Malaysia is a multiracial/ multiethnic country that is rich with many traditional performing arts such as Wayang Kulit and Main Puteri. However, the number of artists practising these traditions is in decline and they are at risk of extinction according to several Malaysian news outlets. This project is dedicated to understanding the challenges faced by these artists and the strategies they use to overcome obstacles to preserve their art form. Tintoy Chuo, a puppeteer of Malay traditional shadow puppet theatre Wayang Kulit, has become my case study in examining Malaysian artists. Through interviews and analyzing Chuo’s online presence, I found some of his challenges include: (1) federal and state funding structures that favour STEM more than the arts; (2) the Malaysian state’s view on arts as tourism; and (3) state government bans on this art form because it is deemed unIslamic. Some of the strategies that Chuo uses to counter these issues are: (1) blending the traditional element with popular culture references, for example, Star Wars; and (2) seeking international recognition of his work through interviews that are published in magazines and aired on news networks. I compare Chuo’s methodology to the transformation Zapin, a Malay traditional dance, underwent in the 1960s when it was blended with Western ballroom dances as documented by Mohd Anis Md Nor. I reveal similarities between the two processes, including: funding, audience, recognition, methodology, inspiration and community. I argue that the concept of “preservation” used in the context of art-making is problematic, and discuss how framing this methodology in terms of “sustainability” can be a model for other artists hoping to see their art form survive into the next generation.

“After a lot of laughter, comes a long cry.” This old Finnish saying was shared with me by a somewhat distant, but beloved, family friend while I was visiting Helsinki last summer. This notion resonated with me and became the root of my current choreographic research. When I returned to the UW for my final year, I knew I wanted to explore the intense emotion this saying evoked in me and how it could propel a choreographic investigation resulting in a dance work. In 2018, I created a dance, Undertow, that explored the broader idea of nostalgia and its often captivating and drowning effects on people. The choreographic process I utilized in creating Undertow, laid a rich foundation for me to expand upon the idea of nostalgia. I began my research on After Everything with the Finnish saying–– looking for meaning, ideas, and images that stood out to me. Coincidentally, on a visit home I discovered an 8mm film reel from 1968 that had hours of candid footage with my mom and uncle as children on it. The first time I watched it, I was brought to tears as I witnessed these young strangers play and explore. The complex emotions of family, personal histories, and longing for a presence in a history that is not directly mine collided with my reaction to the Finnish saying. This intersection became the focus of my research: exploration of familial nostalgia, relationships, and my planned uprooting after graduation. I cast four incredible collaborative dance artists to conduct this dance research with me, inviting them to explore their own reactions to the saying and nostalgia, so we could begin building a communal vessel of knowledge from which to draw movement material. I presented prompts, discussed ideas, and shared sound to facilitate and generate movement material.

Only recently has mainstream American theatre celebrated minority representation on the stage. Small percentages of ethnically diverse actors perform in American theatre due to character-casting limitations and white male playwrights which dominate theatrical history and limit the representation of our diverse and multicultural world. By adapting the Hebrew Book of Ruth to stage, I hope to contribute a female voice and provide opportunities for women of ethnic, cultural, and socio-economic diversity on the stage. Like Shakespeare’s plays, this adaptation will be written in verse; however, "Pilgrims Far from Home" mimics the style of Hebrew and Ottoman poetry to pay homage to the story’s origins and contribute to the diversity of voices outside of the western poetic style. This story emphasizes female relationships and how our sisterhood can unify people of different walks of life. "Pilgrims Far from Home" brings this ancient religious text into the 21st century by addressing contemporary issues such as otherness, particularly in light of refugee crises. My project aims to contribute to the diversity of contemporary American theatre by using a well-known and multi-cultural story to address and ease the suffering relationships between peoples in our present-day society.

Inherently, a work titled Pornocrates exudes scandal, intrigue, and allure. Created in 1878 by Felicien Rops, the gouache and pencil piece was displayed at the Les XX exhibition of 1886. Felicien Rops was a wealthy Belgian-born artist who collaborated alongside the likes of Claude Monet, Odilon Redon, and Pierre-Auguste Renoir. Rops’s works articulated more diabolical motifs and grotesque sexual imagery than both his Impressionist and Symbolist contemporaries. As a result, his artworks are far less circulated and research surrounding Rops is scarce. This does not mean he was the only artist creating taboo media, however. Decadent art which depicted debauchery, excess, and sin was fairly present, especially in France. Rops collaborated with many playwrights, authors, and poets at the end of the 19th century, and was surprisingly popular among wealthy socialites of Paris and Brussels. Rops participated in many coalitions of artists across Europe, but the Les XX society is arguably one of the most significant. Artists, authors, and musicians exhibited their works and celebrated individualized aesthetics rather than a unified, nationalistic style. The group rejected a specific political or regional identity, which contributed to the notion of Belgium as a center for the Avant Garde. Rops’s display of Pornocrates elicited shock and commotion among the artists for its uncensored, sexual critique of high society men and women. This project aims to synthesize some of the scholarship surrounding the display of Pornocrates and serves to further highlight the significance of Decadent art in the Fin de Siècle period.

Between 1527 and 1587, three major pattern books were published in Venice: Giovanni Antonio Tagliente’s (c. 1465-1527) embroidery pattern book, Matthio Pagano’s (1515-1588) cutwork lace pattern book, and Federico de Vinciolo’s (active in Paris c. 1587-99) needlework lace pattern book. These three books represent the first of their respective types, and the beginning of a growing social attention toward lacemaking and its use as textile ornament. Each book provides patterns for different types of needlework: embroidery as applied decoration to fabric, cutwork (punto tagliato) as an evolution of embroidery, which involved cutting away the foundation fabric and filling it in with design, and finally, needlework lace (punto in aria), as freely formed design without a fabric foundation. The Venetian attention to dress and outward physical expression necessitated the use of increasing amounts of ornament—prompting the evolution of needlework into punto in aria designs which the city became famous for. Relevant through its additive aesthetic, public demand and commercial opportunities, lace became an essential export for the city of Venice and many women participated in its production, whether within their private homes, in workshops, or in convents. This essay will explore Venetian negotiations with myth and materiality through lacework as an ascendant medium, examining sixteenth-century pattern books and conduct literature as sources of tension and syncretism between written codification and social practice.

As a result of European colonialism, many objects were taken from the people of Africa. Using two case studies, I have explored the cultural significance of returning objects that were stolen or otherwise taken under questionable circumstances from West Africa during the rise of colonialism. One, an ivory mask of Queen Mother Idia, was taken from the Kingdom of Benin during the British Punitive expedition of 1897. The other one, a sculpture of a man- shark dedicated to King Gbehanizn of Dahomey, was taken by the French under suspicious circumstances. Exploring the cultural and monetary value of these items to the people that made them, as well as to the people that stole them and the value to the people that currently possess the items is something that will be discussed in this paper. I will also include any diplomatic relations that have occurred in reference to the works, citing sources from both sides of the current state of the field on the matter. After evaluating the value of these object to the people involved as well as in their removal from their original African contexts and current diplomatic proceedings, I argue that the return of these items is very important to the cultural heritage, identity and history of Africa and its people and thus should be taken seriously by all parties involved. 

Since the Renaissance, patrons have always held influence on the running of museums and exhibition choices. Increasingly, donors define museums standards, practices, and polices. In many cases, they even define their architectural designs. In this research, I examine all aspects of philanthropic activities, bringing in current debates and controversies associated with Museum finance. In particular, I look at trustees and board members who use their art world cachet to further their business investments and bolster their charismatic personas. In this examination, I consider the funding structure of museums and look at national scandals, such as former vice-chairman Warren B. Kanders at the Whitney in New York. Recently, the vice-chairman was asked to step down after it was reported to the public that Kander’s Company manufactured the tear gas that was used against migrants at the Mexican border. Though protest escalated to the extent of posting personal information on social media, only after artists withdrew their work did Kanders finally step down. I compare this with my own personal experience at the Museum of Northwest Art in La Conner. Obviously, solutions needed to amend this systemic problem, such as better vetting of donors, will requires a major cultural shift that potentially threatens future contributions coming forward. Nonetheless, in this research I highlight how the current system increasingly undermines the public's faith in our cultural institutions. Without any safeguards the current system will most likely lead to further corruption and abuse. Can we afford to have donors associated with public museums and, if not, can art museums continue to exist?

The recent rise of environmentalism encourages more people to learn about the science of climate change, which also extends very specifically to artists and journalists-- myself included. In the past two years at Western Washington University, I have worked with The Planet, an environmental magazine, as well as Klipsun, another Western publication, photographing images around ecological issues. I have also published articles on this topic for The Western Front newspaper. Firsthand, I have experienced the importance of art for the environmental movement, and that has led me to research artists at the forefront of these exciting developments. For example, Isabella Kirkland and Brandon Ballengee stand out with their drive to inform the public on human-induced climate change. For this project, I will analyze Kirkland’s Descendant (from her “Taxa Series”), a painting commemorating animals and plants whose populations are rapidly declining or assumed extinct. Kirkland is also a practicing scientist who extensively researches the subjects of her paintings, before presenting her anatomically-sized works. Concerned with themes regarding the Anthropocene, she creates odes to those biological life forms lost due to climate change. I will also discuss Ballengee’s The Frameworks of Absence, where he collected historical images and carefully cut-out extinct animals from those representations. According to Ballengee, this work is meant to represent the continual decline of biodiversity. Kirkland and Ballengee show their concern for the natural world through their creations. They share the intention to emotionally affect the viewer. In this paper, I am presenting my research on Kirkland and Ballengee’s methods: how they contribute to the growth of environmentalism and how their scientific methods produce startling works of art that compel the viewer to engage with climate change.

 Carpets are keystones in the study of Middle Eastern and Islamic arts. In Eastern display practices, carpets are experienced alongside other culturally specific objects in a rich contextual web of artifacts. Moreover, Eastern cultural sites display carpets on the floor. Yet, when these objects are brought into Western museums, they are hung on barren walls. This presentation draws on global scholarship of Islamic aesthetics to discuss Western practices of exhibiting Islamic carpets. When these textiles are displayed on Western walls rather than on floors, cross-cultural politics come into play and museum norms become suspect. In this presentation, I compare the carpet halls of the Metropolitan Museum of Art to several other carpet installations. The MET’s carpet display, with its vaulted ceilings and accompanying historical text, are a valuable case study in standard museum display techniques. Admittedly, these strategies provide important historical information and supplementary content, but extensive wall text in the museum discourages engagement. Unlike the MET, some institutions utilize period rooms to display their Islamic carpets. An interconnected web of related relics more effectively sparks wonder and facilitates intuitive understanding of other cultures. In this presentation, I argue that Islamic arts deserve to be displayed in a way that simultaneously provides cultural context and promotes cross-cultural understanding. Through comparison of the shows, I demonstrate that museums need immersive, haptic installations that fully transport the viewer and value physical and experiential engagement over mere textual explanations. Carpets must be thoughtfully installed if they are to enable a meaningful degree of knowledge and understanding. Islamic arts are distinctly underrepresented in current art history scholarship, collegiate-level instruction, and institutional exhibitions. In order to produce successful exhibitions of Islamic arts, curators and academics must examine historical failings and pursue innovative display techniques. This presentation attempts to expand that process of critical examination.

The Toba Batak sub-ethnic group in Indonesia is known for their weaving of the traditional textile ulos. At the same time that they uphold ancient Batak traditions of textile weaving, the Toba Batak people are at the nexus of a growing textile industry on national and international scales. As Western interests further influence Indonesia's economy, Indonesian concerns about the production of ulos become more and more urgent to address. The concern that ulos will be replaced by modern, Western styles of clothing and production gives rise to a more pressing fear that weaving traditions will fade from cultural awareness entirely.

In my presentation, I share the uniquely Indonesian strategies utilized by the Toba Batak people to conserve their cultural knowledge and traditional weaving practices. These strategies keep in mind the shifting trends in the textile industry in order to keep ulos relevant, yet are still respectful of Batak tradition. Primary methods of my research include interviews with traditional weavers, with weavers who supervised modern machine looms, the local non-weaving Toba Batak population, Batak culture experts, and Batak culture conservationists. Secondary research includes the work of Sandra Niessen, who has conducted large scale studies on Batak ulos and culture, and news sources such as The Jakarta Post.

This essay is a research project dedicated to the performative viewer in the museum experience. While every public space has an expectation of behavior, spaces set aside to showcase art provide a unique platform and opportunity for the art to attract attention. In viewing these artworks that are presented in a space designed to highlight them, the viewer or audience then performs the ritual that is unique to places of reverence and internal contemplation. Using psychoanalytical and anthropological methodologies along with direct observational data, this essay defines both what a ritual is and how the ritual applies to the participant viewer experience in art museums or art galleries. Pulling from the writings of Carol Duncan, Victor Turner, Helen Rees Leahy, and others; this is an attempt to relate the environment of dedicated art spaces to the environment and settings of the religious temple, and how the actions of the visiting participants mirror each other. In observing Knowledge Bennett’s mixed media triptych, “Comes in Three”, on view in the Western Gallery, and the audience it attracts, the resulting behavior of the participant viewer is recorded and used as empirical data for research. Results of my observations showed that my initial expectation for the amount of participant viewers was higher than what I witnessed, proving that further research and observation is required.
 

Green burial focuses on the interment of the body of a dead person in a fashion that supports decomposition so that the body can be naturally recycled. Green burial has grown exponentially within the past ten years and are now offered by hundreds of providers across the U.S., making it an increasingly popular alternative to conventional funerary customs, such as casket burial and cremation. While recent research has explored the conditions under which the green burial industry has become normalized and competitive, and the types of predictors of green burial support, we know little about the experience of participants and the characteristics of green burial ritual. To what extent have funeral participants internalized the industry’s goals of conservation and fostering an emotional bond with nature? Do green burial rites reinforce community among the living as with conventional funeral rites? Or does the ritual appeal mostly to participants sympathetic to environmentalism? To answer these questions, I conducted ethnographic interviews with green burial workers and participants and observations of green burial rituals. The results of my study furthered our understanding of the green burial as a growing social movement and new meaning making enterprise and revealed how supporters derived distinctive meaning within green burial.

The United States’ major American sports leagues, the NBA, NFL, MLB, and NHL, are turning to franchise expansion to increase their presence in the economic and media markets along the West Coast, including the NHL’s expansion to Las Vegas for the 2017 season and future expansion to Seattle for 2021. However, expansion franchises are very difficult to get off the ground since they require new ownership and management structures, buy-in from the local fan base, and are often subject to much higher scrutiny during their first few years than other franchises. Currently, no universal metric exists to determine whether or not an expansion franchises was successful, and it’s becoming increasingly more difficult to convince leagues to expand when they are unable to evaluate the likelihood of success for potential candidate cities. This research analyzed four major factors I identified that contribute to overall success for a team in any sport: financial prosperity, quality of national media coverage, public and local governmental support, and in-game accolades. Using these metrics as a starting point, I created an index that scored prior expansion franchises using a variety of indicators to quantify their overall success. Additionally, I applied this index to select cities currently under consideration for an expansion franchise to serve as a predictor of their potential success. Going forward, the use of this index as a predictive metric will allow leagues to make more informed decisions about the success of future candidates for expansion.

Over the last decade Seattle has seen the most dramatic population growth of any large American city. The accompanying rapid change in urban form of the city has been called staggering by longtime Seattle natives, causing a state of worry not unlike the feeling of solastalgia, a neologism that describes a form of mental or existential distress caused by environmental change. Longtime citizens can be forgiven for feeling as though their home has been irreparably changed in a way that doesn’t name them as a benefactor. But what has the effect of this rapid change had on the homeless community? Can a stable and strong sense of place have positive influences on the day to day lives of the homeless individuals? Can we facilitate a stronger sense of place as part of our response to the homelessness crisis in Seattle? My study examines the link between sense of place and stability in homeless individuals through interviews with individuals from within the homeless community, hearing perspectives from tiny house villages, tent cities, and the streets of Seattle. Through these conversations I demonstrate the impact that sense of place has on social stability, especially in times of rapid change to the urban environment, and how sense of place differs depending on the method of temporary shelter. The resulting study builds a better understanding of the intersectionality of the existential effects of placelessness in the homeless population, the differences between being homeless in a variety of spaces, and the meaning and significance of place making to people without a place.

Benefits of nature on children’s health and development are becoming increasingly recognized across the globe. Norway is revered for putting this research into practice, centering nature in early childhood education and setting precedents for ways in which preschools and kindergartens can get their children moving beyond the traditional classroom and up into the trees, down into the mud, and everything in between. Norway has taken a progressive stance on multicultural learning as well. The Norwegian Framework Plan for the Content and Tasks of Kindergartens defines education as an inclusive cultural arena to promote respect for the diversities of all children. I explored this intersection as a landscape architecture student. How might nature itself be important to children’s development and expression of cultural values? In what ways was learning facilitated differently outdoors vs indoors, and what implications might that have for the design of outdoor learning environments? Over the course of three weeks in Trondheim, Norway, I visited three barnehage (preschools) and conducted interviews on the connections between the landscape and the Framework Plan’s goal of inclusion. I found that outdoor environments could be less culturally coded than indoor classrooms, creating an unfamiliarity conducive to curiosity. This curiosity, coupled with undefined materials found in nature or man made objects placed outside of typical contexts, encouraged children to use play to design, communicate, and participate in imaginary worlds together, rather than having to rely on language or common frames of reference. Consequently, some Norwegian educators saw nature as a critical component of promoting children’s inclusion, tolerance, respect, and understanding of the diversities among one another, a revelation frequently overlooked in the U.S. More broadly, my findings point to that missed opportunity, where educational goals for children are similar but neglect serious consideration of the landscape as part of the approach.

Legal theorists have studied the advantage third party actors have in courts due to their powerful social and political presence. Third parties are interveners outside of the defendant and plaintiff that can provide comparative research, expertise, or assert a political opinion on a case. This study examines if, how, and why third parties affect international court decisions by expanding the development and protection of rights. I hypothesize that the presence and participation of third parties can lead to an expansion in the scope of treaty rights and findings of a violation. To examine these questions, I utilize the European Court of Human Rights Database (ECHRdb), focusing on cases including third party interveners in a single area of law, Article 8 (right to private and family life) between 1984 and 2014. The analysis codes interpret the types of organizations participating, and the ways in which the decisions expand or redefine convention rights. The preliminary findings of this research explicate the increase of particpation by third parties in international courts over time and how their presence can impact court jurisdiction. The results of this study are relevant for scholars and practitioners concerned with the growing role of the third party interveners in international legal processes and the impact of their actions on rights development and protection. 

In 2016, the Ukrainian Constitutional Court (UCC) went through internal institutional reform where the court amended the constitution to grant additional review powers and changed the procedure for the appointment of judges. These institutional changes were essential for the court to gain independence and legitimacy. This research paper examines how and why these institutional reforms impacted participation in the Ukrainian judicial system. I hypothesize that these reforms expanded societal access to the court which increased participation, accountability and enforcement of constitutional rights. To examine these questions, this analysis engages in a case law analysis of judgments of the UCC between 1997 and 2019. I mainly focus on the judicial review by individual and assess how the court has ruled in these cases. In this research, I expand our understandings of democratic participation to include society engaging with the judicial branch and bringing constitutional rights claims. Preliminary findings suggest that these new institutional reforms will broaden access to the UCC, which will increase participation within the Ukrainian judicial system. Implications of the research arise around the history of corruption and a legacy of distrust towards the Ukrainian government, and the possibility of a new avenue of democratic participation that may enhance and expand societal trust in government.

Recognition of LGBTIQ+ rights has expanded rapidly over the last few decades, yet remain contentious in many countries. Debates over the religious, moral and political stakes of acknowledging these human rights have persisted on a global scale. The purpose of this study is to understand how international legal systems are responsive to LGBTIQ+ legal claims. This is done through examining LGBTIQ+ rights complaint cases before the United Nations (UN) treaty-based bodies to analyze the degree to which LGBTIQ+ rights have expanded in scope and what factors impacted this expansion. The UN complaints procedures allow individuals or groups to assert claims and challenge the denial of rights by the State outside of their own national court systems. While committee decisions are not binding, they can be valuable advocacy tools that can empower domestic actors, validate claims of rights, and provide guidance for State action. The extent to which complaints processes are being used to assert the rights of LGBTIQ+ people has not yet been comprehensively examined. This paper will review decisions and state party reports published in the UN Jurisprudence Database and Treaty Body Database to consider how rights and politics have influenced treaty body verdicts, and then conclude that while broad language and increased political traction can lead to expansions of rights, narrow phrasing and controversial issues, such as family life, can restrict treaty body decision-making. Although the focus of this paper is on LGBTIQ+ rights, this study provides a general framework for utilizing complaint cases to analyze how specific human rights have progressed at the UN.

The Constitutional Court of Taiwan (Court) has been the subject of extensive research in the field of comparative judicial politics, recognized for its protection of civil rights and curtailment of Taiwan’s authoritarian government in the 1990s. However, the literature does not continue its examination of the Court into the 21st-century, and little is known about the Court’s ongoing role within the new democratic government of Taiwan. My research fills this gap in the literature by examining the activity of the Constitutional Court over the last two decades and its engagement with constitutional questions. What role has the Court played in the new, democratic polity of Taiwan? Has the Court progressively developed the body of constitutional law in Taiwan, or has it adjudicated conservatively over political and individual rights issues? To examine these questions, I analyze a number of cases from 1985-2017, coding Court decisions to determine the subject, frequency, and trend of its decisions. I also draw from existing literature to investigate these cases’ context and significance. Preliminary findings suggest that the Court has exercised judicial restraint when faced with constitutional questions involving salient political questions or institutional government powers, and deferred to other state bodies in fear of damaging its own legitimacy. In contrast, the Court has actively developed the constitution in individual rights issues and expanded civil liberties. My findings suggest that constitutional courts in new democracies cannot be categorized simply as either activist or constrained. Instead, judges selectively address constitutional questions and issue judgments that strike a balance between restraint and activism to strengthen their legitimacy.

Courts are a critical institution in democratic societies. Yet their role of protecting minority voices and clarifying executive and legislative power, forever place these institutions in a legitimacy crisis. Place this court in a transitioning democracy and the challenges of balancing political and societal interests are even greater. The Hungarian Constitutional Court (HCC) is my experiment. During the first period of the HCC (1990-1998), the Court relied heavily on activist constitutional interpretation and the use of abstract review through actio popularis participation in order to create legitimacy as a new institution and as a protector of the new Hungarian democracy. Through the following period, 1998-2010, the Court became much more moderate and the use of actio popularis decreased steadily. The purpose of this study is to test how changes in the HCC's methods of constitutional interpretation and access to the HCC affected the legitimacy and power of the HCC and how that legitimacy and power connects to the protection of a new democracy. To answer these questions, I engage in a historical analysis examining the criticism of the court during the first twenty years of the HCC, including an in-depth analysis of cases that use activist or text-based interpretation constitution and their negative impact on the HCC's legitimacy. I also construct a data set of the trends in the use of actio popularis within the court and an analysis of its effect of different levels of access on the Court's perceived legitimacy. Preliminary findings suggest that inconsistencies with the methods of constitutional interpretation used by the Court and decreased access to the HCC have decreased the Court's legitimacy to the point that the Court is no longer a powerful institution and no longer able to protect Hungarian democracy.

With rapid technological advances, robots, unmanned vehicles, and other artificial intelligence (hereby referred to as AI) – simulation of human behavior on a computer - entities proliferate into everyday lives. Increasingly, the military is using AI to put troops out of harm’s way; however, a question of criminal liability arises when certain weapons and drones could cause damage on a grand scale, at a distance, and with higher propensity. Who is to be held responsible for the potentially widespread war offenses of these automated systems when there is not necessarily someone controlling those systems on the ground: the manufacturer, the programmer, or the AI entity itself? While questions of such caliber have fanned the flames of widespread opposition to autonomous weapons, the purpose of this study is to determine ways for adjudicating them as used in war scenes instead of banning them, and to that end, this project turns towards International Laws (specifically Criminal Law) and explores the precedence set by past trialed cases to establish an understanding of responsibility as attached to certain individuals in mass violations. It further moves on to examine the types of adjudicated crimes and review statistical data surrounding the summoning of each of International Criminal Law’s provisions in individual cases as to develop a definition of prosecutable criteria in terms of weapons and destruction in order to find an outlet with jurisdiction to assess the admissibility of autonomous weapons. Preliminary findings bolster a greater demand for the International Criminal Court (that prosecutes grand scale murder and war crimes) as a candidate for trying individuals for AI-related violations. With the presence of such institution’s already well-defined regulations that can be expanded to encompass criminal liability for unlawful use of AI, this paper comes to the conclusion that fully autonomous weapons could be indeed of positive consequences.

The responsibility of the European Court of Human Rights [ECtHR] is to supervise the enforcement of the European Convention on Human Rights. The European Convention on Human Rights is an international convention to protect human rights and political freedoms in Europe. Among the cases that the ECtHR decides are legal framings of women’s rights as human rights. Although the literature on women’s rights in a human rights context is of abundance in the academy, there is a gap in understanding what an international judicial body like the ECtHR does for women’s rights. The purpose of this study is to fill this academic chasm by analyzing if, how, and why decisions made on cases involving violence against women either expand or narrow women’s rights on a global scale. I hypothesize that when the ECtHR decides cases on violence against women, there is a positive relationship between the actions of the ECtHR and the narrowing of women’s rights. To do this, I employ an original data set of compiled case law on violence against women rulings from the ECtHR from 1997-2019. Preliminary findings suggest a dominant understanding that there is a positive relationship between rulings on violence against women and the narrowing of women’s rights. These findings introduce the question of whether the ECtHR exercises a gendered legal consciousness, bringing us to a more robust understanding of how human rights law might overlook violations that disparately impact women.

Seabirds are often used as markers of ecosystem health due to their heavy dependence on the base of the food chain. The Alcidae family consists of small, diving birds who feed exclusively within the water column and rely on the sea year-round. Because of this, many view alcids as the only “true” seabirds. A research apprenticeship at UW’s Friday Harbor Laboratories on the pelagic ecosystem, Pelagic Ecosystem Function, has observed all seabirds since 2004, with consistent data since 2013. Alcids have been largely ignored in previous Pelagic Ecosystem Function studies, as the Common Murre (Uria aalge) artificially inflates the alcid family data due to their high abundance within the San Juan Channel. Upon removal of this species, it is found that non-Common Murre alcids are declining at a higher rate than any other seabird family within the channel, with a near-linear decline since 2013. In order to investigate the leading drivers of population decline, variables regarding food availability and habitat were collected in the form of chlorophyll, photosynthetically active radiation, and sea surface temperature. Compelling correlations were found between non-Common Murre alcid density and photosynthetically active radiation, as well as between chlorophyll and sea surface temperature. The data presented here is important not only for the mitigation of local ecosystem degradation, but also due to the consistency with global trends of seabird populations.

Urban stormwater runoff transports thousands of potentially toxic chemicals, metabolites, and degradation products to fish habitats. While certain contaminants, including metals and petroleum-derived compounds (e.g., polycyclic aromatic hydrocarbons) are known to adversely affect fish health, many others are unidentified and/or uncharacterized. Despite this uncertainty, green stormwater mitigation methods such as bioretention, the filtering of stormwater through a medium such as sand and compost, have recently been shown to be highly effective in terms of both reducing pollutants and improving water quality for the health of fish. However, the scope of this research to date has been limited to a small number of engineered media types. Here we used the embryonic zebrafish (Danio rerio) model to evaluate the impacts of urban arterial runoff, before and after filtration through several novel types of bioretention media. Measured toxicity indicators included eye and pericardial area, body length, and the expression of the biomarker gene cytochrome p450-1A (cyp1a). Untreated runoff from multiple storms was consistently toxic to zebrafish embryos. Conversely, each of the morphological and molecular health indicators was positively influenced by bioretention treatment. Our results indicate that bioretention compositions beyond the sand-compost mixture used in Washington State are promising in terms of reducing or eliminating near-term (i.e., acute) toxicity to fish. This expands the potential toolbox for site-specific pollution removal using green infrastructure methodologies.

In Spring of 2018, I helped collect a 45 cm sediment core from San Blas Basin on the continental shelf of pacific México, 50 km off the coast and north-west of Puerto Vallarta. After noticing that there were large sticks and seeds in the sediment, our Méxican collaborators gifted me the core to analyze lignin, a biomarker for terrestrial organic matter (TOM). Sticks in oceanic mud 50 km off the coast is unusual so we were excited to find out this geological story. I found that the total amount of lignin phenols ranges between 1.5 to 4.5 mg lignin phenol/10 g sediment except for an order of magnitude increase between 18 cm and 26 cm which was 20 to 40 mg lignin phenol/10g sediment. Acid to aldehyde ratios of vanillin-type lignin (Ad/Al (v)), a proxy for degradation, had a range of 0.36 to 0.58 while the section between 18cm to 26cm was fresher with a range of 0.26 to 0.28. S/V to C/V analysis indicated that the lignin is sourced from angiosperms. The section between 18 cm to 26 cm had statistically similar values ranging between S/V of 0.85 to 0.95 and C/V of 0.19 to 0.21. S/V to Ad/Al (v) analysis had a negative slope indicating S-type lignin is preferentially degraded and the background lignin signal could look more like the fresher 18-26 cm section if degradation had not occurred. My lab partners, Méxican collaborators, and I conclude that a large flooding event brought fresh TOM off of the land and the sediment deposition and deposited 8 cm of sediment in one event due to the S/V and C/V values overlapping. The flooding was likely a tsunami, large hurricane, or flash flood from the nearby Santiago river.

Along the shoreline of Possession Sound, located in the southern basin of the Salish Sea are 10 outflows of combined sewage systems. Combined sewage systems collect rainwater, untreated domestic sewage, and industrial wastewater within a single sewer line. When heavy rainfall occurs, these systems overflow and are directed into designated combined sewage outflows (CSOs), which then empty into the estuary, releasing E. coli (Escherichia coli) directly into the estuarine ecosystem. These CSOs, along with other factors, change the pH of the waters within the basin. Preliminary analysis of primary literature suggests a relationship exists between pH and E. coli growth. The pH change affects the enzyme growth within E. coli. As river discharge fluctuates, so does the amount of outflow from the CSOs which then cascades into pH changes at the site closer to the CSOs. The guidelines and regulations in place today allow for significant volumes of sanitary waste to be overflowed into marine systems. When river discharge increases, the overall pH within the Sound decreases. It was hypothesized that when there is a large amount of rainfall that leads to heavy river discharge and low pH, there will be more Escherichia coli growth at all of the sites throughout the Sound. Ocean Research College Academy students collected bacterial sample data at 12 stations in Possession Sound from 2009 to 2019. All data were recorded with distance from a CSO. A Niskin bottle was deployed at the surface and halocline with a YSI 650 testing pH. Samples were tested for bacterial count and compared with other samples taken after heavy rainfalls. Further research will define the trends in river discharge, pH and E.coli for Possession Sound.

As anthropogenic climate change progresses it is drastically altering the health of watersheds globally. In efforts to mitigate changes to marine ecosystems, many studies are using physical and chemical measurements to inform plans and create legislation. The impacts of climate change on local ecological communities are harder to track and take longer to show themselves which is why it is vital that we develop accurate techniques for measuring this kind of change quickly. This project, completed as part of Puget Sound Foraminifera Research Project at the Burke Museum, uses calcareous benthic foraminifera recovered and identified from sediment samples collected by the Washington State Department of Ecology between 2017 and 2018. Benthic foraminifera are marine protists that live on or within sediment and form shells of calcium carbonate or agglutinated sand grains. Foraminifera used in this project were stained with Rose Bengal to ascertain whether they were alive at the time of collection and grouped according to World Registry of Marine Species protocol. Stained and unstained individuals were counted to create living and dead assemblages. The goal of this study is to determine the validity of using total assemblages that include both living and dead foraminifera as a proxy for quantifying the living assemblages in Puget Sound. This is important because previous research has found discordance between living and total assemblages of molluscs, pteropods and ostracods in embayments heavily impacted by anthropogenic activity. This study includes 5 embayments in Puget Sound. Results from Bellingham Bay and Sinclair Inlet suggest that the validity of using total assemblages as a proxy for living assemblages may vary across different areas of Puget Sound; while the total assemblage and living assemblage matched in Bellingham Bay, Sinclair Inlet has been found to have significantly different total and living assemblages.

Eelgrass beds in central Salish Sea are critical components of healthy ecosystems that are vulnerable to anthropogenic impacts. This study utilized two locations in Possession Sound to monitor water chemistry within and near two different eelgrass beds; one unmapped, and one established. The study compared these two locations and investigated the impact of location on water chemistry within the bed. Data-sharing and collaboration with the Samish Indian Nation Department of Natural Resources’ work on eelgrass beds in Fidalgo Bay provided a broader scope into regional differences within the central Salish Sea. This study was conducted by Ocean Research College Academy (ORCA) students at eelgrass beds in Possession Sound located near Mukilteo and Hat Island. The study ran from October 2019 to spring 2020, and utilized background data from past studies to inform studies at the bed near Mukilteo. Data were collected using a combination cast of a camera collecting visual data and a CastAway CTD, which collected vertical profiles of salinity and temperature at recorded geographic coordinates. Data were collected during a free drift across the eelgrass bed. An EXO Sonde was temporarily installed in the bed to collect chlorophyll and turbidity data in a longitudinal manner. The study primarily explored how location impacts the water chemistry eelgrass beds in central Salish Sea both within the bed and within a region. Research going forward could study remote beds more comprehensively using technology previously tested such as drones and SONAR, as well as a longer-term collaboration between the Samish DNR and ORCA.

As climate impacts are amplified in the nearshore regions, the Possession Sound located near the heart of the Salish Sea is a key study area of the local marine ecosystem. Systems such as these are very sensitive to fluctuations in temperature. For example, in 2015, “The Blob,” which was a massive body of abnormally warm water that ranged from California north to Alaska, raised salinity and lowered DO causing high mortality rates in a range of taxa. Events such as these are referred to as Marine Heat Waves (MHW). In 2019 analysis of satellite thermal imagery data concluded that another Marine Heat Wave had struck the West Coast displaying temperatures reaching as high as seven degrees Fahrenheit above average. Additionally, parts of the Salish Sea have observed some possible influence of the MHW from the Pacific. In order to determine the localized impact of these Marine Heat Waves, I used data from a fixed CTD probe from the Ocean Research College Academy were analyzed, focusing on parameters temperature, salinity, and dissolved oxygen. I hypothesized that there is a delay of increased temperature from the Pacific due to possible mixing of coastal currents traveling through the Juan de Fuca Strait into the Salish Sea. When exploring this data set, I have found that since 2009 average water temperatures have risen, 2019 average temperature is .4 degrees higher than in 2009, with 2015 having a peak temperature average of 11.5 degrees Celsius, 1.1 degrees higher than 2009, which aligns with “The Blob” of 2015. Trends have shown an increase in average temperature since 2009, with MHWs event being a prevalent factor in rising average temperatures, and lower dissolved oxygen averages.

Neural tube defects (NTDs) impact 3000 pregnancies a year in the US and are caused by both genetic and environmental factors. NTDs arise from errors in neural progenitor signaling, migration, proliferation, and differentiation during embryonic development. Spina bifida (SB), a prevalent NTD, can reduce the functioning of neural pathways responsible for pain and motor function in the lower body. A recent study discovered a novel variant of the receptor GPR161 present in screened infants with SB, but absent in all screened infants without. GPR161 is a G protein-coupled receptor localized in the primary cilia known to participate in the regulation of the pathways of key stem cell differentiation ligands, sonic hedgehog (Shh) and Wnt. Our study seeks to investigate the molecular mechanisms which connect the novel GPR161 variant p.Trp202Gly to neural tube defects using an in vitro model of neural stem cell differentiation. GPR161 variant and knock-out (K/O) lines are generated using CRISPR Cas9 technology in induced pluripotent stem cells (iPSCs). iPSCs are then guided through neural differentiation and harvested for analysis at multiple key stages of neural progenitor development. Markers of neural differentiation, SB, and downstream GPR161 factors are analyzed using western blot, RT-qPCR, immunostaining, and RNAseq. We expect to see a change in Shh activity in the variant line compared to the WT.

Alzheimer's disease (AD) is the most common cause of dementia, a general term for memory loss and other cognitive abilities. Although this disease has been a major research focus since the 1980s, the pathological mechanisms are still not well understood, and therapeutic interventions have been ineffective. The most definitive method for classifying AD is through identifying accumulations of toxic amyloid-beta (Aβ) and tau proteins in post-mortem brain tissue. Dr. Su-in Lee’s lab has developed a machine learning method that integrates the pathological protein phenotypes with gene expression levels in the same brain tissue. They have highlighted 25 genes with expression level changes that correlate with the tau and Aβ protein aggregation phenotypes. For this proposal, I have integrated these human neuropathology-based phenotypes with the genetic power of Caenorhabditis elegans (C. elegans) to directly test the impact of these candidate genes on the cellular pathology. Previously, all C. elegans tau models had neuronal specific expression. However, neurons are resistant to RNAi. Therefore, I generated a novel transgenic C. elegans tau AD model that has been codon-optimized to express tau in body wall muscles instead of neurons. I measured the animal’s health with age in a series of phenotypic assays: egg-laying, growth, movement, paralysis, and lifespan analysis. This line exhibits premature paralysis and decreased crawling speeds, providing an easy to score phenotype. This new model allows for high-throughput RNAi screening to test the identified 25 genes’ effects on worm health by utilizing the automated worm-movement technology developed in the Matt Kaeberlein lab that can simultaneously determine the rate of paralysis of thousands of worms. The results of my genetic screening will lead to a better understanding of the human genes that are dysregulated in human AD brains, provide a basis for genetically-dissecting the pathways influencing tau toxicity, and suggest new therapeutic targets.

Substance abuse leads to alterations in cognition that affects processes such as impulse control and valuation. Decreased impulse control and aberrant valuation are responsible for continued drug seeking and are thought to be escalated by external stress stimuli. Stress leads to release of an endogenous opioid neuropeptide called dynorphin which binds to the Kappa Opioid Receptor (KOR). Upon KOR binding, dynorphin induces a protein signaling cascade that also promotes drug seeking behavior. In this study, we investigated the dynorphin/KOR system in the medial prefrontal cortex (mPFC) due to its critical role in cognition. We examined the properties of dynorphin release in the mPFC of C57BL/6 mice in response to different external stressors to determine if this nucleus is a potential therapeutic target for stress-induced drug seeking behaviors. Using a pharmacological approach, we first showed that systemic administration of U50,488, a KOR agonist, leads to KOR activation in the mPFC. U50,488 administration also disrupted cognition by impairing performance in a working memory behavioral task. We next tested whether different stress modalities stimulated mPFC dynorphin release and disrupted cognitive performance. Surprisingly, repeated forced swim stress did not cause dynorphin release in the mPFC and did not disrupt cognitive performance although it did activate dynorphin release in the Dorsal Raphe nucleus, as expected. In contrast, different stressors, including repeated foot shock and precipitated morphine withdrawal did effectively lead to KOR activation in the mPFC. This indicates that dynorphin release in the mPFC is dependent on the type of behavioral stress. Future experiments will utilize an in-vivo dynorphin sensor, kLight, to detect dynorphin release in real-time in response to these stressors. Exploration of the differences in dynorphin/KOR system functioning in response to different stress modalities is important for establishing how this system may be targeted to alleviate stress-induced drug seeking behaviors.

Alzheimer’s Disease (AD) is a neurodegenerative disorder characterized by the formation of senile plaques and neurofibrillary tangles through the accumulation of toxic amyloid-beta and Tau protein. There is growing recognition that extracellular vesicles (EVs) can package and transport toxic peptides associated with neurodegenerative disorders – such as AD – to other cells in the brain. Researchers in the Kaeberlein lab have designed methods to isolate these type of vesicles from C. elegans nematodes, a popular invertebrate genetic model. However, current nematode EV purification methods do not permit the following of EV signals from specific tissues when they are under AD proteotoxic-stress. I have generated a transgenic C. elegans AD model that has muscle specific expression of the pathogenic human Tau protein. The protein coding sequence was designed to use optimized codons to ensure high expression of the transgene. I have also generated transgenic nematode lines that express versions of known transmembrane proteins with small affinity tags in a tissue specific manner. The small affinity tags on the proteins make it possible to specifically pull down the EVs from designated tissues through standard immunohistochemistry techniques. The abundance of tissue-specific EV protein and RNA cargos from transgenic lines with or without human Tau have then been quantified using LC-MS-MS and RNAseq analyses, and parsed and condensed into a MySQL database via a C# program. The database allows for simple searching through large amounts of MS data, making data analysis more efficient and effective. Thus the methodology and tools I develop in this project could become a promising new approach for identifying novel therapeutic gene targets and biomarkers of AD stress.

Traumatic spinal cord injuries (SCI) are a catastrophic type of injury that disrupt many functions of daily life such as mobility, temperature regulation, sexual function, and bowel/bladder use. Approximately 80% of individuals who suffer from SCI develop neurogenic bladders, and subsequently require catheterization. Patients with sacral SCI have different neurogenic bladders than those with higher injuries; specifically, severing the peripheral innervation at the sacral level causes a non-contracting and low pressure bladder, without the scar tissue found commonly in higher injuries. This suggests that peripheral denervation decreases the incidence of non-compliant bladder complications. Bladders primarily have two uses - storage and voiding during urination. One common strategy to improve the storage function of neurogenic bladders is to denervate them with direct injections of Onabotulinumtoxin-A (ONA). However, this treatment is only applied after other treatments have failed, leaving time for significant damage to occur. In the current study, we propose to study the effects of direct ONA injections acutely after SCI. Our two aims are 1) to determine the treatment window for chemodenervation that best reduces bladder wall hypertrophy and fibrosis, and 2) to determine whether bladders remain compliant after chemodenervation. Using a rat model, we applied two treatments - SCI + Saline, SCI + ONA, with both saline and ONA directly injected into the bladder wall. A control group received laminectomy without injury. After treatment, we plan to measure bladder function using a urinary incontinence scale, as well as histological measurements of collagen deposition and muscle area to quantify bladder wall hypertrophy and fibrosis. We anticipate that early ONA injection (as defined in Aim 1) will be associated with less fibrous and more compliant bladders in the long term when compared with non-chemodenervated bladders. If our hypotheses are correct, we plan on getting IRB approval for a clinical trial.

Dopamine (DA) neurons found in the ventral tegmental area (VTA) are associated with reward feedback, and dysfunction in DA circuitry is associated with disorders such as Parkinson’s, schizophrenia, bipolar, and addiction to drugs. To adequately treat these diseases, we must have a more complete understanding of how dopamine contributes to emotional processes. This research project addresses this issue by investigating neuropeptide regulation of dopamine neurons in the VTA. The bed nucleus of the stria terminalis (BNST) is a brain region that expresses many neuropeptide genes and sends strong projections to the VTA. We utilized Cre driver lines to isolate neurons that produce the peptides Neurotensin, Neurokinin B, and Corticotropin Releasing Factor. We injected a virus into the BNST that induces the expression of a light activated ion channel and allows us to stimulate axon terminals in the VTA. I then conducted behavioral experiments to assess the effects of activating these peptidergic inputs. Dopamine-dependent behaviors relating to pleasure, reward, and anxiety were measured through the behavioral tests of Real Time Place Preference, operant conditioning, and Open Field respectively. Most likely due to low expression, my behavioral analyses did not yield statistically significant results. Moving forward, it may be necessary to increase viral titer for wider expression. In the future, I intend to use CRISPR/Cas9 technology to isolate neuropeptide function from fast neurotransmitter release in these circuits. This research, by producing findings that help explain how neuropeptides modulate DA neurons, has the potential to generate advances for the understanding and treatment of dopamine-related psychiatric disorders.

With cardiovascular disease as the leading cause of death worldwide, there is a need for improved wearable monitoring tools for assessing the health of the cardiovascular system. Photoplethysmography (PPG) is a continuous, non-invasive measurement that encodes a multitude of informative vital signs, including heart rate, heart rate variability, respiratory rate, cardiac output, and arterial stiffness. Although existing PPG sensing technologies record from the finger or wrist, the face presents a promising and underutilized location for wearable pulse sensing. This work presents a novel wearable PPG sensing system that records at multiple wavelengths and facial locations. As a proof-of-concept, we seek to evaluate a potential application of our system incorporated in a surgical face mask for use in intra-operative hemodynamic monitoring. By collecting data with our system alongside ground truth cardiovascular vital signs, we can build and test non-invasive inference algorithms. After validating our system’s heart rate detection accuracy with a standard error of 2.84 beats per minute, we now proceed to test our device’s ability to infer additional cardiovascular parameters. In addition to showing promise for novel non-invasive, continuous surgical monitoring, this work has broader implications for wearable health applications based on face-worn form factors such as glasses, helmets, and headsets.

Blood pressure (BP) serves as the primary indicator of a patient’s cardiovascular health. Today, cuff-based BP monitors are the gold standard for routine blood pressure monitoring. However, they are prone to inaccuracies and cannot provide continuous readings. Continuously monitoring BP would allow patients to observe their BP fluctuations from eating, medicine intake, and exercise, thus empowering individuals diagnosed with hypertension to make better-informed health decisions. This motivates the need for a non-invasive, continuous blood pressure monitor. Prior studies have already shown the potential for pulse transit time (PTT), which is the time for a pulse wave to travel between two arterial sites, to be used for non-invasively measuring BP. In this work, I hope to improve upon this technique. To do this, I focus on testing and improving the pulse detection accuracy of a system incorporating an optical sensor array in a surgical eye protection face mask. By getting a better resolution of the pulse waves, I believe the estimate of BP will be more accurate and, in turn, provide a valuable dataset to further investigate the relationship between PTT and BP.

Cellular regulation of transcription is a complex phenomenon, with a wide range of biological determinants influencing the binding of transcription factors (TFs). Being able to accurately predict TF binding is important to pinpoint noncoding DNA regions where mutations are likely to cause disease. Existing approaches are able to accurately predict TF binding across the genome, but are not focused on interpretability. Our approach, EXPERT: Explainable Prediction of Transcription Factor Binding based on Histone Modification Data, achieves state-of-the-art predictive performance while simultaneously offering local interpretations that reveal biologically meaningful relationships by using a stage-wise boosting technique and local additive feature attributions. We utilize a performant non-linear model (XGBoost) and an efficient local feature attribution method (TreeSHAP) to demonstrate how to increase the signal learned from a set of biologically meaningful features (histone binding in DNA) using a stage-wise gradient boosting scheme while maintaining high performance. EXPERT enables understanding predictive models through biological explanations and can further our foundational understanding of the epigenome by highlighting novel biological relationships.

The dystrophin protein protects cardiac and skeletal muscle from damage during normal contraction and relaxation by acting as a shock absorber in the cell. Mutations in the dystrophin gene lead to Duchenne muscular dystrophy (DMD), an X-linked recessive disease. Boys suffering from the disease become ventilator dependent at a young age and usually succumb to cardiac failure in their thirties. Other symptoms of DMD include muscle wasting, cardiomyopathy and respiratory failure. Currently there is no cure for DMD and while gene therapy has shown great promise, it still needs to be complemented with additional therapeutic interventions in order to fully address the symptoms of DMD. Previous work by our lab identified several drugs that blocked a certain type of calcium channel in cardiac muscle and protected the cells from damage following injury by correcting calcium movement into and out of the cell during muscle contraction. In this study, the leading drugs will be tested further using a cross-platform approach. The drugs will initially be tested in cardiac and skeletal muscle differentiated from healthy and DMD patient-derived induced pluripotent stem cells (iPSCs). The top three drugs that restore normal contraction and relaxation kinetics in vitro will then be tested in the DMDmdx rat. This novel, small animal model has a similar progression of DMD symptoms to human patients. The rats will be fed the drugs in their chow. Skeletal and cardiac muscle performance will be evaluated to determine whether correcting muscle contraction kinetics ameliorates the symptoms of DMD. Furthermore, we will show whether the same drug is equally effective in treating both cardiac and skeletal muscle. The cross-platform approach may help to better predict drug efficacy, leading to a reduced rate of failure in clinical trials. Moreover, this approach may serve as a benchmark for drug discovery in other neuromuscular diseases.

Blood flow in microcirculation is a significant physiological parameter that reflects the adaptive response of organs to disease, trauma, and cancer. Although ultrasound Doppler imaging was previously unable to assess blood flow in the microvasculature (< 0.5 cm/sec), the introduction of microbubble contrast agents has removed this limitation. However, blood flow of the entire vascular tree is mixed together during imaging. We present a method that segments and visualizes the entire vascular tree, including capillary blood flow, larger sub-spatially resolved vasculature and larger vasculature (>50 µm). In this work, we present an approach that decomposes nonlinear Doppler acquisitions into different groups of velocity projections. We demonstrated the ability to segment these different levels of vasculature in a rat spinal cord injury model where the varying rates of low velocity microbubble decorrelations captured by our high frame rate acquisitions enable us to quantify microvascular blood flow. This approach overcomes limitations encountered in conventional imaging methods by removing tissue signal before Doppler processing by combining high-frame rate plane wave imaging, microbubble nonlinear pulse sequences, and Doppler segmentation of blood flow. Singular value decomposition was used to segment the nonlinear Doppler signal. Our results successfully illustrate the segmentation of lower velocity sub-resolution microvascular flow and higher velocity flow in larger vessels in a rat spinal cord injury model. We isolated low and mid-velocity flow in sub-resolution vasculature (<20 µm). We observed different spatial distribution and bolus kinetics between low- mid- and higher velocity Doppler projections. We are assessing the utility of these different blood flow features for the management of spinal cord injury and other applications (e.g. oncological).

Heart disease is the leading cause of death in the United States, contributing to 1 in every 4 deaths. One of the most common causes of heart failure is dilated cardiomyopathy (DCM), and approximately 35 percent of these cases are caused by mutations in genes encoding for a variety of sarcomeric, cytoskeletal and nuclear envelope proteins. There are nearly 1,500 separate mutations in sarcomere protein genes that can lead to DCM, but patients are not currently treated differently based on these unique mutations. In this study, I use two mouse models of genetic DCM, D230N and I61Q, which have mutations that occur in cardiac Tropomyosin (Tm) and cardiac Troponin C (cTnC), respectively. These mutations result in desensitization of the cardiac myofilament, leading to negative health consequences such as left ventricular dilation and reduced ejection fraction. I am testing the hypothesis that the ionotropic Levosimendan will be more effective in the I61Q cTnC DCM model because the drug binds directly to the N-terminal domain of cTnC, increasing its affinity for Ca2+ which results in increased force generated. At 3-4 months of age, cardiomyocytes are isolated and suspended in Tyrode’s buffer, an isotonic physiological solution. Cell shortening and relaxation was measured using video microscopy (IonOptix, Milton, MA) with pacing at 1.0 Hz and 37°C in presence of Levosimendan ( 1 µM) or control (DMSO). Compared to DMSO, 1 µM Levosimenden increased myocyte shortening in NTG mice (9.36 to 13.02 %, n=5) and I61Q cTnC (7.3 to 8.4 %, n=3) but had no impact on D230N Tm mice (10.4 to 8.2 %, n=5). My results suggests that Levosimendan is more effective in I61Q cTnC compared to D230N Tm model. In future studies I plan to make similar measurements using the myosin activator Omecamtiv Mecarbil.

Humans are incapable of regenerating a majority of their major organs and tissues following traumatic injury, often resulting in an irreversible loss of function. Tadpoles of the frog genus Xenopus can regenerate multiple tissue types in response to injury, however this capability is lost after metamorphosis. This stage-specific regenerative capacity makes Xenopus a uniquely powerful model for studying factors that promote regeneration. Tadpoles develop ex-utero and do not develop mouths until days after fertilization. Before tadpoles are able to ingest exogenous food, they rely instead on maternal yolk stores for sustenance. A regenerative refractory period has been described in tadpoles of Xenopus laevis, in which regenerative capacity is transiently lost. In this study we describe a similar refractory period in the closely related Xenopus tropicalis, and observe that the onset of the refractory period aligns with the transition independent feeding. Based on this observation, we hypothesized that the lapse in regenerative capacity could be due to a lack of metabolic fuel. We used immunohistochemistry (IHC) against the yolk protein vitellogenin (vit) to study the utilization of maternal yolk stores during tadpole development. We find that yolk localization is dynamic over the course of development, and that it is ultimately is depleted by the onset of the refractory period. We additionally used IHC against phospho-Histone 3 (pH3), a marker of mitosis, to study proliferation during development and regeneration. We found that proliferation declines across development heading into the refractory period, in both uninjured and amputated contexts. Lastly, we successfully rescued both regeneration and proliferative rates by feeding tadpoles after they develop the ability to eat. As a whole, this work articulates that nutritive stress may contribute to the loss of regenerative capability in the refractory period, and that alleviation of this stress promotes regenerative ability in this context.

Induced pluripotent stem cells (iPSC) have a high potential, for they can be differentiated into any cell type for regenerative medicine, drug discovery, developmental biology, and disease modeling. However, iPSC’s and their differentiated progeny display an undesired variability in their shape, contractile properties, growth rates, etc. Identifying subsets of phenotypes in iPSCs and their differentiated progeny will allow us to optimize tissue models for research. Here, we generated a rainbow reporter line in iPSCs that can track individual cells as they clonally expand and differentiate while providing phenotypic information. Knocking in four copies of a cassette containing three distinct fluorescent proteins allowed the expression of up to eighteen different colors. However, not all colors were present in equal proportion, increasing the probability that distinct lineages could have the same color. To achieve an equal color distribution, colored cells were isolated by sparsely plating a culture of mixed colored cells. After a week of expansion, individual colonies were picked and imaged under a spinning disk microscope to determine the color of the colony and whether it was single lineage or mixed. Viable cell lines were isolated and frozen in stock. These cells will be examined for markers of cell proliferation, pluripotency, apoptosis and quantitative RNA expression analysis to confirm that the color barcoded iPSCs act the same as non-engineered iPSCs. To date, we were able to create eight color barcoded iPSC lines for further experimentation, increasing the concentration limit of colored cells in non-colored cells by five-fold. The next step will engineer 3D tissues by growing iPSC-derived cardiac cells in a mold to simulate in vivo tissue development. Colored coded cells will allow us to track how the initial location/physical stresses/phenotype of an iPSC-derived cardiac cell in an engineered tissue determines its tissue layer and cell type.

This research focuses on dissecting the molecular mechanism of cardiac regeneration in the animal model, zebrafish, upon a myocardial infarction like injury. Zebrafish are one of the few vertebrates that can fully regenerate their hearts after an injury in 30 days. This phenomenon is not seen in humans, who generate scar tissue after this injury with reduced circulatory efficiency. However, there is evidence that neonatal mice under 7 days old can regenerate their hearts, but this is lost upon adulthood. Determining this pathway is the first step to develop therapeutics in order to provide relief to people suffering from cardiac injuries. In this research, we used chemically ablated transgenic zebrafish to generate a 30% injury. We determined that upon an injury, both the Wnt pathway and the mTOR pathway are sequentially activated and upregulated to restart cardiac proliferation to regenerate the heart. Wnt pathway proteins like Axin and β-catenin are activated 3 days post injury and mTOR proteins like pS6 are activated gradually over 7 days post injury. The inhibition of the Wnt pathway using DKK showed a downregulation of the mTOR pathway and downregulation of cardiomyocyte proliferation. Inhibition of the mTOR pathway using Rapamycin also stopped cardiomyocyte proliferation from occurring. Mass spectrometry data showed a decrease in glutamine and an increase in leucine during the proliferative phase. Since leucine is one of the activators of the mTOR pathway, we see that the glutamine-leucine transporter is also upregulated post-injury. Thus, we show that heart regeneration in adult zebrafish occurs via cardiomyocyte proliferation by using the Wnt and mTOR pathways to upregulate cardiomyocyte proliferation upon injury.

Over the past fifty years or so, scientists have successfully isolated microorganisms, known as endophytes, from inside nearly every region of the plant, including the roots, stems, and leaves. Some of these microorganisms have the ability to fix atmospheric nitrogen, and have even been shown to confer an array of additional benefits to their plant hosts, ranging from fungal pathogen resistance to increased stress tolerance, and more. Recently, many scientists conducting endophyte research have observed their endophytes losing their plant-enhancing activity after extended periods of isolation from their original plant hosts. This makes the plant-enhancing effects of the isolated endophytes impossible to study, and more importantly points to a crucial need for a better understanding of plant-endophyte communication mechanisms. Because reactivation of several endophytes was observed after introducing native plant extract back into the isolated endophytes’ medium, we have embarked on a journey to determine what in the plant medium causes these inactive endophytes to regain their plant-enhancing abilities. To do this, we have first determined the level (transcriptional, translational, or post-translational) at which nitrogen-fixation, a crucial symbiotic activity, is regulated by the presence of plant extract in one of our endophytes. We then designed and built a reporter plasmid to track the expression of key nitrogen-fixing genes, and will next fractionate the native plant extract and use this reporter plasmid to monitor the effects of different plant fractions. This will hopefully lead to key insights into the potential plant signal that initiates many symbiotic endophyte activities. The exploration of the plant-endophyte symbiotic signaling system has the potential to impact the fields of agriculture, forestry, and environmental sciences worldwide.

Helicobacter pylori is a gastric bacterial pathogen that infects more than 50% of the world's population. H. pylori colonizes and persists in the harsh, acidic environment of the human stomach yet it is not an acidophile. The ability of H. pylori to successfully colonize the stomach is dependent on several factors. This includes helical shape that is thought to enhance initial colonization, urease-dependent ammonia production which works to neutralize the pH, and chemotaxis driven motility that enables the bacteria to navigate down the acidic gradient towards the neutral gastric mucosa, its preferred niche. Studies show that flagella-driven motility plays a key role in the pathogenesis of H. pylori; motile strains display higher infection rates and colonize the stomach longer in germ-free piglets compared to non-motile strains. Although the antrum serves as the preferential colonization site due to its lacking acid producing parietal cells, H. pylori can migrate and colonize other sites. In this project, we used a soft-agar assay to characterize the spreading motility of a collection of 42 H. pylori clones isolated from a single host at two different timepoints and from different gastric regions. Our results show that there are quantitative and qualitative differences in the spreading motility of these clinical isolates. The average halo diameters measured are collectively higher in the curved rod isolates from the earlier timepoint compared to helical and rod-shaped isolates from the later timepoint. The clones also display variable chemotactic patterns in soft agar assay with most strains producing concentric rings and a few strains forming uniform halos characteristic of nonchemotactic mutants. By assessing the differences observed, our study explores the factors that impact motility, such as flagellation and cell shape, in order to provide insight into these traits that best equip H. pylori to persist in the stomach. 

HOX genes encode for a group of homeodomain-containing transcription factors essential for patterning the skeleton in the developing embryo.  They are expressed in spatially restricted domains, where they regulate the patterning of specific bones that form within these expression domains.  The expression of Hox genes is continued into adulthood, however their function in the adult skeleton is relatively unknown.  We speculate that HOX genes regulate post-embryonic growth of bony elements which reside within their spatially-restricted expression domains.  In support of this hypothesis, we present three key findings.  First, we identify a subset of vertebrae in zebrafish (V2-V5) whose size and shape are under the control of hox5 genes.  These vertebrae reside in spatial domains associated with hox5 expression in embryonic zebrafish.  Second, we show that severe loss of hox5 induces gross changes in bone morphology, including loss of branching in the fourth ribs and sporadic fusion of the tripus with the fourth ribs.  Third, we note that fish with moderate loss of hox5 exhibit reduced size of bony elements but no obvious changes in bone morphology.  Our studies provide early evidence of the role of HOX genes in mediating post-embryonic growth of the skeleton.  They also help to elucidate potential vertebral homologies in zebrafish- an emerging model for orthapaedic research- and cervicothoracic vertebrae in mammals.

Epigenetic proteins modify the chromatin structure to manipulate gene expression, and dysregulated epigenetic modification in cells has been linked to cancer formation. Previous studies on young female Drosophila have shown that after injury, germline stem cells (GSC) are capable of entering and exiting a protective state called quiescence. When exposed to ionizing radiation (IR), the apoptotic differentiating daughter cells send a protective signal to GSC, resulting in GSC quiescence. This survival behavior of GSCs validates them as a potential model for cancer stem cells, which are a subset of tumor cells that are capable of withstanding traditional chemotherapy through reversible quiescence, resulting in future tumor relapse. To identify genes required for GSC survival, we performed a spatially restricted RNA interference (RNAi) screen. Here we show that two members of the repressive epigenetic regulator complex PRC1, Pc and Sce, are required for entry, while demethylase Utx is required for exit of GSC quiescence. Notably, PRC2 dependent H3K27me3 marks are required for PRC1 function, and Utx is required to erase these PRC2 dependent H3K27me3 marks. Importantly, we detected around a 3-fold increase in H3K27me3 marks in GSC following IR, suggesting that the repressive PRC1-PRC2 dependent complex is critical for entry, and elimination of PRC2 dependent marks is critical for exit from the quiescence state. Furthermore, we show that Trx, a writer enzyme which promotes euchromatin formation through H3K4me1 addition, is required for GSC exit from the quiescent. These data suggest that reversible quiescence in GSC is controlled by specific epigenetic states. In the future, more work is needed to investigate gene specificity of the epigenetic regulation.

The Polycomb Repressive Complex 2 (PRC2) is an important epigenetic remodeler in developmental transitions and cell fate determinations. PRC2 is responsible for the addition of H3K27me3 marks that repress developmental gene expression. The catalytic subunit of PRC2 is the methyltransferase (Enhancer of Zeste 2) EZH2 which binds to EED (Embryonic Ectoderm Development) to methylate H3K27 on gene promoter regions. To investigate the requirement of PRC2 in different developmental transitions, a computationally designed protein was utilized to inhibit EED-EZH2 interaction. The novel designed protein is named EED binder (EB) and competes over endogenous EZH2 on the EED binding cleft with 300 times greater affinity than endogenous EZH2. We cloned EB-GFP under heatshock inducible promoter and injected this construct to one cell zebrafish embryos to generate a germ line transmissible insertion. To study the requirement of PRC2 in early developing embryos (0-3dpf), we applied heatshock (HS) on EB-GFP positive and negative embryos. Western blot analysis revealed global downregulation of EZH2 and H3K27me3 in EB-GFP positive, but not control embryos. Additionally, Co-Immunoprecipitation experiments showed EB-GFP binding to EED. Finally, to test the requirement of PRC2 in caudal fin regeneration, adult (5 month old) EB-GFP positive and negative animals were fin-amputated and the regeneration growth rate was measured for 14 days. Our results show that EB-GFP positive fish were able to regenerate their fins faster, resulting in a large fin size compared to either negative or non-HS clutch mate. Overall, we have developed a computer designed inducible PRC2 inhibitory system to study PRC2 function in Zebrafish, at the whole animal level. In the future, we will utilize EB-GFP to explore PRC2 and other epigenetic modifiers that are required for tissue and organ regeneration before and after injury.

There is currently extensive demand for optical media like CDROM, DVD and Blue ray disks for data storage with computer technologies. Here we combine mathematical modelling studies and photonic laser diffraction experiments to study the optimization of data storage in different types of optical media. Using calculus-based studies, we estimated the data storage capacities in these systems and calculated the CD, DVD and blue ray disk arc length and data storage linear densities. These are in good agreement with reported values. Using red, blue and green laser sources at our photonics lab, we conducted laser diffraction studies and estimated the line spacing of CDROM, DVD and Blue ray disks. The advancement from CDs to DVDs yields higher data storage densities. In the high capacity blue rays disks, because the physical structures called pits that store data on the disks become smaller, there are other challenges in realizing these smaller devices, which make it more expensive. The CD/DVD players' lasers operate at the diffraction limit resolution of light and provide maximum data capacity for their geometry. Magnetic media like floppy disks, hard disk and magnetic tapes are also used for computer data storage. We have estimated the maximum data storage capacity from magnetic floppy discs. We used curve fitting methods to analytically represent the magnetic read-back pulse as Lorentzian functions for data modeling. Our studies provide an integrated STEM learning of data storage in optical and magnetic media.

There are many ways to axiomatize arithmetic. It is natural to ask which axiomatizations place stronger conditions on the structure of arithmetic than others. The language of second-order arithmetic allows for the expression of nearly all results considered arithmetical. The basic axiomatic system expressed in the language of second-order arithmetic is called RCA_0, and the statements provable in it are essentially those that are 'computably' true. The strongest axiomatic system considered is all of second-order arithmetic, denoted Z_2. For any collection S of statements in the language of second-order arithmetic such that each statement in S is provable from the axioms of Z_2, we can consider the axiomatic system consisting of RCA_0 and S; such a system is called a subsystem of second-order arithmetic. The relative strengths of the systems thus obtained is the subject of much research. Here, we examine the strength of the systems obtained when S is a collection of combinatorial results. In particular, we wish to determine whether Hindman's theorem is provable in ACA_0. Hindman's theorem is the result that, in any finite coloring of the natural numbers, there is an infinite set such that any finite sum of elements in the set have the same color. It is stated quite differently from ACA_0, a subsystem of Z_2 that is a natural analogue of Peano arithmetic. However, it is known that ACA_0 can be proved from the subsystem consisting of RCA_0 and Hindman's Theorem. We aim to determine here whether the converse is true. This has implications for the proof-theoretic strength of other 'Ramsey-type' combinatorial results.

Our goal is to predictively engineer bio/nanomaterial hybrid systems with targeted functionality in a wide range of practical, technical, and medical applications. The open literature provides datasets of the functional properties of crystals, aqueous chemicals, and biological macromolecules, but the design of hybrid systems necessitates the modeling of all of these molecular species in a single common framework. Molecular graph convolutional networks and other deep learning methods are capable to train on datasets from multiple disciplines simultaneously, but in order to build these networks, a far-reaching data infrastructure is needed. We have created this infrastructure for three data sets: The Immune Epitope Database (IEDB) of MHC-I binding peptides, the Quantum-Machine.org QM9 dataset (QM9), and results extracted from the Materials Project. The IEDB provides binding affinities between biological macromolecules (peptide sequences in association with multiple MHC-I alleles); QM9 consists of 140,000 small organic molecules encoded as SMILES strings and 17 associated properties (including thermodynamic, energetic, geometric, and electronic information). The Materials Project dataset provides band gaps and formation energies for 70,000 crystal structures. We present a standardized train/test split and machine-learning-ready import interface for each of these datasets, as well as early results on co- and cross-training of deep neural networks across multiple datasets. The framework is expandable to new datasets and provides a strong foundation for ongoing efforts to build universal molecular encoding neural networks.

People living in rural Northern Tanzania are highly dependent on their natural environment for their health and livelihood. Consequently, they are a vulnerable population with regard to the potential negative effects of climate change. In this study, a semi-structured interview questionnaire was used to interview inhabitants of the Laja, Getamock and Kaza Roho villages, and measure their perception of the effect of climate change on their livelihood, health, and gender roles in the Manyara and Arusha regions of rural Tanzania. A total of 226 interviewees were asked about their perceptions of rainfall patterns, natural resource availability, disease rates, gender roles, and birth control use. The majority of respondents perceived a decrease in the frequency of rainfall and an increase in the frequency of drought. Moreover, 89% of the respondents were determined to be pastoralists or farmers that are directly affected by reduced precipitation, which decreases natural resource availability and crop yield, and increases rates of disease in humans and livestock. Respondents also reported shifts in gender roles to accommodate the rise of health, natural resource, and financial burdens stemming from climate change; for example, men are increasingly responsible for gathering drinking water because sources nearby villages have dried up and only men can drive motorbikes and ox carts now required to travel such long distances. Lastly, it was found that contraception usage has increased in younger, more educated groups in conjunction with a decreased number of desired children. Because family planning are associated with girls achieving higher levels of education and economic growth, improved family planning presents a potential solution to the financial hardship imposed by climate change. Considering the challenge of mitigating the inimical effects of climate change to developed nations, solutions to these problems are critically important to vulnerable groups that are highly dependent on natural resources for their health and livelihood.

Oceanic biogeochemical pathways, and particularly nitrogen, play a large role in global carbon cycling. Oxygen Deficient Zones (ODZs) such as the Eastern Tropical North Pacific (ETNP) have microbes that use distinct metabolic pathways for energy and nutrients in the absence of oxygen. These ODZ pathways shape the global inventory of nutrients and are thought to be responsible for around half of marine bioavailable nitrogen losses. While some of these pathways such as canonical denitrification and the more recently discovered anaerobic ammonia oxidation have been studied, there are also other potentially significant pathways. Trimethylamine N-oxide (TMAO), a small nitrogen-containing organic molecule with pathways that connect into other nitrogen cycle pathways, was seen to be diminished to nonexistence in ODZs which suggested it was being utilized. While studied in other environments, microbial utilization of TMAO in ODZs has not been significantly studied. We mapped metagenome reads collected from the ETNP ODZ onto a phylogenetic tree annotated with genes for TMAO degradation found outside the ODZ. We were able to confirm the presence of genes for TMAO degradation pathways and identified a range of organisms involved. We also examined the distribution of these genes and organisms across the different depths we sampled from to create a more complete picture of nitrogen cycling in ODZs.

Sea ice plays a significant role in the global climate, but the details of summer melt processes remain poorly resolved. Sustained low-wind events over the Arctic Ocean have been observed to correspond to high salinity and temperature stratification in sea-ice leads, consequently increasing lateral melt rates during summer. Such an effect is currently unaccounted for in leading climate models, and appropriate parameterization could potentially reduce error in projections. We used a wind reanalysis dataset produced by the National Center for Environmental Prediction (NCEP) and the National Center for Atmospheric Research (NCAR) to study wind patterns in the Arctic. We focused on daily variation in oceanic regions at latitudes above 65 degrees during Summer (May 1 - September 30) over 2010-2019. An index of low-wind events was extracted from the reanalysis data and these events were classified according to length of event and geographical extent. Our results suggest that low wind events are frequent throughout the summer, but are not equally distributed spatially across the basin. Low-wind events were compared to other climate data, including sea-ice extent, on a spatial and temporal basis. Our results were compared to climate model output and differences were examined. We propose a basic parameterization of the effect of low-wind events on sea ice melt, and suggest further collection of observational data to improve physical representation in climate models.

In order to perceive attack by herbivores and pathogens and subsequently mount appropriate defensive regimes, plants rely on a variety of protein receptors that recognize and initiate responses to damage, pathogen, and herbivory associated molecular patterns (DAMPs, PAMPs, and HAMPs, respectively). Although DAMPs and PAMPs have been studied extensively, the first HAMP-receptor pair was only recently discovered. The inceptin receptor (INR) discovered in cowpeas (Vigna unguiculata) binds to inceptin, a small peptide and potent HAMP found in the oral secretions of Lepidopteran caterpillars, promptly eliciting an immune response leading to enhanced resistance against herbivory. In order to further characterize the structure and function of INR, I am developing a fluorescent reporter for use in Nicotiana benthamiana, a model organism that normally lacks INR. Expression of the fluorescent protein mScarlet will be driven by putative promoters of genes found to be upregulated in response to inceptin binding to heterologously expressed INR in N. benthamiana. Transfer of this reporter system into N. benthamiana via Agrobacterium-mediated transformation will allow fluorescence to act as a reporter of INR function by generating quantifiable fluorescence in the presence of inceptin binding functional INR. By simultaneously transforming wild-type N. benthamiana plants with the reporter and mutagenized variants of INR, key domains and residues for the recognition of inceptin by INR and subsequent activation of plant defense will be elucidated. This will identify key structural and functional aspects of INR that will inform engineering practices for enhancing crop resistance to herbivory.

Ceratophora is an endangered genus of lizards endemic to Sri Lanka containing five species. Of them, Ceratophora aspera has the widest distribution spanning the lowland and montane rainforests of Southwestern Sri Lanka. Very little is known about the ecology, population biologyand natural history of C. aspera, although this rainforest dwelling species can serve as a precedent for many more conservation and management projects. The goal of this project is to assess the genetic diversity within and among populations, and determine the phylogenetic relationships among the populations. During the 2018 and 2019 field seasons, tail clippings were collected from lizards that were captured in the wild. In the University of Washington's Biology Department, DNA sequencing of mitochondrial DNA is being used for an initial screening of genetic diversity, followed by the acquisition of genomic information using Single Nucleotide Polymorphism (SNP) data. Together these data are enabling us to model the relationships between geography and genetics to visualize where populations are connected and fragmented. Preliminary analysis suggests that there is low intrapopulation diversity (<0.1%), while the highest interpopulation diversity is 3.8%. Furthermore, the phylogeny supports two main clades indicating that population structuring corresponds to the two main forest complexes, the Sinharaja Forest Complex and the KDN (Kanneliya-Dediyagala-Nakiyadeniya) complex. Each of these forest complexes were historically entirely forested but are now composed of forest patches. Restoring forest landscapes and reestablishing metapopulation and genetic diversity across the landscape and locations of migration barriers and corridors, provides necessary information for effective conservation management of this species and others like it. 

Sex determination is a crucial process in many organisms and there are many ways in which it is regulated, including through genetic and environmental mechanisms. Despite its popularity as a model for development and disease research, the precise mechanisms of zebrafish (Danio rerio) sex determination remain uncertain. Zebrafish lack the typical heteromorphic sex chromosomes present in many other species, and no specific chromosome or gene that determine sex has been conclusively identified. In a preliminary experiment, a genetic analysis found sex was associated with inheritance of single nucleotide polymorphisms (SNPs) from various regions of the genome. To test the reproducibility of this observation, we used PCR and Sanger sequencing to identify SNPs in candidate causal regions. We tested these to obtain genotypes of individual zebrafish. These fish are being crossed to assess whether significant deviations from genotype ratios are found in males and females, which would suggest that loci linked to these SNPs are in fact involved in sex determination. This work attempts to further elucidate the genetic contribution to sex determination in zebrafish.

The direction that plants grow in is dictated by the directions of gravity and light. A mutation in the LAZY-2 (LZ-2) gene in tomato (Solanum lycoperiscum) causes the plant to actively grow downwards, in the direction of gravity. It has been shown that the lz-2 phenotype is dependent on the phytochrome (Phy) protein family; a group of plant proteins that is responsive to red light. In a plant with no functional phytochromes, the lz-2 mutation is restored to wild-type, and the plant will grow upwards as normal. In this project, we discern the roles of three of the five tomato phytochromes, A, B1, and B2, to see if they work together or alone to provide the light cue that initiates the mutant function of lz-2. By gravistimulating combinations of lz-2 and Phy tomato seedlings under red light, we have detetermined that PhyA delays the lz-2 response, while PhyB2 exhibits an additive effect with PhyA and PhyB1 to effectively reverse the lz-2 phenotype. These data expand on previous work to further our knowledge on the relationship between light and gravity sensing in higher plants. 

Genetic data provide important insights into evolutionary relationships within and between species, and are especially useful for studying morphologically cryptic species (species that are visually indistinguishable from one another). I investigated the identity of an unstudied population of Sceloporus lizards from the Laramie mountain range of Wyoming. This population occurs at a boundary between the recorded distributions of S. tristichus and S. consobrinus. These species are morphologically cryptic, so molecular methods are necessary for accurate species determination. In collaboration with the Wyoming Fish and Game Department (WFGD), I conducted molecular genetics work on 10 samples, including extraction of DNA and PCR amplification of the ND1 mitochondrial DNA (mtDNA). Genetic divergence between populations of S. tristichus in western Wyoming (Sweetwater) and the Laramie Range is over 8%. Genetic diversity between a population of S. consobrinus from the Rocky Mountains in Colorado and the Laramie population is low (0.15%). I performed a phylogenetic analysis using maximum likelihood in the program RAxML and the population from the Laramie range is placed in a clade with S. consobrinus. Sceloporus consobrinus is currently not in the State Species Index for Wyoming, yet this result provides support for their addition. Before confirming that this population is S. consobrinus, additional phylogenetic analyses must be done. Previous studies of lizards in this genus have revealed discordance between gene trees made from mtDNA and nuclear DNA (nDNA) for populations occupying boundary zones between species. Therefore, current research aims to check for discordance or agreement between the nuclear genome and mtDNA gene tree.

Pain and unpleasant stimuli carry a negative hedonic valence, indicating an intrinsic aversiveness useful for avoiding harm. There are instances, however, of unpleasant stimuli carrying positive valence – such as the pleasure from spicy food, suggesting that pain and aversion can be decoupled. The Dhaka lab has discovered a small molecule Analgesic Screen 1 (AS1) which reverses the valence of a number of nociceptive and other aversive stimuli, whereby animals prefer normally aversive stimuli. Behavioral studies with zebrafish indicate that AS1 induces preference for noxious heat, painful chemical (AITC) and normally aversive dark environments. As positive valence or reward is often mediated by the neurotransmitter dopamine, we tested for the affects of dopamine antagonism on AS1-evoked behavior and found that the effects of AS1 are reversed by a D1 dopamine receptor antagonist. We currently propose that AS1 potentiates activity in the dopamine reward system in the presence of nociceptive and other aversive stimuli via D1R activation, thereby creating “pleasure from pain.” Understanding these pathways and the mechanisms underlying AS1 action, could provide a path forward for the development of novel therapeutics to treat debilitating pain disorders.

Viral pathogens, especially those that undergo rapid mutagenesis, pose a significant threat to public health. Viruses that exemplify this issue include influenza, HIV, and Ebola. Given the low efficacy of seasonal vaccines for influenza, our project focuses on improving existing influenza vaccinations. Instead of using conventional methods, such as injecting inactivated pathogens or viral subunits, mRNA sequences encoding the viral hemagglutinin (HA) fused to our recently-developed self-assembling I53-dn5 nanoparticle platform will be administered in vivo. The organized array of the protein platform can lead to stronger B-cell crosslinking and a robust immune response. However, characterization of I53-dn5 in vitro is critical before use in vaccination studies. My work focused on optimizing the expression, secretion, and assembly of the I53-dn5 protein platform. To mimic in vivo conditions, I transfected DNA encoding HA-fused I53-dn5 into HEK293F cells. Past experiments have shown that when dn5A and dn5B are transfected separately, they express at disproportional concentrations. To resolve this, we encoded both components onto one DNA plasmid for transfection. However, with this new approach, we also needed to cleave the two components after expression. To do so, we incorporated different cleaving peptides, such as T2A and Furin cleavage sites. Through western blots, SDS page electrophoresis, SCC protein purification, and electron microscopy, I analyzed how these cleaving peptides impacted assembly and secretion of the protein platform. Once we are able to consolidate an effective model, we will be able to start in vivo studies. Furthermore, if effective, our model can be used to create vaccinations against other viral illnesses, including HIV and coronavirus.

Wildfires are known for their destructive capacity towards ecosystems and devastating impacts to human life and property. Regions in the western United States are particularly prone to such events, including large crown fires. However, there is limited research on the potential for smoke from these wildfires to carry and redistribute nutrients in the form of aerosols. Here I show that higher levels of airborne nutrients, specifically iron, phosphorus, and potassium, can be associated both with the timing of active wildfires and a known smoke tracer, elemental carbon. Data from the Interagency Monitoring of Protected Visual Environments (IMPROVE) show a strong correlation with fire presence and local peaks in levels of potassium, with less strong associations with phosphorus and iron. Across a multi-year period, there is also a correlation between the number of acres burned in a particular year and the average concentration of iron and potassium in that region. Using data from the NASA DC-8 airborne laboratory corroborates this information, with calculated potassium concentrations matching those on the ground. These findings indicate the wildfire plumes have a potential to be significant sources of nutrients in the short term. This is especially relevant for areas impacted by wildfires, as these nutrients are key for plant growth and development. These elements may also be carried into the ocean and affect the aquatic biosphere. All of the information gathered will help improve our understanding of the complex networks that make up Earth’s biogeochemical cycles.

My research project involves the AP-3 (adaptor protein) complex, which plays a key role in membrane trafficking within cells. Vesicles (small, membrane bound structures) mediate the transport of proteins and lipids among cellular organelles. These vesicles are created in various ways by proteins throughout the cell, with some including a protein "coat" around the vesicle as it travels to its destination. AP-3 is a coat protein complex that mediates vesicular transport from the trans-Golgi network to the lysosome. We have utilized a gene reporter system called GNSI to analyze AP-3 function in different genetic backgrounds of Saccharomyces cerevisiae (baker’s yeast). One output of the reporter system is a colorimetric assay that measures the intensity of colored halos around yeast colonies grown on an agar plate. We found that targeted truncations of proteins of the AP-3 subunits Apl6 and Apl5 at the C-terminus of AP-3 (specifically the “ear domains”) resulted in increasing defects in AP-3 trafficking ability. However, it is unknown whether the defects are in membrane recruitment to the trans-Golgi network, or whether recruitment occurs but vesicle budding defects arise. Therefore, the current aim of the project is to begin analysis of the truncation defects in AP-3 by fusing Apl5 trunctions with a C-terminal mNeonGreen fluorescent protein and use live cell fluorescence microscopy to further examine AP-3 localization These basic studies will further our understanding of membrane trafficking and may provide insight into diseases linked to AP-3 function, including HIV-1 particle assembly and human genetic disorders.

Natural proteins often assemble into various complex geometric structures based on their interactions with each other. The King Lab at the University of Washington's Institute for Protein Design uses the way these proteins behave to develop computational models that enable the design of novel self-assembling protein cages, or nanoparticles. The designed particles are capable of holding and transporting molecules or displaying antigens on their surface, making them effective vaccine candidates. My project involves recovering the solubility of one of these protein cages known as T33_dn2. T33_dn2 is a tetrahedral protein cage comprised of four copies each of two trimeric components known as T33_dn2A and T33_dn2B. While both components can be expressed individually through E.coli before being assembled in vitro, they can also be expressed bicistronically and assemble in vivo. Currently, the use of T33_dn2 as a vaccine scaffold is limited because T33_dn2B is insoluble, and only seems to be stabilized in solution when associating with T33_dn2A. When expressed bicistronically, however, the cage has an extremely low yield. For a protein to be developed into a vaccine, it must be soluble. To recover the solubility and yield of T33_dn2B, I am testing ten plasmid variants of bicistronic T33_dn2. The “original” plasmid consists of one gene coding for a high-expressing cleavable SUMO protein attached to T33_dn2A and another coding for T33_dn2B. The additional nine variants have single point mutations at specific locations on the T33_dn2A gene intended to affect binding strength. After expression, introducing wildtype T33_dn2A in vitro will allow for the formation of T33_dn2. I will be presenting the results of these expression, purification, and assembly tests.

Newly developed, rational syntheses of the phosphaethynolate anion have taken it from an academic curiosity to a useful synthetic reagent. Since the new state-of-the-art synthetic method was published by Grützmacher et al. in 2014, many studies have emerged investigating the phosphaethynolate anion’s possibilities as a synthetic building block, cycloaddition reagent, and P atom transfer agent. I have performed very preliminary work (cut short by COVID-19) on using this anion to generate phosphide thin film materials. Metal phosphide thin films have many applications in advanced electronics and thus are a valuable manufacturing target. The solution-processed, electrochemical method proposed would have marked advantages over current methods to generate phosphide thin films such as MOCVD and MBE. If realized, this unique method of producing metal phosphide thin films would be entirely unprecedented.

Humans are incapable of regenerating a majority of their major tissues following traumatic injury. Tadpoles from the frog species Xenopus tropicalis have the ability to regenerate lost spinal cord, vasculature, muscle, and cartilage within a few days following injury. The regulatory mechanisms of gene expression necessary for regeneration have not yet been well defined. My primary interest is in understanding the relationship between stress signaling and gene expression during regeneration. The lab has shown that the stress responsive transcription factor Hypoxia Inducible Factor 1α (Hif1α) is necessary for the expression of Wnt target genes, one of the primary signaling processes necessary for regeneration. While we have found that Hif1α is necessary for Wnt target gene expression, we do not know the epistatic relationship between Hif1α and Wnt. In order to test this relationship, I utilized the drug IWR to antagonize Wnt and found that tadpoles treated with IWR have reduced tail regeneration 72 hours post amputation (hpa). I then supplemented these tadpoles with DMOG to stabilize Hif1α and found that DMOG is sufficient to rescue tail regeneration, suggesting that Hif1α is downstream of Wnt. In order to determine if Hif1α is sufficient for Wnt target gene expression, I extracted RNA from regenerating tails 24 hPa and used quantitative PCR (qPCR) to determine relative gene expression. I also utilized in situ hybridization to see if expression of these genes is restricted to regenerating tissues. As Wnt is a known regulator of neural and muscle development, I investigated how inhibiting Hif1α would impact complex tissue regeneration and found that Hif1α is necessary for regeneration of axons and muscle specifically. By determining the epistatic relationship between Hif1α and Wnt through the analysis of specific gene expression, we continue to improve our understanding of how regenerative organisms convert stress signals to cell fate signals.

There is a high prevalence of depression in the United States, and the impairment from depression and the impact it has on an individual’s functioning can also be severe. Thus, it is vital to examine the different causes of depression. In this study, I am investigating the effect a sense of belonging has on depressive symptoms by administering questionnaires to participants from the Midlife in the United States (MIDUS) study and participants from the University of Washington. MIDUS is a national longitudinal study of different aspects of health and well-being. I am also investigating the moderating effect a strong ethnic identity, specifically ethnic identity for individuals of Asian descent, has on the relationship between a sense of belonging and depression through questionnaires that will be completed by participants from the University of Washington. I am predicting that individuals, both from MIDUS and the University of Washington sample, who have a higher sense of belonging will have a lower level of depressive symptoms. Additionally, I am predicting that participants who have a lower sense of belonging but a strong sense of ethnic identity will have a lower level of depressive symptoms. Regardless of the results, the findings of this study will aid in our understanding of the role of sense of belonging for individuals with depression, and will also shed light on the importance of ethnic identity for minority ethnic groups.

Marie Kondo, a Japanese organization expert, talks about objects being able to “spark joy” for those who own them. She says in her best-selling book “The life-changing magic of tidying up: The Japanese art of decluttering and organizing” that we should “keep only the things that spark joy” when advising people as they go through their decluttering journey. Given Kondo’s popularity, this phrase has influenced Western popular culture in the past few years. Still, there is no specific definition of spark joy in her books nor her Netflix show. In Japan, spark joy or tokimeki, is a common phrase that has been used since the 900s, but the phrase has been used more to describe the emotions surrounding romantic relationships rather than for objects. Previous studies on the meaning of objects (largely in the West) have indicated more focus on the self-reflection brought on by an object rather than relational reasons for seeing an object as special. For instance, objects have been found to represent self and achievement. People also give meaning to an object based on memories or identities, such as ties with ones’ family or ethnic background, suggesting some relational ties to an object. In this research, I examine what object sparks joy to people and what “spark joy” means to them. The research was conducted by interviewing people from Japan and the U.S., where popular culture has been influenced by her methods. I also had participants complete assessments of their self-construals and attachment styles. Qualitative data were examined by content analysis where I looked for relationships between self-construals and attachment styles to see if these predict the ways people give meaning to an object. This research deepens knowledge about objects and their meanings as well as provides a cultural comparison of what spark joy means to people.

3% of children live with Amblyopia (‘lazy-eye’), a visual disorder where acuity in one eye is poor even with glasses on. Amblyopia includes (1) reduced sensitivity in the amblyopic eye, and (2) interocular suppression, whereby the good eye suppresses the amblyopic eye. Our goal was to develop a method to efficiently characterize both mechanisms. Participants viewed a Gabor stimulus modulating between 0-100% contrast over time through a stereoscope, that presented a different image to each eye. Participants reported perceived contrast over time using a joystick. On each trial, the initial 14 s consisted of binocularly identical gratings modulating at 1/7 Hz, followed by 48 s where the gratings modulated at 1/8 Hz in one eye, and 1/6 Hz in the other. Separately, we measured visual acuity, stereoacuity, contrast sensitivity, the interocular suppression ratio. This method was highly efficient: only 30 min of data were needed to estimate monocular sensitivity and interocular suppression. Another advantage of this approach is that the stimulus is relatively naturalistic – the images in the two eyes are the same, except for the difference in contrast. This is important due to the nature of its binocularity and naturalistic conditions, as this will be the first of its kind - allowing clinicians a better way to assess those with amblyopia. 

Anxiety symptoms are common in children with attention deficit hyperactivity disorder (ADHD). It is unknown whether the neurobiological origins of comorbid anxiety and ADHD symptoms are shared or distinct. The current study addressed this using an event related electrophysiological potential (ERP) component, the error-related negativity (ERN), which occurs after an individual makes a task error. ERN amplitude has opposite associations with ADHD and anxiety symptoms: it is weaker in association with increased ADHD, but greater in association with increased anxiety. We tested whether 1) anxiety symptoms and ADHD have separate neurobiological origins, indicated by greater anxiety being associated with increased ERN in children with ADHD or 2) anxiety and ADHD symptoms share an origin, as evidenced by no effect of anxiety on ERN in children with ADHD. The current study investigated the association between ERN amplitude and anxiety levels in a sample of 7- to 11-year-olds with ADHD (n = 98) and without (controls; n = 26). Participants completed two ERP tasks of varying difficulty. ERP data were segmented around incorrect task responses, and mean ERN amplitude was extracted. Data on child anxiety and ADHD symptoms was collected via parent report. Linear regression analysis was used to estimate the associations among ERN amplitude, severity of anxiety, and ADHD symptoms. Preliminary results (n = 73) indicated that ADHD symptom severity was associated with smaller ERN amplitude (r =.31, p =.007), but anxiety symptoms were not associated with ERN in the ADHD group. Preliminary results indicated that the ERN is not a marker of anxiety in children with ADHD to the same degree it is in controls. This is consistent with shared neurobiological etiology for ADHD and anxiety symptoms in children, which has clinical implications for conceptualization and treatment of anxiety symptoms in childhood ADHD.

Assumptions about gender and sexuality in the health care system often manifest as structural barriers to accessibility. Stakeholders, such as healthcare program managers, executive directors, practitioners, and other staff members, inform the organization’s structure, which guides providers on the service provision they provide to their LGBTQ+ patients. Thus, understanding health care staff perceptions on how current policies and programs contribute to the health care environment for the LGBTQ+ community will assist efforts for equitable service provision. Semi-structured interviews were conducted on staff from 6 health care organizations. Open-ended questions focused on their perspectives on current practices and policies, organizational areas of growth, as well as barriers, for equitable care. The purpose of the study is to discuss LGBTQ+ health within their organization to elicit staff perceptions on values and perspectives that address LGBTQ+ health disparities. Interviews were transcribed and inductive thematic analysis was done to find common benefits, viewpoints, and areas of improvements. Themes such as lack of knowledge, cultural biases, historical discrimination, and collaborative leadership, emerged. The findings of this study show gaps in delivering coordinated, patient-centered care for LGBTQ+ patients. This suggests greater importance for organizational education and training, particularly related to how historical oppression impacts bias in their setting. The overall implications of the study illustrate the relationship between the themes and organizational impact on service provision, which impacts the accessibility, safety, and quality health care environment for LGBTQ+ patients. This is crucial to giving context and describing the systematic impact on an individuals’ access to and quality of health care for the possible use in future studies.

The relationship between humans and robots is full of feedback loops: how our brain processes what we see, feel, and act affects how the intelligent machine reacts and vice versa. Understanding this feedback loop will enable us to design better day-to-day automated systems. For instance, operations performed with surgical robots are guided with the surgeon’s movement. My research in the UW BioRobotics Laboratory investigates how the human brain adapts to technology. I programmed a haptic paddle to apply timed forces on human subjects to investigate how uncertainty, in the form of a disturbance, affects response over time. Understanding how human behavior changes is crucial for improving automated systems that can be simulated by the haptic paddle such as cable-driven surgical robots. We apply the haptic paddle to model forces a surgeon may experience during surgery, such as tough tissues or other disturbances, while operating a robotic surgical device. As we measure the participant’s learning curve over longer periods and more trials, the participant builds a better understanding of how to react relative to their force exerted on to the paddle. By increasing our understanding of how the human brain works, we can begin to improve precision of surgical operation.

The world’s largest high-energy particle accelerator, the Large Hadron Collider, relies on sub-millisecond processing performed on massive amounts of data coming out of its particle detection system, which is due for a major upgrade in 2024. The old particle detection chips will be upgraded to RD53B chips with faster data transmission, allowing for more complicated data processing. The readout systems that interact with the particle detection chips, YARR and FELIX, need to be tested with the RD53B chips and debugged before the system is put in place. The goal of our research is to create an emulator of the RD53B chip that can produce dynamically generated pseudo-realistic data at the same rates that would be seen in the Large Hadron Collider without the need for heavy radiation. This will allow for readout software to be fully functional and debugged before the actual RD53B chips are fabricated and placed into the Large Hadron Collider. We are using Field-Programmable Gate Arrays (FPGAs) to mimic the hardware inside the real RD53B chips. In place of RD53B’s analog sensors, we have substituted digital logic that generates pseudo-realistic data because FPGAs cannot emulate analog hardware. The alpha version of the RD53B emulator with basic communication and pre-programmed data was completed in February. Recently, the beta version of the emulator with dynamically generated data was completed, and we have been testing communication between the emulator and FELIX. With the beta version of the RD53B emulator tested and verified by us, the developers of YARR and FELIX will use our hardware to help verify that their systems will provide accurate readouts from the real RD53B chips. The next steps for the RD53B emulator include a hardware data decompression accelerator, as well as any additional features requested by the YARR and FELIX teams.

After neurologic injuries like stroke, users must relearn how to interact with devices to achieve different tasks. However, hand dominance before and after neurologic injury could affect motor learning and recovery. Understanding a) whether hand dominance plays an effect on motor learning and adaptation of predictive (i.e. feedforward) and reactive (i.e. feedback) controllers and b) how motor learning is impacted by which side was affected by stroke is crucial for personalizing rehabilitation techniques. We are currently investigating the effect of hand dominance on motor learning during continuous tasks for users without motor impairments. 10 participants were asked to play a simple trajectory-tracking game, first with their non-dominant hand and then with their dominant hand. We found that participants’ feedback controllers improved with practice for their non-dominant hand (p=0.005), but their feedforward controllers were unchanged (p=0.88). Furthermore, the participants transferred their feedback controller from their non-dominant hand to their dominant hand (p=0.33). This suggests that the non-dominant hand may be more specialized for error correction and impedance control, and that rehabilitation for users who had a right-hemisphere stroke should focus on improving reactive control.

Essential tremor (ET) is a movement disorder characterized by kinetic and postural tremor that affects an estimated 7 million people in the U.S. alone. Deep brain stimulation (DBS) of the ventral intermediate (VIM) nucleus of the thalamus is an established medical therapy for the treatment of ET and has been shown to decrease tremor symptoms significantly for patients in which traditional pharmaceutical therapies have proven insufficient. There are limitations to continuous DBS treatment however, with battery replacement surgeries necessary every few years as well as the presence of side effects such as dysarthria, paresthesia, and gait ataxia. Adaptive DBS (aDBS), with its use of internal or external markers as feedback to modulate stimulation parameters, presents a promising avenue through which to mitigate these concerns. However, current aDBS methods typically employ a binary, “on-off,” control system, which may introduce new complications to consider. One such complication is rebound effect, a transient increase in tremor severity immediately following the deactivation of DBS before levelling out to a steady state. However, clinical observations of rebound in ET have been mixed in the literature, which results in an ambiguity about the potential for effect on an aDBS system. To clarify this mixed literature and quantify the potential impact that the rebound effect may have on an aDBS system, we collected inertial measurements from the tremoring arm during the spiral and line drawing tasks of the Fahn-Talosa-Marin tremor rating assessment task. This task was used to characterize rebound over a ~40 minute time window, over which we observed the rebound effect in all three of our patients. Our results indicate that rebound effect should be taken into account when designing aDBS systems, pointing towards the development of a non-binary aDBS system.
 

The mobility of autonomous walking robots is an essential characteristic in their operation. Due to currently imposed constraints in battery technology, the optimization of robotic locomotion for energy efficiency is paramount. Previously, elastic payload suspension has been employed to reduce the cost of transportation in a hexapedal robot. Prior results suggest that the optimal load suspension characteristics are a function of robot morphology and locomotion strategy. A payload suspension system that can be easily adjusted would allow for the accommodation of a variety of these morphologies and locomotion strategies. We have designed a tunable suspension system that will allow for experimentally determining optimal suspension characteristics in a cost-effective manner. The design enables continuous adjustment of the suspension stiffness and damping, so optimal parameters can be determined through hardware experimentation. This hardware experimentation allows for the creation of a numerical model for an oscillating payload’s behavior, which can be compared to simulations. We have completed calculations and design of this hardware, and anticipate seeing that the data collected from its usage will verify the utilization of a haptic testing system in robotics development and allow us to determine methods for calculating the best parameters for elastic payload suspension. This verification of simulated data will allow for further research in developing more efficient methods of payload attachment to legged robots, examination of locomotion when carrying payloads, and design of payload management methods.

Filtering the data produced by the Large Hadron Collider (LHC) is computationally challenging due to the sheer quantity of the data, on the scale of hundreds of terabytes per second. In the coming years, data production for the LHC is projected to increase by a factor of 15 with the high luminosity upgrade. Machine learning algorithms could provide pattern recognition capable of filtering data produced by the LHC. Specialized hardware could increase the throughput to match the data rates required by the LHC. We analyzed several cloud-based specialized hardware solutions including Amazon Web Service FPGAs, Microsoft’s Brainwave Service, Google’s TPU, and NVIDIA GPUs to compare the performance of each of them for particle physics application. The networks accelerated were trained on a variety of data including: Top vs QCD quark classification, Hadron calorimeter data, and electron energy regression. These experiments demonstrate the feasibility of machine learning algorithms in high throughput required situations such as high energy particle physics.

Chronic respiratory infections and diseases are the third leading cause of death globally. The Frevert lab studies the protein versican (Vcan), an extracellular matrix proteoglycan, whose expression is highly upregulated during lung injury and inflammation. However, it is unclear whether this upregulation is an anti-inflammatory or a pro-inflammatory response. We are testing the hypothesis that the cellular source of Vcan determines its inflammatory actions; versican from myeloid cells is anti-inflammatory but versican from stromal cells is pro-inflammatory. Vcan deletion in our cells of interest can test this hypothesis. To do this we have generated Vcan^fl/fl genetically engineered mice, which allow for conditional removal of functional versican with Cre Recombinase. Cre excises versican’s exon 4, recombining exons 3-5 and creating a premature stop codon. This produces a truncated non-functional form of versican. The goal of this research project is the development of cell-specific protocols for in vitro deletion of versican in both myeloid cells (bone marrow-derived macrophages) and stromal cells (lung explant fibroblasts). These protocols will allow for further evaluation of versican’s role in the inflammatory response when different cell types are confronted by a bacterial or viral agonist. So far, I’ve been investigating the dose response and time course for exposure of macrophages to Cre to optimize its efficiency and have been able to demonstrate by qPCR that intact versican decreases and non-functional versican increases. The inflammatory response is quantified through qPCR analysis of the fold increase of Ifn-b compared to the house-keeping gene MRPL32. Ifn-b is a cytokine released by the innate immune system in response to viral pathogens. Next, I will investigate the conditions necessary for efficient Cre deletion of Vcan in fibroblasts. These experiments allow us to investigate the effects of Vcan made by different cell types furthering our understanding Vcan’s function during injury and inflammation.

Capacitation involves biochemical and physiological processes in sperm with motility changes (including hyperactivation) that allow sperm to bind to the zona pellucida, undergo acrosome reaction and penetrate the ovum. Bovine serum albumin (BSA), has been thought to facilitate capacitation in mouse sperm through depleting the sperm membrane of cholesterol. Other albumin preparations including human serum albumin (HSA), fatty acid free (FAFHSA), and recombinant albumin (RHSA) may affect capacitation differently. Moreover, cholesteryl ester transfer protein (CeTP) may facilitate capacitation but it should not be present in yeast-derived RHSA as opposed to human blood-derived HSA; RHSA and HSA may support capacitation at different extents. These preparations were tested in motility experiments after 3-5.75 hr (mean 4 hr) capacitating incubation. Sperm were purified by 40/80% PureSperm gradients and washed in HTF-BSA, then diluted in final albumin to 10-20M/mL. FAFHSA, BSA, HSA, and RHSA had hyperactivation percentages of 2.83%, 3%, 11.3%, and 15.75% respectively. RHSA had a motility of 71.3% on average while the others ranged from 83.6% to 88%, affecting the hyperactivation results. RHSA also caused incubated sperm to agglutinate or aggregate, but its elevated hyperactivation rates relative to HSA are insignificant. These results fit previous data suggesting that BSA and FAFHSA do not support hyperactivation (therefore, capacitation) as both are abnormally below 10% hyperactivation. Since BSA contains low CeTP, this suggests CeTP may help facilitate capacitation. Previous studies demonstrated the amount of CeTP in albumin correlated well with albumin’s ability to support acrosome reaction. The present results suggest that capacitating motility patterns may not necessarily correlate with the ability to acrosome react; this hypothesis could be tested. Since fertilization is largely misunderstood; understanding the roles of Albumin or CeTP may aid in developing infertility treatments, contraceptives and family planning for a growing human populace.

Hemoglobinopathies, a hereditary condition involving abnormalities in the structure of hemoglobin, currently require expensive and highly sophisticated medical facilities to treat using gene therapy. The purpose of this study is to provide a highly portable and scalable approach using in vivo hematopoietic stem cell (HSC) gene therapy to potentially overcome these limitations. The main idea of our in vivo HSC gene therapy approach is to mobilize HSCs from the bone marrow, and while they circulate at high numbers in the periphery, transduce them with an intravenously injected HSC-tropic, helper-dependent adenovirus HDad5/35++ gene transfer vector system. The transduced cells return to the bone marrow where they remain long-term. This study followed one animal for 22 weeks after in vivo HSC transduction with a human-γ-globin expressing HDAd5/35++ vector using Sleeping Beauty transposase 100X for integration. Treatment with G-CSF/AMD3100 (a bone marrow stimulant) resulted in efficient HSC mobilization into the periphery blood circulation. The treatment was well tolerated and after in vivo selection, gamma-globin marking in peripheral red blood cells rose to ~90% and was stable during the duration of the study. Enrichment of gene-modified cells by in vivo selection was also reflected by an increase in mgtm and gamma-globin mRNA levels. Our data suggest that in vivo gene therapy with HDAd5/35++ is feasible and side effects can be minimized or prevented with appropriate pretreatment. This is the first proof-of-concept study that in vivo HSC gene therapy could be feasible in humans and provide the necessary portability and accessibility to reach patients in places with limited medical resources. Future studies will involve the optimization of HSC mobilization, gene transfer vectors and in vivo selection.

Methicillin-resistant Staphylococcus aureus (MRSA) is the leading cause of wound and hospital-acquired infections. Glycopeptides (e.g. vancomycin), lipopeptides (e.g. daptomycin), and lipoglycopeptides (e.g. dalbavancin) are three classes of cell-wall/cell-membrane targeting antimicrobials effective for treating MRSA. Since in S. aureus cell wall synthesis and lipid synthesis are metabolically closely connected, response to cell-wall/cell-membrane targeting antimicrobials is likely to be mediated by lipid synthesis. Indeed, our lab previously demonstrated that the overall lipid abundance decreased in the dalbavancin-resistant S7-D2 strain (vancomycin-intermediate), isolated by in vitro exposure of a clinical MRSA isolate S7 strain (vancomycin-susceptible) to multi-stage escalated concentrations of dalbavancin. Here we examined a lipid synthesis inhibitor, AFN-1252, to determine its potential synergistic effects with vancomycin, daptomycin or dalbavancin, using the S7 and S7-D2 strains. Broth microtiter MIC measurements and static time-kill experiments were performed to evaluate potential synergy. We found that, through the MIC measurements, the pellet sizes in the wells seemed to be reduced when S7 and S7-D2 were treated with AFN-1252 at half the MIC in the presence of vancomycin. Hence, a static time-kill at 2xMIC of AFN-1252 and/or vancomycin was performed to interrogate the growth difference. S7-D2, but not S7, showed increased killing with the combination, suggesting that killing by vancomycin could be aided by AFN-1252 in vancomycin-intermediate strains. The killing by the combination in S7-D2 also approached the level of killing by vancomycin alone in S7, suggesting mechanistically membrane lipid remodeling might have happened that counteracted the slower killing in the vancomycin-intermediate strain. Our preliminary results so far suggest that lipid synthesis inhibitors might be able to enhance the MRSA treatment effects of vancomycin. In future studies, we will evaluate the potential lipid profile changes using comprehensive lipidomics analysis, and further screen and evaluate other lipid synthesis inhibitors and their potential synergistic effects with vancomycin, daptomycin or dalbavancin.

Heart disease is a major health pandemic, and the myocardial infarction (MI), also known as a heart attack, is the leading cause of death. This is because adult cardiomyocytes (CMs) present in the heart cannot divide and proliferate, preventing the heart from regenerating itself and repairing tissue damage. From developmental studies in rodents, we know the Notch signaling pathway is crucial in mediating expansion and proliferation of CMs during development, as impairments in Notch lead to cardiac defects. Notch is inactive in adult CMs, which suggests it plays a role in CM renewal; however, this mechanism is still unknown. Thus, our goal is to investigate the role of Notch in CM proliferation and cell cycle regulation. For the purpose of this study, we are using CMs differentiated from a human embryonic stem cell line, RUES2. Differentiation was completed over a 17-day period by culturing the cells as a monolayer. On Day 0, Chiron 99021 is added to activate Wnt signaling, promoting mesoderm formation. Wnt-C59 is added at Day 2 to inhibit Wnt signaling and differentiate the cells into progenitors, and B27 supplement at Day 6 promotes full differentiation into CMs. This adherent protocol recapitulates every step of natural heart development in vitro. Purity of the cell populations, as assessed by flow cytometry staining for cardiac troponin T (cTnT), was 97.1 ± 0.9% cTnT+. We then determined the proliferative capabilities of CMs in the presence of a Notch inhibitor, DAPT. DAPT inhibits gamma-secretase, a transmembrane protein that normally proteolytically cleaves Notch during signaling, thus inactivating the Notch pathway. Treatment with DAPT significantly decreased cell proliferation by about 50%, confirming that Notch directly affects CM proliferation. The results of this study increase our knowledge of CM physiology and the mechanisms behind cell cycle withdrawal, and provide new insights into improving CM renewal.

X chromosome aneuploidy refers to an atypical number of X chromosomes, differing from two X chromosomes in females, or one X and one Y chromosome in males. Unusual numbers of chromosomes arise from errors in cell division that result in too many or too few chromosomes in a cell, with X aneuploidy reported to occur in around 1 in 1000 births depending on the disorder. X chromosome aneuploidy disorders, such as Klinefelter (47, XXY), Triple X (47, XXX) and Turner (45, XO) syndromes are associated with developmental abnormalities, including cognitive and cardiovascular defects. Our goal is to generate isogenic induced Pluripotency Stem Cells (iPSCs) with different numbers of X chromosomes from patients with X aneuploidy, and subsequently differentiate them into relevant cell types to identify genes affected by aneuploidy. Our lab has previously generated isogenic XXY and XY iPSCs from patients with Klinefelter’s Syndrome by removing the extra inactive X chromosome (Xi). My project is to establish and characterize isogenic lines from mosaic XXY/XY patients, who have a mixture of cell karyotypes. Specifically, I am screening iPSC clones derived from mosaic patients for expression of XIST, a gene expressed only from the Xi, by RT-PCR to determine presence or absence of the Xi. Isogenic control XY iPSC lines derived from the same patient serve as a control to XXY cells, lessening potential for confounding variables due to differences between individuals. Next, I will screen clones with different genotypes for integration of reprogramming vectors by genomic DNA-PCR. Integration-free isogenic XXY and XY clones will be differentiated into cardiomyocytes, neural progenitor cells, and cortical organoids, cell types that are associated with the adverse effects of X aneuploidy. By doing so, we hope to gain insight into gene expression and epigenetic changes associated with X aneuploidy phenotypes in a controlled genetic environment.
 

Midfacial hypoplasia (MFH) is a disorder of severe underdevelopment of the upper face. This problem is significant because MFH can lead to a deformity that impedes chewing, feeding, and breathing. Although it is known that MFH can occur due to gene mutations, teratogens, and trauma, it is unknown how these insults affect anterior growth of the face. Using a novel pig model, this project aims to isolate the location of structural changes to the skull caused by MFH. We first determined if MFH in pigs is isolated to the face, and then identified the specific bones of the face that are altered by MFH. To determine if MFH is isolated to the face, we measured and compared the length of the braincase, mandible, and upper face in pigs with MFH to that of normal pigs. We then measured and compared the dimensions of individual facial bones, including the maxilla, premaxilla, nasal bone, lacrimal bone, and zygoma. All measurements were done on dry skulls with calipers and compared using t-tests. Our initial qualitative comparison of 6 normal and 6 affected pigs has found that MFH is isolated to the face, with the premaxilla, maxilla, nasal bone, and lacrimal bone shortened. The zygoma and palatine bones were normal. We hypothesize that our qualitative comparison of approximately 35 MFH pigs and 115 normal pigs will show similar results. Identifying the bones affected by MFH will help us to hone in on the specific tissue changes and mechanisms that cause MFH, and to discover better treatments for this condition.

Midfacial Hypoplasia (MFH) is an underdevelopment of the upper jaw, cheekbones and eye sockets. MFH sometimes results in difficulty in breathing in children. Severe cases are treated with serious surgeries that often have unsuccessful outcomes. Understanding the consequences of MFH is a critical step in treating this disorder. The goal of my project is to use Yucatan pigs as a model for MFH. MFH is common in Yucatan pigs and resembles MFH in humans which makes it useful for a model. In addition, pigs resemble humans in physiology, anatomy and growth. Humans with MFH have reduced nasal airway capacity, and this is likely true for Yucatan pigs as well. The nature of airway deficiency is not fully understood, but it may be due to reduction of the nasal airway surface area. Reduction in the surface area would hamper the ability of the nasal passage to regulate air temperature and humidity. I hypothesize shortening of the nasal cavity in MFH reduces the surface area inside the cavity. Using CT scans from pigs, I measured the dimensions of the nasal passages via Metamorph and Photoshop software. Based on my hypothesis, I expect to observe lower surface area in pigs with MFH. If a correlation between MFH and surface area is found, this research will provide new insights about the consequences of MFH that will be relevant for better treatment of this disorder.

Breathing rhythms are initiated in the ventrolateral medulla, in a region known as the preBötzinger complex (preBötC). The preBötC generates normal inspiratory activity and breaths of greater volume called sighs. Sighs prevent the spontaneous collapse of alveoli in the lungs, termed atelectasis, and mice that lack the ability to sigh die of atelectasis after birth. For many years, respiratory physiologists have tried to understand how one network, the preBötC, is able to generate two rhythms with distinct timing mechanisms. In order to test this hypothesis that glia drive the sigh rhythm through purinergic signaling, I will make recordings of neuronal population activity from the preBötC in the horizontal brainstem slice preparation, and apply caged ATP. Brief light pulses will uncage the ATP. We predict that ATP and its conversion to ADP will bind to purinergic receptors to facilitate calcium spread and recruit neurons along the inspiratory column to generate these large breaths. Then, I will apply the purinergic receptor antagonist that we have established is effective in blocking sighs. We observed this result in a preliminary experiment, but more data needs to be collected to confirm this relationship. Next, we will confirm that glial increases in intracellular calcium coincide with sigh generation. We will perform calcium imaging using two photon microscopy in the horizontal slice preparation isolated from transgenic cre mice that express GcAMP6f driven by the astrocyte-specific promoter, Aldh1l1. We predict that calcium activity in astrocytes will correspond with the sigh rhythm, and that we can inhibit sighs by applying purinergic antagonists.

 An estimated 50% of patients scheduled to undergo surgical correction of their adult spinal deformity (ASD) are preoperatively managed with chronic opioid therapy. In recent years, preoperative opioid use has been correlated with increased morbidity and mortality following major abdominal, orthopaedic, and spine surgery. Despite high prevalence of chronic opioid consumption in ASD population, limited data is available to guide preoperative opoid tapering to improve postoperatve outcomes. First attempt to identify a morphine equianalgesic (MEA) dose threshold for patients undergoing spine procedures was described in Neurosurgery by Wick and co-workers. After calculating MEA values for 1020 cervical and lumbar spine surgical patients, the group reported MEA dose threshold beyond which patients are less likely to achieve the minimum clinically important difference (MCID). In patients undering a Complex Spine Surgical procedure, we hypothesize that an increased preoperative opioid dose may have a threshold beyond which patients are more likely to suffer from an adverse event. A trained data abstractor will collect data for the analysis during the retrospective chart review. In addition to data collection, abstractor will calculate morphine equivalent dose (MED) values for each patient meeting the inclusion criteria. Statistical analysis will be performed to evaluate association between MED values and postoperative outcomes. Parametic and non-parametic tests will be used as necessary, inclusing but not limited to t-tests for Equality of Means, Fishcer exact test, Chi-square, Independent Samples Mann-Whitney U Test, and logisitic regression. All data will be reported in aggregate. Reporting a defined opioid theshold for ASD patients may aid in developing a standardized tapering protocol aimed at improving postoperative outcomes. 

In inpatient hospital settings, daily interprofessional rounds are when multiple health care professionals come together as an interprofessional team to create a care plan with patients and their families. Despite the promise of coordinated care during interprofessional rounds, poor care coordination and communication is a major contributor to medical errors, which are the third leading cause of death in the United States. A key literature gap is how to implement sustainable observation and feedback processes for interprofessional rounds. To address this challenge, my research focuses on developing and testing an effective process to evaluate communication during interprofessional rounds. I hypothesized that if pre-health undergraduate students can shadow interprofessional rounds with a structured observation tool, there will be benefits for pre-health students, interprofessional teams, and patients and their families. Specifically, pre-health students participate in an active and engaging shadowing experience while providing interprofessional teams with relevant feedback about interprofessional rounds that can be used to improve interprofessional communication. To test my hypothesis, I used a Plan-Do-Study-Act cycle methodology commonly practiced in quality improvement research. The first cycle tested if a single pre-health undergraduate student could train for and use a structured observation tool, leading to a second cycle which expanded that same process to a cohort of pre-health students. The observation data was analyzed into two categories: 1) quantitative tool topics (with inter-rater reliability data) and 2) qualitative comments. When comparing the quantitative versus qualitative data, the main conclusion was that pre-health students offer insightful and relevant qualitative comment feedback that allow interprofessional teams to reflect on ways to improve their communication during interprofessional rounds. This suggests additional cycles are necessary to test a tool that focuses more on qualitative data. The expansion of my research is key to improve interprofessional rounds in order to continually improve the patient care experience.

The 2019 Novel Coronavirus (COVID-19) pandemic has necessitated the implementation of various infectious disease control measures, including the closure of non-essential businesses, social distancing, and the virtualization of schools and universities. As final year nursing students at the University of Washington (UW) adjust to virtual learning and social distancing, certain students working in healthcare may also face the threat of contracting the virus. Little is known about the psychological implications of the COVID-19 pandemic on this population, and there is a need to fill this knowledge gap. This study first aims to capture the perceived stress levels of final year nursing students at the UW amid the COVID-19 pandemic. It secondly aims to explore associations between perceived stress and factors such as COVID-19 testing history, living situation, and healthcare work history. We administered an online survey to final year UW nursing students that inquires about their COVID-19 testing history, living situation, and healthcare work history since March 1, 2020. The survey also includes the 10-item Perceived Stress Scale, which questions students’ feelings and thoughts over the past month. We will analyze the data for associations between scores on the Perceived Stress Scale and students’ testing history, living situation, and work history. Overall, we expect to find moderate to high perceived stress levels among nursing students. We also anticipate that several factors may be associated with higher stress levels among nursing students, including having a history of COVID-19 testing, living with more individuals, and working more hours in healthcare positions. The results of this study may indicate a need for increased psychosocial support in final year nursing students at the UW, as they complete the nursing program and join the nursing workforce to help combat the COVID-19 pandemic.

Fall prevention in hospitals and long-term care facilities in the community setting depends on staff being trained in best practices. However, little is known about the cost, availability, and accuracy of online trainings on fall prevention for nursing staff. The type of education given can be outdated, incorrect or unnecessary depending on what organization that the nursing staff belongs to. The purpose of this research is to identify and evaluate existing online fall prevention training programs and provide the findings to the Washington State Department of Health (DOH) for them to present options to provider organizations as part of the State’s Fall Prevention Strategic Plan. Materials evaluated were previously used for staff education in hospitals and long-term facilities. The criteria for evaluating the websites and educational learning resources (ELRs) were: authority, objectivity, authenticity, reliability, timeliness, relevance and efficiency. Cost was also considered in the evaluation as a factor. Following the evaluation, ELRs were classified by two independent reviewers as either highly recommended, recommended, partially recommended or not recommended for use. From our initial findings, we identified two ELRs that are highly recommended for use, two that were recommended, and three that were partially recommended out of 24 ELRs. After finalizing our results, we will randomly select modules to be evaluated by a group of University of Washington Nursing students for verification of our assessment in reliability and validity of the ratings. We will disseminate findings and recommendations to WA DOH Older Adult Fall Prevention Program and clinical partners through a webinar. This should improve training provided to health providers of care to older adults in hospitals and long-term care facilities. This evaluation method can also be applied to other ELRs to improve the quality of these offerings and assist providers in choosing the best available programs.

Public health nurses and leaders who work for small, rural health departments (LHDs) face unique challenges that prevent the effective use of data to understand and address health equities. Among these is limited access to high-quality training in data utilization. In response, Solutions in Health Analytics for Rural Equity across the Northwest (SHARE-NW) is developing an online learning hub that contains accessible training modules for LHD leaders and practitioners to use and interpret data easier. The purpose of this study is to determine gaps in the quality of the trainings available to rural LHD practitioners in six top health priority areas: obesity, diabetes, tobacco, mental/behavioral health, violence and injury, and oral health, in order to improve population-level health equity. To date, approximately 30 training modules have been evaluated using 25 items from the Quality Standards for Training Design and Delivery tool by the Public Health Learning Network, and approximately 50 webinars have been evaluated using a set of 6 criteria. Next we will identify the patterns of gaps across the evaluation criteria and top health priority areas. Once patterns are identified, we will conduct semi-structured interviews with public health practitioners to assist in the interpretation of identified patterns. Preliminary findings suggest a lack of training activities that assist LHD leaders and practitioners to learn about data utilization in several to health priority areas. By identifying the patterns of gaps in available trainings, we aim to help busy, low-resourced rural LHDs utilize the highest quality trainings to improve their ability to interpret data and address health disparities.

Premature infants are commonly born with an inability to feed properly from their mother’s breast, and enteral (tube) feeding is often too stressful on the still-growing body of a premature infant. A recent intervention called Oropharyngeal Colostrum (OPC) administration aims to safely and effectively strengthen the infant’s digestive and immune system, shortening the time to safe enteral feeding. Colostrum, the milk produced right after delivery, is exceptionally high in antibodies and proteins necessary for an infant’s development, and contains an even higher concentration of protective bodies after delivery of a susceptible premature newborn. With OPC administration, colostrum is swabbed on the infant’s cheek or tongue in small amounts on a regular schedule. When the colostrum comes in contact with the tissue in the mouth, the antibodies are taken in and boost the baby’s immune system, heightening resistance to common infections suffered by premature infants, especially in the gastrointestinal region. In 2018, the Lactation Program of the Hospital de la Madre y el Niño (HMN) in Formosa, Argentina began administering OPC to all infants admitted to the Neonatal Intensive Care Unit who were born <35 weeks gestational age and whose mothers consented to the procedure. This research will examine average time to enteral feeding among 400 premature infants administered OPC between 2018-2019. Differences are examined in average time to enteral feeding by gestational age, sex, Apgar score, birth weight, and colostrum administration. The data will be used to determine what characteristics in premature infants cause the most obstacles to enteral feeding. Results will be compared to other studies of time to enteral feeding with and without OPC administration. Results will be shared with the HMN administrators to inform them of the efficacy of the program.

An estimated 20.6 million people live with Human Immunodeficiency Virus (HIV) in Eastern and Southern Africa. In 2012, the World Health Organization (WHO) announced Option B+ which adopts a single, universal regiment to both treat HIV-infected pregnant women and prevent mother-to-child transmission of HIV. Unlike other HIV treatment options, Option B+ is a lifelong provision of antiretroviral therapy (ART) for all HIV-positive pregnant women regardless of their immune status or viral load. However, I hypothesize that the implementation of prolonged HIV treatment (Option B+) may pose a substantial financial burden for women in very resource poor settings, which could ultimately decrease ART adherence--resulting in no change or even an increase in maternal transmission. This research will examine how economically accessible Option B+ is for expecting mothers in different populations across Eastern and Southern Africa. I am statistically evaluating the cost of ART in relation to household income, living expenses, and cost of childcare at the national level. Future research will use this data to examine maternal transmission risk in relation to economic accessibility, and identify how conceptual frameworks of HIV healthcare are implemented in varied resource settings.

The modern human longitudinal arch is a structural adaptation to bipedal locomotion, rigid enough to be a propulsive lever but flexible enough to stabilize a large body mass over a relatively small base of support. Early hominins appear to have a low or absent longitudinal arch—a condition equivalent to human pes planus. The morphology of the calcaneocuboid joint (CCJ) has been suggested to play a critical role in the stability of the longitudinal arch, but the relationship between articular shape and arch height has yet to be tested empirically. This study examines the covariation of the shape of the CCJ with arch height, via calcaneal inclination angle (CIA). Eleven calcaneal measurements and CIA were measured on weight-bearing radiographs of 103 patients from an urban US trauma center. An equation to predict CIA was determined using stepwise regression analysis using Böehler’s angle, plantar and distal angles of the calcaneal tuber, anterior angle, and the angle of inclination of the posterior talar facet (all p’s<0.01, R2=0.67). Landmark data were obtained from 3D surface scans of the CCJ of 24 calcanei from lower limbs amputated due to infection or acute ischemia. Individuals with bony pathology were excluded from analysis. Data were analyzed in R using Procrustes ANOVA with principal components analysis to explore patterns of variation within the sample. No significant associations were found between CCJ shape and CIA (p=0.51, R2=0.03, F=0.91). Consequently, calcaneocuboid joint shape may not be a useful for interpreting the longitudinal arch morphology of fossil hominin pedal remains.

Disproportionate representation of genders has long existed among different academic fields. Rising interest in gender equality in society, generally, has stimulated concern about how much progress has been made towards gender equality in academic activities. Analysis of peer-reviewed publication authorship patterns shows that archaeology and other academic fields have long been dominated by males. We ask if this gender disparity is evident among archaeologists participating in major conferences, such as the Society of American Archaeology (SAA), European Association of Archaeologists (EAA) and the Computer Applications and Quantitative Methods in Archaeology (CAA) meetings. Do these archaeological conferences show a trend over time towards a more balanced distribution of genders among their participants? We analysed presenter names in the published programs of archaeology conferences to infer gender. We found that the gender distribution from these conferences differs from the distributions of genders in peer-reviewed publication authorship. Recent conferences show approximate gender parity, and some even show more women than men presenters. We also find no relationship between the gender of the conference presenter and the topics presented, meaning that no one gender seems to favor one topic over another topic in their presentations. Our finding demonstrate that women and men are participating in archaeology conferences more equally, compared to journal article authorship. We expect these findings will be similar in other fields especially with increased priority given strong interest in gender equality in many research communities.

Seasonal and weather change on humans have countless effects on mood, sleep, and diet. Seasonal affective disorder, also known as SAD, is a seasonally linked mood and behavior disorder that typically lasts for four months, starting in the autumn. Similar to depression, SAD involves symptoms of lowered mood, energy loss, fatigue and some atypical symptoms such as hypersomnia, and increased appetite and eating. SAD may have a genetic factor related to serotonin metabolism and melatonin secretion, and rates of autumn-winter SAD may increase with latitude because of the increased seasonal differences in daylight hours. Due to genetic variations among certain ethnic groups, SAD may also disproportionately affect populations who are genetically from southern climates but were raised in northern climates. Yet it is not known how prevalent SAD symptoms were historically in populations adapted to northern climates, and what cultural traditions, if any, these cultures may have evolved in response to SAD. To answer this question, I have conducted a systematic search of ethnographies available through the Human Relations Area Files (HRAF) database. The HRAF databases have indexed and coded ethnographic knowledge across over 400 cultures. I examine the prevalence and traditions of SAD in the indexed ethnographies, and apply these insights to current migrant populations in northern latitudes that may experience SAD.

Recent increases in atmospheric carbon dioxide levels have led to a decrease in the pH of the world’s oceans. These changes in seawater chemistry could have severe consequences for calcifying organisms such as corals. Ocean acidification (OA) poses an imminent threat to the world’s coral reef ecosystems, which could lead to massive reductions in global marine biodiversity. Much of the response of corals to OA is still poorly understood from a detailed physiological perspective, particularly the impact of OA on a fundamental process of skeletal growth called nucleation. Nucleation is the process by which calcium and carbonate ions combine in solution to form a new piece of solid calcium carbonate, which corals use to build their skeleton. While calcification has been measured in corals, the specific step of nucleation has not been quantified and has the potential to be more sensitive to ocean acidification. Coral fragments of Stylophera pistillata and Acropora microphthalma were grown on microscope slides secured in petri dishes in normal ocean conditions suitable for growth. Half of the fragments will be placed into another tank that has constant alkalinity and a set pH of about 7.8 controlled with bubbling air at 800ppm carbon dioxide into the water, the predicted level of atmospheric carbon dioxide in 2100 with the current rate of emissions. This will allow for a big enough change in pH to clearly determine if nucleation is affected by lower pH. With an inverted microscope, the calcifying space was visible through the glass slides, and time-lapses were taken to quantify nucleation rates of corals in the normal ocean conditions. Reefs play a huge role in supporting biodiversity, economies, and population health. Understanding how nucleation rates are changing in variable ocean conditions is a key step in developing new ways to conserve reefs.

The Pondaung Formation of central Myanmar consists of fluvial and alluvial sedimentary rocks deposited during the Middle Eocene, 40 million years ago. The formation has yielded fossils of some of the earliest anthropoid primates, coeval with the first primate dispersal from Asia to Africa. Previous sedimentological and paleobotanical studies have shown that the Pondaung Formation was deposited in a landscape of open-forested seasonal wetlands with isolated riparian and monsoonal deciduous forests. However, the spread of these forests and locations of primate habitats within this mosaic landscape remains unclear. This project aims to provide a more precise reconstruction of the landscape at primate-bearing fossil sites using isotopic data gathered from paleosols as proxies for vegetation and climate during its deposition. We propose to use carbon isotope data collected from soil organic matter to produce estimates of precipitation. They are combined with the carbon isotopic composition of pedogenic carbonates to give direct estimates of soil productivity and vegetation cover. Together, these data allow us to decipher the landscape at the fossil sites, providing context for the evolution and dispersal of these primates. Our preliminary results agree with previous studies of the region, showing that the landscape was forested and flooded for considerable periods of time, experiencing strong seasonality in precipitation.

The landscape of southwest Montana is dominated by topography and adjacent sedimentary basins that resulted from deformation associated with the Sevier and Laramide orogenies140-50 million years ago and ~70-40 million years ago, respectively. Stark changes in drainage and deposition from Permian to Miocene (300 to 5 million years ago) have been described from uplifted sedimentary sequences. Despite numerous studies, the precise timing of eastward propagation of the Sevier fold‐thrust belt and its impact on drainage, as well as the timing of sedimentary basin partitioning into smaller intra-basins remain unclear. For this project, I worked under the guidance of Dr. Licht and Megan Mueller to collect field samples in Montana, isolate zircon grains for analysis and synthesize the results. Here we present geochronological and petrographic data that explain the timing of changes in drainage patterns, and place constraints on the chronology of orogenic development from Permian through Miocene sedimentary rocks exposed near Dillon, southwest Montana. The techniques used in this study are zircon geochronology and sandstone petrography. Zircon geochronology determines the age of zircon minerals found in sediment and offers insights into the ages of the sediment source areas. Sandstone petrography is used to classify the mineralogy of grains in sandstones and allow us to determine potential sediment sources. Understanding the evolution of drainages and changes in sediment transportation is critical to reconstruct the complex tectonic history of this region and aid in uncovering the chronology of Sevier and Laramide deformation.

The uplift of the Cascade Range in Washington had a dramatic impact on regional ecosystems and global climate by creating continent-scale rain shadow effects, enhancing regional aridity, and providing an anchor to the North American ice-sheet during glacial ages. Despite their importance, however, the chronology of the Cascades uplift remains poorly understood, with proposed ages for the uplift ranging from Eocene (~40 million years ago) to Pliocene (~4 million years ago). Before the onset of uplift, rivers would have drained westwards across Washington State; the uplift would have disrupted these river systems and separated river drainages on both sides of the mountain range. The goal of this study is to determine when uplift began by determining when this drainage disruption occurred. To do so, we propose to compare the sedimentary provenance of geological units of various ages on both sides of the mountains. Our methods include U-Pb dating of detrital zircons and sedimentary petrography of sandstones. Sediment samples on the west side of the Cascade Mountains have previously been analyzed for these two methods on an earlier research project. We will focus here on new results from multiple samples from the east side of the Cascade Mountains, and compare them with samples from the west side to propose a time window for the onset of uplift.

Tabletop Role-Playing Games (TTRPGs) are an increasingly popular group activity known both for their collaborative nature and creative demands. Despite the growing popularity, TTRPGs have yet to escape their reputation as a medium for social outcasts to construct escapist power fantasies. The intent of my research is to show that these “power-fantasies,” can actually become a valuable way to explore political and personal identity. I constructed a TTRPG called Blank Spaces, designed for exploring identity, and played with 6 different groups for 22 hours. After each session, I interviewed the players and used recordings of these sessions to qualitatively analyze player experiences. Players were often surprised that they had unwittingly performed meaningful self-criticism and world-criticism, all while simply enjoying a game. This has demonstrated that TTRPGs can be incredibly powerful self-exploration, critisim, and development tools.

 After Andrew Wakefield, the physician who claimed a link between the MMR vaccine and autism, was discredited and disbarred, he spent almost a decade in relative obscurity before trying to clear his name and provide a concrete foundation for his pseudoscientific crusade. The method he picked to accomplish that was to publish a movie in 2016, meant to illustrate the supposed dangers of vaccination and the purported concealment of that fact by the Centers for Disease Control and Prevention (CDC) and assorted pharmaceutical corporations. This film, Vaxxed, failed – it was barred from the Tribeca Film Festival, and opened to a crowd of a few dozen. Nonetheless, it is an immensely useful work in understanding both antivaccination ideologies and tactics, as it represents a summation of all their ideas and rhetorical strategies in a single vector. Over the course of this rhetorical analysis, I consider the strategies used by Wakefield and his associates, and find that they focus on anecdotal evidence, a conspiracy narrative, and the manufacturing of doubt, fear and scientific controversy wherever possible. I then discuss the impact of this film, both its reception and its effects as propaganda, and the reasons why it appears to have been such a dramatic failure, when previously, Wakefield’s paper drawing connections between the MMR vaccine and autism stirred such controversy and was so successful. Finally, I offer a few suggestions regarding recommendations for individuals seeking to debate or rebut antivaccination rhetoric, as well as a general perspective on the continuance, such as it is, of the antivaccination movement.

The study of the rhetoric of science reveals the means of persuasion scientists use when communicating their ideas. In the mid-twentieth century, when the controversial oral contraceptive pill was being developed, rhetoric surrounding its biology and morality was cautious and conservative. Some scientists discussed only the revolutionary science behind the pill and others condemned it as socially and scientifically unacceptable, but few walked the middle ground between biology and morality. In his publication “Control of Reproduction in Mammals,” however, Doctor Gregory Pincus integrates science and morality. This rhetorical analysis discusses the techniques Pincus used to communicate his ideas and determine whether or not they successfully persuaded his audience. His diction, figurative language, and argumentative appeals are analyzed and compared to contemporary documents. In assessing his choices, recent studies of the rhetoric of sexuality and medicine are consulted. I find that, while Pincus’ methods may have been unintentional, he successfully communicated to a divided public through contrasting language, varied arguments, and polysemy. This research is valuable because studies of the rhetoric surrounding issues at the intersection of science, politics, and sexuality are surprisingly limited. As medical technology and society progress, it is crucial to learn more about the role of rhetoric in the communication of science in order to preserve scientific integrity, freedom of accurate information, and reproductive rights.

La Llorona's ghostly figure has haunted the pages of Chicanx literature for years as the monstrous woman. While her story shifts forms depending on the cultural context, the essentials remain: she was a woman, wronged by the father of her children, who now wanders the rivers at night wailing for the two children she drowned in anger, grief, or desperation. She has often been considered a monstrous figure whose function has been to regulate female identity. However, authors like Gloria Anzaldúa and Sandra Cisneros have sought to reclaim this ghostly visage from the grasp of patriarchal structures that condemn la Llorona's actions. Anzaldúa's poem "My Black Angelos" and Sandra Cisneros' short story "Woman Hollering Creek" revise la Llorona to acknowledge the female agency she represents. While critics have focused on feminine agency in these works, the function of the monstrous has been overlooked. The monstrous usually refers to something feared or uncanny with women and people of color's bodies representing cultural fears, but in these cases the monstrous is reimagined as a tool for agency. Through the lens of monster theory, and drawing on the theories of Jeffrey Cohen, Cristina Santos, and Luce Irigaray, this paper argues that Anzaldua's and Cisneros' representations of la Llorona develop feminine agency and community just as other critics have mentioned, but they also complicate monster theory by resituating the subjectivity to account for the postive monster of la Llorona. Through this, monster theory's dependence on a self/other dichotomy falls away and, with it, la Llorona's position as only a monster to be feared. Instead, these representations of la Llorona invite Chicanx women into the community of the monstrous, where Cisneros and Anzaldúa transform it from an androcentric space of "othering" and oppression to one of belonging and power. 

British women participated in public political work at a higher rate than ever before during the 1906 United Kingdom general election campaign. Working as fundraisers, door-to-door canvassers, and even orators, women were particularly active in debating the campaign's central economic question: should Britain uphold the free trade policies it had maintained since the 1840s, or should it follow its economic rivals, Germany and the United States, in strengthening tariff protections? Dorothy M. Hunter (1881-1977) rose to prominence during this period as a compelling public speaker who worked on behalf of the Liberal Party and its free trade agenda. Her career serves as a fascinating case study for understanding how some British women established their credibility in traditionally male spheres at the beginning of the twentieth century; nevertheless, historians of the Edwardian era have largely overlooked her work. Based on an analysis of the collection of Hunter's documents held at the Surrey History Centre in Woking, England, this paper argues that Hunter built her authority as an activist in the public sphere upon the conventional understanding of women's power over the private sphere. Following the rhetorical tradition established by Victorian philanthropists, Hunter increased the scope of her influence by extending the definition of "the household" and "domestic duty" to encompass public life and civic virtue. This strategy is present throughout her work from 1900 to 1914, from emotionally persuasive didactic literature written early in her career to economic arguments presented in public meeting halls at the height of her fame.

Members of the LGBT+ community have long been wanting accurate representation on the big screen. Following the outlawing of “sexual perversion” by the Hollywood Production Code of 1930, homosexuality of any form was only allowed if the character was shown as a villain so as not to encourage their behavior. Traces of this restriction still remain today. Bisexual people in particular are even more problematic with their onscreen images often being either negative or absent completely. This is true even of films created by and for members of the LGBT+ community. To illustrate the pervasiveness of these issues, my research has taken a critical look at four films, Gray Matters, Imagine Me & You, I Can’t Think Straight, and Kyss Mig (Kiss Me), that have bisexual themes and have been celebrated for providing LGBT+ characters with rare happy endings. My project is titled “Happy Endings for Whom?” because I argue that these films perpetuate narratives tying bisexuality with deception and infidelity that are harmful to bisexual people and other members of the LGBT+ community. To do so I break down the plot structures and how the main characters are described and portrayed (ex: Who are the heroes? Villains? How do they identify?). The goal of my research is to expose the power structures that exist within media about and for members of the LGBT+ community and encourage individuals to be more thoughtful audience members and content creators. 

The late nineteenth and early twentieth centuries saw a profusion of archaeological discoveries in Egypt which led to the period being dubbed the 'Golden Age' of Egyptology. While much of this archeological activity is tremendously important and well-known, less known yet equally significant are the personal histories of those present in Egypt during this time. The letters of Helen Winlock, wife of the Director of the Metropolitan Museum of Art's Egyptian Expedition, provide an intimate window into daily life and thought in the context of early twentieth century excavations. From bugs in the food and exploding furniture, to racially prejudiced observations of the locals and the difficulty of finding formula for a newborn child in the desert, each of Helen's many letters sheds light on the quotidian and mundane aspects of life in this often-sensationalized period. Her writings demonstrate the intersectiosn of race, gender, and colonial power in this time and place, as Helen's comments about herself, other women, and other peoples around her demonstrate the complicated politics of knowledge in this 'Golden Age' created and maintained. As a research intern in the Emma B. Andrews Diary Project, I have transcribed dozens of Helen Winlock's letters. Working from scans of the original handwritten letters, my role in transcribing these letters is a foundational first step in the process of creating digital editions of these documents, which will include versions encoded in XML, following Text Encoding Initiative (TEI) guidelines. This will facilitate the investigation of the archive using computational methodologies. Our team includes faculty, students and librarians from multiple departments across the University of Washington. This interdisciplinary and collaborative digital humanities project seeks to make a wide range of unpublished or inaccessible primary sources from the period freely available to scholars in an online format. 

This project examines what legal writings from ancient Rome reveal about the political ideology, social values, and power dynamics in Roman history. I focus on these concepts through analyzing selections from two sets of speeches given by Cicero, a politician, loyal proponent of the Roman Republic, and philosopher educated in both Latin and Greek. Additionally, I explore the scholarship on Roman legal history to provide supplementary cultural context. The sociopolitical climate of the late Roman Republic was tumultuous. Near the beginning of Cicero’s political career, he gave a set of orations, In Catalinam, to the Senate that accused Catiline of conspiring against the consuls. Much later, Cicero tried to keep the Republic alive after Caesar’s assassination and accused Mark Antony of being disloyal to Caesar by wanting to create an empire. This urge to defend the Republic prompted Cicero to write his Philippicae to attack Mark Antony. These orations ultimately resulted in Cicero’s death, as Mark Antony wanted, and the Republic ended. My research compares Cicero’s In Catilinam 1, 2, and 4 and Philippicae 4, 5, and 14: both sets involve murder plots, denunciations of powerful men, and the senatus consultum ultimum decree for Republican emergency. Specifically, I analyze how Cicero uses oratory to convince the Senate to declare Catiline and Mark Antony as public enemies. This process reveals elements of Roman sociopolitical culture, such as values, threats, and legal procedures, and follows these differences in this short, but crucial, time period in Roman legal and governmental history. Furthermore, it demonstrates the complexity between the government and the conflicting political ideologies during the late Roman Republic. Thus, through detailed analysis of these selected passages and their wider contexts, I explore how the Catilinarian and Philippic orations use references from Rome’s earlier history to adapt to, and reflect, their particular moments.

Traditionally, scholars of ancient Greco-Roman culture have focused narrowly on the lives of elite Greek and Roman men, failing to capture the experiences of non-elite men, women, slaves, and the indigenous populations that came into contact with Greeks and Romans. In fact, these groups together composed the majority of the ancient populations. This project asks if archaeological evidence related to food can illuminate how indigenous peoples reacted to Greek and Roman colonization. It takes as its case study the ancient site of Jerash (in what is modern-day Jordan), which during the second century BCE to the second century CE experienced a period of rapid growth, development, and colonization from the Greeks and then Romans. To answer the research question, the project collects and analyzes data from existing archaeological reports, focusing on food infrastructure such as market facilities and items associated with feasting and banquets across nearly all social strata. It then puts this archaeological evidence into dialogue with recent scholarship on colonialism in the ancient Greco-Roman world. The preliminary results of this research provide evidence of indigenous populations responding to colonization by assimilation. For example, excavated burial goods related to drinking parties are similar to those found elsewhere in the Greek world, and the food market is similar to Roman food markets of previous centuries. These findings, though not comprehensive, show that Greco-Roman practices related to food, introduced into ancient Jordanian culture during colonization, remained even after the period of colonization. Further work includes expanding the data set, as additional scholarship concerning floral and faunal remains from Jerash is forthcoming. 

Astrology, the prediction of events from the movements of stars and planets, has grown from Mesopotamian omen-literature to the astrology of Hellenistic and Roman periods, and strangely enough it has endured even into the modern day. In this project, by drawing on scholarly analyses of the intellectual and religious context of astrology (Barton) and the worldviews of the non-elite (Toner) I examine how astrology was interwoven into the political, intellectual and social framework of the ancient world; and how the authority of astrologers was both supported and challenged by the polyvalence of their discipline, as well as their ability to connect to individual consultees. As for ancient sources, the Astronomica of Marcus Manilius, composed as a didactic poem in the early first century CE, shows how astrology was understood and used by the elite. It considers astrology in a broad theoretical context, encompassing a wide array of astrological concepts. He also illustrates the historical period in which he writes, under the new Principate of Augustus, by using astrology to legitimate Augustus's self-made status as princeps. On the other hand, other works such as that of Dorotheus of Sidon concern the more practical side of astrology. He is less concerned with theory and more with specific horoscopes, and better depicts the sorts of questions and concerns clients of a variety of social status would have had. Finally, skepticism toward astrology in antiquity is best rendered in Cicero's On Divination and Sextus Empiricus's Against Professors. These critiques and others, however, were directed less at astrological theory and more at its practitioners. By coming to understand how ancient peoples viewed astrology as both an intellectual discipline and a practical tool, we can better understand their cultural conception of the universe and in the process provide a useful comparison to similar institutions today.

As the 2020 presidential election is fast approaching, per our democratic process, Americans will soon be held responsible to fulfill, arguably, their most important political right. Elections are thought of by many as both a mechanism of accountability and a measurement of performance. Although fair elections are a necessity to uphold democracy, it is essential to recognize that this belief is a misconception. The result of a political election is predominantly decided by the culmination of individual voters making decisions based upon their backgrounds, experiences, and beliefs. To examine this notion further, it is our goal to address the question: How does obtaining a college education affect political voting patterns? To accomplish this task, we have obtained data from the American National Election Survey (ANES). The data includes over 50,000 survey responses, ranging in years from 1948 to 2016, covering topics that include, but are not limited to, wealth, background, beliefs, ethnicity, age, gender, and socio-economic status. With this data, we have performed a regression analysis that is weighted and uses the Taylor Series adjustments to compute design-consistent standard errors. Through our research, it can be seen that the trends reveal a widening gap between educational attainment and party affiliation. Furthermore, those with higher education hold more leftward leaning beliefs, and in turn, are more likely vote Democrat when compared to those with less education. Lastly, the trends over-time reveal a dramatic shift; as, non-college-educated Americans used to primarily be Democrat, while college-educated Americans were mostly Republican. We feel that this no longer holds. This research has implications to demonstrate the importance, or lack of importance, of higher education in terms of the effect and influence it has on real life decisions, which in our case is voting patterns for presidential elections.

In this project, I seek to examine the relationship between economic development and Russian influence in the post-Soviet states of Romania and Bulgaria. While scholars have studied the individual paths that these recently formed states have taken since the Soviet Union collapsed in 1991, there has been little inspection of how these nations regard the now Russian Federation. My work studies this relationship at a closer level. Specifically, I examine data on economic development at the district level (standardized by NUTS 3) in these countries and vote share for pro-Russian parties. I run a multivariate regression to examine the effect that level of development has on citizens’ willingness to accept Russian influence in their country, controlling for other relevant factors. I hypothesize that as the amount of development increases, the level of vote share for pro-Russian parties will consequently decrease. This hypothesis is grounded in the well documented positive effect that development has on democracy. In this case, a nation such as Russia, who touts anti-democratic ideals, would be looked upon less favorably by a district that is relatively more developed. Thus, I expect to see an inverse relationship between my variables. This analysis will inform research on the interactions between nations in Eastern Europe and Western Asia, areas of growing importance in this world, and allow us to examine the path forward for other post-Soviet states.

This project seeks to explore the effect of spending on state-level ballot initiative campaigns on voter turnout. Past research into the area of voter turnout has revealed evidence of relationships between campaign spending, voter turnout, and vote outcomes. Differentials in spending by competing campaigns has been demonstrated to have clear effects on vote outcomes in a variety of electoral contexts, and aggregate spending has been shown to affect levels of voter turnout as well. However, no work has in the past sought to look exclusively at the role of spending allocated to ballot initiatives in its ability to affect turnout. I anticipate higher levels of spending by pro and opposition groups correlating to higher overall voter turnout. In addition, I expect spending by opposition groups to have the greatest impact, as these groups are able to allocate funds mainly to media ads which seek to influence voter behavior through psychological aspects like fear. Using publicly disclosed data on ballot initiative spending tied to election year, the relationship between spending and voter turnout for the previous six biannual elections in four states with active ballot measure processes is investigated. Types of election, changes in voter partisanship, voter registration, and quantities and policy areas of ballot measures considered are controlled for to establish a baseline for voter turnout. The influence of campaign spending on turnout is sought after to help bridge-the-gap in relationships between spending and vote outcome, and turnout and vote outcome.

Today, over 50% of the world's population live in cities or urban areas. While cities only count for roughly 2% of global landmass, they contribute over 70% of CO2 emissions globally. With a significant environmental footprint, cities have the opportunity to be integral in climate change action. While many cities globally have begun to implement climate politics and targets, there is still a wide spectrum of engagement. Therefore, it's important to assess when and how cities tend to implement climate policy and why they engage in climate action. Through the Center of Environmental Politics, we are researching what factors lead small to medium sized cities in the state of Washington to implement these climate change policies. Random cities are selected, and data is being gathered largely by looking through the cities website's and surveying for statements of climate change and the resulting policies and goals. Currently, we are in the process of analyzing the data so no set results and conclusions can be resolved yet. However, the results of this research can help us determine the hurdles when bringing climate change rhetoric and action into the political sphere and potentially highlight the political atmosphere around climate change in Washington cities.

The unlimited potential uses of data, coupled with the power of artificial intelligence to find and learn patterns has the ability to transform our world and solve humanities problems. As a result of the abundance of financial data, there is a new opportunity to create models which can learn from patterns and predict future price movements. The goal of this project is to create a portfolio with pure alpha. A portfolio which will produce a consistent stream of returns with little risk. Using data analysis, the portfolio will be constructed with thirty to fifty uncorrelated assets, with the hypothesis that the uncorrelated assets will remove risk due to both general market volatility as well as movements from other assets in the portfolio. Machine learning and artificial intelligence techniques will be incorporated to design an algorithm which will continually learn from itself and optimize the weightings of the assets in the portfolio. To ensure optimal returns, I will use back testing to see how the algorithm would have performed historically in different market conditions, and how the machine would re-learn from past observations to improve in future out of sample testing. While predicting the future is difficult by humans, using machine learning techniques where self-learning is continuous, allows for faster predictions with less lag time and bias than humans are capable of.

Task prioritization, or choosing which order to do tasks, is an essential skill to possess. However, there are no definitive answers to the question of “how” we prioritize tasks. There are numerous factors that are considered in task prioritization, including a vast array of queuing rules (FIFO or First In First Out, EPT or Earliest Processing Time, etc.) However, many of these systematic methods of prioritization don’t account for task complexity. Another common way to prioritize tasks, which will be the focus of this study, is finding the task that seems the closest in time. This perception of Temporal Distance does address task complexity because we balance task complexity and due date when deciding which task is closer or farther away in time. Though individual Temporal Distance Perception is a widely researched topic, there has not been much research conducted on application in a team setting. Prioritizing one’s tasks individually is difficult, but it gets much harder when others’ schedules need to be factored in as well as an individual’s schedule. In addition to this issue of schedule navigation, this research will take place in a multidisciplinary setting, so teammates will not be able to know in detail how long a teammate’s task will take. This too makes it more difficult to choose which task comes first. My research question is this: How does Temporal Distance Perception in a Multidisciplinary Setting affect Task Prioritization in Teams? I am conducting a synthesis of literatures about Task Prioritization, Multidisciplinary Teamwork, and Temporal Distance Perception, as well as creating a data model based off the synthesis. I was sourced the results of team building exercises from NASA’s Human Research Program, which include a variety of real-world examples of how astronaut teams prioritize tasks.

Internet of Things (IoT) devices are increasingly integrated into new construction and legacy buildings. However, the insertion of IoT into buildings increases their cybersecurity vulnerability. While IoT security issues are ubiquitous across devices, a foundational issue for cybersecurity in buildings is the organizational friction due to the increasing integration between Information Technology (IT) and Operational Technology (OT), called IT/OT convergence. These two technological systems are embedded in the siloed institutional histories and work practices that historically managed these technologies: Operations and Management (O&M) and IT. This siloed evolution generated different organizational structures and cultures that facilitate a responsibility vacuum in organizations around IoT cybersecurity. Industry stakeholders suggest that stronger collaboration between O&M and IT could mitigate these challenges. However, improving collaboration requires multi-disciplinary knowledge work and task coordination that fits the policy and organizational contexts of O&M and IT professionals. This project seeks to identify different interactions between macro-level legal policies and formal organizational policies and procedures, and how micro-level daily work practices support the knowledge work and task coordination needed to improve O&M and IT collaboration around IoT cybersecurity. To tackle this subject, our team is undertaking a multi-method qualitative study of three parts: (1) ethnographic study of IOT cybersecurity practices by IT and O&M departments at three small, medium, and large universities in Washington state: UW Bothell, Western Washington University, and UW; (2) 25 expert interviews with IT and O&M cybersecurity professionals working in universities across the United States; (3) 15 case studies of universities in the Pacific Northwest region. The data collection methods for these three parts are observations in the workplace, interviews, and document analysis.The research being presented here will consist of two master inventories of (1) policies addressing cybersecurity, IoT device security, data privacy; and (2) smart city initiatives in the United States.

Cannabinoids, the main constituents of Cannabis, are a class of highly abused compounds of which, Δ-9-tetrahydrocannabinol (THC) is the primary psychoactive molecule. Despite the wide use of THC, its metabolism in humans is still in need of greater understanding. THC is metabolized to 11-OH-THC and sequentially to 11-COOH-THC by cytochrome P450 enzymes (CYPs) 2C9 and 2C19. 11-COOH-THC is then further conjugated to form 11-COOH-THC-glucuronide by UDP-glucuronosyltransferases (UGTs) 1A1 and 1A3. THC and its subsequent metabolites have been shown to bind to liver-type fatty acid binding protein (FABP1). FABPs are intracellular lipid binding proteins (iLBPs) that regulate the homeostasis of their endogenous ligands by solubilizing these hydrophobic compounds in the cytosol. Knockout of FABP1 in mouse hepatocytes was shown to decrease the formation and clearance of 11-OH-THC while the metabolism of 11-COOH-THC appeared to be unaffected. The goal of the current investigation is to translate these results into the human liver. Previously, our lab expressed and purified human FABPs to test their effect on THC metabolism in incubation assays with human liver microsomes (HLMs) and recombinant enzymes. After initiating the THC reaction with the CYP cofactor, NADPH, the 11-OH-THC product was extracted and quantified using LC-MS/MS. We found that both FABP and albumin changed the metabolic rate of THC in an enzyme specific manner. Because 11-OH-THC formation was altered in the presence of FABPs compared to the HSA control, we extend this method to continue our investigation with 11-COOH-THC metabolism. Considering that UGTs are on the luminal rather than the cytosolic side of the endoplasmic reticulum and that 11-COOH-THC has greater water solubility, we expect to observe enzyme and substrate specific effects of FABP. 11-COOH-THC and 11-COOH-THC-glucuronide are biological markers of THC metabolism so understanding this metabolic pathway is important for developing better methods of characterizing THC use in humans.

The link between protein sequence and structure is not always apparent. The dogma is that sequence determines structure, but it is not clear how very different sequences can give rise to the same structure. Here, we employ high temperature molecular dynamics unfolding simulations to probe the pathways and specific interactions that direct the folding and unfolding of the SH3 domain, a family of small proteins consisting of two β-sheets arranged to form a barrel. SH3 domain proteins are involved in various functions including protein binding, cell signaling, and nucleic acid modification. The SH3 metafold consists of 753 proteins with the same structure but varied sequence and function. To investigate the relationship between sequence and structure, we selected 17 SH3 proteins with an average pairwise sequence identity of only 27%. Six unfolding simulations were performed for each protein and unfolding transition states were determined, revealing two unfolding/folding pathways. Transition states were also expressed as mathematical graphs of contacts between chemical groups, and three positions in the transition state structure were consistently more connected to the rest of the graph than other nearby positions. These positions represent a folding hub connecting different portions of the structure in the transition state. Analysis of the multiple sequence alignment and covariation also highlighted positions with high conservation due to packing constraints and long-range contacts. This study demonstrates that the SH3 domain can fold through two distinct pathways, but certain folding/unfolding characteristics are conserved independent of sequence and unfolding pathway. By identifying similar interactions, we demonstrate how different sequences can have the same influence on folding pathway and final structure.

Foldit is a citizen science game developed at the University of Washington that allows people to predict and design protein structures for use in scientific research. The game challenges ordinary people to map the complex energy landscape of a protein sequence to find the lowest energy structure as determined by the protein modeling software Rosetta. The lowest energy structure is the one most likely to fold. While Rosetta’s software uses random sampling to predict a protein’s structure from its sequence, Foldit takes advantage of people’s intuition about things like avoiding steric clashes and large cavities to produce structures that can rival or exceed the accuracy of what Rosetta’s software can produce. However, there are still some flaws in Foldit that can cause a large fraction of player designs to not fold correctly. One of the biggest issues concerns an excess of buried unsatisfied polar atoms in player designs caused by imperfect modelling of the electrical potential energy of these atoms in the game. These atoms can prevent a protein from folding by eliminating the energy gap between folded and unfolded states. We have introduced a score penalty which penalizes designs with buried polar atoms. We are using existing scientific software to determine these atoms, but we have had to modify the relevant code to make it fast enough to be playable while retaining accuracy. We have also tested our calculations against known ways of calculating these atoms, and against previous Foldit puzzles to see if adding the penalty improves designs. We hope to even test this filter on some recent Coronavirus binder puzzles to get better solutions. An increase in design quality would mean that such a score penalty is warranted and it would allow for Foldit structures to be more useful across a range of protein design applications.

RNA-based therapeutics have attracted significant interest as a promising alternative to traditional cancer therapy methods. Coupled with other technologies such as the application of nanocarriers, RNA-based treatments have shown the potential to regulate gene expression in tumor cells with high efficacy and reduced safety risks. Despite these advantages, delivering sufficient amounts of therapeutic cargo while escaping from the proteolytic environment of the endosome has remained a long-standing challenge. This project focuses on addressing this issue by evolving the I53-50-v4 (V4), a self-assembling synthetic protein nanoparticle engineered to encapsulate its own mRNA, to increase RNA packaging and prevent degradation upon endocytosis. With previous data suggesting a decrease in mRNA encapsulation with an increase in mRNA length, additional designs were generated to further validate this finding. Furthermore, different variations of de novo pH-responsive trimers for endosomal escape were fused to the exterior of the V4 and tested for RNA packaging. Using computational tools, algorithms were also developed to model the porosity of the nanoparticle and to label residues for cationic mutations. Protein nanoparticles were expressed and purified via Immobilized Metal Affinity Chromatography (IMAC) and SEC (Size Exclusion Chromatography). Constructs were then analyzed in their mRNA encapsulation levels through RT-qPCR. Success of this project would demonstrate an increase in encapsulation levels with a decrease in mRNA length. As the phenotype of each nanoparticle design is spatially linked to its genotype, the top performing designs can also be selected and further evolved using both experimental and computational approaches. This will allow the development of a novel assay for screening desired proteins and narrow the gap towards achieving an efficacious RNA-delivery system.

 Host restriction factors are components of the innate immune system that play a vital role in inhibiting viral infection. One such example is a family of antiviral proteins, APOBEC3s, that inhibit retroviruses such as HIV by hypermutating the genome through cytidine deaminase activity. There are seven members of the APOBEC3 (A3) family found in primates, A3A-A3H. Each of these APOBEC3s vary in their protein expression levels and antiviral activity Antiviral activity of these proteins depends on expression levels and ability to be packaged in budding virions. A3C, in particular, is of interest because it is highly expressed and packaged but lacks potent antiviral activity when compared to its A3 counterparts. Moreover, when we examined A3C expression across primate evolution, we found that many primates encoded only unstable versions of A3C. Preliminary data, on the other hand, suggests an alternative role of A3C in modulating the effects of the more potent A3 proteins, A3D, A3F, and A3G. Coimmunoprecipitation assays suggest that A3C interacts with other A3 proteins, and subsequently decreases the packaging of the potent A3s into budding virions thus inhibiting their antiviral activity. Future experiments include removing endogenous A3C from cells to determine if the antiviral activity of other A3’s increases in the absence of any A3C. These data suggest that A3C has evolved to fine-tune the amount of antiviral activity of other A3 family members, thus allowing for better control of these potential mutagenic proteins.

When all goes according to plan, newly synthesized proteins within cells fold down an energetic funnel into a functional, minimal energy configuration. If a protein does not fold properly, it is both energetically unfavorable and nonfunctional, often with hydrophobic parts exposed to the aqueous environment. This creates the potential for misfolded proteins to form insoluble aggregates, which are toxic to cells. These aggregates crowd the cellular environment and impair cellular functions, which on a single cell scale, leads to cell death, and on a larger organism scale, leads to diseases like Alzheimer's, Parkinson's, and Huntington's. To deal with this problem, cells have protein quality control (PQC) systems. PQC is composed of two classes of enzymes: chaperones that help proteins to fold properly and ubiquitin-protein ligases that tag misfolded proteins with ubiquitin, leading to degradation in the proteasome. Previous studies concluded that chaperones are required for protein degradation. In this study, we investigated yeast ubiquitin-protein ligase San1, which often requires chaperones, but can recognize substrates independently. The ubiquitin-protein ligase San1 recognizes patches of hydrophobicity on misfolded proteins and is able to accurately tag them for destruction in the proteasome. Primarily through the use of cycloheximide-chase degradation assays and fluorescent microscopy, I worked within our team and found that Hsp70 chaperone dependence is variable along a spectrum of independent to dependent. By studying the interactions of the folding and degradation enzymes, our lab is gaining new insights into the coordination of PQC pathways

Lymphatic malformations (LM) are congenital vascular malformations resulting from dysregulated growth of lymphatic vessels. LM can cause difficulty in breathing, swallowing, or eating, and in rare cases, infection or death. LM are associated with postzygotic somatic mutations in PIK3CA, which encodes for the catalytic subunit of PI3K and regulates cell growth and proliferation. Three hotspot mutations cause the majority of LM: p.E545K, p.E542K, and p.H1047R. These mutations occur at very low level in the affected tissue; usually at <10% variant allele fraction (VAF) which makes diagnosis challenging. We hypothesize that variant allele fraction (VAF) differs throughout the vascular malformation and will correlate with location in the lesion. We used droplet digital polymerase chain reaction to detect PIK3CA mutations in 8 LM subjects with 95 samples including lesion, muscle, skin, and fat. We then mapped the VAF to magnetic resonance imaging to assess genetic heterogeneity by sample location. Globally, no sample had a VAF greater than 10% indicating that assaying multiple samples does not increase the observed VAF compared to other studies where one sample was assayed. Within patients, VAF varies within the lesion, ranging from undetectable to 10%. Non-lesion tissue, including skin, and most muscle samples, had no mutation detected. All four fat samples and two muscle samples were positive for mutation with VAF <2%. We did not identify any correlations between VAF level and location within the lesion. Though we didn’t identify a correlation between VAF and location in our data, these findings will be used for further analysis including understanding how cell type composition within the samples correlates with VAF. Our goal is that this dataset will help us explore the development of LM and eventually provide information useful for the advancement of targeted therapies for future pediatric patients.

The quality of contraceptive counseling has been shown to influence contraceptive use. Minorities and women of lower socio-economic status (SES) report that the contraceptive counseling they received as being lower quality than white, or higher SES women. Additionally, while contraceptive counseling is usually provided through verbal interactions with the clinician, evidence shows patients value the use of visual aids, such as videos, as part of the visit. The extent to which minorities or lower SES patients are satisfied with a contraception counseling video when compared to white or higher SES women is unknown. This project compares satisfaction ratings of a hormonal intrauterine device (IUD) counseling video of minority and lower SES women to white and higher SES women. To date, 93 English-speaking women ages 18+ from family planning clinics in Chicago, Seattle and Los Angeles have been counseled by their clinician and had chosen to receive a hormonal IUD watched a 6-minute counseling video the day they received their IUD. The video showed racially/ethnically diverse clinicians who provided factual information about the IUD, and either white or Asian IUD users who described their personal experiences with the IUD. After watching the video, participants completed a satisfaction survey containing 8 categorical and 2 open ended questions. We will compare the satisfaction scores of the Likert Scale questions and use thematic analysis for the open-ended questions to determine whether attributes of the video differed along demographic lines. Of the 93 women who watched the video, 55% were white, and 45% were African American, Asian, Hispanic, Native American, or multiracial. Approximately 55% had a less than a college degree, while almost 40% reported a yearly income of $25,000 or less. Further analysis may show that certain aspects of the video are perceived differently depending on the demographics of the participants.

 Hoarseness is a common symptom of multiple laryngeal diseases such as inflammation, paralysis, neurologic disease, or laryngeal cancer. Many patients with these diseases are not diagnosed with the correct underlying cause of the hoarseness early enough. Therefore, healthcare providers need better methods to screen for and evaluate different types of hoarseness. Currently, a combination of tools are used to evaluate voice disorders in specialty clinics such as patient history, perceptual voice evaluation, and laryngoscopy. We want to better understand how providers with different medical backgrounds evaluate patients with voice complaints. We are most interested in seeing how history, perceptual voice evaluation, and laryngoscopy impact decision-making and diagnosis. In addition, our group has developed a machine learning algorithm that analyzes voice to detect the presence or absence of a laryngeal mass. We want to see if this algorithm could be clinically useful for generalist providers. To address these questions, a group of clinician evaluators including general practitioners, otolaryngologists, and speech language pathologists, will be recruited remotely. Subjects will be asked to complete an electronic questionnaire with patient case scenarios, asking them to evaluate hoarse voice samples and laryngoscopy exams, with and without case history. For perceptual voice sample evaluations, clinician performance will be compared to the algorithm’s classification of whether a hoarse voice is from someone with a laryngeal mass. From there we will see if clinician detection of laryngeal masses from voice could be improved with this algorithm. If the algorithm has better performance than clinicians, then it may be clinically useful as a screening tool in the future. Our results will help us understand how evaluations for hoarseness are done and can be improved.

In 2018, tuberculosis (TB) was the leading cause of death by a single infectious disease, causing 10 million new cases and 1.5 million deaths. Upon Mycobacterium tuberculosis (Mtb) infection most people develop non-transmissible latent TB, and some develop active TB disease. Healthy hosts have a 5-15% lifetime risk of progressing from latent to active TB, and prior studies have demonstrated that a type I interferon-stimulated gene (ISG) signature can predict progression to active disease. Type I interferon (IFN) and ISG expression is induced by DNA-sensing pathways and has anti-viral functions. However, the innate immune mechanisms and genetics controlling type I IFN responses following Mtb infection are not well understood. We hypothesize that genetically regulated higher type 1 IFN responses are associated with lower anti-microbial responses and higher Mtb replication in macrophages, as well as increased risk of TB disease. To test this hypothesis, blood was collected from 40 healthy donors. Donors were genotyped using Illumina MEGA^EX SNP Array. Monocytes were isolated, differentiated into monocyte-derived macrophages (MDMs), and stimulated with 3 ligands (supercoiled plasmid DNA, cyclic guanosine monophosphate–adenosine monophosphate, and sheared calf thymus DNA) to activate DNA-sensing pathways and induce a type I IFN response. RNA was isolated at 4 and 24 hours. Interferon-beta (IFN-β) and interleukin-6 (IL-6) gene expression were quantified using Real-Time PCR. Donors had highly variable IFN-β induction upon ligand stimulation. Donors’ genotypes will be linked to these in vitro phenotypes to identify expression quantitative trait loci (eQTLs) that regulate IFN-β expression. We will assess if these functional polymorphisms in genes of interest are associated with TB disease using patient samples from a Brazilian cohort. The results of this investigation will identify novel pathways that control TB progression that can inform vaccine development and host-directed therapeutic approaches.

yEvo (“yeast evolution lab”) provides experimental evolution teaching modules to high school classrooms to bring genuine research experiences to students. Our goal is to show students how evolution can be applied to solve biological problems by asking them to carry out their own evolution experiments. Recently, we developed an experiment where students evolve yeast in wood hydrolysate media, which comes from lumber waste and contains plant sugars that yeast can convert to biofuel. Wood hydrolysate is a stressful condition for yeasts, as it contains acetic acid and harsh phenolic compounds, therefore yeast struggle to grow in this media. Mutations that yeast acquire while evolving in this media could be used to engineer a better biofuel-producing yeast. Students at Foster High (Tukwila, WA) evolved yeast in this media for seven weeks with transfers to fresh media every week. We retrieved the student’s evolved yeast and they showed striking new characteristics. We then used whole genome sequencing to identify underlying mutations. Some yeasts developed ring-like growths above the wood hydrolysate media and whole genome sequencing of two of these clones revealed mutations in regulators of cell-cell adhesion. Other yeasts reached very high population densities but whole genome sequencing revealed no novel mutations. I then utilized DNA staining and flow cytometry to check the DNA content of these evolved yeasts to see if changes in ploidy caused their new characteristics. The results showed that these yeast effectively became diploid, which has been shown by other labs to increase fitness in sugar-rich environments. yEvo is currently designing future experiments to further expand upon these interesting new results so that we can further understand mechanisms of adaptation to wood hydrolysate.

Of the estimated 38 million HIV infections globally, approximately 1-2 million are people living with HIV-2 (PLHIV-2). Infection with HIV-2, which is endemic in West Africa, is characterized by lower viral loads (VL) and slower disease progression to AIDS than HIV-1. However, many PLHIV-2 eventually progress to AIDS and death if left untreated. Antiretroviral therapy (ART) for HIV-2 is complicated by fewer effective drugs and significant challenges in drug resistance testing to identify appropriate therapy. This study aims to validate a novel method of HIV-2 drug resistance testing using nucleic acid from dried blood spots (DBS). One hundred and fifty DBS have been collected as part of two clinical studies of ART for HIV-2 in Senegal. HIV viral nucleic acid (DNA/RNA) is extracted and the regions encoding protease (PR) and reverse transcriptase (RT), which are the most common drug targets, are amplified by PCR, then sequenced. Sequence data are examined for evidence of drug resistance-associated mutations. To date, we have tested 115 samples with a median viral load of 84 (range: 0-24,000) and obtained drug resistance data from 43 (37.3%). Testing was most commonly successful from samples with higher viral loads; samples with viral loads >250 copies/ml were successful 75.7% of the time. We observed several drug resistance mutations, including PR V47A (5 patients), I50V (3 patients), and L90M (2 patients), and RT K65R (4 patients), Q151M (2 patients), and M184V (10 patients). Eight patients had no known resistance mutations. Once DBS testing is complete, we will select a subset of sequences to compare to genotypic resistance testing from corresponding plasma samples to evaluate the sensitivity of DBS-based testing vs. standard methods. DBS-based resistance testing has the potential to revolutionize ART for HIV-2 by allowing faster, easier, and cheaper assessment of drug resistance to optimize second-line therapy for HIV-2-infected patients.

Embodiments of additive manufacturing that utilize hydrogels as building materials have recently received much attention for their ability to construct biological systems in vitro. These embodiments, often referred to as 3D bioprinters, build up layers of cell-laden hydrogel by rasterizing two dimensional patterns of material deposited in a layer-by-layer fashion; the most common mechanisms of pattern deposition include extrusion, where a shear-thinning gel is forced through a thin nozzle, and light-induced polymerization, where a laser polymerizes material from a vat of liquid hydrogel precursor solution. These methods of material deposition work well for hydrogels which are designed and optimized for their respective deposition method, however, many unique and useful designer hydrogels cannot be printed using conventional 3D bioprinters. Herein, we describe a novel method for layer-by-layer fabrication of hydrogel structures using open microfluidic patterning. For each layer of a printed hydrogel structure, a hydrophilic track or “rail” is manually placed parallel to and several hundred micrometers above the previously patterned layer. Hydrogel precursor solution is then introduced between the underlying layer and the rail, and flows along the rail via capillary action. The cross-sectional geometry of the rail constrains fluid flow to the space directly under the rail, meaning that the pattern of each layer of hydrogel is defined by the pattern of the rail corresponding to that layer. We show that this methodology can be used to fabricate relatively large (1 cm^3) structures of agarose gel, as well as cell laden structures of collagen and a novel peptide-based synthetic hydrogel. Finally, we show that the patterning rail can constrain fluid flow via differential surface chemistry, opening up the possibility for an automated 3D printer and advancing the commercial viability of the method.

Native mass spectrometry (MS) experiments provide direct mass measurements of intact proteins and protein complexes. Protein samples for native MS are prepared in solutions that mimic physiological conditions, which maintain a protein’s native folded state before entering the gas phase of the mass spectrometer. Ammonium acetate solution is typically used due to its volatility and relevant ionic strength. However, protein purification protocols typically require inorganic salts and detergents to maintain protein stability. Native MS experiments can be hindered or made uninterpretable by those salts and detergents. Furthermore, the presence of protein modifications or multiple proteins can make native mass spectra difficult to interpret. Anion exchange chromatography (AEX) is well suited for the requirements of native MS, as it can simultaneously desalt, remove non-ionic detergents, and separate proteins or proteoforms directly into an ammonium acetate solution. This project seeks to develop a comprehensive method for desalting, removing non-ionic detergents, and separating proteins through an ammonium acetate-based anion exchange chromatography method. Preliminary experiments in egg whites, a complex matrix with a high sodium concentration, showed separation and four distinct proteins using an AEX pH gradient from pH 10 100 mM ammonium acetate to pH 4 100 mM ammonium acetate. Native MS analysis showed low interference from sodium or other contaminants and the various modified forms of those proteins were identified. Refinement of this preparation technique can result in the improvement and efficiency of native MS analysis of proteins.

Mutations in Complex I (NADH:Ubiquinone Oxidoreductase) of the mitochondrial electron transport chain have been reported in up to 30% of pediatric mitochondrial diseases (MDs) and affect 1 in 5000 live births. One of such pathologies, Leigh Syndrome, is often fatal in the first three years of life and has no known cure. Knock-out (KO) of Ndufs4 in mice recapitulates several aspects of the disease, including lethargy, encephalopathy and retarded growth. Acarbose delays the onset of neurological symptoms and prolongs the lifespan of both Ndufs4−/− mice and heterogeneous wild type mice. Acarbose is a type 2 diabetes drug that inhibits alpha glucosidases, resulting in delayed absorption of complex carbohydrates and increased activity of the intestinal flora, including increased levels of short-chain fatty acids (SCFAs) and other fermentation products. No formal study has been conducted to determine whether increased SCFA concentration mediates the effects of acarbose on longevity and MD suppression. Our goal is to elucidate the mechanistic basis of acarbose. We hypothesize that acarbose delays MD progression in Nduf4-/- mice by increasing circulating levels of SCFAs. Thus, SCFAs supplementation should recapitulate the effects of acarbose. We will feed chow containing tributyrin to control and mutant mice from weaning until humane endpoint. We will measure body weight, observe the incidence of forelimb clasping behavior (a widely tested neurological symptom), and record percent survival daily. We expect to observe a delay in clasping behavior and prolonged survival in acarbose-treated KO mice compared to that of untreated KO mice, and that these effects are recapitulated more pronouncedly with higher tributyrin doses. SCFAs are promising therapeutics because they are natural metabolites that are inexpensive and can be manufactured into easy consumable pills. A successful outcome of this study will help progress therapies for patients with MDs and for anti-aging purposes.

Cells use a variety of mechanisms to regulate gene expression. One way cells control gene expression is by forming structures such as DNA loops to position genes in 3D space near regulators. My lab is developing tools to synthetically control 3D genome structure and assess the relationship between positioning and expression. Our strategy is to engineer DNA loops by fusing DNA binding domains to targeting domains that dimerize and bring the two sites together. To ensure the binding domains bind to each other to form a loop, we are developing an allosteric sensor of DNA binding where the dimerization motifs are only active when bound to DNA. Without this switch, the looping interaction would be outcompeted by free binding domains that are not attached to DNA. We used LOCKR, a bioactive protein switch comprised of a protein cage that switches to an ON-state in the presence of a key protein. These proteins are tethered to DNA using dCas9 as a programmable binding domain. I have tested different system parameters, including the length of the linker between dCas9 and the key protein, for increased activation of green fluorescent protein, a reporter gene that is expressed when the switch is activated. My data show that the switch is effective with a variety of linker lengths. I am also exploring other parameters for optimization, such as changing the relative orientation of the dCas9 complexes. Exploring these parameters is important because the switch might be sensitive to small changes in structure or orientation, and we want to identify the optimal arrangement of dCas9 and switch proteins for effective switch function. After this system is optimized, we will be able to direct our efforts toward looping DNA, which will allow us to address broad questions about the relationship between gene position and expression.

A wide range of diseases and disabilities are associated with mitochondrial dysfunction including diabetes, cancer, Alzheimer’s disease, cardiovascular disease, and Parkinson’s disease. Mitochondrial dysfunction triggers activation of a novel mitochondrial retrograde response pathway in Caenorhabditis elegans termed the PMK-3 Retrograde Response, which upon activation leads to a reduction in mitochondrial stress and extension of lifespan. A mitogen-activated protein kinase (MAPK) cascade comprised of DLK-1, SEK-3, and PMK-3 forms the signaling core of the PMK-3 Retrograde Response. In this presentation, I will outline my attempts to develop photo-regulatable PMK-3 and DLK-1 kinases which are tools that can aid in the identification of other proteins involved in this pathway. These photo-regulatable kinases were made by engineering a recombinant form of the fluorescent protein Dronpa into two discrete sites of DLK-1 and PMK-3 to confer photosensitivity of the kinases. Photoregulation of the kinase occurs through dimerization of Dronpa in violet light (inactive kinase) and dissociation in cyan light (active kinase). After determining the identities of phosphorylated proteins purified from nematodes exposed to cyan and violet light using mass spectroscopy, a comparative analysis between the two datasets will suggest which proteins are involved in the PMK-3 Retrograde Response. Further investigation into these proteins could elucidate the role of these proteins in mitigating mitochondrial stress.

Traumatic spinal cord and brain injuries may lead to a devastating loss of neurological function. After the traumatic primary insult, a secondary injury phase ensues, which greatly increases the extent of the injury. Inflammation is a dominant component of secondary injury, which includes the immune response in which free radicals and proinflammatory cytokines are released that induce the death of surrounding neurons. Dexamethasone, a commonly used corticosteroid, has been shown to produce a neuroprotective effect by inhibiting inflammation and reducing cytokine release. However, the clinical application of large systemic doses of steroids is limited by side effects, such as sepsis and pneumonia. For this reason, a localized microneedle delivery system may be favored to allow for a therapeutic dose at the injury site, with reduced side effects. The goal of this work is to design a local, controlled drug release system, in which the drug could be given for a sustained duration at a therapeutic dose at the site of injury. Using a mixture of polyvinylpyrrolidone (PVP) and poly (ethylene glycol) diacrylate (PEG-DA), a biodegradable microneedle array can be formed that can encapsulate dexamethasone that then slowly release into the tissue over 72 hours. With a 3D printed system, various sizes can be quickly developed and produced to create customizable arrays for various target locations. Using a 2-photon polymerization laser, Nanoscribe, arrays can be formed on a scale of <300 μm as well as larger arrays using traditional printers. The needles are developed using a three-stage molding method, casting customized 3D printed arrays in a silicon elastomer, PDMS, and polymerizing our monomer-drug solution to the desired shape. Current results indicate sustained release over the target window and are currently branching into trans-membrane in-vitro studies.

Quantum confined nanomaterials have become an important field of study with many applications from color displays to low-energy alternative lighting sources. Discovered in the early 1980s, these semiconducting nanocrystals continue to draw attention; their unique properties differ from their bulk counterpart’s due to a quantum confinement effect rising from their small nanometer-scale size. Indium phosphide (InP), a group III-V semiconductor, is a promising nontoxic, environmentally innocuous material. My research is currently focused on investigating the implications of destabilizing kinetic InP intermediates. I explore how creating small nuclei with the same zinc-blende structure as quantum dots (QDs) can potentially lead to unique shelling applications to increase photoluminescence (PL) and color purity. I have developed a synthesis employing a destabilizing additive to yield the small nuclei, which I then shell with zinc chalcogenides to produce luminescent core-shell QDs. I characterize all material via UV-vis and emission spectroscopy as well as powder X-ray diffraction. My goal is to optimize the shelling procedure to produce QDs suitable for use in the market, particularly as blue emitters with a narrow emission range and high PL quantum yield. In optimizing this process, I can contribute meaningfully to the nanocrystal field and the display industry by presenting a unique strategy for making small materials through careful control over crystal phases.

Over recent decades, scholars have noted declining trends in perceptions of legitimacy in high-income, democratic countries. Many theories offer competing explanations for this decline, including immigration, polarization, and neoliberalism, but scholars have yet to connect declining perceptions of legitimacy to stagnant wages. I argue that within the Group of 7 (G7) countries, wage stagnation triggers a crisis of legitimacy because ordinary citizens’ living standards worsen as wages remain low and the cost of living rises, resulting in the belief that their governments are unresponsive and illegitimate. I hypothesize that wage stagnation plays into a legitimacy crisis in domestic democratic institutions, because stagnant wages increase economic inequality and generate perceptions of an illegitimate government. To test this claim, I create a legitimacy index to compare trends in perceptions of legitimacy cross-nationally. I also run a multivariate regression test to assess the strength of the relationship between stagnant wages and government legitimacy, controlling for other relevant factors. This project may reveal that if a productive economy fails to benefit its people, there is the possibility of democratic decline.

The implications in the connection between the Fifth Amendment and Miranda warnings have been well explored in Miranda v Arizona, highlighting that understanding such rights are relevant during police interrogations. The standardization of administering Miranda warnings before police interrogation becomes a mechanism in which to legitimize the State as well as the system of justice, since doing so becomes a form of checking and balancing the power dynamic between State and individual. The question then becomes, is there an effective way to clearly communicate an individual’s protection under the Fifth Amendment through the Miranda warnings in cases where the individual in question may be inherently more vulnerable, such as a child or a person with intellectual disabilities. In 2016, King County implemented a different set of Miranda warnings catered to juvenile competency. It is also crucial to examine how police are trained to understand the difference in the language in which they are mandated to employ. In this research, I aim to find out what and how the different sets of language of Miranda warnings influence the police’s state of mind. I anticipate on obtaining qualitative data by interviewing the police and other legal actors. First, I will ask police to recall what they are thinking after I have given a word cue in a randomized order to see if they are able to give the same meaning in both the adult and youth version. This should yield a baseline towards how the police are thinking about language. Secondly, my questions will also pertain to the ways in which the police are trained to interact with juveniles. I expect to find from interviews that the component of training is what will change how police perceive juveniles differently, rather than administering the warning.

After Saudi Arabia announced its VISION2030 plan to reduce its oil dependence through economic diversification, a wide-scale crackdown on corruption was carried out. Following that was the imposition of the first tax policy in the Kingdom, a 5% value-added tax. These reforms strike a delicate balance; to develop infrastructure and new industries, particularly for an oil-dependent economy (ODE), the country must sustain high levels of foreign direct investment (FDI) among other sources of funding. Nonetheless, attracting FDI requires transparent and resilient fiscal institutions, and the presence of corruption negatively impacts FDI by conveying uncertainty and increasing costs of conducting business. ODEs may also seek to enhance non-resource taxation, providing another stable revenue stream. FDI enlarges the non-resource tax base by including new firms and increasing employment opportunities. By relying on FDI as an indicator of institutional quality, this paper tests the interaction of corruption and FDI and its impact on levels of non-oil tax revenue. I employ regression models to conduct a cross-national study of 17 ODEs, controlling for population, GDP per capita, government expenditure, oil sector ownership and oil price. I expect to find that higher levels of corruption lead to lower levels of FDI, which in turn decreases non-oil tax revenue collection. Additionally, I will use a Fiscal Reliance Measure (Haber and Menaldo, 2011) to test the same interaction’s effect on the ratio of hydrocarbon and oil revenue to total government revenue as a second proxy for economic diversification. This research contributes to the growing field of oil-dependence and economic diversification in two ways: it rejects the presence of a “resource curse” and examines the relationship between corruption and non-resource tax revenue by studying institutional state structures, and it explores the causal mechanism running from FDI to tax revenue, whereas previous literature has tested the inverse relationship.

One of the Ultracold Atoms Group’s themes is to study the interaction between trapped ultracold atom mixtures. Certain experiments, such as the study of spin-dependent Feshbach resonance, requires us to select one specific nuclear spin state of the atom from the mixture. This process is achieved by the optical Stern-Gerlach technique, where we use the laser to produce the magnetic field gradient. However, this technique requires us to use the imaging path with relatively poor imaging quality due to, for example, vibrations of the optics. When we normalize these atom images, these vibrations introduce misalignment between images and thus unwanted noises. Hence my project is focused on stabilizing the imaging process by a software implementation of imaging alignment scheme. The code I developed can be seen as the analog to the inner product of two vectors, which characterizes the extent of misalignment between two vectors. This scheme has increased the efficiency of the experimental procedure and at least doubled the signal-to-noise ratio. Its easy implementation provides another route to reduce noises in the data of similar experiments. 

HIT-SI3 is a plasma physics experiment built for studying magnetic confinement of a fusion plasma for eventual clean energy production. HIT-SI3 utilizes steady inductive helicity injection to form and sustain spheromak equilibria, a stable arrangement of plasma. A tomography system has been installed to assess the symmetry of plasma density in the HIT-SI3 spheromak plasmas. The tomography diagnostic consists of four toroidal chord fans and three sets of three poloidal fans that provide 3D plasma emission information. Each fan expands from a wide-angle lens with a 130 degree field of view coupled to bundles of fiber optics. The light collected by the fiber optics is split into two paths, filtered at 668 nm and 728 nm HeI emission lines, and imaged by a high-speed camera. Since the ratio of the 668/728 nm emission has a strong plasma density dependence within the range of typical HIT-SI3 plasma parameters, the 3D emissivity profiles constructed by inverting line-averaged emissivity along chords can be related to the plasma density profiles. The reconstruction of emissivity profiles constitutes a highly underdetermined and ill-posed inversion problem and the maximum entropy method was chosen to find the most physically informed solution. My work in this project is to develop an algorithm that performs plasma density reconstruction accurately on HIT-SI3 to facilitate the study of plasma dynamics and comparisons to HIT-SI3 simulations. According to the NIMROD simulation, the HIT-SI3 plasma has lower density at the center of the flux conserver and higher density around the edge of the wall. We expect that the plasma density profiles reconstructed by the tomography system will capture this "hollow" feature.  

In this project, we consider two particle scattering with an arbitrary finite-range potential interaction, contained in a volume by a harmonic trap. The properties of this scattering system are then studied through Effective Field Theory. From this analysis, we attain general relationships among effective range parameters in various dimensions. This project is an extension of confinement induced through periodic boundary conditions studied previously. From this work, calculations can then be made to deduce properties of multi-body condensates. Currently, we already have shown the general relationships between effective range paramaters in toroidal compactified spaces. 

Recent studies reveal differences in the number of calories consumed by women residing in rural communities compared to individuals living in urban areas. Research suggests that there are many physical activity (PA) barriers for these women. Certain environmental and social factors hinder PA engagement, including environmental safety concerns as well as lack of PA facilities. For the current study, survey administration commenced on September 2018 and concluded October 2018. Researchers partnered with Grays Harbor Community Hospital to administer a 32-item electronic survey to N= 237 female employees. Inclusionary criteria included, English-speaking and self-identify as overweight/obese. My research team conducted a thematic analysis of a single open-ended question, “Are there any barriers to you engaging in any type of physical activity?” and the respondent was asked to explain the barriers as a follow-up. In total, 60 women completed the health surveys, (25% response rate). The mean age of respondents was 53.2 years of age, with a mean BMI of 32.2 (obese status), and majority were healthcare professionals in the hospital. We analyzed the data by conducting iterated evaluations of responses to the question and then deriving themes to characterize respondents’ perspectives. In response to the question of barriers to PA engagement two emergent themes were evident (1) daily life obligations (family and career hinderances) and (2) acute/chronic health concerns that limited physical activity engagement (arthritis, fibromyalgia and back pain concerns). In view of our findings, rural women appear to experience barriers to physical activity engagement. These barriers can help us to identify strategies that specifically target the health needs of residents in rural communities.

Social determinates of health are such factors as lack of education, low income and low social support that can lead to health disparities. Presence of health disparities can lead to poor communication between patients and providers and consequent poor health decisions among patients. Marginalized populations such as racial/cultural minorities, elderly and the mentally ill are shown to face many of these unique challenges when accessing care. This study aims to determine the ways in which the healthcare system can better support the needs of these safety-net patients. In Washington state, electronic surveys were administered to patients accessing services at a community health center that offers healthcare to disadvantaged and low-resourced communities. The questionnaire was a 47-item instrument that evaluated patients' perception of health services recieved. We were tasked with conducting a qualitative evaluation of a single item on the questionnaire that asked about patients' perceptions of how the healthcare system can best meet their healthcare needs. Using a thematic analyses approach, we evaluated survey responses from 61 patients by reviewing responses for codewords, codeword clusters, and finally, themes that accurately captured perceptions. Two themes were derived in response to the query of how the healthcare system can best support healthcare needs. (1) Accessibilty of affordable resources. (2) Availability of healhcare resources that can support decision making. These finding show that even with access to health resources, low-recourced populations still face barriers to fully gaining access to affordable care to meet healthcare needs.

Although curable, tuberculosis (TB), remains a global health threat. The long (6-9 months) daily antibiotic regimen is one of many challenges for patients and providers in completing treatment. Patient-centered innovative strategies are recommended that promote self-management and support during TB treatment. The World Health Organization endorses mHealth, mobile devices facilitating health information/services, as an under-explored area of potential patient-centered strategies. To help address this gap, Dr. Iribarren with her team iteratively developed a mobile application (app) with end-users and experts. 42 patients underwent pilot testing in Argentina, randomly assigned to use the app or not. The app includes daily medication tracking, symptom tracking, a paper based test-strip to detect the presence of drug metabolites in urine and resources surrounding treatment and disease processes. In addition, the intervention supports interactive communication with the participant and the treatment coordinator, a registered nurse. As an honors student, my goal is to gain insight on how the 20 patients assigned to use app are engaging through qualitative analysis of the interactive communication within the text-intervention using Atlas.ti. A codebook was iteratively built to classify the types of interactions and define with representative quotes. The main participant themes surrounded questions on side effects and medication, reinforcement of good compliance and reasons for not reporting. The relationship between codes are described, for example, the nurse inquiring about missed report(s) triggers a patient response. To understand how experience in treatment affects usage quantitative analysis compared codes by month in treatment showing higher occurrence trends during first two months. This research will inform further refinement of app, with stakeholders in mind. The goal of understanding the communication themes is to suggest areas where adaption to the app could improve ease of use and support for participant while decreasing the burden to treatment supporter through automated responses where appropriate. 
 

Safety-net systems in the United States are crucial in ensuring that low-income and low-resourced, marginalized communities have access to healthcare. Many of the individuals accessing safety-net clinics within their communities are transgender, people of color, and/or Spanish speakers who often-times have unique healthcare needs compared to the general population. The aim of this study is to understand which needs of safety-net patients are not currently being supported by community health clinics, so that they can be considered and implemented in future healthcare frameworks. Our study team reviewed data from a sample of 61 patients from community and human services centers in Washington, DC. The patients who agreed to participate and met the eligibility criteria were taken to a private room where they completed a 47-item self-administered questionnaire. Using thematic qualitative analyses, we evaluated the responses to a single item addressing patients’ perceptions of how the healthcare system can address their healthcare needs. Themes that emerged from our analyses of the single item include the following: (1) safety-net patients seek more resources to support decision-making, (2) patients require accessible and affordable healthcare services, (3) transgender patients require improved gender-related healthcare, and (4) Spanish-speaking patients require more attention and compassion from clinical staff. These findings imply that even with access to healthcare, certain populations continue to face significant barriers to achieving good health outcomes within the safety-net system.

In the United States, women in rural communities face a lot of barriers to healthy eating due to their food environments. Many barriers are associated with busy lifestyles, cost of purchasing healthy foods, lack of healthy food options, and lack of will-power which can lead women to eat less fruits and vegetables and consume more unhealthy foods. The purpose of my project is to evaluate the barriers to unhealthy eating among a sample of rural middle-aged women. Partnering with Grays Harbor Community Hospital, a 32-item electronic questionnaire was administered to N = 237 female employees and 60 surveys were received back (25% response rate). Inclusionary criteria included, English-speaking and self-identifying as overweight/obese. My research team did a thematic analysis of a single open-ended question from the questionnaire, “Are there any barriers to you eating healthy (at least 2 servings of fruits and 3 servings of vegetables each day)?” The respondent was also asked to explain what the barriers were. The mean age of respondents was 53.2 years, they reported a mean BMI of 32.2 (obese status), and majority were healthcare professionals in the hospital. We recorded the responses to the question of barriers to healthy eating and conducted a thematic analysis by deriving codes, codeword clusters and themes. Emergent themes indicated that the barriers to healthy eating among rural obese women were time constraints, affordability, dietary restrictions and nutritional preferences. These findings suggest that perceived barriers are strongly related to their dietary behaviors and this information can offer important insight about helping women to manage their health behaviors.

Parents’ use of early math talk with young children is important for later math outcomes. Existing research focuses on monolingual English-speaking populations and provides limited data on Dual Language Learners (DLLs). As the DLL population continues to grow, outcomes are important to investigate due to a possibly higher risk for academic disadvantage in comparison to their monolingual peers. The purpose of this project was to evaluate early maternal math language during mother-child play-based interactions in a primarily Spanish-speaking sample of low-income Mexican-American families. Specifically, this study investigated: (1) the type of math talk mothers used with their 3.5-year-old children in the context of semi-naturalistic play (2) how early math talk in Spanish predicted kindergarten math skills measured with standardized assessments. Mothers and children (3.5 years) played with toys (a bug-building kit and a pizza-making kit) for 15 minutes. Conversations between the mother and child were recorded and transcribed using Systematic Analysis of Language Transcripts (SALT) software. Transcripts were coded for instances of math talk, including numeracy, cardinality, quantity, spatial terms, and sequencing terms. We expected that preschool-aged children who were exposed to a greater quantity and diversity of numerical language during play would demonstrate more robust math skills in kindergarten measured with standardized academic assessments. We conducted descriptive analyses of mothers’ math talk and children’s language development (e.g., vocabulary and grammar) and used correlational analyses to see if mothers’ math talk was related to children’s language development. Predictive analyses assessed whether math talk predicted children's math outcomes in kindergarten. Findings from this work may have implications for parents, educators, and Speech-Language Pathologists in understanding how to support learning and academic success before entry to Kindergarten for the growing population of DLLs. Further research will help establish a better understanding of socioeconomic impacts on math talk.

This study investigates the ways in which parents consider and rate their child’s temperament in relation to the ways in which their child interacts with them during play. Participants were typically developing children (TD, n = 26) ages 2-5, children with autism spectrum disorder (ASD, n = 25) ages 2-12, and children with fetal alcohol spectrum disorder (FASD, n = 18) ages 4-9. Attentional focusing was assessed with a parent-report child temperament questionnaire; child’s age determined which Rothbart Childhood Temperament Questionnaire was given (for example, the Children’s Behavior Questionnaire for ages 3-7). Play sessions were unscripted, free play time between the parent and child with a consistent set of toys (M = 14.4 minutes, SD = 2.8). The number of child attention switches (to a different toy, person, or object) was coded from video. Preliminary results indicate that parents rated TD children as more attentive in comparison to children with ASD and FASD based on the attentional focusing score from the Child Temperament Questionnaire (p< .001). There were fewer attention switches during the play sessions by TD children in comparison to children with ASD and FASD (p<.05). There is no significant difference in either of these measures between children with ASD and FASD. There is also no correlation between parent-reported attentional focusing scores and the observed number of attention switches. These results suggest that TD children demonstrate more attentiveness in both parent-reported and examiner-rated measures. Children with ASD and FASD were not distinguishable from each other based on parent report or direct observation of attention. Overall, attention should be considered as a contributor in a child’s interactions with their environment, relationships to others, and further growth and development.

The International Panel on Climate Change report of Global Warming of 1.5°C (October 2018) strongly recommends unprecedented scale of systems transitions to ensure little to no overshoot of global warming of 1.5°C. In the interest of examining educational efforts towards this reality, this qualitative study is designed to understand how the field of early childhood education is adapting to living with climate change and preparing early learners for their future under the guiding principle of the required systematic change that must be met to mitigate the effects of the human ecological footprint on the environment. This qualitative study features a cross-cultural comparison of early childhood professionals’ ideas about the role that early childhood teachers play in helping children learn foundational ideas about climate change. Three participants from Washington state and 4 participants from Trondheim, Norway were interviewed and the content of the interviews was analyzed and coded for themes. Participants all had early learning teaching experience, but their roles varied from State policymaking, teacher educators, and early learning educators. Participants described barriers in supporting early learners’ understanding of, relationship with, and caring for the environment and how the field of early education can be intentional in supporting these goals. Participants identified lack of educator knowledge, lack of access to nature, family emphasis on spending time in nature, the risk of scaring children, emphasis on academic readiness, and cultural emphasis on environment as the main barriers to effectively preparing early learners for their complex future. All the participants felt children were capable of understanding topics related to sustainable practices, cause-effect, and co-existing with the other life on this planet. The next steps of this work is to determine how to best prepare early educators in supporting the goals identified by participants through professional development or teacher preparatory programs.

Aphasia is a language impairment that is often acquired due to left hemisphere stroke. Although current treatment for aphasia is successful, it is often limited to what is explicitly trained during treatment. That is, when conducting a single-word treatment protocol, persons with aphasia tend to only improve on naming the words given during treatment and show little to no improvement on untrained words. This study is focused on identifying what conditions improve naming untrained words after receiving aphasia treatment. For 8 individuals with aphasia, we assessed change in naming 120 pictures after undergoing a repetition priming task for 30 of those words. For a given “trained” word, its corresponding picture, printed name, and spoken name were presented to the participant four times, followed by an opportunity for the participant to verbally produce the name. For pre- and post- test naming, we implemented a standard coding method to ensure specific and consistent scoring, where responses were only scored correct if the participant verbally produced the entire picture name. We found that all participants improved in naming the 30 trained words. However, our results were mixed regarding improvement on naming the untrained words. Critically, we noticed two participants whose responses were not always verbal productions of the word, but rather spelled words (in one case verbally and in the other case through writing). This led us to question whether these coding parameters allow us to accurately identify if an individual is benefiting from treatment. We argue that incremental gains in productions (e.g., through spelling) also provide evidence of improved access to words. We focus this presentation on how alternative coding schemes for these two participants might change our interpretation of their outcomes, and discuss how future research should consider more inclusive scoring methods.

In 2016, women earned 57% of bachelor degrees awarded in the U.S. but only 42% of bachelor’s degrees in the field of mathematics and statistics (U.S. Department of Education, 2018). Math-related careers tend to have higher salaries than other careers (Hira, 2010), meaning that the lower proportion of women receiving mathematics and statistics bachelor’s degrees may be contributing to the gender wage gap. One potential explanation for this lower proportion may be the perception of math as valuing speed over accuracy. Societally, many speed-focused characteristics and activities, such as impulsivity and racing, are associated with masculinity. If this leads women to believe that they are less speed-focused and more accuracy-focused than men, women may avoid a field they view as emphasizing speed more than accuracy. In an ongoing study (target N = 100), undergraduate women and men will be presented with two different student reviews for a math class - one emphasizing the importance of having speed and the other the importance of having accuracy. We predict that women, but not men, will have greater interest in the class emphasizing accuracy than the class emphasizing speed. We also predict that when asked to make a choice between which class to take, women will be more likely to choose the class emphasizing accuracy and men will be more likely to choose the class emphasizing speed. Results will have important implications about the causes of gender gaps in participation in mathematics and possible interventions to reduce these gaps. Future research should investigate the impacts of the perceived value of speed and accuracy on gender gaps in other STEM fields.

Racial microaggressions are subtle manifestations of racial bias. Though they are subtle, they cause tremendous psychological strain for minoritized populations. Prior research from our lab has found four main types of microaggressions: intergroup anxiety, color blindness, objectifying, and negative stereotypes. Intergroup anxiety refers to nervousness about interracial communication, such as being perceived as racist, and often manifests into avoidance of such interactions. Color blindness is the denial of racial identity, saying “I don’t see color” for example, while objectifying is being preoccupied by differences, such as fixating on a Black woman’s hair. Lastly, negative stereotypes are predetermined racial judgements based on known labels like “welfare queen.” In the research study, we sought to understand whether women of color experienced these or other microaggressions while seeking services following domestic violence. A focus group was conducted with advocates, people who provide emotional support and resources for domestic violence survivors, to gather information about their clients’ experiences. From advocate responses, we identified themes that fell into one of our four microaggression categories. In addition, we uncovered a fifth form of microaggression, and found that many survivors reported macroaggressions. This research could inform future training strategies for service providers, and help domestic violence survivors access services without encountering microaggressive racial bias.

Prescribed opioids are the most common analgesic used for alleviating acute and chronic pain. Despite this positive attribute, opioids are also highly abused drugs that can lead to tolerance and dependence. This abuse has caused a dramatic increase in opioid overdose-related deaths over the past couple of decades, deeming it a crisis. However, cessation of opioid use in tolerant individuals who wish to detoxify can precipitate severe withdrawal symptoms, often leading to relapse in order to avoid experiencing these negative symptoms. In recent studies, modulation of neuropathic and neuroinflammatory responses have been linked to withdrawal symptoms. As a result, we hypothesized that microglia, the resident immune cell of the central nervous system, serve as a potential target for withdrawal treatment. In order to test this, we reduced microglial activity by inhibiting the purinergic signaling pathway. This was achieved by first exposing mice to escalating doses of fentanyl over the course of a few days to create tolerance. Then, we administered clopidogrel, a selective antagonist of the P2Y12 receptors which are expressed in microglia, before inducing withdrawal using naloxone. Subsequently, in order to quantify whether inhibition of microglial P2Y12 receptors mitigated naloxone-precipitated withdrawal in fentanyl-tolerant mice, we measured avoidance of the withdrawal context with the conditioned place aversion (CPA) test, and evaluated somatic signs of withdrawal with EthoVision video analysis. Avoidance of the negative emotional and physical symptoms of withdrawal is a key driver of relapse, therefore the results from this experiment can provide prospective molecular pathways to target for future studies in treating opioid withdrawal symptoms. Reducing the severity of withdrawal would thus allow ease in discontinuing opioid use and diminish relapse.

Opioid abuse leads to over 40,000 annual deaths in the United States; even more individuals are impacted by anticipatory withdrawal anxiety and subsequent treatment avoidance. Despite the magnitude of this issue, there is a lack of effective treatments that address opioid dependence and withdrawal. Molecular responses to opioids have traditionally been linked to neuronal activity, but recent literature suggests that microglia also play a role in opioid addiction. Recent experiments we conducted reveal that opioid dependence and withdrawal have inverse effects on the microglia translatome, with morphine treatment correlating to decreased gene expression and withdrawal correlating to increased gene expression. We found a dramatic change in genes relating to cyclic AMP signaling during withdrawal, which has been shown to modulate microglia motility and potentially their interactions with nearby neurons. From this, we sought to further investigate the molecular basis of microglia morphology during opioid tolerance and withdrawal. To do so, we constructed four experimental groups consisting of mice who received saline followed by saline, saline followed by naloxone, morphine followed by saline, and morphine followed by naloxone. After obtaining the mouse brains through perfusion, we took sections of the striatum. In order to visualize and quantify microglia morphology, we performed 1ba1 immunohistochemistry to stain the slices, then imaged mounted slices on a confocal microscope to acquire confocal stacks of the striatum. This was followed by 3D reconstruction of individual microglia for analysis using 3DMorph software. The results of this experiment are a step towards clarification of molecular mechanisms behind opioid dependence and withdrawal for future work on mitigating the effects of opioid addiction. Alleviating withdrawal symptoms through translational research would allow users to more easily cease opioid use and therefore reduce opioid abuse mortality.

Pituitary adenylate cyclase-activating polypeptide (PACAP) is an excitatory neuropeptide which has been associated previously with stress, fear, and post-traumatic stress disorder (PTSD). One region that strongly expresses Adcyap1, the gene encoding PACAP, is the lateral habenula (LHb), a node of stress in the brain. Studying PACAP and its role within the LHb provide insight into the aid and treatment of a variety of stress disorders by understanding the mechanisms of these conditions. In order to investigate the function of PACAP within the LHb we used a 2x2 experimental design. We injected a virus expressing either a Cre-inducible excitatory Designer Receptor Exclusively Activated by Designer Drugs (DREADD) hM3Dq and RiboTag or Cre-inducible RiboTag alone into the LHb of Adcyap1-2a-Cre mice to activate and quantify gene expression in these neurons specifically. Mice were injected with either the DREADD-specific drug clozapine-N-oxide (CNO) or vehicle just prior to contextual fear conditioning, a behavioral procedure in which mice are placed in a novel chamber and given repeated foot shocks in order to elicit a fear memory. The following day, mice were placed in the same chamber without CNO and their time spent freezing indicated the strength of their fear memory. We hypothesized that mice which have their LHb PACAP neurons activated will have increased time spent freezing within the contextual fear chamber, indicating they have a stronger fear memory than the control groups. This study could shed light on the mechanisms of PTSD and other stress disorders.

Sexual harassment engenders post-traumatic stress disorder, anxiety, and depression in targets, and disproportionally affects women (vs. men) in the United States. Critically, many harassment cases in the U.S go underreported. For instance, only 25% of women formally report instances of harassment to their employer and less than 20% of women describe sexually harassing behavior at work as “sexual harassment.” The current project explores whether women’s self-perceptions of gender-prototypicality impacts the reporting of sexual harassment. Specifically, it explores whether the extent to which a woman views herself as prototypically feminine promotes the reporting of harassment after it occurs. To begin to explore this hypothesis, we first test whether women can be primed to feel prototypically feminine (vs. masculine; Study 1). Second, we describe our experimental methodology, and predicted results, for testing whether feelings of sexual harassment can be engendered in women in the laboratory. Implications for the reporting and reduction of sexual harassment are discussed, along with theoretical and empirical extensions related women’s individual differences.

The lack of intimacy has long been known to contribute significantly to our mental health and psychosocial adjustment. Empirical research has substantiated strong connections between the fear of intimacy and other mental health problems such as depression. One could expect that the fear of intimacy poses as an obstacle to forming intimate relationships. The lack of such intimate relationships causes the individual to feel lonely and precedes a negative cognition as seen in depressed patients. Many other psychotherapy patients, too, experience intimacy deficits. Not much research has delineated the mechanisms underlying fear of intimacy. Specifically, when we say that we fear intimacy, what do we really mean? In this exploratory research, we are investigating how dyads with different levels of intimacy can possibly interact with each other to derive a certain level of connectedness within the relationship. For example, it is possible that both individuals with high fear of intimacy can still connect well with each other and form a close relationship. What then, in this scenario, is causing the relationship to work? For individuals with different levels of fear of intimacy (i.e., one high, one low), what should we expect to see in terms of connectedness within the dyadic relationship? In this research, we investigate these issues in a sample of 35 dyads (people in ongoing relationships, including friendships, family, and romantic partners) who were recruited for a larger intervention study to improve relationships. We attempt to integrate our findings with what is known from social psychology about relational functioning to explain the interaction of different levels of fear of intimacy within the dyadic relationship. This research advocates the importance of early screening of individuals' fear of intimacy and raising awareness for those at risk of difficult dyadic relationships.

Ever since they first appeared during the Late Cretaceous, members of the palm family (Arecaceae) have been ubiquitous in the fossil record. Traditionally, palms have been considered a key indicator of warm climates. In addition to leaf macrofossils, fruit, and pollen, palm phytoliths have gained utility as paleoecological indicators. Phytoliths are microscopic silica bodies accumulated in the tissues of many plants. Different plant taxa have unique phytolith morphologies, making them useful diagnostic tools. However, palm phytoliths currently lack diagnostic resolution below the family level, limiting our ability to fully utilize these powerful tools. The goal of our project was to increase this resolution by analyzing the morphology of phytoliths from across the entire Arecaceae family in more detail than has been possible before. We used confocal microscopy to take sharp, high-resolution images of palm phytoliths. Using these images, we took several key measurements, to which we applied multivariate ordination methods. Our analysis allowed us to test how well we can differentiate palm subclades within Arecaceae based on phytolith morphology. Ultimately, we hope to use this information to determine when and where specific clades of palms appeared in the fossil record, increasing our understanding of the evolution of the palm family. This will also allow us to describe past environments in more detail based on palm phytoliths, including estimating more specific climate parameter ranges, and characterizing particular biomes and habitats.

Microplastics (plastic < 5mm) has become a prominent research topic over the last 15 years. Microplastic research papers are published around the world, using a variety of organisms, environments, and interactions as study systems. Findings from these papers suggest that microplastics have negative effects, like behavior disruption and cell death, on marine organisms. This research project uses scientometrics to analyze if trends in regional plastic policies are correlated with marine microplastic research papers. Utilizing spatial analysis, we compared the rate and spread of microplastic research and plastic policies across the globe to identify a statistical relationship between them. We then further explored this relationship by determining if the distribution of study systems in a region’s research affected the quantities of policies in that region. We used the Web of Sciences database to obtain data on microplastics papers from 2006 to 2018. Our data has revealed that specific species are being used in research more than others and that there are large concentrations of microplastics papers in areas like Europe and China. Countries with large amounts of plastic research also have the most plastic policies. The results of this project help decipher how marine microplastics research can have an impact on plastic policy.

Chondricthyes such as sharks, skates, rays, and chimeras share the slow maturation and low fecundity traits, thus making them susceptible to overexploitation. To ensure the survival of these vulnerable species it is necessary to protect the habitat important to their recruitment. The purpose of the present study was to determine the sediment preference of juvenile Beringraja binoculata to gain evidence as to where their nursery grounds may be located in the Salish Sea. To do this we conducted one-hour filming trials of three four-month old skates in a tank sectioned off into four different sediment size classes. Skate 1 died one-third of the way through the trials so was removed from statistical analysis. However, we found that Skate 2 and 3 preferred to bury and rest in the smallest sediment size of 0.125-0.3 mm grain size with p-values of 0.00126 and 0.0814 respectively. This is consistent with literature on different species of skates around the world. Ongoing research would be valuable to determining the reason behind this preference and to use the information to locate the Salish Sea nurseries.

CD8+ T-cell exhaustion during chronic human immunodeficiency virus (HIV) infection is characterized by increasing expression of exhaustion markers on the cell surface that lead to decreased function, greatly diminishing immune responses and therapeutic vaccination efficacy. Blocking the activation of one such marker (PD-1) using a monoclonal antibody (aPD-1) can help reverse exhaustion. We hypothesized that dosing with aPD-1 would boost the immune system and decrease expression of certain exhaustion markers on CD8+ T-cells throughout infection in order to improve therapeutic vaccine responses. To investigate this hypothesis, we studied how exhaustion changed over time in simian-human immunodeficiency virus (SHIV)-infected rhesus macaques treated with a combinatorial therapeutic regimen consisting of a conserved-elements vaccine to circumvent viral mutants, GS986 to reverse latency, CCR5 gene editing to prevent viral entry, aPD-1 to reverse T-cell exhaustion, and antiretroviral treatment. We characterized exhaustion in peripheral blood mononuclear cells (PBMC) with surface staining and flow cytometry, focusing on markers of exhaustion including PD-1, CTLA-4, LAG-3, TIGIT, and TIM-3 at various timepoints throughout SHIV infection and therapeutic vaccination. We observed higher CD8+ T-cell vaccine responses in animals receiving aPD-1, compared to animals that were vaccinated but not given aPD-1, suggesting aPD-1 dosing improved vaccine responses. We did not observe any significant correlation between PD-1 signaling and expression of other exhaustion markers, suggesting that blocking PD-1 does not reduce exhaustion by decreasing the expression of other exhaustion markers. However, we found a significant negative correlation between CD8+ T-cell vaccine responses and the expression of TIGIT (Spearman r= -0.75, p=0.007). As reduced TIGIT on CD8+ T-cells correlated with increased vaccine responses, there may be a role for dual TIGIT and PD-1 blockade in future studies. Defining the role of CD8+ T-cell exhaustion in therapeutic vaccine immunogenicity and efficacy is crucial to improving combinatorial immunotherapy towards a cure for HIV.

The spread of grasslands 26-22 million years ago has been linked to global climate changes in the late Oligocene to early Miocene. The pattern of vegetation change was established analyzing assemblages of microscopic plant silica (phytoliths) extracted from sediment samples from the Central Great Plains of North America. It is often presumed that as open-habitat grasses became abundant, vegetation structure concurrently transitioned from closed forests to open landscapes. However, recent work in the Cenozoic of Patagonia has pointed to a decoupling of grass dominance and habitat openness, each independently driven by climatic conditions. We set out to test if a similar decoupling occurred in the Central Great Plains by means of an a-taxonomic phytolith proxy using phytoliths produced in non-grass epidermal cells. Work in modern plants and soil assemblages has shown that the size and degree of undulation in these phytoliths (quantified by, respectively, Phytolith Area, PA, and the Phytolith Undulation Index, PUI) is correlated with the amount of light in the environment, reflecting habitat openness (measured as Leaf Area Index, LAI). We measure the PA and PUI of phytolith samples from Nebraska, dating 35 to 17 Ma, to reconstruct the regional LAI over time and place time constraints on the opening of habitats. By comparing this timeline to that of the rise to dominance of grasses, we hope to better understand changing vegetation and linked climatic conditions in Cenozoic North America.

During the Pleistocene Epoch approximately 13,000 years ago, retreating glaciers deposited several stranded icebergs where the wetlands on the north end of the North Seattle College campus lie today. We collected and analyzed several borehole samples within the Meridian Avenue Basin to determine the stratigraphy and identify organic material found within the basin. The stratigraphy of the basin revealed that these features are a result of Vashon Glaciation during the end of the Pleistocene Epoch. Our subsurface core samples exposed a glacial till-lined basin ranging from 3-9 meters deep with overlying layers of gravelly clay, silt, sand and peat. A carbon date we retrieved from organic material within the peat indicates the sediment was deposited during the last glacial recession. The fill of the Meridian Avenue Basin and the carbon date retrieved indicate that it was created by an iceberg stranded in a shallow, post-glacial lake during the last ice age. Unlike typical kettle basins, load-induced subsidence of this iceberg into saturated lake-floor sediments created its shape and depth. The basin lacks the characteristic outwash depositions seen in kettle lakes and appears to be unique, suggesting the geologic processes which may have created the basin seldom occur and are poorly understood. Researching the origins and stratigraphy of the Meridian Ave basin will help advance knowledge of this possibly rare periglacial phenomenon.

Microplastic (plastic < 5mm) pollution is increasing at an exponential rate in marine environments, leading to increased contamination of marine organisms. Previously, it was thought that the majority of microplastics were found in surface waters due to their positive buoyancy; however, recent studies show pollution exists throughout the water column. Mussels, suspension feeding bivalves, can be catalysts to the spread of microplastic pollution to different trophic levels through filtration, ingestion, and egestion of microplastics. Mussels are prominent benthic-pelagic coupling organisms in marine ecosystems, moving particles and nutrients between habitats through different types of biodeposits. Mussels produce feces and pseudofeces, where feces is the digested biodeposit and the pseudofeces are the rejected materials. Accumulation of microplastics in their biodeposits may interfere with this coupling by altering sinking rates, giving other organisms more opportunity to feed on deposits and promote the spread of microplastics through the food chain.The purpose of our project is to test how different types of microplastics affect sinking rates of mussel biodeposits. Our hypothesis is that mussels filter microplastic particles differently.The more readily ingested microplastic will lower the sinking rate of the feces more drastically. In our experiment we fed mussels either polystyrene or polyethylene microspheres along with algae, collected their biodeposits, and measured sinking rate. Further, we quantified microplastic and algae found in each type of biodeposit. The selectivity of mussels toward a particular microplastic was determined by the amount of a microplastic present in the feces vs the pseudofeces. The results from this study can help us understand the impact of microplastic pollution and how mussels play a major role in the spread of microplastics. They may also provide insight into which types of microplastic are more readily spread, potentially providing information on how to distribute microplastic waste.

A cardinal feature of Post-Traumatic Stress Disorder (PTSD) is avoidance of trauma-related stimuli. This avoidance is implicated in the development of PTSD. Other common features of PTSD include anhedonia, a loss of pleasure, as well as anxiety, and problematic substance use. Anhedonia may impact avoidance by decreasing motivation to approach and substance use is conceptualized as a form of avoidance. Trait anxiety is defined as a predisposition to response with feelings of apprehension, dread, and tension. Higher levels of trait anxiety have been found to predict avoidant decisions. The current study investigates the relationship between self-report avoidance and anhedonia, substance use, and trait anxiety. Participants will be recruited through community advertisement as a part of a larger experimental study. Participants will complete self-reports to assess constructs of interest, including the Snaith Hamilton Pleasure Scale (SHAPS), Multidimensional Experiential Avoidance Questionnaire (MEAQ), the Cannabis Use Disorder Identification Test (CUDIT), the Alcohol Use Disorders Identification Test (AUDIT), and the State Trait Anxiety Inventory (STAI). We hypothesize higher self-report avoidance will be associated with higher levels of anhedonia, higher substance use, and higher trait anxiety. Through their impact on avoidance, these constructs may help explain why some individuals are more likely to develop psychopathology following trauma. Targeting anhedonia, substance use, and anxiety-related cognitions after traumatic events could lead to decreased avoidance behavior and lower rates of PTSD development overall. Additionally, these constructs could prove to be important secondary clinical targets which help to attenuate avoidance behavior in those with PTSD.

Early home life and classroom environments are crucial to lifelong success. Family structure, opportunities for learning, and socioeconomic status (SES) influence experiences within the home. Further, SES impacts the resources that a student has access to, which can shape academic support and learning opportunities. Factors such as family climate, caregiver occupation and education, and parenting style have been shown to influence academic self-regulation and achievement. Literature has also pointed to the importance of relationships with peers and teachers on academic and social success. Few studies have examined how both of these environments impact student life into adulthood and litte research has explored social success in terms of feelings within interpersonal relationships and self-efficacy. The goal of the present study was to retrospectively explore experiences in the home and school environments throughout formative, developmental years and success later in life. Participants (n = 149) were asked about their family structure, SES, home learning environment (HLE), experiences in three separate academic eras, academic success, feelings in relationships, and self-efficacy. This study fills a paucity in the discussion of academic success by exploring a new measurement tool: the academic ladder, which examines a student's own perception of their achievement in relation to their classmates. Results confirm the impact of early experiences in the home and at school on later academic and social success. Interestingly, caregiver factors do not significantly impact students' interpersonal success as much as relationships with teachers and peers. Results also show that SES does not have an effect on opportunities for learning in the home; however, caregiver educational attainment affects those opportunities. Both HLE and caregiver variables have an effect on self-perceived academic success. Continuing to examine how educational experiences and environmental factors impact students' well-being will help develop intervention programs that best prepare youth to succeed throughout life.

Animals in the natural world rely on sensory information in order to make quick decisions to stay safe. Sometimes a stimulus is paired to a predictable outcome, allowing the animal to make the appropriate response when encountering that stimulus later. However, it seems unlikely that individuals would have to learn, de novo, those stimuli which might signal danger during the first learning opportunity. Laboratory studies of animal fear, however, have largely been done using Pavlovian Fear Conditioning; generating fear by pairing a tone or other benign stimulus with an aversive one—typically a painful footshock. This is problematic. An owl does not hoot three times before it strikes, allowing the prey to form a conditioned response. To explore this innate behavior, we developed a novel, naturalistic approach to fear conditioning that uses a pseudo-predator paired with physical pain to induce fear responses. In this experiment, I trained male and female Long-Evans rats to forage for food pellets in an open space. After training, a fake owl plunged towards the foraging rats while a shock was delivered through subcutaneously implanted wires. Certain groups were presented with a tone or light to pair with the owl and shock, while others received no other predictive stimuli. Additionally, to test the effects of environmental context on their behavior, animals were tested in two chambers that differed in size, lighting, shape, etc. Even after changing foraging contexts, some rats that were never conditioned to fear the tone fled upon its presentation. The visual cue also induced a fear response in rats that were never previously conditioned to fear the light stimulus, but only fled when the shock was presented with the owl. Experiments performed under naturalistic conditions utilizing animals’ innate behaviors will better aid in our understanding of how the brain makes decisions in complex environments

Homelessness rates among the adolescent population have steadily risen over the past decade, and homelessness is shown to be correlated with a variety of detrimental outcomes. Though prior studies of this population have focused on participant recruitment as well as the representativeness of samples, there has been little work regarding designing survey content to optimize participant honesty, retention, and willingness to divulge often sensitive details about themselves. As a part of a larger investigation on youth/family engagement with homelessness prevention services, this research project uses youth focus group responses to explore how, in conducting surveys, questions can be worded to increase the likelihood of obtaining an earnest and accurate response. We used data from six focus groups across two locations to gather information from housing instable adolescents. The focus groups presented example research surveys and asked participants to provide responses on whether the survey items were well-received and easy to understand, among other factors. Through a grounded qualitative analysis of the responses provided, we determined that there are topics that have a high probability of causing confusion and therefore reducing construct validity, or that may cause participants to become guarded because of their sensitive nature. To address these issues, we recommend phrasing questions to be unambiguous by being hyperspecific—for example, asking how often caretakers give praise for good deeds rather than a general question about pride in their children. We also suggest phrasing topics likely to evoke either a strong positive or negative emotional response in a more neutral manner or, when they are not strictly relevant, eliminating the questions altogether—for instance, instead of asking about whether someone is seeing a counselor directly, which could be a source of stigma and shame, asking instead about whether they have a professional that they can trust with their personal issues.

When a child is diagnosed with pediatric cancer, it puts a strain on the whole family, including the marital relationship. However, there is little consensus as to the factors that predict marital adjustment. This proposal examines the relationship between parental emotional styles and marital adjustment in the first year following a pediatric cancer diagnosis. I propose that poorer marital adjustment as well as declines in marital adjustment over the first year of treatment will be predicted by 1) emotion-dismissing philosophies in both men and women and 2) a mismatch in couples' emotion philosophies. These findings would provide insight into the factors that may put the marital dyad at risk, which could inform clinical interventions to better support the families after diagnosis.

I began this project last year as an honors student in the Psychology Department Honors program and under supervision of Dr. Lynn Fainsilber Katz and clinical graduate student, Laina Keim. I independenty discovered a gap in the literature, developed my hypothesis, performed an extensive literature review and wrote a proposal for this this honors thesis. I became a trained and reliable coder for the internationally recognized coding system that is being utilized in this study. This year I have focused on coding our data with our lab team, data analysis with Laina, and now I am writing up results. 

Chronic myeloid leukemia is among myeloproliferative neoplasms that cause an increase of myeloid cells. All CML cases have a reciprocal translocation causing the novel fusion BCR-ABL gene causing the disease. BCR-ABL protein is a drug target and BCR-ABL mRNA is a diagnostic target. The polymerase chain reaction (PCR) is a technique used to diagnose and monitor treatment response in CML. Tyrosine kinase inhibitors (TKIs) have raised the survival rate of CML patients from an average lifespan of 6 years to a normal lifespan. However, monitoring and diagnosing CML patients requires expensive equipment and technical expertise. Medical testing and therapy of CML in lower to middle income countries is difficult, because drugs are expensive and access to sophistical equipment rare. However, when the disease is confirmed by testing, a non-profit organization (the Max Foundation) can provide TKIs free for life through partnerships with pharmaceutical companies. Dried blood spots (DBS) can potential stabilize nucleic acid for long periods of time. This approach has facilitated retrieval of the genetic material on the DBS that can be used to analyze more genetic analysis related to CML response and progression and has reduced transportation costs. In this study, we isolated DNA & RNA from DBS, quantified the genetic material, performed quality control, and carried out DNA & RNA sequencing-based on gene mutation assays. Tapestation and qPCR were performed to check the quality of the genetic material. DNA samples showed a higher DNA integrity number with an average of 7.7. However, the RNA samples demonstrated a lower RNA integrity number with an average of 1.8. The DNA samples had excellent quality whereas the RNA samples had poor quality but were acceptable enough to carry on with our study. These results will help us determine if genetic material on DBS can be used for more genetic analysis.

Rhabdomyosarcoma (RMS) is a devastating pediatric soft tissue cancer. Currently, the standard treatment regimen for RMS patients remains relatively unchanged. Conventional treatment of RMS includes a combination of chemotherapy, radiation, and surgical tumor resection. Unfortunately, with the heterogeneity of cancer between patients, these therapies are not always effective and can cause undesired health issues for the patient due to their non-specific effects. Targeted drug therapy can help patients live more normal lives. The angiotensin II receptor type 1 (AGTR1) and 2 (AGTR2) are potential targeted therapy targets that can inhibit RMS cancer growth and cause less side effects compared to conventional therapies. AGTR1/2 are the main effectors in the renin angiotensin system regulating cardiovascular health. While there are Federal Drug Administration (FDA) drugs blocking these receptors to treat high blood pressure (Irbesartan), AGTR1 and AGTR2 have yet to be investigated for their role in RMS. Tumor propagating cells (TPC), which function as tumor stem cells in RMS, are proposed to drive tumor metastasis and relapse through a process called self-renewal. Preliminary disruption of AGTR1 and AGTR2 in the two major subtypes of RMS (embryonal and alveolar) with the CRISPR/Cas9 gene editing system resulted in decreased tumor cell growth and self-renewal capabilities. RMS cell lines treated with Irbesartan also decreased in viability compared to untreated cells. Based on our preliminary results, I propose that AGTR1/2 plays a role in regulating RMS cell growth and self-renewal. Further functional characterization of AGTR1/2 and investigation of the cellular mechanism by which AGTR1/2 regulates RMS tumor cell growth and self-renewal can provide a strong rationale for prioritizing AGTR1/2 as targets for drug therapies to slow the progression of RMS without greatly compromising more of the patient's health.

Fanconi Anemia (FA) is a recessively inherited genetic disorder resulting from a loss-of-function mutation in any of the 23 genes that comprise the Fanconi (FANC) gene family. FA is characterized by congenital abnormalities, bone marrow failure, and a predisposition to hematologic and solid cancers. The elevated risk of cancer in FA is largely limited to the squamous mucosa lining of the oropharynx, esophagus and vulva, and is not ameliorated by successful prior bone marrow transplantation. Apart from surgery, effective treatment of these cancers is limited by patient hypersensitivity to standard-of-care therapies that include ionizing radiation and DNA cross-linking drugs. The pathogenesis of FA-derived cancers is still not well understood. As a result, we are developing a “Fanconi Anemia Cancer Cell Line Resource” to provide preclinical models for research on FA-derived head and neck squamous cell carcinoma. The resource will include genomically well-defined, isogenic HNSCC cell line pairs (FA-patient/complemented cells) from patients with FA and isogenic cell line trios (wild-type/FANCA-deficient/FANCA-complemented cells) from control sporadic HNSCC cells. FANCA-deficient control cells were generated by targeted Cas9-mediated deletions in both FANCA alleles. Complementation of the resulting FANCA-deficient cells and patient cells was accomplished by targeted FANC- transgene insertion at a chromosome 4 ‘safe harbor site’. The molecular and cellular phenotype of the clonally-derived knockout and knockout/complemented cell lines was verified by PCR analyses; by Western blot and Multiple Reaction Monitoring (MRM) mass spectrometry analysis of FANCA and FANCD₂ protein expression/modification ± DNA damage; and by cell survival as a function of mitomycin-C (MMC) dose. The FA-HNSCC and sporadic HNSCC cell lines will be available for investigators via the FARF- sponsored Fanconi Anemia Cancer Cell Line Repository. We anticipate this versatile resource will be valuable in the development of novel treatment and faciliate in improving our understanding of FA cancers.
 

Nicotine is the addictive substance found in various tobacco products. CYP2A13 is an enzyme localized in the lungs, metabolizes tobacco-specific nitrosamine carcinogens that contribute to lung cancer. Therefore, pinpointing CYP2A13 inhibitors is an approach to lower tobacco-based lung cancer risk. Cinnamaldehyde is a common flavoring agent in the fluids of electronic nicotine vaping devices. Cinnamaldehyde was found to be a potent inhibitor of CYP2A6, another enzyme that metabolizes nicotine. Because CYP2A13 and CYP2A6 exhibit overlap in substrate/inhibitor selectivity, the goal here was to evaluate the inhibition of CYP2A13 by cinnamaldehyde. A time-dependent inhibition coumarin assay was performed to determine the kinetic parameters for cinnamaldehyde in recombinant CYP2A13. Primary incubations contained cinnamaldehyde, CYP2A13 Supersomes, and potassium phosphate buffer. Incubations were initiated with NADPH. Secondary incubations contained coumarin, NADPH, and potassium phosphate buffer. At selected time points, an aliquot of the primary incubation mixture was transferred to the secondary incubation tubes, which were terminated with trichloroacetic acid after heating at 37°C for 5.5 minutes. A linearity study was conducted to determine the appropriate termination time. CYP2A13 activity was measured by the detection of hydroxycoumarin using high-performance liquid chromatography (HPLC) and a fluorescence detector. Hydroxycoumarin formation decreased with time and inhibitor concentrations. Maximal inhibition following an 18-minute incubation was 38.3 ± 1.6 and 4.0 ± 0.6%. The maximal rate of inhibition was 0.109 per minute. The results provide evidence that cinnamaldehyde is a time-dependent inhibitor of CYP2A13. Furthermore, cinnamaldehyde appears to be a more potent inhibitor of CYP2A13 than CYP2A6, based on the maximal rate of inhibition. The results imply that cinnamaldehyde could interfere with the bioactivation of nitrosamine lung carcinogens. Additional kinetic studies are needed to confirm the results of this study and to evaluate the safety and toxicity profiles of cinnamaldehyde in complex physiological models.

Currently, there is no clinical best practice in place to support and manage follow up care for previvors, individuals who have tested positive for one or more mutations that increase cancer risk. Patients and providers need support in making complex decisions about follow-up. This project aims to design ‘previvor plans’, modeled after survivorship care plans received by cancer survivors, to help inform and support previvors after they receive a positive test result. Content of the previvor plans includes recommendations for screening, prophylactic surgery, pharmacological interventions, and lifestyle changes based on national guidelines as well as resources to receive further education and support. This project is a part of a larger study which evaluates two different methods of engaging patients in genetic testing for cancer risk across 12 different primary care clinics. Each patient in the study who tests positive for a mutation that increases cancer risk receives a previvor plan tailored with resources and recommendations specific to their test results and location. Use of the plans will be evaluated through surveys and interviews of previvor patients. If successful, the creation of previvor plans will provide a tool which can be used by patients and their providers to manage follow up care after receiving a positive test result.

Infection is the most common cause of mortality among patients with multiple myeloma undergoing autologous stem-cell transplant. Understanding the risk factors that lead to higher probability of infection is instrumental in their prevention. Although some risk factors are known, there are no robust predictive models for infection among myeloma patients receiving a hematopoietic stem-cell transplant. In this research study, we compared a variety of machine learning algorithms to reveal clinical features pre-transplant that are most predictive of infection post-transplant. Using features extracted from the electronic health record by means of natural language processing and manual abstraction, we trained our model using patients diagnosed with multiple myeloma who received an autologous stem-cell transplant at the Seattle Cancer Care Alliance. Along with being robust to missing data, the model is capable of processing diverse, heterogeneous data and identifying a class of predictive factors that can guide treatment decisions. Preliminary results point to lab tests such as lymphocyte, neutrophil, and platelet counts as the strongest predictive features for infection, and further exploration is underway to understand the full impact of these and other features.

Liver cancer is the world's second most deadly malignancy with a five-year survival rate of just 18%. Hepatocellular Carcinoma (HCC) accounts for most of the cases and its incidence is projected to rise to one million deaths per year by 2030. Unfortunately, HCC is extremely difficult to treat because several molecular pathways that promote drug resistance are upregulated in tumors, the most important of these being the epithelial-to-mesenchymal transition (EMT). Under physiological conditions the EMT regulates embryonic development, wound healing and tissue repair. However, cancer cells can hijack EMT signaling pathways to acquire a metastatic and drug resistant phenotype. Therefore, cell signaling enzymes that promote the cancer cell EMT are an attractive target for pharmacological intervention. Recently, we discovered that 71 protein kinases are highly enriched in mesenchymal HCC cells. To determine if these signaling enzymes are bona fide drivers of the EMT and drug resistance, we generated kinase RNAi knockout cell lines, quantified differences in EMT marker mRNA expression by qPCR, determined EMT pathway activation using quantitative proteomics, and tested differences in drug sensitivity. Here we demonstrate that inhibition of several of the 71 candidate EMT kinases reverses the phenotypic transition and sensitizes drug resistant HCC cells to chemotherapy. We conclude that these kinases could present attractive targets for the development of novel drugs that block cancer metastasis and overcome therapy resistance.

Embryonal rhabdomyosarcoma (ERMS) is a common pediatric cancer that has poor prognosis for patients with relapsed disease. ERMS typically harbors mutations in one of 3 RAS proteins. Mutations in NRAS, a member of the RAS family, have been shown to be a driver for many different cancers, including ERMS. The Chen lab previously demonstrated that genetic disruption of the NRAS gene by the CRISPR/Cas9 technique successfully reduced ERMS tumor growth in a human xenograft mouse model. However, these mice also experienced disease relapse. I was able to confirm successful targeting of at least one copy of NRAS in ERMS cells. I have subsequently isolated clones of ERMS cells that continued to grow despite the presence of NRAS gene disruption. In my investigation of candidate genes and pathways that might be responsible for driving continued ERMS tumor cell growth, I saw an increase in the level of YAP1 being produced in NRAS-targeted ERMS cells when compared to the control cells. Based on my preliminary findings, once NRAS is successfully disrupted in ERMS cells, tumor relapse is then driven instead by YAP1. This study could provide novel insight into the mechanisms underlying cancer relapse in response to NRAS targeting and promise alternative treatment plans for ERMS patients.

Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by the presence of repetitive patterns of behaviors and deficits in social communication and interaction. Attention to social information is a key component of the development of social communication. Previous studies used eye tracking to examine visual scanning patterns associated with social attention in children with ASD and neurotypical children. Eye tracking is used to both identify the atypical patterns of social attention and to predict clinical outcomes in ASD. Although atypical eye gazing patterns are considered as potential biomarkers, researchers commonly consider data loss in eye tracking as error or noise, and rarely investigate it more thoroughly. In this proposal, we hypothesize that loss of data is a potential signature of core social motivation issue when a social video is playing, and, rather than being a nuisance variable, which reflects the broader continuum of social attentional-motivational challenges faced by individuals with ASD. We used eye tracking to confirm previous findings on atypical attention patterns, and further utilize behavior coding to examine the three types of causes of data loss including blinking, non-compliant behaviors, and technical error. We hypothesize that data loss due to blinking is associated with a lack of social motivation and that data loss due to non-compliant behaviors is associated with executive function. Social motivation and executive function were measured by parent reports. Exploring data loss in eye tracking may help reveal comprehensive and fundamental factors of diminished social motivation and neurocognition in ASD.

Hoarseness is a common symptom reported to generalist healthcare providers, with approximately 1% of the clinical population being affected by it each year. It can be caused by multiple etiologies, such as hoarseness due to a cold, acid reflux, or laryngeal cancer. Perceptual evaluation of the voice is inaccurate, and it is therefore difficult to differentiate between hoarseness requiring urgent referral for specialty evaluation (i.e. laryngeal cancer) versus a disorder that could be managed without specialty care (i.e. acute laryngitis). The current gold standard of diagnosis for hoarseness is laryngoscopy, an in-clinic endoscopy recording of the larynx performed by an otolaryngologist specialist. Our research team seeks to improve perceptual voice evaluation by developing and testing machine learning algorithms which analyze voice for underlying pathology, beginning with an algorithm which screens voice for laryngeal masses. We hypothesize that our algorithm will have greater than 80% sensitivity and specificity in the classification of voice samples from patients with laryngeal masses. To test this, we are developing a large, prospective database of voice samples from a laryngology clinic using a smartphone application. Subjects are adult patients presenting to the laryngology clinic, with and without voice disorders, who have had a recent laryngoscopy exam and no laryngeal surgery within the past three months. We are collecting patient history which could influence voice quality, such as age, gender, alcohol use, smoking history, and subject-perceived voice disorder impact. After collection of the voice sample and patient history, cases are classified into underlying pathologic categories. We see recruitment of a well-classified and prospective patient population with a range of voice disorders. This work could lead to improved screening of patients with hoarseness in underserved and primary care settings, and more appropriate and timelier specialist referrals and treatment.

Autism Spectrum Disorder (ASD) is a chronic developmental disorder classified by deficits in social communication and restricted or repetitive patterns of behavior. A highly comorbid disorder with ASD is Attention Deficit Hyperactivity Disorder (ADHD) characterized by inattention, hyperactivity, and impulsivity. Deficits in empathy and facial affect recognition have been observed in children with ASD and ADHD. Through the APEX 5-week intensive Summer Treatment Program (STP), the UW Autism Center integrates naturalistic therapy for children diagnosed with ASD and/or ADHD aged 6 to 12. Within this evidence-based program, children develop social and behavioral skills through structured recreational and learning activities. During the summer of 2019, we collected the Positive and Negative Affect Schedule for Children (PANAS-C) from 120 participants ages 6-12 weekly. The PANAS-C measures youth anxiety and mood by asking participants to rate each emotion, five positive and five negative, on a point scale from 1 (not at all) to 5 (very much). Through multiple regression analyses, we aim to examine child affect through the PANAS-C within the context of variables such as age, gender, and diagnosis. We hypothesize that during participation in the summer treatment program, positive affect ratings will increase as negative affect ratings decrease. We expect that the combination of integrative social skills training, positive reinforcement, and acquisition of behavioral skills, will increase positive affect for children in this setting.

Current estimates by the centers for disease control (CDC) suggests that roughly 1 in 59 children in the US are affected by Autism Spectrum disorder (ASD). Despite its high prevalence, our current understanding on the biological etiology of ASD is limited. However, an ASD associated gene, TAOK2, which encodes a serine/threonine protein kinase, has been found to mediate neurodevelopmental disorders like ASD. Mutations within TAOK2 are associated with atypical neural connectivity in different brain regions, abnormal synapse formation, reduced cortical layering, and increases in brain size. Though TAOK2 plays a crucial role in neuronal development through phosphorylation of its substrate proteins, catalyzing or inhibiting their activity to regulate cellular processes, substrates of TAOK2 have not been fully elucidated and several candidate substrates have yet to be explored. Through my research, I explore TAOK2's interaction with putative substrate HDAC6, a histone deacetylase enzyme that functions to mediate a variety of cellular processes such as protein degradation, transcription, and the ability to regulate α-tubulin to mediate microtubule dependent cell motility. HDAC6 has been identified as a putative substrate of TAOK2 through mass spectrometry. Here, I present my findings regarding TAOK2's regulation of HDAC6 deacetylase activity and the potential it has for expanding our understanding of the molecular mechanisms underlying ASD.

The maternal bond is an intimate attachment between a primary caregiver (PC) and their infant which provides the infant with security, facilitating physical, social, and emotional development. A sensitive and responsive environment, such as the presence of healthy maternal bonds, guides an infant’s neurodevelopment. Changes in mood and emotional state can alter the care a PC provides and cause difficulties in bonding with their infant, impacting the baby’s psychological and physical development. In infants, the mechanisms by which development may be impacted are unknown. Recently, research has shown early atypical attention to visual pop-out in autism spectrum disorders. Attention to visual pop-out describes our cognitive ability to quickly identify differing objects presented among similar looking ones. In this project, we investigated whether PC mental health affects attention to visual pop-out in infants. Participants included 50 infants who were assessed at 6 and 12 months of age. PCs completed the Beck Depression Inventory (BDI) and Beck Anxiety Inventory (BAI) at both timepoints. Infants watched a 4-minute visual pop-out paradigm, which included social (face) and non-social (shape) trials. We assessed responses to visual pop-outs and explored whether BDI and BAI scores correlated with visual pop-out performance at 6 and 12 months. We also investigated whether BDI and BAI scores influenced the development of attention to visual pop-out between 6 and 12 months. We hypothesized infants of primary caregivers who report more (a) depressive and (b) anxious symptoms will demonstrate weaker identification of the pop-out during social trials compared to their peers but be unaffected during nonsocial trials. This study will help deepen our understanding of the impact of maternal depression and anxiety on infant development and help health providers identify and support families.

Recent work published by my colleagues and I showed that diagnostic ultrasound, applied directly over the visual cortex of human participants, increased the likelihood that they would observe visual effects while looking at a visual target, with that likelihood increasing over the course of the experiment. However due to a lack of EEG data, it is impossible to know the biophysical mechanisms and neural pathways that generate the observed effects on brain function. To better understand the full effect and mechanism of our initial findings, I helped develop a surgical protocol for a mouse model allowing for the collection of EEG data while exposing the animal to a combination of light and diagnostic ultrasound stimuli. EEG data was collected from 5 mice using a stimulus paradigm that we believed would increase the rodent's susceptibility to light stimulus. I utilized Matlab for processing, visualization, and statistical analysis of the data to determine the effects and hypothesize potential biophysical mechanisms of the stimuli. My analysis focused on determining whether the ultrasound stimulus successfully increased the susceptibility of the visual cortex, and which brain frequency bands were modulated by the stimulation.

As information processing machines approach the nanoscale level, DNA has emerged as a powerful tool in molecular engineering systems. The specificity and programmability of its hybridization interactions offer flexible and fine-tuned control over reacting species. Among the DNA computing techniques used today, strand displacement circuits are highly popular, with potential applications ranging from disease diagnostics to DNA-based artificial neural networks. The fundamental mechanism of these circuits is the hybridization of a single-stranded DNA input strand to a double-stranded complex which triggers the release of a prehybridized output strand. When released, this output can be detected and used to characterize circuit behavior. The output strands of strand displacement circuits are typically read out using fluorescence spectroscopy. However, due to spectral overlap of traditional reporters (e.g. FAM, TAMRA, Cy5), the number of outputs that can be detected in parallel is severely limited. To address this, we present the use of nanopore sensing technology as an alternative readout device that enables highly scalable, real-time detection and quantification of DNA strand displacement circuits. We demonstrate dynamic sensing of an operating circuit within the flow cell of a commercially-available high-throughput nanopore sensor array (Oxford Nanopore Technologies’ MinION device) and show that strand capture frequency can be correlated to concentration, allowing for direct quantification of desired circuit elements. To investigate this reporter strategy’s multiplexing potential, we present a collection of ten orthogonal circuit output sequences (barcodes) that can be classified at the single-molecule level from raw nanopore signal data using machine learning, with the potential to scale to larger barcode sets. We conclude that nanopore-based detection of strand displacement circuits holds key advantages over fluorescence-based methods for real-time, multiplexed circuit readout on an inexpensive, portable sensor device.

The goal of this research is to bridge the gap of stereotypes between homeless adults and other community members. In order to do this, we propose a pen pal application where homeless adults can write and exchange letters with adults who are not homeless. The application provides unique card templates and a pen pal request system that matches two users together based on common interests. By building relationships through customizable letters, our goal is to create understanding and reduce stereotypes between these groups of people. The method we are using to create this application is Value Sensitive Design. Specifically, we are seeking to address the values of storytelling and community, while supporting the interests of the key stakeholders, namely homeless adults, other community members, and Operation Nightwatch. Because we are partnering with Operation Nightwatch, a nonprofit organization that assists the homeless, we are constantly in contact with them, seeking to develop the pen pal application for their organization. However, the main focus are the users of the application so we are making sure to include them in the design process allowing them to share their stories and build relationships through this application. We have interviewed the stakeholders with semi-structured interviews, completed data analysis, analyzed potential stakeholder value tensions and the challenges that may come up, designed wireframes, created a high fidelity prototype, and conducted formative user testing. The main features of the app are: (1) Getting matched to a pen pal based on a profile; (2) Choosing a letter template and writing a letter; and (3) Receiving mail in the inbox. We have kept this application very simple: The focus is on sharing stories and catalyzing new understandings. We are designing and coding the application of the working prototype for the research symposium.

Software engineering is a known difficult task that spans a wide variety of problems; as such, an important skill for seasoned software developers is problem solving. Current software engineering research focuses on building tools that support software development processes, but very little research has been done to assist developers with learning programming strategies. Yet, previous research has shown that developers using explicit programming strategies (i.e. procedures in problem solving that were verbally described) were objectively more successful at code design and debugging tasks. In this research project, we extend the previous work in understanding how programming strategies may be used at scale, and whether it is a potentially effective way of improving developer productivity. We propose a novel platform composed of a repository of explicit programming strategies across various programming activities, such as debugging, design, and testing. Developers will be able to search, use, create, and provide feedback on programming strategies on this platform, which will require innovations in defining how explicit programming strategies are searched and indexed. With this explicit strategy sharing platform, we want to understand the experiences of developers who are strategy seekers or givers as well as their motivations using the platform. For strategy seekers, we would like to understand their experience in using strategies, as well as how the feedback process may evolve strategies. For strategy givers, we would like to understand their experience in writing strategies and why they do it. After building the platform, we will deploy the platform in a classroom setting that relates to software engineering, and allow students to use the platform organically. Then, we will perform user interviews and data analysis to evaluate our research question.

In the general population, adherence to a daily routine is linked with well-being.This appears to be the case to an equal if not greater extent among individuals with schizophrenia. Individuals with schizophrenia-spectrum disorders who consistently engage in activities that typically occur in a routine – e.g. employment, education, healthy sleep, social connections, and physical activity – enjoy an array of physical and mental health benefits. The study of adherence to routine has been limited by the use of retrospective scales, which are common in clinical research. These measures require respondents to provide estimates of the amount and frequency of behaviors over weeks or months. Such estimates are insufficiently granular to assess adherence to routine. The present study aims to examine the relationship between behavioral stability and symptoms in schizophrenia. Our team previously deployed a multi-modal mobile assessment system in a sample of individuals with schizophrenia for twelve months. In this study we revisit those data to develop models that quantify within-day adherence to routine among individuals with schizophrenia. We operationalize adherence to routine as defined in an individual’s behavioral stability, or the extent to which their behaviors detected by sensors stay stable from one day to the next during the study period. The present study has four main aims: whether (1) passively sensed behavioral stability can be quantified, (2) whether it is associated with symptoms and dysfunction in schizophrenia, (3) whether the addition of behavioral stability improves predictions of symptoms and dysfunction, and (4) whether this behavioral stability metric might be associated with future risk of poor outcomes.

Automatic gender recognition (AGR) is a subfield of facial recognition that has recently been scrutinized for bias in the form of misgendering and erasure against various identity groups in our society. Recent studies have found that several commercial AGR classifiers (from Microsoft, IMB, Face++) are biased against women and darker-skinned people as well as gender non-binary people. In this work, we investigate and quantify AGR classifier bias against transgender people by developing and evaluating three different convolutional neural networks (CNN): using images of cisgender individuals, using images of transgender individuals, and using images of both cisgender and transgender individuals. We find that the cisgender trained classifier is 91.7% accurate when evaluated on cisgender people, but only 68.9% accurate when evaluated on transgender people, with the worst performance on trans men with 38.6% precision. We also find that the classifier trained on the combined dataset performs nearly as well as and occasionally outperforms both other classifiers when evaluated on their own datasets, highlighting potential methods for avoiding overfitting. Additionally, we visualize how the classifiers differ by obscuring different parts of the face. Overall, the disparities of accuracy between each classifier demonstrate the degree to which they are impacted by the composition on their dataset and highlight the possibility for commercial AGR classifiers potential to misgender trans people, in particular, transgender men, at a high rate.

 The internet has changed the norms for how we write and communicate. Many major technology companies communicate with their users in much more casual and conversational language than the formal written English taught in schools. For instance, contractions and sentence fragments are common, the word “like” often used in place of “such as”, and “info” often used instead of “information”. This casual writing style may help users view a company as more approachable, but they may also perceive the casualness as lacking professionality and trustworthiness. Trust and taking the right action are especially important in security-related interfaces, as a user’s security and privacy may be compromised if an interface fails to educate users on secure behaviors. Through an online quantitative study we will explore the effects that formality of language has in security-related prompts. These effects include: how a user understands a prompt, their perception of the prompt’s formality, and how likely they are to take the action the prompt suggests. We will also investigate how users perceive the formality of various major technology companies and whether these perceptions match how the companies actually communicate with users. As average-level formality is different for everyone, we will analyze our results across different demographics, such as education-level, age, and the country the person grew up in. Our goal is to measure the likelihood of a person to take an action based on the wording of a security prompt, the person’s sentiments towards the prompt, and whether these depend on the person’s demographics and the company with which they are interacting. Our work serves a practical purpose, that is, helping technology companies decide what tone they want to address users with to accomplish their goals. It also serves a more theoretical purpose, in furthering understanding in the intersection of human-computer interaction and security.

We are studying the effects of Nuclear Fermi Beta Decay. Beta decay occurs when a proton in the nucleus of an atom decays to a neutron or a neutron decays to a proton. Protons and neutrons are in the class of particles known as “baryons”, as they each consist of three quarks, one of the most fundamental types of particles in our universe. They are both made of two types of quarks, “up” quarks, and “down” quarks. Neutrons consist of two down quarks and one up quark, while protons consist of one down quark and two up quarks. Beta decay specifically takes place when an up quark in a proton decays to a down quark, turning a proton into a neutron, or a down quark in a neutron decays to an up quark, turning a neutron into a proton. Our project studies the radial wave function overlap of these two particles. Particles have wave functions associated with them that describe the probability of measuring them in any given state. The overlap that we study is between the wave functions of the proton and neutron in the nucleus of the atom, as this overlap describes how Beta decay occurs. Particles have an intrinsic quantity known as isospin, which is very often conserved in particle interactions, but is not an exact symmetry. We analytically calculate how isospin symmetry breaking influences beta decay. Studying beta decay is useful for understanding solar radiation and energy, as the sun’s fusion reaction causes protons to decay into a shower of particles which are detectable from Earth. However, we are more interested in beta decay for its applications to testing the validity of assumptions of the standard model of particle physics.

From a materials design perspective, phosphates offer nearly limitless possibilities for various applications, chelating agents, synthetic replacements for bone and teeth, phosphors, detergents, and fertilisers. As a consequence of its thermal and chemical properties, Barium Hydrogen Phosphate (BaHPO₄) plays an important role in catalytic chemistry, industrial paint manufacturing, and ink-related charge direction. Because hydrogen and hydrogen bonds significantly affect these properties, studying proton mechanisms such as proton diffusive motions and jumps is critical to developing a comprehensive understanding pf the compound, allowing further determination of potential applications. Here, we utilise incoherent Quasi-Elastic Neutron Scattering (QENS). QENS is a useful technique to determine diffusive motions in the 10^-12 to 10^-9 second time range at length scales from 3Å to 60Å, which apply to hydrogen ion diffusion and hydrogenous species. Protons have a substantial neutron cross section, which subsequently enchances the associated QENS signal, and permits studies for low-proton systems such as BaHPO₄. The QENS investigation was conducted using 7 discrete temperatures ranging from 293 to 572 Kelvin, while subsequent fitting and analysis revealed the diffusion coefficient as a function of temperature. Obtained results for BaHPO₄ were compared to Monetite (CaHPO₄). In addition to QENS, Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM) provided support by giving insight to the surface topography of the sample. Structural and topographic results were compared with similar studies of other relevant phosphates.

In recent decades, researchers have begun to learn about exoplanets, which are planets that orbit a star outside of our solar system. Exoplanets are very diverse in their properties, for example, their masses, periods, radii, average temperature, and average densities. They also show a variety of elements and molecules in their atmospheres. One of the ways that we use to analyze the composition of their atmospheres is by using transmission spectroscopy. In our research group we are interested in learning whether there are any trends between the physical and orbital properties of the exoplanets, the host stars, and the atmospheric composition of the exoplanets. Our study centers on exoplanets with up to 3.5-days orbits and radii between 1 to 2 times the radius of Jupiter, the so called “hot Jupiters”. The Habitable Zone Gallery is a website which provides information about planetary parameters and how much time each planet spends in its stars habitable zone. The habitable zone is the region in which exoplanets can be found where they have the ability to hold liquid water on their surface. This region is a specific range of circumstellar distance from the host star depending on the host star. We will present the results of this study, utilizing data from the Habitable Zone Gallery, Astrophysics Data System for published data on each exoplanet, and NASA Exoplanet data archive for additional information. We have focused on studying the wavelength range from 3000 to 17000 Å, which is where absorption by sodium, potassium, and water can be found. Any trends between physical, orbital and atmospheric properties will be useful for future selection of targets.

In recent years, gravitational waves have yielded insights across many fields, from the expansion of the universe and black hole populations to the origin of heavy elements and nuclear physics. The Laser Interferometer Gravitational-Wave Observatories (LIGO) are michelson-type interferometers formed by two, 4-kilometer-long arms with suspended test masses (mirrors) at the ends. The test masses reflect high-power lasers to be combined at the intersection of the two arms and create an interference pattern, which is sensitive to gravitational waves passing through the interferometer. For a stable interference pattern, the test masses must be oriented precisely; the low frequency orienting of the test masses is done using optical levers (OpLevs). They consist of optics that launch light from a diode laser which reflects off the test mass and hit a quadrant photodiode, which measures the tip and twist of the test mass as a function of beam spot displacement. However, this measurement has elevated noise at low frequencies, limiting its accuracy. The reduction of this noise by a factor of 10-100, would allow for a more accurate orienting of the test mass. The source of noise is likely to be found in the fiber optics connected to the launching telescope. To isolate this noise I recreated parts of the LIGO OpLev setup with a level of positional noise well below LIGO's current OpLev noise at 0.1 Hz (below a nanometer per square root hertz). To achieve noise at this level I have designed a low noise pre-amplifier, improved the physical stability of the setup, and analyzed data to identify the noise sources. With this reduced noise setup we plan to search for the source of LIGO's OPLEV low frequency noise. Reduced noise would increase the uptime of the observatories thus increasing the number of gravitational wave detections.

Since the discovery of extrasolar planets (planets orbiting another star) in the early 1990’s, more than 4,000 exoplanets have been confirmed to exist by January 2020 according to the NASA Exoplanet Archive. We are searching for trends between exoplanets’ atmospheric compositions and their physical and orbital properties. To do so, we gather and analyze numerous publications of transmission spectroscopy data on the atmospheres of these planets. The focus parameter space of each search we conduct is expanded incrementally throughout the research process. This expansion requires remaking plots and reanalyzing data, which is a step that has the potential to be simplified. Another problem was needing to input data that would later be used for representation manually. This allowed for the possibility of errors in the data. We also were not able to easily represent all the aspects of the exoplanets we desired to in our graphs and plots, such as stellar type and atmospheric absorption of elements. In order to accomplish these tasks in a more effective and efficient way, the team is automating the data collection, expansion, and representation processes through developing computer programs that are used alongside database queries. This includes developing code that will reduce the amount of human interaction with the data aggregation and representation steps. We will present the improvements introduced with the SQL Server database to store our large data intake and query relationships between planetary properties. Python code is used in SQL Server Management Studio to visually represent these relationships in plots and graphs. This makes for a more efficient pipeline from information intake to representation, which can then be used for planetary analysis. These results will be included in the Habitable Zone Gallery, making it accessible for the community of researchers who wish to use the information as well.

Just as the Schrodinger equation is the pivotal theoretical tool for analyzing atomic systems, the application of light front dynamics to derive light front wave functions provides a powerful point of leverage for analyzing the structure of hadrons. The governing theory for hadronic systems (systems composed of quarks and gluons) is Quantum Chromodynamics (QCD), and the expression of this theory in light front variables presents a mathematically-convenient format. In this project we construct a light front Hamiltonian for a 2-parton hadronic system and solve the resulting differential equation to derive a basis of light front wave functions (LFWFs) representing the corresponding bound states. We subsequently modify the Hamiltonian to include terms characterizing effects such as momentum transfer, and attempt to solve the resulting system non-perturbatively. The ultimate goal is to apply our LFWFs to analyze structures in problems important to nuclear physics, such as deep inelastic scattering.

Circumbinary planets, those that orbit two stars, are a unique challenge for traditional ideas of planetary habitability. Due to orbital considerations and different stellar properties for the two host stars, these planets receive sunlight that continuously varies in time and as a function of wavelength. These changes could result in time-dependent changes to atmospheric composition and climate. Earlier theoretical studies of circumbinary planets indicate that there is likely a habitable zone where liquid water could exist on these planets, and that climate variations may not be extreme. However, these models did not take into account the effects of atmospheric photochemical reactions, which can alter planetary composition, including the abundance of greenhouse gases, and so also the climate as well. We have developed a new time-dependent coupled photochemical-climate model which allows for exploration of these effects. Here we describe the model, validate it against prior results, and present initial results on the impact of photochemistry in binary star systems.

This paper argues that a discourse of “commercial colonization” permeated the writings of early European explorers of the Pacific Northwest, which fundamentally shaped how colonizers understood the spaces and peoples with whom they interacted. In doing so, I have examined the writings of several late eighteenth century European and Creole American colonizers of the region, especially Juan Perez, James Cook, and Robert Gray. From this basis, I was able to explicate the content and function of the economic discourse in these colonial texts as well as the colonial process more broadly. The discourse of commercial colonization in this region emphasized reciprocity in all encounters yet consigned Indigenous peoples to inherently inferior status in relation to colonizers and was used to justify colonial violence. My work with these texts shows that when the idealized practice of European commerce was challenged, instead of revising their guiding ideals, colonizers relegated those who challenged their theories to subordination. I illuminate how Cook, Gray, and others portray relationships in transactional terms, motivated by profit above all else. Thus, this paper argues that the tensions in these colonial texts are examples of an emerging Capitalist worldview that links the European colonial project in the Pacific Northwest to modern theories of race, commodification, and exploration, allowing us to better understand the relationships between each

 Contemporary scholarship on the transition of immigrant churches from the language of their country of origin to the one of their new country has often focused on it as a process of concessions undertaken by a reluctant first-generation aiming to keep a partly assimilated third-generation in the church. This functionalist view of religion sees the church primarily as a transmitter of culture, and the immigrant church as a tool for ethnic preservation. By contrast, historians studying ethnic minority groups have warned against treating them as monoliths pursuing a single trajectory towards cultural accommodation. In line with this criticism, I study the Swedish-American church ‘’Swedish Evangelical Mission Covenant of America’’ and its transition from using Swedish to English in the 1920s. As with the ''Augustana Church'', which was the largest Swedish-American church in the U.S, The Covenant Church had its theological origins in Lutheran pietism. However, The Covenant came to develop differing views on the role of the church in relationship to Swedish culture, which was reflected in its openness towards the use of English and American theological concepts. My research draws on Swedish-American newspapers, the Nordic Museums collection of information on Scandinavian-American churches that I have assisted in digitalizing this quarter, and secondary literature to show that the Covenant’s adoption of English as its main language instead of Swedish was based on the denomination’s historical and theological background and its perceived duty to spread the gospel beyond the Swedish-American community. This case study illuminates the complex motivations that drive the process of cultural accommodation in immigrant groups, and offers a counternarrative to the portrayal of immigrant churches as helpless victims of assimilation, instead coming to the conclusion that the Covenant members actively and counsciously created a new form of ethnic identity.

In 2017, Israeli artist Shahak Shapira released his online art installation, “Yolocaust.” In this installation, he downloaded tactless selfies and artsy photographs from people’s public social media accounts which were taken in front of the “Memorial to the Murdered Jews of Europe” in Berlin. He then replaced the background with real images of the Holocaust - inmates on bunks, corpses stacked in piles, etc. His mission was to call attention to the thoughtless and insensitive photographs that people take in front of this key memorial to the Holocaust. In the age of social media, the internet is littered with tweets, Instagram pictures, Facebook statuses, and Tumblr posts about the Holocaust. It begs the question: has social media chipped away at the authenticity at the core of Holocaust remembrance? In my research, I argue that many aspects of social media have been detrimental to Holocaust memory. It has helped to commercialize and trivialize the Holocaust in a new way; however, this phenomenon is not new. For years, the film industry has “dumbed down” the Holocaust in order to make it easy to digest for the average viewer and has greatly romanticized the event in many instances. My project analyzes the Holocaust in film, as well as in Social media in order to argue trends of commercialization and simplification regarding Holocaust memory did not develop newly in the last 15 years. With more digital technology, how we remember this event has been greatly shifted, often at the expense of historical truth, but not as a rule. This project hopes to warn viewers about the price of conveying Holocaust memory through such digital mediums as social media and film, but concedes that the wide-reaching arm of such sources is not always inherently negative and may be exploited as a memory tool in the future.

  Throughout his administration from 1998-2002, Colombian president Andrés Pastrana collaborated with the United States in Plan Colombia, a foreign aid program whose stated purpose was to eradicate the drug trade in Colombia through US military assistance and social development programs. Plan Colombia has been predominantly studied through the exploration of the economic reasons that have prompted US officials to acknowledge the Plan as a successful policy, regardless of its exacerbation of violence in Colombia and its failure to eradicate the drug trade. The lack of scholarship on Plan Colombia from the Colombian perspective fosters a narrative of US imperial imposition. Although this narrative is clearly justifiable, it disregards Colombian agency, and it obfuscates the internal racial and social hierarchies that sustained the Plan's implementation. Therefore, to explore the Colombian State’s project behind Plan Colombia, this research analyzed the Plan through a framework of intervention by invitation. Furthermore, it investigated how the Colombian government utilized hegemonic racial and social hierarchies to facilitate said intervention. Through detailed analysis of Pastrana's presidential speeches, I propose that through Plan Colombia his administration conflated a rhetoric of peace with participation in a modern neoliberal extractive economy. Colombia invited US intervention because it needed its aid funds to build the infrastructure and train the labor force necessary to bring Colombia to the fore of neoliberalism, which would consequently resolve the armed conflict. Furthermore, I argue that through such a conflation, his government instrumentalized Colombian social and racial hierarchies to mobilize impoverished campesinos to become the labor force necessary to sustain a licit neoliberal extractive economy that could attract foreign direct investment (FDI) to Colombia's agricultural and mining sectors. Therefore, this research illuminates the rearticulation of 19th century land struggles and racial and social hierarchies that have historically subjected—and continue to subject—campesinos to political violence.

This research project relates to my faculty mentor Dr. Michael Honey's involvement in deep south Civil Liberties / Rights organizing from late 1969 until roughly 1976. This activity got him on the radar of the Federal Bureau of Investigation's "Counter-Intelligence Program" (COINTELPRO); a series of covert and, at times, illegal projects aimed at surveilling, infiltrating, discrediting, and disrupting American political organizations and people deemed subversive such as feminist organizations, anti–Vietnam War organizers, activists of the civil rights movement, and a variety of organizations that were part of the broader New Left. Although Dr. Honey was a known subscriber to MLK's non-violence doctrine and his political activities were primarily constrained to participating in marches, fundraising, and distributing pamphlets, this still garnered him a 800 page FBI file. This includes agents illegally wiretapping his home, going through his mail, and writing detailed notes of every civil liberties meeting he attended. What made this situation remarkable was that it was not a unique experience. Mass surveillance on Americans was mundane in the FBI at that time. This research is in support of a full length book project underway by Dr. Honey documenting his experiences through this time, using it as a framework to tell the history of the period. Currently, the main goal of my research is to analyze this 800 page file in addition to several boxes of miscellaneous, period specific, primary documents. This includes decoding FBI serial numbers, cross-referencing known agent handles, and pulling information applicable to the book. In addition to this, I plan to create a table of contents for the file and fit the surveillance record into the timeline I have been organizing of the other documents. This project contributes to the mass of literature on the period by demonstrating how this federal operation effected everyday Americans.

Despite research supporting the efficacy of certain mental health practices, many mental health care providers in community mental health institutions are not utilizing evidence-based practices (EBPs) consistently as their method towards patients receiving mental health care. This is a problem because the patients in need do not benefit from therapies intended to help them. Clinical supervisors (clinicians who provide clinical direction and guidance to less experienced providers) play an important role in how often and how effective clinicians are at implementing EBP’s in their treatments. The purpose of our project is to examine the feasibility and self-reported usefulness of providing case-based consultation to clinical supervisors in supporting the implementation of EBP. We expect that clinical supervisors will report that they benefited from EBP case-based consultations by having an increase in confidence and competence in supervising mental health care providers in their efforts to use EBP. So far, we have successfully recruited 9 clinical supervisors from 9 community mental health organizations. Clinical supervisors completed a pre-survey that tested their comfortability in supervising mental health care providers implementing EBP. In addition to gathering quantitative data via the survey, we are also analyzing qualitative data from transcripts of the consultation calls with the clinical supervisors. After the 6 consultation calls, we expect that supervisors will rate the calls as high in usefulness. We also plan to learn about any barriers or challenges that the clinical supervisors experienced. With the data that we have collected during this pilot test, we hope to conduct further research testing if EBP implementation to clinical supervisors through case-based consultations creates a downstream effect by resulting in an increased level of competency and comfortability for clinicians.  

There is a demand for on-going training resources for American Indian and Alaskan Native (AI/AN) resource parents caring for older youth (12-20 years) in the foster care system. This exploratory, qualitative study evaluated the effectiveness of a resource parent on-going training curriculum, Critical On-Going Resource Family Education (CORE Teen), which was developed in 2016 to increase the stability and longevity of resource parents facing the unique challenges of caring for older youth. CORE Teen was implemented with a tribe located in the south east region of the United States for a pilot study. Two semi-structured focus groups were conducted with a total of 22 parents that participated in the sample: 1) with resource parents that have received CORE Teen, and 2) with those that did not. Interview questions focused on differences in the experiences of resource parents who have received teen-specific, on-going training compared to foster parents without access to resources that specifically target their parenting practices. A thematic content analysis produced these ten themes: (1) desire for information; (2) lack of preparation; (3) normal teen behavior; (4) difficult transitions; (5) trauma; (6) mental health; (7) importance of kinship ties; (8) training outcomes; (9) training recommendations and (10) policy recommendations. Themes indicated that the intervention group articulated a greater comfort level in parenting teens than the parents in the comparison group. Findings highlight the importance of training to increase caregiving capacity of resource parents caring for American Indian youth, who are at the intersection of lacking developmentally appropriate support and historical marginalization.

Racial stereotypes can promote and reinforce racial discrimination. While racial stereotypes are often conceptualized along an inferior-to-superior axis, research suggests that in addition to this axis, perceptions of racial groups can also be placed along a foreign-to-American axis, with different racial groups at different levels on each axis. If stereotypes of foreignness are viewed differently than those of inferiority, this model may explain why views on discrimination against racial groups vary based on the specific racial group. We hypothesize that Americans will view stereotypes of perceived cultural foreignness as more acceptable and less discriminatory than stereotypes of perceived inferiority. In two studies (N = 70, 103), UW undergraduates reported that stereotyping someone as foreign because of their race is viewed as more acceptable than stereotyping them as inferior (ps < .05). However, undergraduates did not report discrimination based on perceived foreignness as less discriminatory than discrimination based on perceived inferiority. These results suggest that while racial stereotypes about foreignness are considered more acceptable than racial stereotypes about inferiority, they are viewed as equally discriminatory. However, minority groups may view racial stereotypes differently than Whites, skewing these results. A third study (N = 100) found Whites rated racial stereotypes of foreignness as more acceptable (p < .001), less harmful (p < .001), and more fair (p < .001) than racial stereotypes of inferiority. Understanding how inferior- and foreign-based stereotypes are viewed differently from each other may allow us to understand why some forms of racial discrimination might be viewed as more acceptable to engage in.

There is a long-standing concern that in American society, historical roots, structural racism, and systems of power have perpetuated the spectrum of negative health outcomes in communities of color. As the status quo is maintained, policy and public opinion reflect the continual oppression of minority families. Through my research, I created a visual and multimedia project that will identify the relationship between public policy and the social determinants of health, and how these factors have affected different communities of color within Washington state and the national level. I conducted extensive library research, analyzed and interpreted data, and utilized GIS technologies to visualize the placement of different communities. This work includes identifying historical background information, past U.S. policies, and relevant literature. In this visual and multimedia project, I displayed four puzzle pieces representing different communities (Asian, Native American, Hispanic, and African Americans) on a map, and examined public policies implemented by European colonizers that racialized minorities in unique ways. This chronological project displayed contemporary policies in housing, economic, employment, education, and criminal justice. Overall, I looked at each group's distinct experience of racial health disparities and will use this platform for dialogue to emerge for students and community members on these topics to prioritize the needs, barriers and solutions to confront racism.

Design awareness is the practice of being cognizant of the steps one takes in the design process and how one moves through them. Design awareness can help designers make more informed choices and find the best pathway through their design processes. In a quarter-long seminar, students explored the concepts of design awareness and created prototypes for a design awareness tracking tool to help them stay aware of the design process while fully immersed in it. The goals of this research are to explore methods of teaching students design awareness and to determine the effectiveness of these methods at demonstrating concepts of design awareness and the tools that can help them stay aware. The seminar began with students keeping a journal as they engaged in a design project over the spring break. The students closely examined their own design processes while exploring other methodologies of design. The seminar culminated in student presentations of their sketched prototypes for an artifact designed to increase their personal design awareness. Students reflected on their experiences and were found to have new definitions and understandings of the design process and what it meant to them. Follow-up research explored one design awareness tracking tool and the development of a physical working prototype. The design awareness tracker could record a design process through the different design activities that were engaged in. Data from paper prototypes of the device demonstrated that using the device increased users awareness as engaged in the design process. Further work is being done to widen the scope of design awareness teachings to reach a broader audience.