historic town of Silver City. Altogether, the SEIA process revealed that perceptions of project scale varied among participants, highlighting the need for specificity about spatial and temporal scales. Overall, the SEIA generated substantial information concerning potential impacts associated with habitat treatments for Greater Sage-Grouse. The SEIA is transferable to other land management and conservation contexts because it supports holistic understanding and framing of connections between humans and ecosystems. By applying this SEIA framework, land managers and affected people have an opportunity to fulfill NEPA requirements and develop more comprehensive management plans that better reflect the linkages of social-ecological systems.

Older Adult Injury Risk Assessment in the Driving and Occupational Environments

Thesis (Ph.D.)--University of Washington, 2016-08

Economic Evaluations of State Policies

Thesis (Ph.D.)--University of Washington, 2016-08