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.

The questions of equality, decolonization, and anti-imperialism have rarely been so acutely felt as they are today, in a period of rising global fascism, repression, and radical social movements. Looking back to the 1950’s and 1960’s, we see another such turning point in world history. Analyzing the historical documents from global revolutionary movements, Marxist-Leninist and Maoist theory, and locally-situated knowledge, this project examines how a future that had diverged in the 1950’s towards a communist world could have manifested for the both the international community and the Pacific northwest through the medium of a fictional newspaper published in Seattle. Ultimately, this project hopes to examine and critique our current moment of potential change, show the interpretive and political power of creative fiction, and stir interest in historical materialist analyses of similar moments of world history.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.
 

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.

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.

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.

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.

 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.

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.

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.

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.

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 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.

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.

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.
 

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.

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.

The Indian education system, like many countries, focuses strongly on annual national exams. This results in an emphasis on rote learning, discourages student participation and engagement in lessons, and yields a lack of motivation to learn. Prioritization of rote learning is ubiquitous across India, but is especially detrimental for students from adverse backgrounds and resource-poor settings. Parikrma Humanity Foundation is a non-profit school in Bangalore India that serves students from “the poorest of poor” backgrounds. The school integrates a “360-degree” model that prioritizes student happiness, while emphasizing student and family health as central to learning. Despite these well-established values, students at Parikrma are not exempt from national exam requirements and are also subject to the implications of the heightened importance of exam results. We sought to understand whether key stakeholders perceived that exclusive focus on rote learning hinders development of social-emotional life skills. We conducted over 70 interviews with students, teachers, school faculty, and alumni and analyzed these qualitative results through a method of conceptualization. From this we developed a scale to determine the degree to which the absence of explicit instruction of psychosocial skills impacts overall well-being. Respondents overwhelmingly reported that the lack of material to develop psychosocial skills such as teamwork and active listening results in students with less established life skills and limits access to future opportunities. We aim to develop a curriculum that promotes psychosocial skills key to professional success and overall happiness to be integrated within the context of Parikrma by establishing a cross-aged peer mentorship program that encourages students accountability to each other and themselves. We seek to engage teachers and students through the administration of comprehensive surveys that empowers individuals to self- report the impact of the program in order to assess results.
 

Social robots may be an effective and appropriate technology to help reduce stress in adolescents, thereby improving mental health. Previously, we designed a social robot that teens found engaging and desirable to help them with stress. Moving forward, we were curious about how different robot platforms affected adolescents’ stress levels and robot preferences. We conducted an exploratory study with seventy-one US adolescents (ages 14-21) comparing interactions on three different technology platforms: computer screen, physical robot, and immersive, virtual reality. Participant stress levels and attitudes towards robots were measured along with their momentary ratings of stress and levels of verbal engagement. Finally, participants completed an exit survey and interview in which they identified and discussed their preferred platform. The physical robot was rated most comfortable and resulted in significantly reduced momentary stress. However, after experiencing all three platforms, most adolescents preferred the virtual reality environment, with significant gender differences among preferences. 

As the Latinx student population continues to grow in public K-12 institutions, it is increasingly important to center this student group within educational research studies. Existing research shows that Latinx students face challenges that discourage them from continuing on to higher education, such as apathetic teaching, language barriers, and institutionalized cultural erosion. The bulk of such findings come from studies conducted in larger urban cities, thus excluding students from the many small farmworker communities that exist in rural agricultural areas such as those in eastern Washington. My research aims to include such populations in the educational discourse, using an asset-based model. Instead of asking students from these rural populations what went wrong for them, the current research project focuses on what went right for them in their paths to higher education. The present study asks the following question: What familial, social, and aspirational forms of cultural capital aid Latinx students in their transition from rural school districts to higher education institutions? To answer this question, I analyzed the retrospective accounts of first year Latinx students from small schools in central and eastern Washington state. Following the Grounded Theory approach, I used semi-structured interviews to collect data, which will later undergo transcription and analysis. I anticipate that results will highlight cultural resources such as community bonds, familial support, and encouragement as helpful resources in the pursuit for higher education. The findings of this research will aid rural secondary-school educators and staff to provide resources that support the educational attainment and success of Latinx students.

In cities throughout the developed world the dizzying rate of urbanization has led to a growing instability in housing fueled by a lack of affordability. A survey of 200 global cities by the Lincoln Institute of Land Policy revealed that 90% were considered unaffordable, based on the standard of a house price being more than three times the median income. This paper will analyze and compare factors impacting affordability in residential real estate markets in Singapore, Vancouver B.C., the Seattle-Bellevue-Tacoma MSA, and Copenhagen. Preliminary research indicates that the primary drivers leading to a lack of affordability are an insufficient supply to meet demand, geographic limitations, household costs rising faster than incomes, demographic impacts from population growth, ageing, and shifts in household composition. The financialization of housing has resulted in its commodification, a means of accumulating wealth and as the security for financial instruments traded on global markets, which has in turn decoupled housing from its primary connection to local markets. The primary intent of this project is to advocate for long term strategic policy changes to affect affordable housing solutions that address both the supply and demand side of housing markets, as well as advocating for a unified national housing policy that places housing as a human right.

Treatment of neurological disease has made little progress due to the inability of many therapeutics to access the brain environment. However, delivery vehicles like nanoparticles can allow therapeutics to overcome brain-specific biological barriers including the blood-brain barrier (BBB), the dense extracellular space (ECS), and cellular targeting. The ability of nanoparticles to overcome these barriers is influenced by surface properties which can be modified through the formulation process. One understudied parameter is the choice of surfactant, molecules which stabilize nanoparticle formation and likely form an interface between the nanoparticle and brain environment. First, we investigated the potential toxicity of several commonly used surfactants on brain cells and slices. We added surfactant solutions to mouse microglial cells (BV2) or cultured brain slices and assessed cell viability two days later with colorimetric assays. Our results showed that while surfactants cholic acid (CHA) and polysorbate 80 (P80) caused toxicity at high doses, they were nontoxic at the low doses involved with nanoparticle formulation. Other surfactants, including Pluronic® F127 (F127) and poly(vinyl alcohol) (PVA), were nontoxic throughout the tested dose range. Interestingly, although the F127 compound is nontoxic on its own, nanoparticles formulated with F127 reduced cell viability. This result was not observed with any other nanoparticle-surfactant combination. Confocal microscopy indicated higher intracellular accumulation of the nanoparticles formulated with F127 compared to all other formulations, suggesting that toxicity is mediated by nanoparticle internalization and surfactant choice. Finally, we used a live cell imaging technique to capture videos of the nanoparticle internalization process. Building off these results, ongoing experiments will evaluate several nanoparticle-surfactant formulations on their ability to accumulate within brain tissue after in vivo administration. Findings from this work will guide nanoparticle design for future clinical translation.

Understanding cause-and-effect is key for informed decision-making. The gold standard in causal inference is performing controlled experiments, which may not always be feasible due to ethical, legal, or cost constraints. As an alternative, inferring causality from observational data has been extensively used in statistics and social sciences. However, the existing methods critically rely on a restrictive assumption that the population of study consists of homogeneous units that can be represented as a single flat table. In contrast, in many real-world settings, the study domain consists of heterogeneous units that are best represented using relational databases. We propose and demonstrate CaRL: an end-to-end system for drawing causal inference from relational data. In addition, we built a visual interface to wrap around CaRL. In the demonstration, I will use CaRL, which I have implemented, to show a live investigation of causal inference from real academic and medical relational databases.

Immunological memory prevents reinfection by a pathogen. This protection is accomplished by memory T cells expressing T cell receptors (TCR) specific for previously encountered pathogen-derived peptides (antigens). Conventionally, memory T cells are thought to be inert during novel infections because there is no interaction between their TCRs with their specific antigens. Despite this, we and others have demonstrated that these T cells (here termed “bystanders”) can be activated by inflammatory signals alone and gain cytotoxic effector function in the absence of TCR-antigen interaction. This study aims to determine how inflammation regulates and attenuates bystander responses and how we can leverage these cells therapeutically. Using in vitro cell stimulations, we found that the inhibitory receptor, programmed cell death protein 1 (PD-1), is strongly upregulated by bystanders after exposure to certain inflammatory cytokines. This finding is unique because the current paradigm is that PD-1 expression is caused by TCR stimulation and PD-1 represents a target to manipulate bystander responses. Further, in mouse models of vaccination, we found that bystander-mediated killing can limit vaccine antigen. We believe that interfering with bystander T cell effector functionality could be targeted to improve antigen-specific vaccine responses. Through understanding the mechanisms that dictate bystander function, we may better modulate bystander T cells function during infection, vaccination, and cancer to improve patient outcomes.

Early diagnosis is one of the key factors to decrease disease-related mortality of advanced prostate cancer. Prostate-specific antigen (PSA) has been utilized as a diagnostic marker for prostate cancer. However, PSA test can cause false positive because enlarged or inflamed prostates also cause increased PSA level. Therefore, new markers are needed to improve the diagnostic accuracy. To discovery novel markers for advanced prostate cancer, we analyzed prostate fluids by mass spectrometry proteomic analysis. We discovered that the regenerating islet-derived protein 3 gamma (Reg3g) is specifically and highly expressed in the prostatic fluids of prostate cancer bearing mice. Reg3g has been shown upregulated in pancreatic cancer and can promote pancreatic cancer progression via STAT3 pathways. Additionally, overexpression of Reg3g induced immunosuppressive tumor microenvironment by recruiting myeloid-derived suppressor cells and enhancing CTLA-4 and PD-1 levels in T cells. Based on these observations, we hypothesize that Reg3g is a novel diagnostic marker and therapeutic target for advanced prostate cancer. We seek to 1) Determine whether overexpression of Reg3g affects prostate tumor cell proliferation and metastasis in vitro. 2) generate a prostate specific Reg3g overexpression transgenic mouse model to investigate whether Reg3g promotes prostate cancer progression and how it affects immune cells infiltration into tumor. Therefore, if we demonstrate that Reg3g expression is closely correlated prostate cancer progression and metastasis, this study will have an immediate impact on diagnosis and treatment of prostate cancer. This proposal addresses two overarching challenges: 1) Develop treatments that improve outcomes for men with lethal prostate cancer and 2) Define the biology of lethal prostate cancer to reduce death.

The ease of data retrieval, analysis, and distribution can accelerate the pace of scientific research. A difference in philosophies and methodologies in the manner of data-collection and storage leads to a lack of semantic-consistency in naming-conventions across disciplines. Previous studies have opened this discussion within various research disciplines, however there has yet to be a study accomplished within a Convergence Science Lab such ours, Genetically Engineered Materials Science and Engineering Center (GEMSEC). Well-defined naming-conventions, clear-cut data standards, and set programmatic interfaces are required to provide improved data access to researchers and the public. This consistency is necessary to create a robust database with the ability to query large amounts of related information. The key point is that integrating design (of variables, research questions, etc.) with traditional, empirical research approaches in the natural sciences will result in more robust data and clearer analysis. Here we discuss the need for standardized protocol and metadata standards for data-collection and storage. We began by creating a data analysis and storage pipeline for two computationally involved experiments, Molecular Dynamics (MD) and Next Generation Sequencing (NGS), within GEMSEC. A database schema storing metadata associated with raw, cleaned, and analyzed files was created. We found homogenous collections of metadata with inconsistencies, especially in peptide naming conventions, between experiments. The target-database includes those from other labs some of which may store unconventional file types, use other naming schemes for key variables such as sequences, and have different standards for what metadata is stored and what is computationally retrieved at a later time. This schema is formed via interviewing various other researchers, determining similarities and differences within metadata, and creating a standard for all to use based on this collected information.

Cannabidiol (CBD), an active compound in marijuana that provides diverse health benefits such as treating epilepsy, anxiety, inflammation, and chronic pain, is increasingly used in the United States. However, little is known about the pharmacological effects of CBD on neurological diseases. Although the chemical-induced dimerization (CID) system, in which dimerization binder and anchor binder dimerize only in the presence of small molecules, has been well established, very few studies have applied it as a biosensor, especially to detect CBD. This hinders our exploration of the medical values of CBD. Here we utilize the nanobody-based CID system to develop a novel CBD biosensor, consisting of split nano luciferase - a reporter protein, CBD induced CID - a sensor protein, and glycine-serine linker - a protein linker, which ensures the biosensor efficiency. We validated the biosensor between Dimerization binder 1(DB1) and CBD anchor 14 (CA-14) both in vitro and in vivo. From our previous in vitro data, we predict that the biosensor containing Dimerization binder 4 (DB4), an analog to DB1, can detect CBD at a higher concentration in vivo than DB1, which detects better at lower CBD concentrations. We use split-luciferase assays to test the binding affinities of DB4 for a better biosensor with a broader range of drug concentration-response evaluation. This study demonstrates an effective method to maximize the CBD biosensor system, extending the applications onto detecting drugs in several brain regions, different cell populations, and even subcellular components, thus, furthering the understanding of physiological mechanisms and therapeutic potentials of CBD.

 Chemically induced dimerization (CID) systems, in which a pair of proteins only dimerizes in the presence of a specific ligand, have wide use for the biosensing of small molecules, such as drugs, metabolites, and signaling molecules. However, few CID systems are currently available, and thus, until recently, little work has been done to design effective and highly specific CID-based biosensors, limiting the diversity of our small molecule detection toolkit. Previously, we established a highly efficient and generalizable method for de novo engineering of new CID systems, and demonstrated its effectiveness by designing a CID system specific to cannabidiol (CBD). Here, we engineer circularly permutated green fluorescent proteins (cpGFP’s) to have the cannabidiol CID system flanking it, allowing for small molecule detection via protein dimerization to be translated into measurable changes in fluorescent signals. As an experimental platform to screen high-performance designs, we employ the yeast display technique, where yeast cells are engineered to display on their surfaces any cpGFP-CID biosensor constructs we wish to screen. The lengths and compositions of the linker regions connecting the cpGFP and the dimerization proteins play a crucial role in the efficiency of the biosensor. Currently, we seek to understand what linker lengths and contributions help optimize biosensor performance. Initially, we introduced linkers with lengths of about 20 residues that are composed of alternating glycine and serine residues. Biosensor constructs with these linkers exhibited minimal, baseline performance. We generate biosensor construct libraries by mutating the linker regions, displaying this library on yeast cells, and performing high-throughput screening to identify optimal linker lengths and compositions for our biosensor constructs. Afterwards, we will validate the biosensor’s performance in vivo and in vitro. Our research will not only contribute to fluorescent and CID-based biosensor design and study, but will also aid in the expansion of clinical small-molecule detection toolkits.

Nondestructive 3D pathology is poised to play a transformative role in biomedical research and precision medicine in the decades to come, helping to usher pathology into a digital 3D era. Recent improvements in high-throughput volumetric microscopy, including light-sheet microscopy, have made it feasible for large pre-clinical and clinical specimens to be imaged in toto within reasonable time frames [Glaser, et al., Nature BME, 2017; Glaser, et al., Nature Communications, 2019]. However, these imaging methods depend upon the quality and reproducibility with which fluorescent labeling and optical clearing of thick tissue specimens is performed. In particular, while high-quality volumetric datasets can be acquired with pain-staking optimization and iteration of manual tissue-preparation protocols, high-throughput imaging assays demand that these methods be highly consistent and require minimal labor. We developed a protocol for automated micro-controller-based labeling and clearing of clinical specimens in order to generate volumetric imaging datasets that consistently mimic the appearance of “gold-standard” H&E histology. Archived formalin-fixed paraffin-embedded (FFPE) tissue blocks are first de-paraffinized with a combination of heat and xylene removal of paraffin wax. Next, specimens are put in an acidic, ethanol and water solution so that an aqueous nuclear and eosin labeling step can be achieved. This otherwise labor-intensive, two-day procedure is a critical step for automation since manual processing can lead to variabilities that will affect downstream labeling and clearing performance. Finally, specimens are cleared with a non-toxic clearing agent for refractive index-matching and 3D microscopy. By using automated micro-controller-based buffer exchange hardware, we demonstrate the reliability of these low-cost and convenient methods for imaging a diverse range of tissues. These methods will facilitate pre-clinical and clinical studies with large numbers of tissue specimens, such as those needed to validate the benefits of 3D pathology for clinical decision support.

Judaism has a long history in Yemen and the Horn of Africa, with community folklores and oral histories often tracing their origins to Solomon and the Queen of Sheba and a textual history which highlights unique cultural and religious practices and communal resilience. However, the Jewish communities of Yemen and the Horn of Africa are often underrepresented in sources on Jewish history or presented with sensationalist, Orientalist tropes. Similarly, Ge’ez, Amharic, and Judeo-Yemeni Arabic are significantly understudied in Jewish Studies and in mainstream linguistics and literary studies more generally. In this project, I will develop a digital exhibit using Google Sites or Omeka about the history and culture of the Yemenite Jewish community and the Ethiopian Jewish community known as Beta Israel. I will also discuss the Jewish-origin foundation myths for the Somali Yibir clan and the implications of this legend within broader Somali and Jewish contexts. In the online exhibit, I will incorporate multilingual and multimedia elements, including historical work on Yemenite Judeo-Arabic and Beta Israel Ge’ez scriptures and texts, trade documents showing these communities in context, and modern cultural projects with which Yemeni and Ethiopian Jews, the vast majority of whom emigrated to Israel in the second half of the twentieth century, have navigated their often-marginalized status and moves toward integration/assimilation or cultural preservation. In this interdisciplinary project drawing on the tools of the digital humanities while using language as a lens for historical study, I hope to present an innovative perspective on the role of oral and written literatures in the formation of peoplehood and of Jewish identity specifically, and to contribute an underrepresented angle to the broader body of digital Jewish history.

Humanity is on the verge of a biotechnological epoch. What this will entail is a union of opposites: biological humans merging with artificial machines. Unity of opposites, or non-duality, is a timeless theme, one that is not only found in the ancient teachings of Heraclitus or in Greek mythology, but also in that of the I-Ching, and the Tao Te Ching. The yin-yang symbol is one of the earliest visual depictions of this. Support for a unified theory of the universe is now being widely embraced; hence the shift from the old Standard Model of particle physics to the new Core Theory. Modern interpretation of what Laozi referred to as, Tao, suggests a process that is characteristic of a double torus, with a cuboctahedron at its heart. Such a synergic principle would not only be observable in the nature of matter, but also in that of the mind. It is of no coincidence that humanity is beginning to integrate with arguably its finest of achievements—tools. Synergic inquiry is the method by which this literature review will be conducted, so as to build the argument that all things are interdependently connected, distinguishable sub-systems of a larger system. This implies that the convergence of human and machine is simply the beginning of a brand new, distinguishable part in that whole. The study of synergetics, in its wider applications, proves to be an invaluable tool to understand the macrocosm and microcosm relationship, and it will no doubt contribute greatly to the symbiotic relationship between nanotechnology and molecular biology, as humanity seeks to build a better world.

Long-term exposure to traffic-related air pollutants (TRAPs) has been associated with multiple adverse health effects. However, many TRAPs, such as ultrafine particles, are poorly measured and thus their association with health outcomes is difficult to characterize. Our objective is to identify geographic characteristics that distinguish diurnal trends in TRAP concentrations at monitoring sites in order to estimate spatial contrasts in long-term average concentrations. Mobile monitoring (driving to many locations with multiple instruments) permits us to measure many TRAPs at many locations, but these measurements have short durations and may not capture the pollutant’s underlying trends. Therefore, these measurements may not reflect a site’s true long-term average due to our inability to fully sample the diurnal trend of the pollutant. In order to quantify the role of diurnal trends on annual averages, we use hourly measurements of CO, NO₂, and PM2.5 from California Environmental Protection Agency’s (CalEPA) monitoring sites where the true long-term averages are known. Using Principal Component Analysis (PCA) to reduce the number of dimensions, we will regress the pollutant levels against the time of day, the physical covariates and their interaction terms. The geographical variables include, but are not limited to, distance to a highway, distance to bodies of water, Normalized Difference Vegetation Index (NDVI), and population density. We utilize Akaike Information Criterion (AIC) and Bayesian Information Criterion (BIC) as our model selection criteria and we assess model performance via leave-one-out cross-validation. We identify the most influential geographic factors that are associated with two to three categories of similar diurnal trends for each pollutant. With these variables, we will be able to group Seattle monitoring sites and distinguish their diurnal trends. Appropriate adjustment for diurnal trends in our mobile measurements will permit us to estimate spatial contrasts more accurately.

Human immunodeficiency virus (HIV) infection is a significant global health issue, with approximately 75 million infections, and over 35 million deaths, since the beginning of the AIDS pandemic. The majority of these are attributable to HIV type 1 (HIV-1). A second form of HIV – HIV type 2 (HIV-2) – is endemic in West Africa and has spread to other areas with socioeconomic ties to the region. Historically, regimens for first-line treatment of HIV-2 have differed from those used in HIV-1-infected patients due to the intrinsic resistance of HIV-2 to nonnucleoside reverse transcriptase inhibitors. This distinction is coming to an end, as countries throughout West Africa are implementing a new WHO-recommended treatment regimen for first-line treatment of all HIV-infected patients, including those with HIV-2. The regimen, known as TLD, is comprised of the nucleoside reverse transcriptase inhibitors tenofovir and lamivudine and the integrase inhibitor, dolutegravir that has potent activity against both HIV-1 and HIV-2. Although treatment-emergent drug resistance has been well characterized for HIV-2 patients receiving tenofovir and lamivudine, there are few data regarding resistance mechanisms in patients receiving the third component of TLD, dolutegravir. The two objectives of my project are: (1) to construct a system for generating recombinant HIV-2 clones that encode and express integrase sequences from TLD-treated HIV-2 patients, and (2) to determine the in vitro susceptibility of viruses produced from the patient-derived clones to the integrase inhibitor dolutegravir. Specifically, I am engineering a plasmid vector into which patient-derived integrase sequences can be ligated for virus production and drug resistance testing in culture. The plasmid vector produced in this study will be used to characterize novel genetic pathways to dolutegravir resistance in HIV-2 and will help identify patients who are failing TLD treatment due to drug resistance. This information is crucial for improving treatment outcomes in HIV-2-infected individuals worldwide.

Gene therapy may be an effective way to treat vascular disease. However, current approaches to achieve efficient gene delivery to the blood vessel wall require invasive surgery. Others have reported that high-concentration lentiviral vectors injected into the jugular vein preferentially transduce cells in the vascular wall of mice. If reproducible, this approach would be a major advance in vascular gene therapy. To test reproducibility, we injected high-concentration lentivirus into mouse jugular veins and measured transduction 4 days later. Eight mice received 10e8 particles of a green-fluorescent protein-expressing lentivirus; six mice received vehicle only. Mice were euthanized and their aortas, hearts, lungs, livers, kidneys, spleens, quadriceps and gastrocnemius muscles, carotid and femoral arteries were excised. These tissues were prepared for DNA analysis (to measure transduction) and histology (to detect GFP expression). Tissues used for DNA analysis were frozen in liquid nitrogen and DNA was extracted. We used quantitative PCR targeting the GFP gene to measure integrated proviral DNA, which we quantified as proviral copies per cell. We found a mean of 3 proviral copies (range 1-6) per cell in the liver samples and 13 proviral copies (range 4-28) per cell in the spleen samples. Essentially no proviral DNA was detected in the liver or spleen of vehicle-injected mice (p<0.003 for both; limit of detection 0.3 copies per cell). Proviral DNA was not reproducibly detected in the other organs/tissues listed above. Tissue for histology was fixed and embedded in frozen blocks for immunostaining. We anticipate that immunostaining will enable detection or exclusion of lower levels of transduction. Intravenously injected lentiviral vectors seem to efficiently transduce the spleen and liver and do not efficiently transduce cells in the blood vessel wall. Our data suggest that better vector targeting is needed to deliver gene therapy to the vessel wall by IV injection.

Human Immunodeficiency Virus (HIV) remains on the forefront of research due to the ongoing global epidemic. HIV is comprised of two genetically different types, HIV-1 and HIV-2. HIV-2 is inherently resistant to some classes of antiretroviral drugs, and many HIV-2 patients develop drug resistance to first-line and subsequent regimens. HIV-2 can further be divided into two distinct genetic groups: A and B. While both are endemic to West Africa, group A accounts for the majority of infections and remains the most studied of the two groups. In-depth knowledge of drug resistance in HIV-2 group B is lacking, as only a few patients with drug-resistant virus are described in the literature and there have been no systematic efforts to characterize the drug resistance patterns of HIV-2 group B isolates in cell culture. My project's goal is to build drug resistance mutations, documented in literature, for HIV-2 group A into a full-length HIV-2 group B infectious molecular clone. Those results are used to compare the relative drug resistance conferred by those mutations to the phenotypes observed for equivalent mutants of HIV-2 group A. More specifically, common drug resistance mutations are introduced into the pol gene of a group B clone, individual mutant clones are isolated, and these are used to transfect replication-competent cells for virus production and drug susceptibility testing. Inhibitors targeting the reverse transcriptase, protease and integrase targets of HIV-2 are evaluated. The resultant drug resistance profiles are then compared to those found in published datasets for HIV-2 group A to determine how HIV-2 group A and group B mutants differ in terms of the magnitude and/or scope of drug resistance. These data are essential for developing evidence-based treatment guidelines for HIV-2–infected patients that harbor drug-resistant group B strains.

 Seed dispersal is a crucial phase of plant lifecycles. Effective dispersal is important to the ecosystem as a whole because it affects composition of the community, ecological succession, and response to climate change. Given the importance of seed dispersal, understanding the factors that contribute to the evolution of varied dispersal modes and promote convergence on specific dispersal strategies is particularly important to understanding grass ecology because it may allow us to understand the relationship between dispersal mode and habitat. In this study, we are interested in dispersal modes within the onion grasses (Melica), a small genus of perennial grasses, primarily distributed in temperate regions. The onion grasses are found in a wide variety of habitats and possess a remarkable diversity of seed dispersal strategies. These traits make them a useful case study for better understanding the factors that influence the evolution of dispersal strategies in grasses. We are testing the hypothesis that evolution in traits associated with seed dispersal is correlated with changes in habitat. In particular, we hypothesize that the evolution of wind dispersed seeds follows transitions into open habitats. Seed dispersal structures (diaspores) were collected from 46 grass species (35 Melica and 11 outgroup). To assess wind dispersal potential, we quantify falling velocity by filming seed descent at 1000 fps. Lower falling velocities are associated with higher wind dispersal potential. Diaspores were photographed and the images were used to measure surface roughness, which is associated with adhesive dispersal potential. These data, along with diaspore mass and plant height, were mapped onto the evolutionary tree of the onion grasses. We then ran tests of correlated evolution between seed dispersal traits and habitat type. Initial results indicate that convergence upon wind dispersal may be in part driven by convergence upon disturbed habitat types.

Extracellular Vesicles (EVs) are group of cell-derived structures including exosomes, microvesicles, and apoptotic bodies, which have been found to play a key role in intercellular communication, through the biological cargo that these EVs can carry. Their ability to deliver proteins and nucleic acids from donor cells to their target cells has led to growing interest in the potential of EVs being used as biomarkers for disease. But, a comprehensive understanding of EVs behavior is lacking, especially in neuroscience, which may hinder the development for further application of the EVs. Thus, we are interested in investigating the effect of brain-derived EVs (bEVs) on brain cells, especially microglia, the brain’s primary resident immune cells. To do this, we first extracted the bEVs from the rat brain through ultracentrifugation and purified them through size exclusion chromatography (SEC). We then applied the bEVs to cultured mouse BV-2 microglial cells and incubated for 24 hours before performing quantitative reverse transcription PCR (RT-qPCR) on the treated BV-2 cells to explore any bEV induced inflammation response. Preliminary results have shown that bEVs play a role in inducing both pro and anti-inflammatory responses in microglial cells, both to varying degrees in the cytokine markers expressed after incubation for 24 hrs. Furthermore, to better understand the interaction between bEVs and microglial cells, we labeled bEVs with fluorescent nano-sized semiconductor quantum dots (QDs). Through fluorescent confocal microscopy and time-lapse imaging, we were able to explore the time-dependent interaction of bEVs and BV-2 cells at high resolution. Our study can provide insights into bEV behavior, which can be used to better understand their potential use as biomarkers for specific brain disease models. 

Brain extracellular matrix (ECM) structure mediates many aspects of neuronal function. When ECM structure becomes dysregulated in neurological disease, one resulting impact is impaired neuronal function. Therefore, probing changes in ECM structure could provide insights into disease mechanisms and expose potential therapeutic pathways. Previous work in our group determined that degrading neural ECM structures, including perineuronal nets (PNNs), leads to a significant increase in the diffusive ability of nanoparticles navigating the brain extracellular space. However, this diffusion-based analysis provides little insight into changes in PNN-specific morphology or structure; it only predicts whether or not they are present and the degree to which they may be altered from normal. With this project, we aim to quantify changes in PNN structure with high spatial resolution. PNNs are stained using a fluorescently labeled lectin (Wisteria floribunda agglutinin) and images are acquired via confocal microscopy. Using Python, a coding language, we developed an automated image processing workflow to characterize morphological and structural features associated with PNNs, including total number of branches, average branch length, average mesh size of the net, and the areal density of fluorescence. This approach was applied to brains that span a range of chronological ages, from 14 days old to adult. PNNs are known to increase in counts early on in life, so this age-based study served as a proof of concept for our methodology. This same approach can be applied to study the effect of various neurological diseases on PNN structure. Collectively, this work aims to enhance our understanding of neurological disease mechanisms and open new avenues of therapeutic intervention.

Executive function (EF), a broad term for an individual’s higher-order cognitive abilities, has been shown to be an important factor in proper development of play in childhood. Children with autism spectrum disorder (ASD) have been noted to score significantly lower on tasks requiring EF and are often noted to engage in more simplistic levels of play compared to typically developing peers. We investigated within-group associations of average play level for children with ASD, as observed during parent-child play sessions, in relation to parent-reported EF scores, as measured by the Behavior Rating Inventory of Executive Function (BRIEF). Participants with ASD (n = 28, age = 3-11 years) and participants with typical development (n = 27, age = 2-7 years) engaged in a video-recorded, 15-minute unscripted parent-child play session. Blind coders determined the child’s level of play, ranging from object manipulations to pretend play, on a numeric scale of 1-13. The highest level of play was coded at each one-minute epoch of engagement using Behavior Observation Research Interactive Software. Participant’s play scores were averaged and analyzed with their BRIEF scores using Pearson’s correlations. Results indicated no significant correlation between average play level and the BRIEF working memory, planning, and inhibition subscales, with Pearson’s correlations ranging from less than .01 to 0.03 (p > 0.8). Likewise, for participants with typical development, there was no correlation between average play level and BRIEF global composite scores, with Pearson’s correlations less than 0.01 (p > 0.9). Our current analysis did not account for parental support of the child’s play, which may contribute to why parent-reported EF scores did not relate to child play level in these unscripted parent-child play sessions. Future directions include examining the relationship between EF and play in children with other developmental disabilities.

Childhood obesity rates in rural Lower Yakima Valley of Washington State are alarming at 67% which is three times that of the state average of 23%. Multi-level interventions focused on increasing nutrition and physical activity among children, their families, schools and the community can help curb obesity rates. Ciclovía is a community event that promotes cycling and physical activity on streets closed to motorized traffic for a day. The purpose of this study was to assess the community's engagement with Ciclovía during two years (2018 to 2019). We defined engagement as attendance and participation in the activities provided at the Ciclovía. We used three assessment tools to evaluate engagement including survey, participants’ observation count, and participants’ activity count. Our findings show an increase in attendance in both children and adults from 2018 to 2019. There were 361 individuals (275 children and 86 adults) in 2018 to 394 individuals (277 children and 117 adults) in 2019. The number of children on wheels (bicycles, scooters, skateboards) increased nearly fourfold from 11 children (in 2018) to 44 children (in 2019). The number of parents on wheels remained mostly the same from 2 (in 2018) to 11 (in 2019). Activities with high engagement of children were corn hole, jump rope, hula hoops, and obstacle courses in both years; approximately 80 percent of children were engaged in these activities. Parents were mostly engaged doing Zumba, aerobics, yoga, and walking. Ciclovía is a community event that promotes physical activity and shows high engagement of rural communities. Future study will examine the adoption of Ciclovía and long term sustainability.

Creative assignments like zines are accessible forms of knowledge that challenge traditional barriers to information, narratives, and histories of marginalized people. In my work, I helped craft a class that gave undergraduate students the opportunity to produce research on women from the Pacific Northwest whose stories deserved to be heard. Combining multiple disciplines like art, poetry, and videography, students were able to connect their work to their own personal narratives in meaningful and lasting ways. Based on their reflections, employing non-traditional forms of knowledge production like that of zines strengthens connection to course material and empowers students to see themselves as both a researcher and subject.

Although transcriptomes are highly tissue and cell type specific, curated gene set collections are not constructed or analyzed with recognition of this bias despite much of transcriptomic analysis depending on curated gene set collections. Prior work has recognized the potential for gene bias due to the variable nature of manually curating gene set collections, but coverage has yet to be characterized across all tissues and in all commonly used gene set collections. The goal of this study was to perform a comprehensive analysis of curated gene set collections from the data repository Molecular Signatures Database (MSigDB) based upon tissue specific expression. We analyzed KEGG, REACTOME, BIOCARTA, and Gene Ontology (GO) including Biological Processes, Cellular Components and Molecular Function gene set collections available on MSigDB. We curated lists of enriched and elevated genes as defined by Human Protein Atlas for 36 tissues. Analyses, visualization, and statistical analyses were performed using the R statistical programming language. We revealed that the MSigDB gene set collections differ among themselves in the fraction of tissue genes covered. GO Biological Processes has the highest gene coverage. BIOCARTA has the lowest gene coverage. Additionally, each collection differs among tissues in the fraction of genes covered. We also showed differential gene coverage among tissues even when collections are combined. Within elevated tissues, the liver has the highest and the fallopian tube has the lowest gene coverage. Within enriched tissues, the lymphoid has the highest and the testis has the lowest gene coverage. We created a database describing the presence or absence of tissue specific genes for each tissue with which researchers can elect the most appropriate gene set collection to use for analysis of a specific tissue. This increases the utility of our findings and creates a direct resource for researchers in the field.

Despite containing over two meters of DNA, the human nucleus amazingly is still able to fit said quantity of genetic material into a space only ten micrometers in diameter. To explore this phenomenon, we first utilize computational tools to find optimal locale on strands of DNA for the deployment of synthetic oligonucleotide (oligo) hybridization probes that, when hybridized, allow us to visualize the structure of chromosomes in the nucleus using microscopy. To find these optimal regions, we use a set of programs known as OligoMiner to parse through sections of DNA, considering certain parameter restrains like melting temperature and nucleotide content ratios. One area we explore is how altering the acceptable level of the nucleobase guanine in target regions affects the number of optimal sites located on a given strand, as well as whether these findings are relevant to the reverse strand. We hope that pinpointing ratios of only guanine will allow for better hybridization efficiency, which in turn would result in increased probe effectiveness overall. To accomplish this, we first encode the aforementioned checks into OligoMiner since, in its original form, this set of programs only verifies the combined ratio of guanine and cytosine. Following the creation of these additional tools, we run OligoMiner on both random and specific sections of the human X-chromosome. We’ve seen that guanine-rich regions of DNA will see a greater concentration of probe sites on the reverse strand under our new parameter involving solely this nucleobase, compared to that of the combined cytosine ratio, due to the guanine-cytosine pairing ensuring the ratio stays constant between strand compliments. The future of this research lies both in further updating and expanding OligoMiner while also examining probe density in regions beyond just those of the X-chromosome.

The blood-brain barrier (BBB) is a layer of tight-junction endothelial cells that make up the capillaries in the brain and strictly regulate what molecules can pass from the blood into the brain. Many molecules, including insulin, cannot passively cross this barrier but require an active transport system at the surface of the BBB. Once in the brain, insulin plays a role in memory and cognition. Indeed, Alzheimer’s disease is characterized by decreased sensitivity to insulin, which could be explained by a malfunctioning insulin receptor (IR) or impaired transport at the BBB. However, before we can begin to investigate the IR under disease conditions, we must first understand its standard regulation and function in a healthy system. Specifically, we aim to determine what factors mediate the endocytosis of insulin into the endothelial cells of the BBB. To do this, we focused on clathrin and caveolin, two proteins involved in different endocytic pathways. We performed cardiac perfusions on mice, where we first administered a drug to inhibit either clathrin or caveolin, and then we perfused with radiolabeled insulin. Afterwards, brains were collected and dissected into regions. Radioactivity was measured in the hypothalamus, olfactory bulbs, and whole brain, and the data was graphed over time to determine if there were changes in insulin binding or transport rates. Our results help elucidate the molecular processes necessary for insulin transport and binding at the BBB, which can ultimately help us understand how IR uptake and insulin transport may go awry in Alzheimer’s disease.

Sleep deprivation (SD) is a major health concern in developed countries, especially in the elderly, and is associated with physiological disturbances including cognitive impairment, increased risk of dementia, and leads to the acceleration of neurodegenerative disorders such as Alzheimer’s disease. A logical question is whether cognitive impairment associated with SD can be prevented. To address this question, 22-month-old C57BL/6 mice were sleep deprived during the middle of their sleep cycle for four hours each day for two days. Three days before the SD procedure, treatment was started with the anti-aging peptide glycyl-L-histidyl-L-lysine (GHK) 15 mg/kg per day for five days through the two-day SD procedure. GHK has wound healing and anti-inflammatory properties and readily passes through the blood-brain barrier. Immediately following the last day of SD and GHK treatment, mice were tested for learning impairment using a spatial learning activity which requires them to find the escape hole. Tissues were then collected for neuropathology assessment. Sleep deprived mice treated with GHK consistently found the escape hole more quickly than sleep deprived mice treated with saline suggesting that GHK can prevent the learning impairment associated with short term sleep deprivation. This preliminary observation provides the rationale to investigate the neuro-molecular and neuropathological pathways targeted by GHK including inflammation, oxidative stress and vascular dysfunction.

Geoarchaeology is a field in anthropology that furthers archaeological research by employing and borrowing techniques from several Earth sciences disciplines. Our research focuses on the site of Prasat Basaet, northwest Cambodia. This is a small provincial temple in Angkor, one of the largest preindustrial settlements in the world. Archaeological excavations at Prasat Basaet reveal an occupational history that includes the Pre-Angkorian (6-8th centuries), Angkorian (8-15th centuries CE), and Post-Angkorian (15-17th centuries CE) periods. Our research aims to investigate the relationship between urban expansion, temple construction, and the physical environment that people lived in. Our results will provide environmental context to help understand how provincial populations were involved in the rise and demise of the Angkorian state, and the importance of climate change in these processes. We are using sediment samples from archaeological excavations to extract lipid residues using Gas Chromatography and Mass Spectrometry (GC-MS) to reconstruct ancient vegetation and faunal communities. We are also using particle size analysis to understand the environment of deposit formation at the time of human habitation. We hope that our research will provide insights into human-environmental relationships at key moments in the history of one of the world’s largest ancient empires.

The viability of Greek communities and their place on college campuses has been a subject of intense debate in recent years. Researchers and critics of traditional Greek communities on U.S. college campuses have argued that rigid gender segregation and rules that give fraternities control over the party scene creates a gendered imbalance which favors men which, in turn, perpetuates the incidence of sexual assault and other harmful behaviors. But do the students that participate in Greek life agree? Do sorority and fraternity men also draw a connection between harmful behaviors and the formal rules and regulations that govern Greek life? Would structural changes to the Greek community alleviate its social risks like sexual assault, alcohol abuse, and hazing, or do they persist in the absence of gender segregation? This study explores how gender-segregated household regulations perpetuate these norms and further perpetuate social risks such as sexual assault, hazing, or alcohol abuse. Within Greek Communities, access to parties, drugs, and sex are the effective social currency that is exchanged within the Greek microsociety. Formal, gender-segregated household regulations give control of the party scene. Research has established widespread knowledge of how this control creates gender-based power dynamics and courtship patterns that contribute to a social culture with higher rates of sexual assault perpetration and victimization within its population. However, the University of Washington Panhellenic and Inter-Fraternity councils allow sorority women to live with fraternity men in their chapter houses during the summer. By interviewing a mix of 20 sorority women and fraternity men who from the University of Washington, Seattle Campus we can better understand if these gendered power dynamics are alleviated when men and women are regulated equally within their household.

Understanding complex population dynamics between species is key for guiding environmental and wildlife management decisions. Accurately identifying the diet of various predator species across northeastern Washington (NEWA) and central Washington (CWA) can provide comprehensive insight into these relationships in terms of predator-predator and predator-prey dynamics. DNA metabarcoding can identify species-specific DNA within a sample and presents an ideal way to perform diet analysis in this context. In a previous NEWA study, the diet profiles of a range of predators were fully resolved, but for the American black bear (Ursus americanus), approximately 80% of its diet composition was undetermined. For increased understanding of the black bear’s diet in Washington, prey species must be identified across a range of geographic areas. This study compares the prey components of the black bear’s diet in both NEWA and CWA in order to provide a comprehensive analysis of its role in the predator-prey community. DNA samples used for analysis were from scat collected by detection dogs during a 2015-2016 NEWA and 2018 CWA field term. Of the 12 bear samples from CWA,9 samples had identified prey and of the 15 bear samples from NEWA, 6 had identified prey. These results add valuable information about prey species composition in a key predator’s diet across a wide geographic region, as well as seasonal shifts in diet composition in relation to other carnivores in the NEWA community. Future research will be conducted on the plant portion of the black bear’s omnivorous seasonal diet. Data collected from this project will provide valuable information that must be considered for further studies on the Washington black bear population and the food groups it consumes.

Over the last decade, wolves have been naturally returning to Washington state. Mapping the population growth and reproductive activity of wolves across Washington is key to understanding their recovery and to assisting wildlife conservation management. Accurate identification of the number of pregnant wolves per pack during the breeding season could help. Progesterone levels excreted in feces provide a reliable index of pregnancy in most mammals; progesterone rises post-ovulation but only remains elevated above a “pregnancy-threshold” among females that become pregnant. Unfortunately, this pregnancy-threshold metric is less reliable in canids because progesterone levels often remain above this threshold during the typical gestation period among all post-ovulatory females, regardless of whether the females become pregnant. Since gut microbiome diversity has also been shown to differ between pregnant and non-pregnant mammals, this study examined whether the combination of progesterone levels and gut microbiome diversity can refine pregnancy diagnosis in free-ranging wolves. Five high progesterone and five low progesterone fecal samples from ten unique female wolves were provided by the Center for Conservation Biology from the 2015-2017 study in Northeast Washington. Gut microbiome profiles were generated by sequencing the V4 16S rRNA gene region in each sample and analyzed using Qiime 2 and R with the Silva reference database for microbial taxonomy classification. Principal coordinates analysis of Bray-Curtis distance between samples at the microbiome phylum level showed separate clusters among high versus low progesterone samples, with one exception. The microbiome community of one high progesterone sample clustered with the low progesterone samples. This sample also had the lowest progesterone concentration among the high progesterone samples and may thus be from a non-pregnant post-ovulatory female. These initial findings suggest that the combination of progesterone levels and microbiome diversity show promise as a pregnancy diagnostic tool that may be able to distinguish pregnant from non-pregnant wolves.

More than 442 million people worldwide have been diagnosed with diabetes, many of which regulate their glucose levels using the pump/catheter system. However, just 2-3 days after the catheter is inserted into the body, the tissue clogs due to the foreign body reaction (FBR), an immune reaction elicited by the body in response to any foreign material injected in the body. At this point, the patient must remove the catheter and insert a new device into fresh skin elsewhere, resulting in excess scar tissue. Our project focuses on preventing the FBR by reducing its triggering event--protein attachment--so that insulin catheters can last longer (2-3 weeks) and can reduce fibrotic accumulation in patients. To combat the frequency of delivery site changes, we have designed a nonfouling zwitterionic polymeric brush coating for the surface of the catheter to reduce protein attachment. For the coating, zwitterionic sulfobetaine methacrylate (SBMA) was surface-polymerized onto the catheter using atom transfer radical polymerization (ATRP). SBMA has been shown to resist protein adsorption down to less than 1ng/cm². The ATRP initiator was plasma-deposited to robustly adhere to the unique geometry of the catheter. In this work, we used a full factorial design of experiment (DOE) to determine significant experimental factors to the polymerization protocol to maximize the amount of SBMA on the surface. The coating was characterized using x-ray photoelectron spectroscopy (XPS) to confirm the presence of SBMA and the radiolabeled protein adsorption assay to measure the amount of protein adsorbed to the coating. We plan to use the results of the DOE screening to further optimize the nonfouling coating and ultimately plan to test this coating on insulin-delivering catheters in a diabetic mouse model to observe sustained lowered blood sugar levels and histologically review the extent of the FBR through collagen accrual.

Cancers are broadly characterized by changes in cell metabolism. Tumor cells typically exhibit functional respiration and inhibition of electron transport chain can impair cancer cell proliferation. However, certain neuroendocrine cancers can arise from loss of function (LOF) mutations in succinate dehydrogenase (SDHx/complex II), which plays a key role in the TCA cycle and in mitochondrial respiration. SDH, which catalyzes the conversion of succinate to fumarate, comprises four subunits: A, B, C and D. LOF mutations in subunits B, C, and D can promote tumorigenesis and mutations in subunit B (SDHB) are particularly associated with malignant and metastatic neoplasms. Interestingly, SDHB impaired cells show an accompanied loss of activity in complex I, implying that unlike the majority of cancer cells, respiration is not essential and may even be antagonistic for SDHB mutant cancer cell proliferation. Indeed, preliminary experiments indicate that inhibition of complex I can restore proliferation to cells treated with an SDH/complex II inhibitor. However, the molecular mechanisms behind this phenomenon are not well understood. We aim to investigate the metabolic mechanisms by which dysfunctional respiration is essential for the proliferation of SDH impaired cells. We hypothesize that inhibition of respiration in these cells can prevent oxidation of NADH to NAD+ at complex I and alter the redox homeostasis in the mitochondria to support proliferation. Specifically, we will test to see if increasing the NADH/NAD+ ratio is the required function of complex I inhibition that rescues cell proliferation in SDH impaired cells. In addition, we will characterize the metabolic consequences of specific alterations SDH, complex I, and mitochondrial redox state. Results from this study should allow us to delineate the importance of metabolic alterations in SDH mutant cancer cells and potentially help identify metabolic vulnerabilities for treatment of SDH impaired cancers.

 Cancerous cells have a modified metabolism that supports their demands for increased proliferation. One of the essential molecules in cancer cell metabolism and proliferation is the amino acid aspartate. Aspartate is not only incorporated into proteins, but is also a substrate for nucleotides and other amino acids, including asparagine. Aspartate availability can constrain tumor growth rate, and the consumption of aspartate to generate downstream products can alter aspartate levels. One gene that draws from the aspartate pool is asparagine synthetase (ASNS). ASNS converts aspartate into asparagine, which is used in the production of proteins, but does not increase cell proliferation. Thus we hypothesize that ASNS expression and activity can affect aspartate levels. With this, we aimed to determine if ASNS expression could alter aspartate availability and change sensitivity to aspartate suppressing therapies. Since cancer cells express ASNS to varying degrees, this project sought to determine if ASNS expression could be used to identify those cancers that are most amenable to aspartate suppression therapies. This research sought to better understand the conditions that determine aspartate levels, and how to exploit those conditions to inhibit tumor growth in association with asparagine synthetase.

Conjugated polymers (CPs) are used in a wide variety of organic electronics such as photovoltaics, organic thin-film transistors (OFET), and flexible displays because of the increased flexibility of polymer-based electronics. However, CPs often have significant downsides such as being prone to environmental degradation, lacking mechanical robustness, and being overall very expensive to create and use. To combat these limitations, CPs can be blended with lower-cost commodity engineering plastics (CEP), such as polystyrene, to create a blended composite that forms nanoscale structures of CP in a CEP matrix. To characterize the blends, we use small-angle neutron scattering (SANS) and wide-angle x-ray scattering (WAXS), which are techniques that provide information in the form of a scattering pattern. After data reduction and background removal, SANS data can then be modeled to extract information about the structures that develop from 1 nm – 1000 nm. We have decided to focus on using the sphere, fractal, and parallelepiped models since those geometries often form from self-assembled CPs. The scattering pattern from WAXS can be used to determine the preferential growth direction of CP self-assembly, either along the pi-pi stacking or lamellar directions. Through the combined use of these techniques, we are able to characterize structural dependence on the choice of solvent including both moderate and good solvents for the CPs. We also tested different side-chain lengths on the CP which will affect solubility and the ability to self-assemble. From these experiments, we found that a moderate solvent (such as toluene) will encourage nanofiber formation growth at lower concentrations of CP. Control over nanofiber formation could potentially lead to more favorable electrical performance for these materials. Conductivity and rheology tests will be conducted which will allow us to determine how much of an effect solvent choice has on the mechanical and conductive properties of these blends.

 Over 30% of patients with epilepsy remain resistant to antiseizure drugs (ASD), despite over 20 therapies available. Individuals that do not achieve seizure control with two or more ASDs are considered “pharmacoresistant”. Limited preclinical studies have suggested that the early chronic administration of ASDs that selectively inactivate sodium channels may dramatically alter subsequent pharmacoresistance in chronic seizure models of epilepsy. However, it is unknown whether inhibition of slow inactivation state of sodium channels specifically leads to pharmacoresistance. Therefore, we sought to determine whether the chronic administration of lacosamide (LCM), which inhibits the slow-inactivation state of sodium channels, would lead to a similarly pharmacoresistant chronic seizure model. Male CF-1 mice were treated with an anticonvulsant dose of LCM (4.5 mg/kg, ip) or vehicle one hour prior to each corneal kindling session twice/day for three weeks until animals achieved the fully kindled state of five consecutive Racine stage 5 seizures. Once mice had achieved kindling criterion, they were given a two week stimulation-free period. We then tested the sensitivity of kindled seizures of both treatment groups to 9 mg/kg (ip) lacosamide. There were 7/9 vehicle-kindled mice (77.7%) versus 3/10 LCM-kindled mice that achieved kindling criterion (p=0.0698). There was also a significant time x treatment effect on kindled seizure severity (F (23, 391) = 2.169, p<0.002), demonstrating that chronic LCM administration significantly delayed acquisition of the fully kindled state. Further, administration of 9 mg/kg LCM reduced seizure severity in both LCM- and VEH-kindled mice, indicating that LCM-kindled mice are not resistant to escalating doses of LCM. To assess the mechanism of pharmacoresistance, immunohistochemistry data will be further discussed, specifically the differences in sodium channel subunit expression and neuroinflammation in LCM- versus VEH-kindled mice. This study provides insight into how ASD monotherapy early in the epileptogenesis process may influence pharmacoresistance and the development of the epilepsy.

Martian regolith is very different from Earth soil. In order to colonize, or have an extended stay on Mars, agriculture must be established. The purpose of this experiment was to investigate growing plants in Martian regolith in a manner that would be fuel-efficient, by using the existing soil of Mars with minimal interference and minimal materials brought from Earth. The regolith used was Mojave Mars Simulant-2 (MMS-2), developed by The Martian Garden. MMS-2 is more than a 90% match to the chemical composition of the regolith on Mars. Plant growth was compared between Earth soil (control), 50% Martian regolith MMS-2/50% Earth soil mixture (Mars Mix A), and 50% Mars regolith MMS-2/25% coffee grounds/12.5% Earth soil/12.5% vermiculite mixture (Mars Mix B). Plants were grown in all three mixtures and growth was measured during three month cycles. Although several plant species were planted, only kale produced any significant measurable data. Plant growth decreased with decreased percentages of Earth compost additive as measured by plant length and robustness. Efforts to reduce the mass of additives required to support plant growth include an exploration of acidifying Martian regolith MMS-2 prior to planting. Acids have been chosen for their ability to add critical nutrients of nitrogen and phosphorus. Nitric acid and phosphoric acid have both effectively lowered the pH to 6, similar to the optimal pH range for plant growth. The implications of this study indicate that Martian regolith and Earth soils on their own will not be sufficient to begin agriculture on Mars. Further research on chemical soil amendments will be needed for sustainable agricultural development on Mars.

Growing sustainable crops on Mars is an important aspect of developing a colony on the red planet. The goal of this research is to modify Martian regolith simulant to support plant growth. Results will be presented for the readjustment of the pH of Mars soil (pH 8) simulant to match that of typical fertile earth soil (pH 6) using nitric and phosphoric acid. The acids used were chosen based on their viability for transport to Mars and their ability to add crucial nutrients for plant growth in the unfertile soil. During the project, both acids effectively lowered the regolith pH, but in the hours and days following the pH increased significantly, which has motivated testing the buffer capacity of Mars soil simulant. The data collected was used to prepare three samples of Mars soil simulant; the first was modified with phosphoric acid, the second with nitric acid, and the third was also modified with nitric acid and had a buffer of dihydrogen phosphate added. The growth of kale was measured in the three modified soils, each mixed with equal parts potting soil. Kale growth was compared to trials performed without the acidification or buffering of Mars simulant soil.  Our research presents progress towards growing food in Mars regolith to sustain colonization efforts on the planet. This work can also be applied to the potential need to grow food in adverse conditions on Earth as the human population increases and the impacts of climate change advance.

Disability research in the Black community is limited nationally and absent for Seattle area residents. This study is an effort to center the narratives of Black Seattle area residents who live with a disability and to identify commonalities among these individuals. This study used qualitative interviews (n=5) as the methodology to identify three key themes across the study participants' lives. The themes identified are: (1) Fear and Shame, (2) Resilience, and (3) Consequences. Based on disability critical race theory, my analysis aimed to understand how participants navigate dominant society with intersecting identities. I found that they often experience adverse outcomes which continue to remain invisible and unacknowledged throughout political, social, and judicial realms of society. This study illuminates the need for state institutions and nonprofits that serve clients with disabilities to expand their reach to include more people of color. Additional research on outcomes experienced by African Americans with disability is needed to combat the institutional invisibility that this population faces.

Major Histocompatibility Complexes (MHC) are transmembrane proteins that utilize a selective binding domain to recognize peptide fragments in the cell environment and display these antigens on the cell surface. This selectivity of binding to different substrates is a feature that would be highly useful to mimic in the realm of genetically engineered peptide-solid surface binding, with broad implications applicable to engineered biomimetic systems. Our goal is to engineer selective binding biological molecules by mimicking the characteristics of the MHC-1 protein binding slot. The conventional approach to this problem applies directed evolution in a lab setting, selecting mutant MHCs with higher binding affinity against an antigen of interest. This method is resource and time intensive. Instead, we propose a machine learning approach to this problem via molecular graph convolutional neural networks (MGCNs) which are structured just like the connected atoms of input molecules. To explicitly model the MHC-peptide binding event as a graph, it is necessary to find computationally tenable representations of the MHC binding site. Prior attempts represented MHC binding alleles using only the critical contact residue positions of the MHC. This method omits the protein architecture, making it untenable as a graph encoding strategy. In this study, we evaluate alternate approaches to generate graph encodings of the two actively-binding alpha helices in the MHC-I complex and evaluate their performance on the task of predicting antigen binding affinity. We present an open Python toolset for generating graphs of MHC-I alpha helices and preliminary evaluation of their performance on a regression task on the Immune Epitope Database MHC-I dataset. The ability to generate de novo binding molecules for unique surfaces such as cancer surface proteins, viral spike proteins, metallic surfaces etc. has various use cases in: diagnostics, therapeutics, and engineered biomimetic systems.

Congenital heart defects have been linked to numerous genes, but many of the genes responsible are not yet identified. The purpose of this research is to identify the unknown genetic causes of human congenital heart defects, utilizing zebrafish as a model organism. Using CRISPR-Cas9 to edit the genome of zebrafish, we are creating mutations in genes we predict are involved in human congenital heart defects. Our lab has used a CRISPR-based screen in zebrafish to identify new genes that, when knocked out, lead to defective heart development in zebrafish embryos. For this research project, our questions are: Can we associate specific, CRISPR-induced genetic mutations in these genes with our heart-defective zebrafish embryos? And, can we genetically engineer heritable mutations in these respective genes in zebrafish? The methods used in this project involve using several sets of DNA oligonucleotide primers to assess where and how the CRISPR reagents have altered the screened candidate genes. We have analyzed three genes—grpel1, pomp, and psmd6—each with four CRISPR target sites. The primer testing and animal genotyping have been done using PCR, gel electrophoresis, and gel imaging, with genotyping also requiring restriction digests. Our results have been promising. First, we determined which CRISPR target sites are effective for each gene. Second, we successfully identified CRISPR-induced mutations in F0-generation animals for each of these three genes. Third, for the pomp gene, we identified germline-transmission of a specific CRISPR mutation corresponding with heart-defective embryos. This result identifies pomp as a new candidate gene for heart defects. A key implication of these findings is that we can successfully create lineages of zebrafish carrying mutations in these new heart defect genes. Our work will allow for further testing and a better understanding of the genetics behind heart development.

Joubert syndrome (JS) is a genetic neurodevelopmental disorder that affects ~1 in 100,000 live births. JS is diagnosed by a distinctive hindbrain malformation that manifests as the “molar tooth sign” on axial brain imaging. Remarkably, >40 genes have been associated with JS, making it one of the most genetically heterogeneous Mendelian conditions. The clinical and brain imaging features of people with JS display a broad range of severity. In fact, we have identified a substantial number of individuals without the molar tooth sign but that have imaging features suggestive of JS. It is not known whether these “JS-like” patients represent the mild end of the phenotypic spectrum associated with variants in JS genes or a different set of genetic disorders. It is also not known whether these JS-like patients are at risk for the progressive retinal, kidney and liver disease seen in some JS patients. To answer these questions, I performed targeted DNA sequencing of the JS genes in JS-like patients, and I used an in-house bioinformatics pipeline to identify predicted-pathogenic variants. We hypothesize that a large subset of JS-like patients will have genetic causes in JS genes. If this hypothesis is supported, we will expand the phenotypic spectrum associated with variants in JS genes and improve the medical care of JS-like patients by supporting monitoring of JS-associated progressive features and sequencing of JS genes in these patients. This will also be proof of concept for evaluating mild clinical presentations of other conditions to determine if they share the same genetic causes.

Euler integration is the simplest, most versatile, and underappreciated method of integrating partial differential equations (PDEs) that only involves repeated addition. Evaluating the evolution of connected dynamical systems is critical to fundamental research as well as to students’ understanding of the physical world and their classwork. The purpose of this research is to design and implement an educational tool that empowers students and faculty to understand the beautiful simplicity of the most applicable method of evaluating PDEs on a computer. Physical law is always written in the form of a PDE. Traditional physics education does not emphasize this technique. This is largely due to the lack of computers over the last six hundred years. But now, we have super computational ability at our fingertips, and it is time that everyone in the field of physics has access to this versatile and simple tool. The first educational tool is complete and has already helped students learn about this technique. Over the next few months this tool and ones like it will be sewn into existing curricula in the physics department. This technique applies to an enormous range of disciplines from fungal growth to fluid dynamics and will be a skill at every student’s disposal.

Spinal cord injury (SCI) is a debilitating condition that impairs motor function and overall quality of life. We have previously shown that Targeted Activity-Dependent Spinal Stimulation (TADSS) improves motor function in a rodent cervical SCI model. The hypothesized mechanism underlying TADSS is Spike-Timing Dependent Plasticity (STDP). During STDP, the strength of the synapse, or connection, between two neurons depends on the spiking behavior of a presynaptic neuron (A) relative to the postsynaptic neuron (B) within an optimal delay window. A synapse strengthens if A spikes less than 50 ms before B. Conversely, a synapse weakens if A fires less than 50 ms after B. It is currently unclear if TADSS strengthens corticospinal tract (CST) input into cervical spinal cord. In this project, we investigated the efficacy of TADSS therapy in inducing STDP in the rodent CST. TADSS therapy involved behavioral retraining of injured animals, with treated animals receiving concurrent spinal stimulation and control animals receiving no stimulation. In separate weekly sessions, the synaptic strength between motor cortex and spinal cord was assessed by measuring spinal cord evoked potentials (EPs) during test electrical stimulation. Test electrical stimulation involved current application to the forelimb region of motor cortex and recording of the EP response in cervical spinal cord caudal to the site of injury. After 3 weeks of TADSS, we observed larger EPs in TADSS animals and smaller EPs in injured control animals. In the weeks that followed, TADSS animals exhibited improved motor function while control animals exhibited declined motor function. Our results indicate increased connectivity between the motor cortex and spinal cord which precedes behavioral improvement -- this possibly suggests that strengthening the synaptic connectivity of the descending CST input to spinal cord is incorporated in the mechanism of TADSS-induced motor recovery.

The network model of memory proposes that concepts are linked together in the mind. When a concept is activated in the mind (e.g., “ocean”), this activation spreads to semantically, or meaning, related concepts (e.g., “boat”), making them easier to recognize. Expanding on this model, the present study investigated whether concepts presented without conscious awareness could still facilitate faster recognition of semantically related stimuli. Participants first completed a rating task, where they rated how much they liked a word, and then completed a lexical decision task, where they decided if a string of letters was a real word or not. In both tasks, some words were related to the concept “school”. Thus, the goal of the rating task was to implicitly prime, or unconsciously activate, the concept and make recognition of “school”-related words in the lexical decision task easier. Participants were not informed of the relationship between the tasks. We found that implicitly primed words (words related to the concept “school”) had faster reaction times and higher accuracy in the lexical decision task than words that were not primed (not related to the concept “school”). Our results expand on the network model, providing evidence that priming can elicit easier word recognition even when a participant lacks conscious awareness of the priming and when there is a time delay between the prime and target. This provides a potential mechanism for how our behavior is affected by the stimuli we encounter in everyday life, without us knowing it.

Systematic underrepresentation and the predominance of cisgender male voices in STEM fields put women and queer students at disproportionate risk and in need of support. Addressing these communities and redesigning access to resources is vital to their continued growth and development. This project aims to address the mental health of undergraduate students and the resources available at the UW’s Human-Centered Design and Engineering (HCDE) department. The project utilizes a mixed-method survey and interview protocol to understand students’ experience with their mental health in HCDE and their awareness of available resources. With this data, I will analyze trends amongst students to identify disparities and key issues for students and then create a speculative design for how the HCDE department can better support their students. Speculative design is a framework for redesigning thought patterns and systems. I am currently in the data-collection phase and have released the survey. Based on first-hand observations and literature review, I expect to see disproportionately poor mental health outcomes in queer and women students. I also expect that a majority of participants are unaware of the existence/extent of many of UW’s current resources. Diversity is vital to a department’s success, especially because HCDE prides itself on its inclusivity. This research will illuminate how certain groups of students are supported more than others. These voices are often not recorded and demographic details not asked for beyond age and sex, meaning researchers cannot directly address specific populations that are historically underrepresented and create a working framework to better support them.

Alcohol use disorder (AUD) is associated with severe alcohol-related harm, especially in vulnerable populations. People experiencing homelessness comprise one such population that is disproportionately affected by AUD and its sequelae. Although traditional abstinence-based treatment does not adequately reach or engage this population, a growing body of research has indicated that harm-reduction treatment may present an efficacious alternative. Harm-reduction treatment uses a compassionate and pragmatic approach to help people who use substances and their communities reduce substance-related harm and improve quality of life without requiring abstinence or even use reduction. Primary components of harm-reduction treatment for AUD can include medication-assisted treatment alongside counseling, in which interventionists assess and track harm-reduction metrics, help patients set harm-reduction goals, and discuss safer-use strategies. Studies that have implemented safer-use strategies, or means of staying safer and healthier even if patients are drinking, have shown efficacy in reducing alcohol-related harm. However, no studies to date have explored the associations between implementation of safer-use strategies and alcohol-related harm in people experiencing homelessness and AUD. This secondary study (N=213) was conducted in the context of a larger randomized controlled trial (N=308) of harm-reduction pharmacological and behavioral treatments with people experiencing homelessness and AUD. We aim to quantitatively and qualitatively describe treated patients’ engagement with safer-use strategies over the three-month treatment course and examine the longitudinal association between safer-use strategies and alcohol-related harm. We expect to see inverse associations between the application of safer-use strategies and alcohol-related harm. These findings may help clinicians and counselors better understand what kinds of safer-use strategies are most commonly endorsed by this population and their relative contribution to the reductions in alcohol-related harm and improvements in quality of life observed in the context of harm-reduction treatment with people experiencing homelessness and AUD.

For many years, the primary mode of treatment for people experiencing alcohol use disorder (AUD) has been abstinence-based treatment. Research has indicated, however, that abstinence-based treatment does not optimally engage or treat more severely affected populations, such as people experiencing AUD and homelessness. Instead, harm-reduction treatment approaches are more desirable for this population and can serve as an effective treatment alternative for people experiencing AUD and homelessness. Harm-reduction treatment entails a set of compassionate and pragmatic strategies to emphasize client autonomy, mitigate substance-related harm, and promote quality of life (QoL) without the need for abstinence or use-reduction. Specific components include assessment and tracking of harm-reduction metrics, harm-reduction goal-setting, and implementation of safer-use strategies. This secondary study (N = 213) served to qualitatively and quantitatively explore harm-reduction goals generated by participants in a larger, 4-arm randomized control trial of harm-reduction treatment for people experiencing homelessness and AUD. The three treatment groups included in this secondary study received: a) harm-reduction counseling only, b) harm-reduction counseling + medication assisted treatment (i.e., extended-release naltrexone), and c) harm-reduction counseling + placebo. Participant goals were recorded using the Safer Drinking and Harm Reduction Efforts (SHaRE) scale at baseline assessments and weeks 4, 8, and 12. Qualitative analyses will be conducted to determine the kinds of goals participants generated throughout the 12-week treatment period. Additional descriptive, quantitative analyses will establish quantity of participant goals set at each time point. Finally, inferential statistics will be used to test harm-reduction goals as correlates of alcohol outcomes across the 12-week treatment period. It is expected that a) the combined pharmacotherapy group will generate, progress toward, and achieve more goals than other study conditions; and b) quality-of-life goals will be more strongly associated with reduced alcohol-related harm than drinking-related goals.

Mental health and wellbeing of undergraduates in STEM is an important and pressing issue in college that needs to be addressed both culturally and institutionally. The UW College of Engineering (COE) is a high-stress environment, facilitated in part by the college’s competitive admission process, making it an interesting case study to look at this question. The purpose of this research project is to understand the characteristics of mental health of undergraduates in the COE and determine if there is a noticeable difference between mental health before and after studying engineering. I hypothesize that is an overall poor quality in mental health outcomes across all groups. The study utilizes a mixed-method design, incorporating a survey and qualitative data. Survey results recorded numerical data and free response questions to assess mental health. The data was analyzed and broken down for further analysis by gender, race, etc. Preliminary results suggest disparities between groups based on social characteristics. This research is important because it will help inform the further study of mental health in engineering environments and provide direction for the COE to address these issues.

Pain catastrophizing (PC) and pain-related self-efficacy (PSE) have both been shown to be associated with patient function in individuals with chronic pain, but the extent to which they may contribute independent variance to the prediction of pain and pain interference has been rarely examined. We conducted a secondary analysis of baseline data from a randomized controlled clinical trial with 177 individuals with chronic low back pain and/ or chronic pain associated with multiple sclerosis, muscular dystrophy, acquired amputation, and/ or spinal cord injury. We hypothesized PSE and PC would each be associated with pain interference (BPI), over and above the variance they share with each other and with a measure of pain intensity (0-10 NRS). Linear regression analyses revealed PC and PSE were each uniquely associated with BPI, after accounting for their shared variance and NRS. PC and PSE together accounted for substantial variance in BPI, over and above pain intensity, ΔR² = .20, F(1,170) = 59.74, p < .001, PC (B = 0.35, p < .001) and PSE (B = -0.20, p < .01). The findings indicate PSE and PC may play unique roles in adjustment to chronic pain, although PC may have larger effects. Conclusions regarding the causal role of PSE and PC on patient function cannot be determined from this cross-section study. However, future research should evaluate temporal and possible causal associations between PC, PSE, and subsequent changes in BPI and other important pain-related domains. Findings from such research would inform the potential importance of targeting these variables to maximize treatment benefits in individuals with chronic pain.

Adolescence alcohol use and opioid addiction in adults are systemic issues afflicting the world, and thus, it is important to elucidate the long-term individual and relational consequences of both substance abuse disorders. The purpose of this experiment was to examine the long-term consequences of voluntary adolescent alcohol use on morphine tolerance, fentanyl self-administration, and the effects of previous opioid exposure on fentanyl self-administration in adulthood. Using a preclinical model to examine this hypothesis, adolescent rats had access to alcohol in gelatin form for twenty days, after which a three week withdrawal period occured. Morphine was then administered intraperitoneally for five days and morphine tolerance was measured by a tail-flick test for those five days. Finally, fentanyl self-administration occured in an operant chamber and self-administration will be measure by the amount of fentanyl consumed. My expected results were that adolescent alcohol use will increase morphine tolerance as evidenced by decreased tail-flick time, and fentanyl self-administration will also be increased. Additionally, I expect that previous opioid exposure will increase fentanyl self-administration. Future research should examine the neurobiological mechanisms by which adolescent alcohol use increases morphine tolerance and fentanyl self-administration and how previous opioid exposure increases fentanyl self-administration since these biological mechanism are not well understood.

Diets high in fat and sugar have increasingly adverse effects with increasing age because of generally decreasing activity and energy needs. This nutritional overload results in an increase in body fat mass and an increase in risk for metabolic and cardiovascular disease and possibly other chronic diseases such as cancer. However, the role of fat mass in age-related pathology and survival over an extended period of time is controversial. To address this issue, C57BL/6 mice, 18 months of age, were started on a high caloric (HC) diet consisting of balanced protein, lard, and sucrose. A second cohort was maintained on a standard rodent caloric (RC) diet consisting of balanced protein, wheat, corn, and soybean oil. Fat mass was measured by quantitative magnetic resonance imaging (QMRI) monthly for 10 months (28 months of age) at which time mice were evaluated for physiological performance, and tissues collected for geropathology. Both diet groups started with an average 5 percent fat mass. Fat mass increased to 15 percent in the HC diet group over the next 3-5 months, then gradually decreased to 9 percent after 10 months on the diet. Fat mass gradually decreased to 3 percent in the RC diet group over 10 months. Mice fed the RC diet had increased paw grip strength and were able to stay on a rotating rod longer than mice fed the HC diet, but there was no difference in survival between the two diet cohorts over the 10-month trial period. These preliminary observations suggest that percent body fat mass generated by a diet high in animal fat and table sugar may be associated with unhealthy aging, but not survival. This provides rationale for subsequent investigations into the pathological consequences of high caloric diets with increasing age.

Abnormal activity in the extended face processing system has been implicated in face processing challenges in autism spectrum disorder (ASD). However, the impact of comorbid attention deficit hyperactivity disorder in individuals with ASD (ASD-ADHD) on social impairment and the neural substrates underlying face processing has not been investigated. To address this, we conducted an fMRI study of emotional face processing in participants with ASD-ADHD, ADHD and significant sensory processing challenges (ADHD), ASD, and typically developing children (TD). After excluding for motion, 16 children with ASD (Age M (SD) = 9.57 (0.06)), 16 children with ASD-ADHD (Age M (SD) = 10.08 (0.07)), 20 ADHD (Age M (SD) = 9.46 (0.06) and 40 TD controls (M (SD) = 10.04 (0.06)) were included. Social functioning between autism groups were compared using the Autism Diagnostic Interview-Revised (ADI-R). MR data were collected on a 3T Philips Achieva system. For the fMRI task, 54 volumes of high resolution data (2.3mm3) were collected. Participants were shown blocks of rapidly-presented (150 ms) fearful faces, houses and scrambled images. fMRI data were processed in FSL using standard processing methods. We tested group differences in the contrasts faces > houses and faces > scramble. The ASD participants were rated significantly higher than the ASD-ADHD group on the ADI-R social domain (ASD M=17.88, SD=6.18, ASD-ADHD M=12.33, SD=6.29, p<0.05). Children with ASD-ADHD exhibited reduced left amygdala (p = .025) and left fusiform (p = .03) activity compared to children with ADHD for faces > scramble contrast. However, activation in these areas did not significantly differ between the ADHD and ASD groups. These preliminary results indicate significantly altered brain activation during face processing in children with comorbid ASD and ADHD when compared to children with ASD alone, suggesting a possible additive effect of comorbidity on social difficulties.