Kelp forests are hotspots for productivity and biodiversity in marine ecosystems. Currently along the West Coast, many kelp forests face significant threats due to climate change. Several studies have looked at the effects of ocean warming on kelp, but less is known about the effects of ocean acidification (OA), and how OA and warming in conjunction impact kelp. With climate change continuing to worsen, it is crucial to understand how these drivers may contribute to kelp’s decline. We examined the effects of warming and acidification, both separately and concurrently, on bull kelp (Nereocystis luetkeana) from two populations in Puget Sound, WA. Bull kelp, the primary canopy-forming kelp species in Puget Sound, has a heteromorphic life history with macroscopic (sporophyte) and microscopic (gametophyte) stages. Little is known regarding how the microscopic stages will be impacted by multiple stressors in Puget Sound. This study aims to better characterize the effects of warming and OA on Puget Sound bull kelp throughout its microscopic life stages. We exposed kelp gametophytes from two genetically distinct populations to elevated levels of temperature and pCO2 in laboratory conditions for five weeks, monitoring their survival, development, and reproductive success from spore settlement to the juvenile sporophyte stage. Our collection sites differed in the overall temperature, urbanization level, and state of the kelp forests. Previous studies have shown declines in reproductive metrics when subjected to thermal stress, and an increased thermal tolerance when subjected to acidic conditions, which is what we expect to find within our experiment. Understanding local adaptability is critical in predicting future population responses to climate change. The potential range shifts or local extinctions that could occur because of these stressors will need to be understood to plan for conservation, restoration, and recovery of healthy kelp forests.

The seeds and fruits of all flowering plants derive from female reproductive structures called pistils. At the tip of a pistil is an area called the stigma that is the site of pollen reception, making the structure of stigmas critical for ensuring plants are pollinated and produce seeds. In the genus Thalictrum, stigmas vary widely: short stigmas are present in insect-pollinated Thalictrum species to facilitate closer contact with visiting insects and ensure successful pollen deposition, while longer, more feathery stigmas have evolved from these to take advantage of wind for pollination. However, the evolutionary and genetic basis for this transition is not known. I aim to explore the genetic mechanisms that facilitate the transition from insect-mediated to wind-mediated pollination using Thalictrum thalictroides as a model species. Specifically, I am testing the function of two candidate genes in T. thalictroides stigma development, STYLISH1 and STYLISH2 (STY1/2), which are known to be necessary for stigma development in other plant species. To do so, STY1/2 genes have been silenced and overexpressed by virus-induced gene silencing and virus-mediated overexpression. I collected and imaged floral tissue from transgenic plants and validated them by qPCR. The effects of silencing and overexpressing STY1/2 are then determined using scanning electron microscopy to visualize cellular changes to the pistil and stigma. I predict shortened or loss of stigmas in silenced flowers and elongated stigmas in overexpressed flowers, which would support my hypothesis that STY1/2 are necessary for stigma formation in T. thalictroides. Not only do these findings enhance our knowledge of plant adaptation to environmental changes, but they also create implications for efforts toward conservation and sustainable agriculture, offering potential insights into the prospect of engineering wind-pollinated plants in a changing climate with dwindling pollinator populations.

The emergence and spread of antimicrobial resistance (AMR) in bacterial populations pose a significant threat to global health. A key factor in this process is the horizontal gene transfer (HGT) of resistance genes carried by mobile genetic elements (MGEs). This study focuses on understanding how mutations in plasmids influence the rate of HGT and its implications for the spread of AMR. Plasmids are small, extrachromosomal DNA elements that can transfer between different bacterial cells, a process crucial in spreading antibiotic resistance genes. These plasmids often use a mechanism known as conjugation for transfer, a process encoded by approximately 30 genes in the transfer (tra) operon. This research aims to understand how changes in the tra operon affect the plasmid's ability to move between bacteria. This movement can be either horizontal (between different cells) or vertical (from parent to offspring during cell division). Our project is divided into two main aims. The first is creating a library of mutated plasmids. I modified a large, well-studied conjugative plasmid, the F plasmid, and introduced specific mutations into the tra gene. This step is achieved using a method called Lambda Red recombination, a technique suitable for large plasmids where standard cloning methods fall short. The second aim is to measure the rates of vertical and horizontal gene transfer (VGT and HGT) in these mutated plasmids. We employed the Luria-Delbrück method for precise estimation of these transfer rates. Understanding the trade-off between VGT and HGT is crucial in predicting and controlling the spread of AMR. By mapping the relationship between specific mutations in the tra operon and their impact on gene transfer rates, our study contributes to a broader understanding of how antibiotic resistance spreads through bacterial populations. This knowledge is vital for developing strategies to combat the increasing threat of antibiotic-resistant bacteria.

Bilingualism has been shown to have numerous cognitive benefits, ranging from improved executive function to possible protection against neurodegenerative diseases. Structurally, early childhood bilingualism correlates with expanded subcortical regions, specifically the basal ganglia (BG), an area most famously involved in functions such as motor control. In this project, we explore possible domain-general effects of infant and early childhood bilingualism. Specifically, we investigate the effect of early childhood language environments on motor skills in the form of rhythmically controlled babbling. Audio recordings were made in infants’ natural environments using the LENA system. Infants’ own utterances were captured and preprocessed. From these data, I first calculated the ratios of language use in the environment (English versus Spanish), and from these ratios, I categorized infants into monolingual English, monolingual Spanish, and bilingual groups. To determine the rhythmicity of babbling, I isolated subjects’ utterances from the audio data based on criteria that define canonical babbling (e.g., CV syllable structure where C is supraglottal, 4 or more syllables, and so on). For the purposes of this project, we defined the perceptual center as the peak intensity of each syllable and used it to determine the duration between syllables, or inter-syllable duration. The variation of syllable duration was subsequently calculated. We anticipate more rhythmic (i.e. lower variability across inter-syllable duration) babbling in bilingual subjects than in monolingual English and Spanish subjects. However, in accordance with previous findings on early childhood language development, we do not necessarily expect to see differences in whether or not infants are in the canonical babbling stage based on language group. We expect these findings to contribute to the theory of domain-general benefits of early life bilingualism through the quantification of language development milestones.

The National Associations of the Deaf astimates there are 308,648 deaf and hard of hearing D/HH students in the US, 79.2% of D/HH students attend mainstream school. D/HH students tend to stuggle academically and socially in high school which leads to low self-esteem. This project uses a Critical Youth Participatory Action Research (YPAR) ethic, framework, and methodology to put power of research and advocary into the hands of the D/HH youth at a PNW high school. YPAR is an epistemological framework in response to the historical dehumanization of marginalized communities. It democratizes ad decoonizes research by privileging the knowledge production of vulnerable communities and foregrounding justive and change. In practice, YPAR develops research skills of typically non-academic co-researchers who drive the research question, design, and analysis. The aim of this project is to guid the students and build their skills to name the challenges they face as minoritized D/HH students, to collect and analyze relevant data, and to use the data to drive policy. the signifiicance of this project is the methodology thatdemands youth voices to be centered in policy making and cultural change. The expected results are findings that speak to the challenges of D/HH students at a PNW high school. The hope is for the D/HH students to present their finingds to policy and decision makers. There is limited research on D/HH students using YPAR methodology and this research will add the perspectives of D/HH students of the discourse.

In recent years, the United States has seen an increasing number of opioid overdoses, causing many research studies to focus on opioid drugs of abuse. However, we still lack a fundamental understanding of the opioid receptor that these drugs bind to. The mu-opioid receptor (MOR) is involved in both the pain-relieving and euphoric effects of opioid drugs. The Ventral Tegmental Area (VTA), known to be a major source of dopamine, contains a significant amount of inhibitory gamma-aminobutyric acid (GABA) neurons that express MORs. Previous research has shown that activity at the MOR in the VTA is rewarding and that dopamine projections from the VTA to the Nucleus Accumbens (NAc) are activated to reinforce behavior. However, whether the expression of MORs on GABA neurons in the VTA is important for influencing VTA to NAc dopamine activity during reward seeking behavior is unclear. To investigate the role of the MORs in reward seeking behaviors, I knocked out MORs in the VTA and used dLight, a fluorescent dopamine sensor, to measure dopaminergic release from synapses in the NAc during Operant and Pavlovian conditioning tasks in the absence of these receptors. Pavlovian conditioning utilizes chambers in which a house light turns on and a sucrose pellet is delivered via food hopper to food-restricted mice. In the operant conditioning task, mice must learn to complete a nose poke in order to receive the sucrose pellet reward. These tasks allow me to gain a more complete understanding of how the absence of MORs in the VTA changes behavior and dopamine activity during reward-seeking. This research furthers our understanding of how the MOR affects natural reward and motivated behaviors and is crucial in helping us understand how opioids of abuse alter existing brain circuitry to cause opioid use disorder.

I am studying the phonetic effects of childhood exposure to a secondary language on a speaker’s adult primary language. Although some studies explore the effects of secondary language exposure during childhood on the ability to learn that language later in life, there are no studies on the effects of secondary language exposure during childhood on a speaker’s primary language. This study of two English speakers from the Boston area aged 18 and 20 compares the Voice Onset Time of a speaker exposed to Gujarati during childhood to a speaker from a monolingual English household. Voice Onset Time (VOT) is the amount of time between the release burst of a stop phone and the onset of voicing. While American English has a 2 way VOT contrast with no negative VOT (voicing before the release burst), Gujarati has a 4 way VOT contrast with contrastive negative VOT. I recorded the two participants reading aloud English words containing the sounds /p t k b d g/ as the initial consonant.To assess the extent to which sound patterns might differ between participants, I analyzed these sound files in Praat for VOT, for the F1 and F2 at the 25%, 50%, and 75% point of the following vowels, and for the duration of the syllable. The positive hypothesis posits that the speaker with Gujarati exposure would demonstrate significantly more of the negative VOT characteristics typically found in Gujarati speakers than the speaker only exposed to English. These findings may provide insight into both the process of bilingual language acquisition and the long term effects of childhood exposure to secondary languages. In the future, I’d like to expand the study to include more participants and additional speaking or listening tasks.

Neuroscience research relies heavily on cell counting to assess brain injury and evaluate neuroprotective treatments. While manual methods (i.e. hand counting) have been used traditionally, automated programs offer the potential for standardized and error-free data analysis. In the context of studying hypoxic-ischemic encephalopathy (HIE), a prevalent brain injury in infants, we aimed to assess the accuracy of an automated cell counting program in an in vitro slice culture brain injury model of injury and treatment. Code templates from ImageJ/Fiji were taken and modified using ChatGPT, and other snippets were used and modified from online forums like GitHub. The program searches through folders/subfolders for images, converts them into a binary image based on fluorescence threshold data (used to stain cell nuclei), applies the ImageJ function “watershed” that breaks larger groups into smaller groups, runs the “analyze particles” function which outputs a total cell count based on the size and circularity of the cells, and then saves the final image. Fluorescence threshold, cellular size, and circularity values were determined before data collection by adjusting the values to best fit the final image of a random slice. The settings were then kept consistent within studies. Preliminary results show the program’s high accuracy and precision, with consistent results across caffeine and Azithromycin treatments in our in vitro injury model. Despite yielding higher counts than manual methods, the program remained consistent across models. Validating this automated method represents a significant advancement in research methodology. These programs offer standardized data collection, error elimination, and faster analysis compared to manual counting, potentially saving time and resources for labs. Current limitations in our research involve differentiating between healthy and dead or dying cells, which would be an important future step for automated cell counting.

Games are engaging for children, and educational games make learning fun. Scholars have conducted research on how games help kids learn, but little is known about the specific ways that children’s mental models of real world concepts change through playing games. This study specifically investigates the process of how children learn STEM principles through exploring real world STEM concepts integrated into games. This ongoing research is being conducted with cohorts of middle schoolers in the King County area. Current games include Dragonwood, a card game with elements of probability, a Dungeons and Dragons campaign structured with elements of physics, and a STEM systems game called Oxygen Not Included. As the participants play these games, we ask them questions about their actions, interpretation of the game space, and why they make certain choices within the games. In doing so, we hope to understand the specific learning processes and parts of the game that change their understanding. Through qualitative analysis we are identifying trends in the students’ responses. We expect to find insight into learning through games and particular components in games that are beneficial for learning STEM concepts. This research has implications for educational game design and delivery. Our findings could impact the use of games in classrooms and provide insight for the development of educational games. This research could further make STEM learning more accessible and engaging for children.

Plants grow in designated areas known as meristems that contain stem cells. The gene LEAFY (LFY) is a flower meristem identity gene that initiates flower development in flowering plants and its homologs are also found in plants that do not produce flowers. Ferns are a lineage of land plants phylogenetically midway between the non-vascular bryophytes (such as mosses) and the seed plants. The model fern Ceratopteris richardii (Ceratopteris) contains two LFY homologs (CrLFY1/2). Given the described meristematic function of LFY in flowering plants, I am investigating whether CrLFY overexpression affects the development of the multicellular lateral meristem known as the “notch meristem”, the main site of cell division for the heart-shaped Ceratopteris gametophytes (the haploid stage of a plant’s life cycle). It has previously been established that CrLFY maintains cellular division in the apical cell of Ceratopteris gametophytes (a unicellular meristem that initiates development), but it is unknown if CrLFY also plays a role in notch meristem division once the apical cell terminates early in development. Consistent with our hypothesis, qualitative observations suggested that plants over-expressing CrLFY have larger and more U-shaped meristems, whereas wild type meristems appear more V-shaped. I am working to quantify the meristematic region within the notch in wild type compared to CrLFY over-expressing gametophytes using scanning electron microscopy and cell hand-tracing. I measure each traced cell and identify meristematic cells as those thinner and longer than the surrounding cells, with a length-to-width ratio of 3 or above. Additionally, I analyze the meristem angle to determine if there is a correlation between CrLFY expression and notch meristem shape. Understanding the role of LFY in plant meristems has important implications for the overall evolution of land plants, as master regulator genes such as LFY emerged in early plant lineages and evolved their functions over time.

This study seeks to explore the perspectives of Latine students at the University of Washington (UW) regarding racial representation within the institution. Do Latine students see their experiences and identities represented and reflected in university life? How do Latine students experience and perceive Latine representation (or lack thereof) in terms of the composition of faculty, student body, and community spaces at UW? This research project aims to uncover the realities, challenges, and promise of support and community on campus. Through interviews with Latine students, faculty, and staff, this inquiry will describe how students find support at UW and navigate their academic environment. To attain a comprehensive understanding, the research utilizes a combination of primary and secondary sources, incorporating interviews with Latine students, staff, and faculty at the UW to capture personalized and nuanced perspectives. By examining the lived experiences and perspectives of individuals directly affected, the study aims to thoroughly examine the complex dynamics at play. Exploring the lived experiences of Latine students, the study will contribute to the discourse on racial representation in academia and its impact on student well-being and academic success. The findings will inform discussions on how institutions can foster an inclusive environment that recognizes and supports the diverse backgrounds of all students. This research project has emerged from ongoing conversation and collaboration with the Washington State Commission on Hispanic Affairs members. This research project will inform the community report that the Commission is preparing.

Panãra, an Indigenous language native to Brazil, is currently the focus of Dr. Myriam Lapierre, Sunkulp Ananthanarayan, Ella De Falco, and Jessamine Jeter as some of the only linguists to document and conduct a comprehensive study on this language. Our research focuses on streamlining the process of organizing and analyzing field data – specifically in the context of Panãra, though generally applicable to other Indigenous and/or under-researched languages  – for use in future research by Dr. Lapierre and other scholars in the field of linguistics as it applies to Indigenous and minoritized languages. We have digitized the data from the field journals of Dr. Lapierre and the graduate students working with her, and our current focus is on the analysis of verb and sentence construction, via this digitized data, to organize grammatical paradigms into efficient and accessible indexes. We are also compiling and organizing PDF, image, sound, video, and experimental data for use on the California Language Archive (CLA) with a similar focus on efficiency and accessibility. The completion of this research entails the more complex understanding and organization of Panãra sentence and word structure for use in future research, both by Dr. Lapierre and by other scholars, as well as for usage in a Panãra dictionary. Our expected results also involve the creation and organization of the CLA page dedicated to Panãra, with a transparent structure making this data available to a wider audience of both linguists and non-linguists interested in learning more about the language.

Mutations in phosphodiesterase 6 (PDE6) underlie photoreceptor degeneration through cyclic guanosine monophosphate (cGMP) accumulation, triggering a series of down-stream processes, which eventually kill photoreceptors. We hypothesize that knocking out inosine monophosphate dehydrogenase 1 (IMPDH1), the rate-limiting enzyme in de novo guanine synthesis, will rescue cell death caused by PDE6 mutations. Supporting evidence from a mouse mutant model (rd10) suggests that inhibiting IMPDH1 pharmacologically delays photoreceptor degeneration (Yang, 2020). Our procedure for this experiment is as follows. Fish heterozygous for impdh1a and pde6c mutations are mated to produce fish that are double homozygotes. Fish homozygous for mutations in both the cone-specific pde6c and the impdh1a genes are genotyped and embedded for histological analysis of the retina. Histology is examined on days 3,5, and 7 post-fertilization (dpf) for cone degeneration. To date, we have genotyped our two mutant lines. Normally, pde6c^-/- fish have severe cone photoreceptor degeneration at 5 dpf and impdh1a^-/- fish show no signs of photoreceptor degeneration even as adults. If degeneration is rescued, the double knockout larvae should retain similar photoreceptor nuclei counts to wildtype fish at all time points. Demonstrating that IMPDH1 inhibition rescues PDE6 deficiency would provide proof-of-concept for the therapeutic potential of IMPDH1 targeted inhibition for the treatment of photoreceptor degeneration due to cGMP imbalance.

A product we consume daily, such as honey, may contain heavy metals like copper (Cu). Copper can get into our environment and food from urban sources such as tires, roofing materials, and generators. We therefore expect food products from more urbanized areas to show a higher concentration of copper. To test this hypothesis we evaluated copper levels in raw honey from different regions with different levels of urbanization. The honeys we tested were divided into three different categories according to their source: rural, suburban, and urban. The samples were digested by reflux with HNO₃ and H₂O₂ then it was diluted and filtered in preparation for analysis by Atomic Absorption Spectroscopy. The results suggest that the honey with the highest concentration of Cu was from an urban area, and the honey from a rural setting had the least amount of Cu. A one-way ANOVA analysis confirmed a statistically significant difference in the copper levels in raw honey from the regions (p < 0.01). The concentration of Cu in the samples of honey we analyzed ranged from 0.2 ug/g to 0.4 ug/g which is well within the recommended upper limit of 300 ug/g. Since bees collect pollen and honey from plants within roughly one mile of their hive, evaluation of the levels of copper and other metals in honey can provide a snapshot of the background levels of exposure in that area.

Bees collect a variety of pollen as a protein source to feed their brood. Any decline in the concentration of protein in pollen can be detrimental to the survival of younger bees, and any environmental factors that decrease the amount of protein in pollen can therefore have downstream effects on the overall health of honeybee colonies. As part of a longer longitudinal study, this study explored the difference in protein concentration in pollen collected in 2022 and 2023. This short term study is part of a larger project in which we seek to discover if there is a correlation between the changing climate and the changes in pollen protein concentration. We used a Bradford Assay to determine the protein concentration in pollen collected from hives at Edmonds College during equivalent time periods in 2022 and 2023 . We also subjected the pollen samples to microscopic analysis following an acetolysis procedure, in order to view the pollen's characteristics and floral type. Our results showed a statistically significant drop in protein concentration (two sample t-test, p< 0.05) from 2022 to 2023. As the climate continues to rapidly change around us there could be unforeseen impacts on bee health and populations. As this experimental project continues throughout the years, we hope to understand the full impact of climate change on the concentration of protein in pollen.

The inner ear is crucial for both hearing and balance and undergoes rapid changes during embryonic, fetal, and postnatal stages. Its complex structure poses challenges in studying auditory systems, especially the organ of Corti responsible for sound perception. Hair cells (HC’s) within the cochlea are essential for hearing, and their loss in mammals results in permanent deafness due to their inability to regenerate. We recently established Early B cell factor 1 (Ebf1) as an important factor for the development of the cochlea. Our lab’s work with conditional knockout (cKO) mouse models revealed that Ebf1 restricts sensory development within the cochlear duct. The Slc26a9-Cre Ebf1cKO model deletes on embryonic day (E) 9.5. We have seen over 2-fold increases in HC’s and their associated support cells (SCs). We have developed a tamoxifen-inducible Sox2Cre recombinase mice model for precise timing of genetic manipulation within the cochlea. Specific temporal activation of tamoxifen-inducible Sox2Cre recombinase in the cochlea will uncover critical regulatory time periods for the establishment of the sensory domain. Activation of tamoxifen-inducible Sox2Cre at embryonic day 11/12 only shows an increase in inner hair cells. These findings lead us to ask, what’s the critical window for Ebf1’s regulatory role in cochlear development? To determine the optimal window for Ebf1's regulatory role, I will activate the Cre expressed in Sox2 Ebf1-cKO mice with tamoxifen at different embryonic days (9-14) via oral gavage. Immunostaining experiments utilizing HC markers (Myo7a), inner HC markers (Vglut3), and SC markers (Sox2) will be conducted. I will quantify HC densities and cochlear length of Sox2 Ebf1-cKO and littermate control mice. Due to tamoxifen toxicity, samples will consist of embryonic day E18 specimens. Anticipated results include varying HC numbers, innervation and the presence of ectopic sensory patches. This study will offer valuable insight into the temporal dynamics of Ebf1's regulatory role.

Marine sediments are a critical carbon reservoir, locking both inorganic and organic carbon in sediments for thousands to millions of years. This carbon is released in part through the production of methane at methane seeps, places where methane gas is emitted from the seafloor. In this study, I characterized source differences and quantified the amount of inorganic and organic carbon stored in Puget Sound sediments at a methane seep site. A multicorer collected sediment samples at two sites: one at the Alki Point methane vent field and a control site within the Main Basin of Puget Sound. The sediment cores were subsampled in 2 cm sections downcore and stored for later analysis. Analyses included physical parameters (grain size, percent loss on ignition (% LOI), and 210Pb sediment dating) and geochemical parameters (Total Organic Carbon (TOC), Total Carbon (TC), del 13C TOC, del 13C TC). Both sites were silt-dominated (Alki = 37 microns, Control = 30 microns), with a well-mixed layer 15 cm thick. Organic content (% LOI) was high at both sites (Alki = 7.5%, Control = 6.9%). Likewise, TOC and TC showed trends of increased organic material at the methane seep site. Isotope data indicate that methane-influenced sites have lighter del 13C. Given the amount of carbon they store and the potential for these reservoirs to be disturbed by bottom trawling, deep sea mining, and other invasive human activities, understanding how carbon is cycled through marine sediments is critical for preserving these reservoirs and adequately factoring their role in the carbon cycle into global climate models.

Air pollution is a key component to understanding the Public Health of populations globally, with Diesel Exhaust Particles (DEP) being a significant contributor to traffic-related air pollution. Exposure to DEPs varies across populations and is therefore crucial to understanding the continual impacts of traffic-related air pollution on the public. Prior research has indicated that the formation of Amyloid-𝛽 (A-𝛽) plaques and activation of the  nucleotide-binding domain, leucine-rich–containing family, pyrin domain–containing-3 (NLRP3) inflammasome is linked with the development of Alzheimer’s disease (AD) later in life. AD is a form of progressive disease that impairs memory and other cognitive functions and impacts the lives of tens of millions of people globally. This study aims to confirm the linkage between exposure to DEP and memory impairment through NLRP3 inflammasome activation, utilizing an animal model to investigate a potential increase in AD later in life. We exposed male and female low-density lipoprotein receptor knockout (LDLR KO) mice chronically to inhaled DEP or filtered air as a control for 18 weeks. We then utilized the Object Location Memory (OLM) and Object Recognition Memory (ORM) behavioral tests to investigate the immediate impact of multi-week DEP exposure on short-term memory, another indicator in AD progression. Afterward, we sacrificed the mice and harvested a variety of tissues, including the brain. I conducted Immunohistochemistry (IHC) on cryosections of the exposed and non-exposed brain to assess DEP-induced AD-like brain architectural changes and to quantify the impact of DEP exposure in activating the NLRP3 inflammasome, ultimately leading to neurotoxicity, and to the development and progression of AD. Confirming the association between diesel exhaust and the NLRP3 pathway provides a potential therapeutic target in populations at an elevated risk for AD.

Compassion fatigue, also known as secondary traumatic stress or vicarious trauma, is a condition characterized by emotional and physical exhaustion, often experienced by individuals who provide care, support, or assistance to people who are suffering or traumatized. This phenomenon commonly affects professionals in helping and caregiving roles such as healthcare workers, social workers, counselors, and first responders as they’re constantly exposed to secondary traumas. This study investigates five domains of worker well-being of social services employees at a local Homeless Services Agency in King County. Seventy-five social workers and case managers were invited to participate in the online, anonymous survey that contains the National Institute for Occupational Safety and Health (NIOSH) Worker Well-being Questionnaire. This questionnaire consists of sixty-eight questions related to work evaluation and experience, workplace policies and culture, workplace physical environment and safety climate, health status, and home, community, and society. We hypothesize that those who have been employed at the Compass Housing Alliance for more than 2 years will exhibit higher compassion fatigue through their survey responses. The insights gathered will aid in understanding the working environment on service employees’ well-being. Results will offer areas to support workers and hopefully serve to mitigate the potential development of compassion fatigue among staff members.

Benthic foraminifera are single-celled organisms that are a useful proxy of environmental conditions. They can provide insights into water quality, sediment composition, and ecological changes. In the past, urban bays of Puget Sound developed chemical toxicity in sediments surpassing acceptable state limits. Recent mitigation efforts have brought most chemicals within limits. However, foraminifera health parameters continue to show signs of poor environmental conditions including increased test dissolution and decreased species richness and density. Studies on individual embayments are necessary to understand local variations and utilize foraminifera in an effective monitoring program. In this study, we investigated how the percentage of dissolved foraminifera varies based on distance from shore in Elliot Bay, Seattle. We obtained grab samples through the Washington Department of Ecology collected in 2021 from multiple sites in Elliot Bay. We processed the samples and picked up to 300 foraminifera from each sample then calculated the percentage of dissolved Elphedilla hannai, the most common calcareous species in Elliot Bay. Additionally, we reviewed foraminiferal densities, species richness, and diversity for distance from shore. We found that the percentage of dissolved Elphedilla hannai increased closer to shore and with decreased depth. These results suggest a spatial influence on the health of foraminifera.  As Elliot Bay is a major maritime hub, its shorelines are heavily industrialized, and areas both shallower and closer to shore may be experiencing higher levels of anthropogenic activities. This study highlights potential water quality gradients in urbanized embayments such as Elliot Bay, contributing to our overarching aim of understanding interactions between foraminiferal health and environmental factors in urbanized embayments.

Over the past several years, the city council of Oxford, England has developed an environmental policy project named the Zero Emissions Zone (ZEZ) which seeks to establish an area in the city center where vehicles will be either prohibited or fined for through travel. The council conducted many studies on where the policy would most effectively reduce carbon emissions from private vehicle use and considered existing pedestrian-only areas for potential use. The policy has proven controversial in the community. There has not been significant research on the popularity of the policy and how environmental policies like the ZEZ affect the political beliefs of Oxford community members. I hypothesized that the ZEZ would be a popular policy on the whole with the primary opposition coming from those who utilize private vehicles as their primary mode of transportation in Oxford and that approval or disapproval of this policy will significantly affect one’s perceptions of the city council and political parties. Through an anonymous online survey, I collected data from respondents over the age of 18 who either work, live, or study in Oxford or the surrounding suburbs. Participants answered questions regarding their political opinions, views on the ZEZ policy, and how the policy has affected their politics. These questions sought to illuminate a connection between views on environmental planning policies and political perceptions in general. I expect the results to show that younger respondents, particularly those whose political beliefs are to the left of center, will approve of this policy, and by extension the city council, more so than older respondents. This research has implications for Oxford City Council and any others interested or intending to install or expand similar zones so that they may understand what impact it may have on local politics. 

The composition of metropolitan governance has many effects on land use decisions, budget allocations, housing development, transportation planning, and racial, economic, and social equity in urban areas. However, there has been little academic inquiry into the effect of regional governance structure on transportation accessibility. This paper seeks to examine statistical linkages between regional governance fragmentation and trends toward and away from greater transportation accessibility in metropolitan areas. I perform a comparative statistical analysis of 47 of the 50 largest Metropolitan Statistical Areas, examining census data from 2002 to 2022 and transit accessibility data from the University of Minnesota Accessibility Observatory from 2014 to 2021 to examine this relationship. The causal factor I investigate is metropolitan governance fragmentation, which I capture through a Governance Fragmentation Index (GFI). The dependent variable, transportation accessibility, is captured through an Accessibility Gap Index, which categorizes transportation access through accessibility levels throughout each Metropolitan Statistical Area, utilizing data from the Accessibility Observatory from 2014 to 2021. My analysis controls for potential confounding variables, such as geographic area, population size, poverty levels, and region. I expect to find that lower levels of governance fragmentation in a Metropolitan Statistical Area will be associated with greater gains in transportation accessibility. Whether or not a significant relationship is identified, the research conducted will contribute to literature and ongoing research surrounding metropolitan governance and transportation accessibility.

The world’s oceans are witnessing a surge in plastic pollution, a consequence of human activities and the growing urbanization of coastal regions. Urban estuaries are complex habitats that are especially good at trapping sediment, carbon, and pollutants, such as plastics. However, our understanding of the extant of plastic accumulation within estuarine sediments remains limited. I determined the first quantification of the total amount of microplastics (>5 mm) in Puget Sound, WA – a heavily urbanized estuary – and identified deposition hotspots related to current hydrodynamics. To measure plastic concentrations, I collected both shoreline and shipboard sediment samples and density extracted microplastics using an NaI solution. Extracted plastics were counted, sized, and categorized under a microscope. To complement these plastic analyses, energy of the environment was determined using both grain size analysis and extraction of current velocities from LiveOcean, a hydrodynamic model of Puget Sound. I found that plastic concentrations are the highest near land-water interfaces, which are correlated with human population. The dominant source of microplastics came from fibers shed from clothing, giving a well-sorted particle size distribution. Furthermore, using the plastic concentration data I developed a predictive model of plastic distribution that relies on Puget Sound currents and could be adapted for other estuarine systems. Providing a comprehensive analysis of the sources and sinks of microplastics in main basin Puget Sound that can be used to inform preventative management on the negative impacts of urban waste.

Foraminifera, a diverse and widespread order of shelled marine protists, can be utilized as a reliable measure of marine ecosystem health due to their innate sensitivity to environmental changes. This study takes advantage of foraminifera to investigate the health of Commencement Bay, which is located in the South Puget Sound and encompasses the city of Tacoma. It is a highly industrialized embayment that features many man made inlets. Commencement Bay now hosts numerous proposed and implemented clean-up efforts, our primary focus being the U. S. Environmental Protection Agency Superfund sites put in place in 1992. This study, part of the Puget Sound Foraminifera Project at the Burke Museum, aims to build upon previous efforts to monitor the health of Commencement Bay and determine the effect of these clean up efforts. Sediments were collected by the Washington Department of Ecology from multiple sites in Commencement Bay and brought to the Burke Museum, where they have been washed, dried, floated, and picked to collect the foraminifera. Where possible, a minimum of 300 individuals was picked for analysis of assemblages. Results varied by location, with low diversity assemblages near the shore and higher diversity assemblages deeper into the bay. This indicates despite restoration efforts, waterways close to shore are still impacted by anthropogenic influences despite mitigation efforts.

As the percentage of individuals facing homelessness rises, many organizations working closely with unhoused populations aim to secure consistent housing and essential resources for those who lack this necessity. There are roughly 14,000 individuals facing homelessness in the greater Seattle area. Previous studies have shown positive correlations between employee well-being and job performance. Our research aims to investigate worker well-being among staff members who are currently employed at the Compass Housing Alliance, an organization working closely with unhoused populations to provide both housing and access to other essential needs. This research utilizes the National Institute for Occupational Safety and Health (NIOSH) worker well-being survey. We collected data on five factors that influence the perceptions of 75 staff members on their overall well-being including workplace safety, mental wellness, leadership support, coping strategies and job satisfaction. Expected results from our survey indicate that staff members' perceptions significantly impact their overall well-being. Collecting data using quantitative methods enables us to gain a deeper understanding of these current perceptions in order to utilize relevant resource implementation in areas where gaps are identified to support the needs of staff and improve their well-being.

Rare Intracranial Tumors (RIT) are a heterogeneous group with unmet medical needs. Although infrequent in individuals, RIT affect millions of people who lack effective disease monitoring and treatment. Frequent chromosome gains and losses are common in cancer, leading to the upregulation of oncogenes and downregulation of tumor-suppressor genes, respectively. Somatic Copy Number Variations (CNVs) affect a greater fraction of the genome than single nucleotide polymorphisms (SNPs) and have been correlated to drug resistance and tumor progression, highlighting a potential prognostic value. Cell-free circulating tumor DNA (ctDNA), which are DNA fragments released by necrotic or apoptotic tumor cells can act as a noninvasive cancer biomarker, offering a potential alternative to invasive tissue biopsies. In the present work, we aim to establish the somatic CNV signature of tumor and matched ctDNA to identify non-invasive tumor-related CNVs that may serve as biomarkers for use in liquid biopsy. We performed Whole Exome Sequencing (WES) in gDNA isolated from tumor tissue and matching ctDNA of 15 patients with RITs (pituitary tumor (n=9), craniopharyngioma (n=2), and meningioma (n=4). Raw reads were assessed for quality (Trimmomatic, FastQC), following alignment against human reference genome GRCh38 (BWA). Aligned reads were sorted and subject to duplicate removal using Picard. CNV profiles will be generated using CNVkit tool. Data analysis and visualization will be performed using R and python. The most promising aspects of liquid biopsy in cancer applications are cancer screening and early diagnosis because they can lead to better survival results and less disease burden. At the end of this work, we hope to identify the CNV signatures shared between tumor tissue and ctDNA, provide novel insights into the pathophysiology of these RITs and ultimately, suggest promising biomarker candidates for liquid biopsy.

Pediatric high-grade gliomas (pHGG) are highly aggressive brain tumors with limited therapies and extremely poor clinical outcomes. pHGG and many other cancers are often driven by genetic fusion events, which produce oncogenic fusion proteins such as TPM3-NTRK1. NTRK1 regulates signaling pathways promoting cell survival and differentiation. In cancer, NTRK fusions lead to constitutive activation of these pathways, resulting in uncontrolled cell growth. The purpose of my project is to validate TPM3-NTRK1 as a potential drug target for a novel therapeutic strategy known as targeted protein degradation and to clarify the role of TPM3-NTRK1 in cancer signaling. I hypothesize that degrading the TPM3-NTRK1 protein will decrease the activity of downstream signaling pathways that lead to pHGG. To model pharmacological degradation of TPM3-NTRK1, I applied the degradation tag (dTAG) system to rapidly and reversibly degrade a target protein fused with an FKBP12^F36V-tag. In this approach, dTAG molecules bind to the FKBP12^F36V-tag and recruit an E3 ubiquitin ligase to ubiquitinate the target protein, leading to its degradation by the proteasome. In my project, I used Gateway cloning to generate a lentiviral expression plasmid containing FKBP12^F36V-TPM3-NTRK1. I then produced lentiviruses, which I used to transduce NIH/3T3 cells and express FKBP12^F36V-TPM3-NTRK1. To test whether the FKBP12^F36V-TPM3-NTRK1 protein can be successfully degraded, I plan to treat the cells with dTAG molecules and use Western blotting at various time points to evaluate levels of FKBP12^F36V-TPM3-NTRK1 and known proteins involved in downstream signaling pathways, including pERK and pAKT. I expect that samples treated with dTAG molecules will show significantly decreased FKBP12^F36V-TPM3-NTRK1, pERK and pAKT. Importantly, these results will justify the development of small-molecule degraders to target TPM3-NTRK1, a promising therapeutic approach for pHGG and other hard-to-treat cancers. Additionally, a cell line expressing degradable TPM3-NTRK1 will serve as a powerful model system to further explore its role in cancer.

The fallopian tube function is central to several pathophysiological processes from facilitating fertilization to initiating ovarian cancer. However, existing models to study these functions are limited in relevance to the human condition due to the available conventional cell sources and culture systems. Many make use of mouse models that do not naturally develop pathologies such as ovarian cancer and lack the ability to replicate human genetic variation. Others use traditional 2D culture systems that have been shown to be poor in maintaining fallopian tube epithelium (FTE) secretory and ciliated function. Here, we propose to develop a novel in vitro model of FTE that is easy to manipulate and better mimics the in vivo environment through use of induced pluripotent stem cells (iPSC) as a cell source and a scaffold-based microfluidic culture system. We have modified an existing protocol and found that 3D culture allows for generation of organoids expressing FTE markers faster and at a higher rate. Additionally, I have developed a prototype device with a lumen and assessed cell viability and behavior of seeded immortalized FTE to establish the device. Further work can be done to elucidate drivers in FTE maturation through assessing primary FTE function in the device, and incorporation of iPSC-derived FTE cells into the device for studies on functional maturation.

The expression of prosocial behavior holds significance across various species, and its disruption stands out as a characteristic symptom in numerous neurodevelopmental and neuropsychiatric disorders such as autism, schizophrenia, and depression. Neuronal activity in the nucleus accumbens, particularly modulated by the neurotransmitter- dopamine, plays a pivotal role in regulating reward-motivated behavior towards both food and social interaction. Although the influence of dopamine on social behavior is extensively researched, there exists a notable gap in our understanding of neuronal activity within the nucleus accumbens during social interactions. In this study, a viral vector was employed to induce the expression of a genetically encoded fluorescent calcium sensor, coupled with mini-scope imaging, to observe calcium dynamics in the nucleus accumbens. The recording of calcium dynamics occurred during an operant task where an experimental mouse pressed a lever to interact with a social partner or in separate sessions to receive a food reward. Our investigation revealed distinct subpopulations of neurons within the striatum that specifically encode food reward, as opposed to other discrete cues or actions associated with the task. The insights gained from this research contribute to a more comprehensive understanding of how signals related to general reward-motivated behavior and prosocial behavior may contribute to disruptions observed in social behavior, particularly in disorders characterized by impaired prosocial behavior.

It is well established that formerly incarcerated individuals face stigma upon their return to society, negatively impacting reintegration efforts and life outcomes. Existing literature highlights the significance of stigma in influencing reentry outcomes, emphasizing gender-specific challenges faced by formerly incarcerated individuals, but fails to quantitatively study how these gender stereotypes impact public stigma. Although some research does address variations in the formerly incarcerated population and their influence on public opinion, these studies rarely focus on gender. My study directly explores how gender and stigma interact throughout reentry, by quantitatively investigating public perceptions of formerly incarcerated individuals in Washington State. I develop vignettes about hypothetical formerly incarcerated individuals that vary key facts between participants to examine the extent to which gender, offense type, and post-release behavior influence the public stigma of incarceration. Respondents are randomly assigned these vignettes to evaluate their varying perceptions, unveiling whether gender leads to differential perceptions and therefore, differential treatment in the daily lives of formerly incarcerated individuals. My preliminary expectations are that the public will value relationship-building (a traditionally feminine ideal) more for formerly incarcerated women than for formerly incarcerated men. Additionally, I hypothesize that respondents will exhibit a greater willingness to interact with formerly incarcerated women as compared to their male counterparts, due to negative stereotypes about male aggression and criminality. The implications of this research extend to a broader understanding of the obstacles faced by individuals with criminal records in achieving successful rehabilitation. My analysis will help create more targeted and effective reintegration support policies by uncovering how public stigma impacts the reentry outcomes for women differently than for men. Addressing these dynamics is crucial for fostering inclusivity and dismantling barriers that impede the social and economic participation of formerly incarcerated individuals.

Autism is a developmental disorder that involves differences in social communication skills and the presence of restricted or repetitive interests. One way children learn language is fast mapping: the process of forming associations between new words and their meanings. Growth in language production and comprehension is predictable by 2.5 years of age, and autistic children who learn more slowly during fast mapping tend to score lower in receptive language measures. However, there has not been extensive research on the differences in language learning during fast mapping between autistic and neurotypical children. Therefore, this study addresses fast mapping in one particular learning context and tests whether receptive language learning in a storybook-listening task differs between autistic and neurotypical children. There are 47 participants aged 2.7 to 11 years (mean age = 5.8 years, SD = 2.0), of which 17 are autistic (mean age = 7.8 years, SD = 1.6) and 30 are neurotypical (mean age = 4.7 years, SD = 1.2). I code videos of participants completing the storybook listening task for demonstrations of receptive language skills for novel vocabulary words — namely, the accuracy of participants' responses to prompts to look at named novel objects. I use t-tests to measure the statistical significance of the differences between the groups. Additionally, I conduct group matching based on participants' Growth Scale Value (GSV) scores on the Peabody Picture Vocabulary Test-5 (PPVT), a standardized measure of receptive vocabulary (autistic participants’ mean PPVT GSV score = 488.1, SD = 18.5; neurotypical participants’ mean PPVT GSV score = 479.2, SD = 12.2). I hypothesize that the autistic group of participants has lower receptive language learning scores. I hope the findings of this study can provide insights to help professionals in the field of communication sciences and disorders in their work with autistic children.

Vocabulary acquisition in children involves many cognitive processes, with semantic descriptions playing a potentially crucial role in word learning. Broader literature points to a positive impact of metacognitive strategies early in children’s learning. Additionally, studies have highlighted semantic descriptors in facilitating word acquisition. However, there is a lack of literature regarding the direct correlation between children's productions of semantic descriptions and success in word learning, which is the current focus. We examined the number of semantic descriptions produced by children alongside their corresponding word-learning performance to determine whether a relationship exists. We analyzed data from 33 participants ages 2.7 - 6.7 years (mean = 4.6 years, SD = 1.2) with an average Expressive Vocabulary Test-3 standard score of 120. Our team coded a total of 165 word-learning assessments from videos of participants engaging in a storybook listening activity where participants were exposed to novel words paired with objects, assessing their receptive and expressive word learning. Participants were asked to look at the objects, imitate the novel words in real-time, and both point to and label the associated objects after a delay. We recorded the number of times children made semantic descriptions (ex. “The gek is spiky”), total correct identifications (receptive performance), and total phonemes correctly imitated and labeled (expressive performance). For analysis, we tested the correlation between semantic descriptions and both receptive and expressive word learning scores. I hypothesize that a positive correlation between these scores will indicate that children who produce semantic descriptions find more success in word learning. I anticipate that our study will contribute to a better understanding of how semantic descriptions play a role in receptive and expressive word learning outcomes. Also, study findings could serve as a foundation for future research on how parents and educators might better support children’s vocabulary acquisition.

The invasion of Indo-Pacific lionfish (Pterois volitans) throughout the western Atlantic, the Gulf of Mexico, and the Caribbean has led to declines in the biodiversity, biomass, and abundance of native coral reef fishes. These ecological consequences have been thoroughly documented in shallow zones of the reef; however, little is currently known about how lionfish interact with deep-reef ecosystems, and which native fish are the most vulnerable to predation. Deep-reefs may serve as refuges for lionfish, as they would avoid being speared by SCUBA divers in the top 30 meters. Thus, deep lionfish populations could potentially serve as a near-infinite source of replenishment for shallow populations. The microchemistry of calcium carbonate fish otoliths has emerged as a powerful tool in examining fish ecology, including their vertical movement. The elemental compositions of these “ear stones” reflect the chemistry of the water in the fish’s environment, ultimately providing insight into natal origin and dispersal events. I am analyzing the oxygen isotopic signatures in a sample of 10 lionfish otoliths collected across the entire depth range of reefs in Curaçao using micromilling and isotope ratio mass spectrometry (IRMS). Specifically, variations in δ18O from the core to the rim of the otolith indicate changes in water temperature, and presumably depth, from settlement to adulthood. This approach provides insight into whether lionfish migrate between deep and shallow reefs throughout their lives or remain in their deep refuge, how frequent and long these migrations are, and ultimately whether they are susceptible to surface removal by spearfishing. Effective management plans will require a holistic understanding of this invasive predator’s vertical migration patterns and where they are on the reef at various life history stages, and a deepened understanding of the connectivity between deep and shallow populations can inform future research to control mesophotic populations.

An important part of addiction recovery is degrading high value associations between drug cues and the drugs themselves. Dopamine plays a crucial role in learning, and is specifically implicated in the prefrontal cortex (PFC) and reversal learning - learning to update and change behavior when it is no longer being rewarded. Past studies have reported elevations in dopamine during contingency reversal, but the timescale of how activity of PFC dopamine neurons maps to reversal learning remains unclear. Here we investigated the activity of PFC dopamine during reversal learning in a longitudinal fiber photometry study, recording dopamine signal on a timescale of seconds. Mice were trained on a reversal learning task where they initially learned that two of four presented odors precipitated a sucrose reward in 85% of the trials while the remaining two odors precipitated the reward for only 15% of the trials. Once the learning was stable, reward probability flipped for two odors (one 85% odor and one 15% odor) and the mice had to update their behavior to the new odor/reward structure. Fiber photometry recordings were conducted during pre-reversal, reversal, and post-reversal stages of the study. Our data replicate findings demonstrating elevated dopamine release during the reversal period, centered around the 15-85 cue. Analysis of the relationship between the dopamine signal and behavior also revealed significant cue, reward prediction error, and 15-85 reversal coding in the majority of animals, suggesting a multi-faceted role for dopamine in the PFC. Given this, dopamine in the PFC may play an important mediating role in the enhancement of associations between drugs and drug cues, but does not play a clear role in contingency degradation.

One of the most common types of vaccines used today are subunit vaccines. Subunit vaccines consist of an antigen that triggers the adaptive immune system to create antibodies but also require a separately added adjuvant, which is a substance that induces longer-term immunity by stimulating the immune system to pay attention to the antigen. Current adjuvants are non-specific - often things like oil-water emulsions that irritate the immune system and cause inflammation in unknown ways to draw attention to the antigen. This project aims to create a more specific adjuvant by directly stimulating B cells. In order for B cells to replicate antibodies, they need a primary signal from the antigen and a secondary signal that certain ligands on T-cells can initiate. We decided to investigate whether CD40Ligand (CD40L), an immune protein present on T cells that works to signal B cells to either replicate or create antibodies, could be used to achieve this goal. The idea behind the project is to co-display CD40L with antigen on the nanoparticle in hopes of creating a more specific adjuvant. We designed 10 different versions of this nanoparticle, where we tested two versions of CD40L, the placement of CD40L, and the linker length between CD40L and the nanoparticle surface. Our preliminary results also show that our cages retain their ability to bind both antibodies and CD40 as well as activate NFkB transcription - a proxy for B cell activation. We expect CD40L-displaying nanoparticles will promote B-cell proliferation to a greater extent than the nanoparticle vaccine displaying only hemagglutinin (flu) antigen accompanied with an adjuvant like Addavax. Ultimately, we hope to examine how co-display of CD40L with antigen will change the quality of immune response and memory in-vivo in comparison to currently used vaccine adjuvants, and begin testing in-vivo in the coming quarters.

Religious nationalism is receiving growing attention because of its current influence in democracies, but it manifests itself differently across countries. Previous research has identified socio-historical characteristics that impact the salience of religion as a factor in an individual’s conception of national identity. However, scholars of these studies have mainly used International Social Survey Programme data to analyze European nations. My study uses World Values Survey (WVS) data from 1981 to 2022 to evaluate democracies from a global perspective. I conduct a multilevel analysis to determine the salience of contextual characteristics that prime religion to be used as a vector through which nationalism is mobilized among individuals. Accounting for individual-level factors, I identify which country-level factors influence the relationship between religion and support for nationalism among individuals surveyed. I determine an individual’s support for religious nationalism using the beliefs and groups they express to support in the WVS. I then compare these results to events of national stress to determine if a relationship can be identified. My results suggest that religions favored in church-state relations are more likely to be included in conceptions of national identity. Additionally, affiliates of favored religions are more likely to support nationalism during times of outgroup threat, such as increases in immigration, socio-economic turmoil, and political/cultural shifts. By completing a global analysis of these phenomena, I am able to identify a more comprehensive pattern, something less expansive studies struggle to achieve due to hyper-partisanship debates that can overshadow case studies and regional analyses. Furthermore, by determining which socio-historical characteristics have the greatest impact on conceptions of a religious national identity, I provide a framework to develop a predictive theory on the circumstances under which a religion is not only primed to mobilize nationalist movements, but then comes to be employed to mobilize these movements.

On February 24, 2022, Russia began its full-scale invasion of Ukraine. Washington state was already the home of over 42,000 Ukrainian immigrants, and since the invasion, upwards of 22,000 Ukrainian refugees have arrived. Refugees face significant challenges due to obstacles stemming from the trauma of war, displacement, resettlement, and the lack of access to essential health and social services. The Nashi Immigrants Health Board is a non-profit, community-based organization founded by public health professionals. Nashi partners with the Ukrainian community via health promotion, education, and advocacy programs and provides culturally and linguistically appropriate public and mental health services. Nashi’s warm-line was created as a caller line for Ukrainian-speaking communities, primarily refugees fleeing Ukraine to provide psychosocial support and connection to community resources. The purpose of this study is to examine the data collected during warm-line calls to identify the greatest needs and streamline resources for the community. Warm-line operators collect anonymous information such as age, zip code, primary language, the purpose of the call, and action taken are recorded. Caller data are entered into a database with specific categories and analyzed for trends and patterns of needs, which will help Nashi refine the pathways to interventions and resource allocation. We hypothesize the callers will be mainly calling about needs such as housing, health insurance, and job opportunities. We expect the navigators to provide emotional support in addition to resources, even if callers are not expecting or requesting it. The findings would help Nashi inform the Office of Refugee and Immigrant Assistance (ORIA) for future programming and quality improvement interventions. The study results are not inclusive of every need in the Ukrainian refugee population. Ongoing efforts of resource allocation would continue to evolve based on the changing needs of the Ukrainian community.

DNA-binding proteins contain DNA-binding domains with specific or general affinity to DNA strands and play a crucial role in multiple cellular functions, including regulation of gene expression and DNA repair. Their ability to interact with specific DNA sequences makes them attractive engineering targets for therapeutic intervention in various genetic diseases. In this project, we employed deep learning models to design novel DNA-binding proteins tailored to recognize and bind to four specific DNA sequences associated with three different genetic diseases: neurofibromatosis, sickle cell disease, and Caffey disease. These DNA sequences served as input for RFDiffusion and LigandMPNN to generate a diverse set of theoretical DNA-binding protein designs, which were then filtered with the goal of having high specificity and affinity for the target DNA sequences. These optimal designs will be expressed using yeast surface display and assayed for binding in the presence of their DNA targets, which assists in quantifying the efficacy of the deep learning models. This project aims to evaluate deep learning algorithms and to further optimize these models, as well as characterize the proteins generated with the protein design pipeline. Our designs demonstrate evidence of the feasibility of protein design through deep learning models, providing valuable data for future refinement of the algorithm. Moreover, the de novo design of DNA-binding proteins targeting DNA sequences related to genetic diseases has transformative implications for novel therapeutic interventions.

This essay dives into three key stories: the myth of Pygmalion and his statue from Ovid's Latin epic Metamorphoses, the story of Acis, Galatea, and Polyphemus also from Metamorphoses, and Madeline Miller's novella Galatea. The major factor that these stories have in common is the overlying theme of male desire, and the consequences of not being able to attain that desire, typically in the form of control or violence. I analyze the literary choices made by both Ovid and Miller, and how they characterize their male characters in juxtaposition with their female characters. I take these analyses and connect them to the modern phenomenon of the Internet "Incel", which is a colloquial term for "involuntary celibate"— an online community of avid misogynists. My main methodology is utilizing critical sources by feminist classicists such as Donna Zuckerberg in conjunction with research studies done on male-dominated online forums to understand how misogynistic rhetoric is formed out of male desire. My essay focuses on Pygmalion and Polyphemus as prototypical Incels, and how this sort of misogyny and desire for control over women has a long history in popular media, and emphasizes the role that the classics play in modern day sexism.

DNA-Binding Proteins (DBPs) hold a strong affinity to interact with the major grooves of DNA for the purposes of transcription, translation, and repair. Although DBPs are found in nature, their specificity is difficult to predict and their production expends excessive resources. Therefore, our project’s goal is to efficiently generate DBPs that allow us to enable exact processes to occur. This is promising for future use in treating genetic diseases. In our research, we studied point mutations in the hexosaminidase subunit alpha (HEXA) gene and adenosine deaminase (ADA) gene, which can lead to Tay-Sachs disease (TSD) and Severe Combined Immunodeficiency (SCID), respectively. TSD is a rare genetic disease that affects infants and ultimately leads to brain damage, resulting in these children not making it to grade school age. Similarly, SCID is genetic, where children lack a strong immune system. This increases their susceptibility to infections, especially during their first year of life. Targeting the HEXA and ADA genes, we first developed designs utilizing computational software like RosettaFold, RFdiffusion, x3DNA, and LigandMPNN, followed by rigorous filtering via RosettaFold Nucleic Acid. Afterwards, we tested the final designs using yeast cultures and fluorescence-activated cell sorting (FACS) in the laboratory to determine which bind best to their generated DNA template sequences. Overall, we expect to find a few specific DBPs that bind effectively as predicted during the computational pipeline. These successful designs can be utilized as genome-editing proteins, correcting their target DNA sequences and restoring normal function.

Plants, like other multicellular organisms, possess a complex set of defensive measures deployed upon stimulation of the immune system. In the case of insect herbivory, these defensive measures depend upon specific pattern recognition receptors (PRRs). These PRRs enable the plant to detect herbivore associated molecular patterns (HAMPs). The Inceptin Receptor (INR) is a known herbivore receptor found in legumes, which detects Inceptin 11 (In11), a peptide fragment unique to the oral secretion of Spodoptera exigua. Beyond the plasma membrane much of the pathway from In11 detection to immune response is unknown. This research investigates possible signaling kinases downstream from INR. Receptor-Like Cytoplasmic Kinases (RLCKs) are known to mediate both specific immune and developmental signaling cascades in Arabidopsis thaliana. This suggests that they may play a role in the signaling network of herbivore induced immune responses. In this work we systematically investigate seven RLCKs of interest from three clades within the legume RLCK protein family which are upregulated by In11 treatment in the common bean (Phaseolus vulgaris). To better understand their effects, selected RLCK’s are transformed into the plant model organism Arabidopsis thaliana via Agrobacterium tumefaciens, and homozygous transgenic lines overexpressing the RLCKs are isolated. To examine the effects of over-expressing RLCKs on immunity, classic immune assays will measure their effects on Reactive Oxygen Species (ROS) and the phytohormone Ethylene production during immune activation. This research provides insights into plant immune signaling specific to caterpillar herbivory. In the future, this research may enable the intentional amplification or modulation of plant immune response to reduce losses in crops susceptible to herbivore infestation, a growing concern for food security in the near future.

Protists are single-celled eukaryotic plankton that are abundant and diverse in the surface ocean. Many of these protist species are motile often through the use of flagella or cilia. The gene expression of protists is closely synchronized with the daily cycle of light (diel), particularly through the activity of photosynthesis-related genes. However, there has been little investigation of which motility-related genes are expressed in open-ocean protists, among which species, and whether motility-related gene expression is also coupled with the diel cycle. This study aims to investigate the hypotheses that motility-related genes are highly expressed in protist communities, particularly among mixotrophs, and that their expression is synchronized with daytime light. To investigate motility-related gene expression of open-ocean protists throughout the diel cycle, triplicate eukaryotic metatranscriptomes, within the 0.2 – 100 μm size fraction, were collected from 15-m depth approximately every four hours for three days at 158 °W, 41.6 °N, located in the North Pacific Ocean. We annotated protein families (Pfams) present in the metatranscriptomes with Gene Ontology (GO) terms. Pfams were selected based on GO terms related to motility, such as flagella, motility, and cilia. This approach resulted in the examination of the expressions of 21 motility-related Pfams.

H3K9me3 heterochromatin regulates gene silencing and plays key roles in genome stability and cellular identity. During cell reprogramming, H3K9me3 has been shown to block transcription factor binding and changes to cell identity. Recently, enhancer of rudimentary homolog (ERH) was identified as a master regulator of H3K9me3 and a repressor of reprogramming in humans. However, the roles that specific ERH protein amino acids play in its function remain unknown. We hypothesize that elucidating the role of ERH and identifying pivotal amino acids will enhance our understanding of its molecular mechanisms. We are targeting amino acids which are evolutionarily conserved as these are more likely to be important for ERH function. We have shown that the ERH knockdown phenotype of low proliferation and decreased H3K9me3 can be rescued by exogenously expressing a wild-type copy of ERH. Our approach has involved generating mutant variants of ERH, introducing them into cells where the endogenous ERH is depleted and then assessing whether they rescue the phenotype. To efficiently assess many amino acid substitutions in the ERH protein simultaneously we are developing a multiplexed alanine scanning approach with Nanopore readout in a functional screen. In this approach, multiple ERH mutants will be simultaneously introduced and integrated into the genomes of a population of cells with the endogenous ERH depleted. Mutant forms of ERH which do not rescue the proliferation phenotype are expected to decrease in abundance over subsequent proliferation. The ratio of exogenous ERH sequences will be assessed upon initial introduction and after 2 weeks by targeted long read DNA sequencing. Identifying which amino acids are important for ERH function will allow us to design further experiments, investigating their role in localization, protein-protein interactions and H3K9me3 regulation. Understanding the regulation of H3K9me3 heterochromatin will be essential to improve cell reprogramming, enabling autologous transplantation and regenerative therapies.

The sensation of acute pain is fundamental to survival, indicating tissue damage that motivates an animal to engage in adaptive protective behaviors. Chronic pain, however, is pain that persists beyond the typical recovery window and serves little adaptive function. Due to the negative affective component inherent in chronic pain, it has been shown to contribute to the development of comorbid psychiatric disorders such as depression, social aggression, and social withdrawal. Our research aims to understand how acute and chronic pain affects social reward, motivation, and behavior. Our experiments used social self-administration (SSA), where pair-housed mice were placed in operant chambers and underwent voluntary lever press trials for the reward of social interaction with their cagemate. Following the acquisition of the SSA task, male and female mice then received a spared nerve injury (SNI) to induce neuropathic pain. Following recovery from the SNI surgery, experimental mice were returned to the lever press trials at different post-operative windows to simulate acute and persistent pain windows. Changes in social behavior were measured via changes in lever press frequency and interactive behavior during trials. Our data shows that male and female mice who underwent SSA trials immediately following nerve ligation maintained normal levels of social interaction. In contrast, female experimental animals that began SSA at a delayed time point, simulating persistent pain, lever pressed significantly less for a social partner compared to their respective control group. Conversely, male experimental animals at the delayed time point displayed high lever pressing behavior for a social partner. This data points towards divergent social adaptations following pain due to the sex of the experimental mice. Future experiments will further delineate these sex-specific adaptations following a traumatic injury. This research can inform social intervention as an adjunct or alternative treatment to pharmacological pain intervention and its comorbidities.

Including undergraduate peer facilitators (UPFs) in lectures can support students in large and often challenging introductory science courses. Our team decided to increase UPF involvement by creating BIOL113, an undergraduate-led companion course to BIOL180, the largest introductory Biology course at the University of Washington. This study is part of a larger project that assessed the overall experience of both UPFs and students, and this particular work focuses on the former. I investigated how BIOL113 contributed to the experience and development of UPFs.This study incorporated qualitative data from graduated and enrolled UPFs alongside UPF applicants. Through surveys, I explored personal, professional, and academic growth, leadership skills, program expectations, and a sense of belonging. My findings underscore the distinctive role of undergraduate-led programs in enhancing UPF experiences. UPFs’ expectations for the program aligned with their experience and enriched their sense of belonging within the biology department, STEM, and the broader University of Washington community. Furthermore, engaging in UPF roles developed individual leadership skills while also contributing to their personal, professional, and academic development. In addition, the survey highlighted an important area for improvement: tailored professional development for UPFs should focus on marketing the skills gained in teaching and leadership as particularly valuable in the job market, graduate school, or professional school. Understanding the impact of BIOL113 on UPF experience can inform instructional approaches, with the possibility of expanding such initiatives university-wide.

The crustaceous zooplankton Daphnia magna usually reproduce asexually but are also capable of sexual reproduction. Daphnia populations are generally all female; however, they occasionally produce male offspring for sexual reproduction. In lakes, the production of males and sexual eggs (ephippia) usually occurs in autumn, allowing Daphnia populations to reestablish in the spring. However, in our research, introducing males into some replicates but not others increases variability and decreases the ability to determine treatment effects. There are many hypotheses for what causes female Daphnia to have male offspring – the most common being alterations in light cycles or temperature – but we have encountered male offspring despite controlling these variables. Another hypothesis is that crowding leads to behavioral changes that cause female Daphnia to produce males. I am exploring this hypothesis to determine if the production of males is a behavioral response to crowding or to a lack of food availability. My experiments consist of two treatment groups of four replicates each: crowded and uncrowded. These groups are then fed proportionally upon creation, and reproductive behaviors are measured. The Daphnia cultures are fed with a mixture of three species of algae – Ankistrodesmus sp., Scenedesmus acutus, and Selenastrum capricornutum – to maintain a consistent level of food availability based on measured algal cell density estimated from in vivo fluorescence. If the hypothesis that density causes a behavioral change in reproduction is correct, the crowded treatment will produce more male offspring despite having the same amount of food. If food availability is the major factor impacting reproduction, the number of male offspring between the two groups will be constant. Eliminating sporadic male production would improve reproducibility within and between experiments.

Plants utilize cell surface protein receptors to recognize insect herbivory through the detection of Herbivore Associated Molecular Patterns (HAMPs) Following the detection of HAMPs, plants initiate specific immune responses, often measured by the increased production of the hormone ethylene gas and by Reactive Oxygen Species (ROS). The Inceptin Receptor (INR), which is specific to legume plants, recognizes the HAMP Inceptin11 (In11). The binding of In11 to INR initiates a signaling cascade, leading to an immune response. However, the signaling mechanism activated by INR is unknown. The Receptor-Like Cytoplasmic Kinase (RLCK) gene Herbivore Induced Kinase 1 (HIK1) is upregulated by In11 treatment in the bean species Phaseolus vulgaris. The goal of this research is to determine if HIK1 and other RLCKs are downstream proteins required for INR signaling. Because of the genetic intractability of P. vulgaris, I transform Arabidopsis thaliana with RLCK genes using Agrobacterium tumefaciens infiltration. Isolated genetic lines are then used to analyze the effect each RLCK has on immune signaling. Transgenic plants are treated with bacterial associated molecular patterns to trigger an immune response, then tissue samples of the leaves are measured for ROS and ethylene gas production. Results are then compared with ROS and ethylene gas production of wildtype plants. If the studied RLCKs are involved in downstream INR signaling, the transgenic plants will have increased ROS and ethylene gas production. I anticipate HIK1 to have the strongest increase in ROS and ethylene gas production due to the upregulation of HIK1 after In11 treatment in P. vulgaris. Understanding the INR signaling pathway is vital for engineering of plants that are resistant to insect herbivory without the use of pesticides.

Plants' defense mechanism against herbivory and disease is integral to both natural ecological balance as well as global food supply. Induced plant responses to these threats are often triggered by specific molecules such as Herbivore Associated Molecular Patterns (HAMPS) and Pathogen Associated Molecular Patterns (PAMPS). In this project, we are refining a Luciferase Reporter Assay (LRA) and then using that assay to categorize novel HAMPS and PAMPS. This assay’s first main part is the HAMP/PAMP Receptor (HPR). By inoculating a Nicotiana benthamiana (NB) leaf with an Agrobacterium containing a Plasmid with an HPR, we are expressing specific HPRs. We are then using the second part of the LRA, a Luciferase Reporter, whose promoter is tied to an immune response related gene to measure levels of immune activity without the need for Transcriptomic Analysis. After we have infiltrated the NB leaf with the vector containing Agrobacterium and induced a response by adding a HAMP/PAMP, we are measuring Luminescence as a proxy for immune response via an imaging machine, then using R to run an analysis on the levels of immune response, helping us characterize an HPR and its signal pathway. We recently optimized time points for HPR and Luciferase imaging and have found that infiltration by Agrobacterium only takes 24 hours for sufficient plasmid integration. As such we are using this assay to run an experiment on the molecular mechanisms of an HPR in only a few days. Refining this LRA and the information we have gathered has helped shed light on the underlying mechanisms used in induced plant defense. In the future, we are planning on expressing genes of interest using this technique to seek novel activators and suppressors helping us further understand mechanisms of plant defense.

Evidence-based changes to instruction can lead to better student outcomes and performance. For this reason, instructors are interested in collecting data about student experience and outcomes. However, the process of data collection and analysis can be time-intensive for instructors, making it challenging to gather data necessary to improve their classes. Moreover, when data collection does happen, it often centers quantitative data, but this systematically devalues students' experiences. Therefore, we sought to develop a tool that aids instructors in processing and analyzing qualitative data pertaining to students’ class experiences. As part of our research project, we have developed an R-shiny based data processing app that integrates Artificial Intelligence to summarize findings from open response questions on student surveys. This tool is intended to alleviate the time-intensive nature of analyzing student responses to open-ended survey questions. In order to validate the accuracy of AI-generated summaries, we compared them to manual summaries generated using in vivo qualitative coding methods. We find that our app generates responses that are comparable to the manually generated summaries. The summaries include prominent themes along with details about student experiences within those themes. With this tool, instructors can gather real-time data, even in large classes, eliminating the concern of the time-intensive process of manually reviewing responses. By developing this tool, we hope to empower instructors to explore diverse questions that provide them with valuable insights on how to optimize the structure of their classes to improve student experience and outcomes.

Pembrolizumab (brand name KEYTRUDA), is a monoclonal antibody therapy classified as an immune checkpoint inhibitor, a type of immunotherapy that works by facilitating the detection of cancer cells by T cells. Specifically, it blocks the PD-1 pathway that cancer cells use to suppress and evade the immune system. With neoadjuvant-adjuvant treatment, patients receive systemic therapy both before and after the surgical resection of their tumor(s). There is limited research evaluating the association between tumor responses (changes in tumor measurements) and survival time with neoadjuvant anti-PD-1 treatment. In this study, I analyzed the association between tumor measurements and event-free and overall survival in a clinical trial of patients receiving neoadjuvant-adjuvant anti-PD1-therapy. Event-free and overall survival were estimated using the Kaplan-Meier method, and associations with RECIST tumor size were evaluated using martingale residual plots and Cox proportional hazards regression models. In this study, I found significant associations between event-free survival and baseline tumor burden and changes (quantitative and categorized) in tumor size between baseline and the end of neoadjuvant therapy. Increases in tumor size were associated with a lower 2-year event-free survival. Larger values of baseline tumor burden were significantly associated with event-free survival but were not significantly associated with overall survival. The findings from this study may serve to better inform clinicians on making prognoses for patients on anti-PD-1 therapy and indicate a need for more research to be done on associations between tumor size and overall survival outcomes.

Antarctic blue whales (Balaenoptera musculus intermedia) are the largest animal on Earth, but much of their life history remains a mystery, including mating behavior, gestation periods, and early development. Antarctic blue whales were hunted excessively throughout the 20th century and are currently listed as endangered due to this exploitation. More than 340,000 blue whales were killed which led to an immense amount of data regarding the length, sex, and reproductive status of individuals. Using this data, compiled by the International Whaling Commission, I will build a model of Antarctic blue whale fetal growth and calf development. I will fit this model to the fetal and calf catch data to predict fetal and calf lengths in each month of the year. This model will result in predictions of the time of year that Antarctic blue whale calves are born and weaned as well as their growth rates. Determining the trends of fetal and calf growth in Antarctic blue whales contributes to the knowledge of reproductive behavior and growth trajectories. This is especially important for Antarctic blue whales, where no breeding areas have been identified. Understanding when calves are born and weaned can be combined with other data, such as satellite tracking, to provide insight into breeding areas.

Advances in language models have made diverse applications in social and health sciences possible. With many models emerging daily, it is important to understand general principles behind how language models represent text data. The study aims to investigate the effects of text manipulation on the numerical representation of meaning, or embeddings, through language models. The research is focused on numerous text manipulations (ex. changing direction of sentiment, sentence incompleteness, object of reference, intensity of statement). The research employs SentenceTransformer for embedding generation and UMAP for the visualization of semantic shifts. A comprehensive analysis of over 50 sentence manipulations provides a preliminary basis for understanding the nuances of semantic space organization. The core hypothesis suggests that incomplete and intensity based modifications lead to greater shifts compared to negative and referencing modifications. This outcome is expected to be substantiated through measures of embedding displacement from original text, and generalized through statistical validation, with tests accounting for multiple comparisons. This research is positioned to scale towards publishable results, aiming to systematically quantify the effects of semantic modifications across a larger dataset. The anticipated contributions of this work are twofold. It offers insights into the organization of semantic space as influenced by sentence modification, providing implications for the development of more nuanced NLP (Natural Language Processing) applications. And crucially, it lays a foundational basis for understanding how large language models (LLMs) process psychological statements and terms, an area of increasing importance as these models become more integrated into applications involving psychological analysis. Overall, the outcomes of this research are poised to deepen the understanding of how semantic alterations impact sentence embeddings, with potential applications in enhancing NLP models' interpretability in processing complex linguistic constructs.

Cells' ability to efficiently replicate their genomes is essential for regulating chromosomal division and maintaining chromosome integrity.  Defects in any of these cellular processes may cause genomic instability, potentially leading to cancer.  The Chaos3 allele in the yeast Saccharomyces cerevisiae is a single base pair change causing an amino acid substitution in the Mcm4 protein.  Mcm4, a component of the replicative helicase, is recruited to replication origins to unwind double stranded DNA and initiate replication.  Chaos3 is in a region of MCM4 that is highly conserved across eukaryotes; while mutations in conserved regions are generally non-viable, Chaos3 is a viable allele that causes genomic instability, leading to elevated cancer rates in mice.  In S. cerevisiae, Chaos3 decreases early firing of the autonomously replicating sequences (ARS) where DNA replication begins.  Chaos3 does not affect all early firing ARSs in the genome; rather, a large proportion of origins near centromeres, thereby delaying replication of those centromeres.  Essential for chromosome segregation, the centromere is the location where spindle fibers attach to pull apart sister chromatids during cell division.  I hypothesize that this delay in centromere replication results in chromosomal instability, including the loss of a chromosome.  I am using CRISPR guided cutting directed by a customizable guide RNA to replace centromeric adjacent ARS510, that has decreased firing levels in Chaos3, with unaffected, early firing ARS305, to see if firing levels in the mutants are affected based on ARS chromosome location (i.e., proximity to a centromere) or ARS sequence.  If replacing ARS510 with ARS305 restores early origin firing in this region this will confirm the Chaos3 mutation affects specific ARS sequences rather than ARS location on the chromosome.  Furthermore, if centromere replication delays are the cause of genomic instability in Chaos3, this ARS replacement should rescue chromosome loss.

Advances in language models have opened up endless applications in the social and health sciences. A key to fulfilling this promise is the development of rigorous practices for gathering data used for model training. This research project reports the development of standardized protocols for building databases for self-reported psychological questionnaires that can be used to fine-tune language models. These questionnaires cover a spectrum from childhood experience to altered states of consciousness and beyond. We systematically review the literature on appropriate database-building strategies, use combined data collection techniques, and instill specific criteria into our collection process to help facilitate the research’s computational pipelines on evaluating the relationship between semantics and psychological factor membership. Using a 2D semantic map of the PsycTests database as a reference, we can further evaluate the effectiveness of our protocols by developing quantitative measures to assess comprehensiveness and objectivity in our database building. Our initial explorations involve measuring data variance and density of data distribution. This research provides a general toolkit for building text databases for appropriate and effective model training purposes across various disciplines.

Individuals exposed to chronic, severe, and/or inescapable stress may adopt dissociation as a coping mechanism to alleviate suffering. In dissociation, individuals report losing connection to their bodies, reality, memories, etc. Dissociative experiences are hallmark symptoms of psychiatric disorders such as Post-Traumatic Stress Disorders (PTSD) and Dissociative Identity Disorder (DID), which has high comorbidity with personality disorders. Maladaptive personality patterns can be thought of as chronic patterns of behavior that are resistant to change. Brain systems are known to underlie these sorts of habitual learning. The interactions between dissociative experiences and action-learning mechanisms are still poorly understood. In this project, we review the history of dissociation and action-learning research and synthesize a model of how dissociative mechanisms may interact with action-learning mechanisms to develop pathological behavioral patterns and even multiple personalities. We propose employing the PRISMA method to select relevant literature from PubMed or other medical databases systematically and conduct a comprehensive review across the spectrum of dissociative disorders. By utilizing the Dissociative Experiences Scale (DES) scores as a measure method, we aim to identify action-learning brain region activity (BOLD signal) and volume changes and explore how these changes correlate with dissociative symptoms; and we’ve found that DID, Dissociative Disorders, and PTSD has the highest DES mean scores. Beyond this, our findings show that in these patients with different kinds of dissociative disorders, for the patients with different kinds of dissociative disorders, their Hippocampus and Amygdala are smaller while their Pallidum Volumes in their brains become typically larger. Despite this relationship having elementary processes, combining with future findings and discoveries, the relationships between dissociative experiences and action-learning mechanisms will be better understood.

Family businesses are some of the most important economic contributors in the United States, accounting for approximately 64% of the U.S. GDP. The family business model, which refers to any business with two or more family members on the board or in ownership, is a crucial and enduring part of business in the Seattle area and abroad. Historians have often pointed out that the family business model seems to be the base model for business and has thus been present since the beginning of organized business, often in the form of farms, merchant companies, banks, and other small businesses. Despite its prevalence, the family business model is far from perfect because of its numerous commonly encountered limitations. One of the limitations family businesses face is the challenge of succession, as only about 30% are able to succeed from the first generation to the second. Other limitations relate to growth, sustainability, and qualification problems. This study, conducted as a literature review, uses a combination of peer-reviewed articles and popular sources (chosen based on criteria of relevancy and prominence) as quantitative data to examine the consensus of family businesses in Seattle and the solutions that have been proposed to address these limitations. Interviews with family business owners in the Seattle area were also conducted to provide qualitative data and to highlight specific opinions. The economic and historical implications of Seattle family business are also discussed. This research aims to provide insight into otherwise costly financial, succession, and leadership difficulties in order to ensure that the family business model is an enduring contributor to the Seattle economy. Having the proper knowledge on how to approach these difficulties and reconcile with their seemingly conflicting nature can help family businesses in the Seattle area thrive while working through complicated business situations.

As healthcare advances, older people with HIV (PWH) have a longer life expectancy. Biotechnological advancements, such as antiretroviral therapy (ART), suppress HIV replication, resulting in decreased mortality among PWH, while increasing quality of life. However, despite these advances, older PWH generally experience an earlier onset of comorbidities due to increasing age, increased adiposity related to HIV medications and sedentary lifestyle, poorer physical function, and high symptom burden. Physical activity is a significant non-pharmacological approach to mitigate chronic disease in older adults and promote healthy living. Among different physical activity types, exercise can be defined as structured and repetitive bodily movement, objectively improving or maintaining physical function. Exercise can take many forms and be performed at different intensities. HEALTH Study participants perform continuous moderate exercise (CME) of walking or jogging at 50% of heart rate (HR) or high-intensity interval training (HIIT) of alternating short intervals of high- and low-intensity aerobic exercise at 80-90% and 50% HRmax. Healthy aging among PWH includes decreased fatigue, greater functional mobility, and fewer symptoms of HIV and its related treatments. What is unknown, and currently investigated in the HEALTH Study, are exercise types to improve physical health outcomes in older PWH. This study aims to explore the relationship and associations between exercise interventions on body composition across older PWH participants. In this year-long undergraduate research project, we conducted a secondary data analysis to compare the effectiveness of exercise interventions, HIIT versus CME, on DEXA scan outcomes of body fat and bone density in older PWH. DEXA scans are medical imaging x-ray tests to quantify body composition. We anticipate that with increased physical activity, especially in HIIT participants, body fat will decrease and bone density will increase. These findings may indicate how exercise interventions can mitigate fatigue and provide recommendations to prompt healthy aging in older PWH.

Every year, substance abuse, mental disorders, and psychotic behavior become more and more severe, especially in the local communities of Seattle. Our research explores the interconnection between psychology and the utilization of modern computer vision and emotion recognition tools to analyze human semantics/behavioral patterns in order to comprehend the underlying dynamics that lead to these behaviors and ultimately, spearhead efforts for prevention. As a computer science researcher, I employ advanced machine learning algorithms built upon open source programs to track and interpret gestures, facial expressions, and body language in video data. By filtering the data through a database and comparing emotions through statistical analysis in similar individuals, I am able to uncover nuanced patterns in behavior and emotion that a human researcher would otherwise not notice. While the study is ongoing, our current data illustrates correlations between specific gestures, facial expressions, and emotional states in hallucinogenic use. Though these individuals exhibit typical behavior of substance use, they have different backgrounds and contexts that lead to unique obscure changes in emotion and behavior, and our program allows us to identify and analyze these transformations that current medical knowledge overlooks. These findings hold promising implications for informing future research directions and intervention strategies across multiple domains, from understanding human behavior in social interactions to enhancing human-computer interfaces.

Individuals exposed to chronic, severe, and/or inescapable stress may adopt dissociation as a coping mechanism to alleviate suffering. In dissociation, individuals report losing connection to their bodies, reality, memories, and more. Dissociative experiences are hallmark symptoms of psychiatric disorders such as Post-Traumatic Stress Disorders (PTSD) and Dissociative Identity Disorder (DID), which have a high comorbidity with personality disorders. Maladaptive personality patterns can be thought of as chronic patterns of behavior that are resistant to change, which often are heavily present in brain systems that focus on habitual learning. However, the interactions between dissociative experiences and action-learning mechanisms are still poorly understood. In this project, we review the history of dissociation and action-learning research to synthesize a model of how dissociative mechanisms may interact with action-learning mechanisms as a cause for the development of pathological behavioral patterns and even multiple personalities. We propose employing the PRISMA method to select relevant literature from PubMed or other medical databases systematically and conduct a comprehensive review across the spectrum of dissociative disorders. By utilizing the Dissociative Experiences Scale (DES) scores as a measure method, we aim to identify action-learning brain region changes and explore how these changes correlate with dissociative symptoms; thus far, we’ve found that DID, dissociative disorders, and PTSD have the highest DES mean scores. Beyond this, our findings show that in these patients with different kinds of dissociative disorders, the hippocampus and amygdala are smaller and the volume of the palladium is typically larger. When combined with future findings and discoveries, the relationships between dissociative experiences and action-learning mechanisms will be better understood.

The Type 2 inflammatory response occurs during intestinal parasitic helminth infection that affects over a billion people worldwide. Type 2 inflammation involves the activation of intestinal epithelial stem cells (ISCs) in the remodeling of the intestinal epithelium. ISCs differentiate into effector-like cell lineages such as goblet cells and tuft cells that lead to the production of mucus that flushes the helminth out of the intestines. However, the mechanisms that promote and regulate this epithelial remodeling and its long-term consequences remain unclear. For this project, I am analyzing the role of the bioactive lipid prostaglandin D₂ (PGD₂) in the regulation of goblet and tuft cell differentiation by creating enteroids that carry fluorescent reporters that mark ISCs. Enteroids are 3-D cell cultures formed from stem cells that model the intestinal epithelium without the presence of immune cells or other mediators; thus they allow me to hone in on the epithelial intrinsic factors that impact the immunological response in vivo. I hypothesize that PGD₂, and its receptor CRTH2, is necessary for the resolution of the Type 2 epithelial response during helminth infection. By treating enteroids with PGD₂, a PDG₂ antagonist, and inflammatory type 2 interleukins (IL) such as IL-4 and IL-13, I can analyze the role of these elements in the initiation and termination of the Type 2 epithelial response. This research is impactful because understanding the basic principles of intestinal epithelial regulation will be key to furthering the field of therapeutics and treatments for intestinal inflammatory diseases.

 The cGAS-STING pathway is a critical component of the innate immune system responsible for the detection of cytosolic DNA, which is key in identifying viral or bacterial infection. Activation results in the transcription of type I interferons (IFN), including IFN-β, and other proinflammatory cytokines that lead to the recruitment of immune cells to help control the infection. To explore this pathway, we use a system of conditionally immortalized macrophages (CIMs) derived from Cas9-transgenic i-TOMCAT mice. The Cas9 transgene allows us to make gene-specific knockout (KO) CIMs using CRISPR, while the i-TOMCAT allele, which drives expression of a fluorescent reporter (TdTomato) from the endogenous Ifnb1 locus, allows us to quantify type I interferon expression using flow cytometry. Using the STING agonist DMXAA, I found that CIMs in the progenitor state undergo rapid cell death upon STING activation, while terminally differentiated CIMs were resistant to cell death and produced high levels of IFN-β. Further experiments using the pan-caspase inhibitor Z-VAD-FMK and the necroptosis inhibitor necrostatin-1 showed that cell death in progenitor CIMs following STING activation is independent of apoptosis and necroptosis. This project aims to explore the signaling pathways downstream of STING that dictate either a cell death of IFN-β response. While considerable progress has been made in understanding how cGAS-STING signaling leads to IFN-β secretion, the cell death pathways associated with this cascade remain enigmatic. With recent studies showing that cGAS-STING activation can lead to tumor suppression by preventing cell division, promoting immunosurveillance, and triggering the release of tumor-specific antigens, research into this pathway may help guide the development of improved cancer immunotherapies.

Germany's Energiewende, a pioneering transition to renewable energy, has drawn global attention. Yet, despite initial enthusiasm, disillusionment has grown among energy sector workers and civil society. This study aims to uncover the factors driving this disillusionment. The primary research question is: What contributes to disillusionment amidst the Energiewende transition? Through qualitative analysis of data from the IGBCE union, a key stakeholder in the transition, this research examines the gap between policy objectives and worker concerns. Hypothesizing that a lack of legal support fuels dissatisfaction, the study contextualizes evolving worker attitudes through literature review. Concerns over job insecurity and transition policy inadequacies have shifted sentiment despite initial optimism. Using a comparative case study approach and narrative analysis, I explore how the IGBCE's legal narrative reflects worker grievances. By examining documents from 2015 to 2023, including union materials, recurring themes and narratives are identified. This illuminates the complex relationship between government policies, and worker discontent amidst energy transition. Employing sociological theories such as the "treadmill of production" and "creative destruction," I examine barriers to a just and sustainable transition, questioning whether there are other growth factors that leads to advanced economies being stuck on a "treadmill". Analysis reveals that the Energiewende is primarily driven by societal pressure, prompting political and economic transitions. This shift also influences the stances of labor unions, exemplified by IGBCE's evolution from advocating for fossil fuels to embracing renewables. However, union interests persist in advocating for members. Consequently, with only half of energy jobs governed by collective bargaining agreements aimed at ensuring fair wages, dissatisfaction among workers intensifies. Therefore, workers find themselves vulnerable to precarious conditions and insufficient wages. Result also shows notable lack of government support, further exacerbating worker concerns. This deficiency underscores the importance of unions as effective representatives of workers' interests, with perceived benefits such as collective bargaining power and legal representation, which contribute to the increasing appeal of unions for workers seeking support and protection.

Approaches to learning new languages and cultures, among other subjects, are ever-changing alongside advancements in technology. In this study, I examined the informal self-regulated English as a foreign language (L2) and intercultural learning of a native Japanese (L1) speaker through their interactions with social media platforms such as YouTube, Discord, and X (formerly known as Twitter). In an online semi-structured retrospective interview with "Yuki," a Japanese woman in her thirties, I queried three areas of informal online language and intercultural learning: 1) her language learning background and the various media she interacts with; 2) her perspective and attitude towards self-regulated learning, her motivations, and how this compares to formal learning; and 3) how her connections to multiple communities on various platforms have influenced her learning and outlook on life. This interview was conducted in Japanese, to allow Yuki to describe her thoughts and experiences as clearly as possible. Findings show the strong impact of online communities on Yuki's motivations and objectives. The global community she encountered offered plentiful opportunities to expand her outlook on life, from her views on pronunciation to the importance of identities. Through the internet, Yuki found others with similar learning goals and began hosting weekly online book club sessions, where individuals of varying English levels are able to develop skills in collaboration with one another despite never having met in person. Along with the possibility for self-directed informal learning, other forms of online content, such as podcasts, allow Yuki to freely teach and challenge herself to interact with everyday spoken English, such as slang and diverse dialects. Although this study only consists of one participant, findings show the tremendous developmental potential of online spaces for language learners. 

Resilience in Alzheimer's Disease (AD) is defined by the difference between a person's expected and actual rate of cognitive decline given the severity of their disease. However, the mechanisms behind resilience are still unclear and I wanted to see if the anatomy of the brain over the course of AD could offer any clues. To accomplish this, cognitive tests and brain scans were obtained from patient data taken at Harborview Medical Center. Brain tissue atrophy in regions of interest were defined and combined into two measures. The first--biological subtypes--is whether the disease primarily affected the limbic regions or the cortical regions, while the second is left-right asymmetry. I found that resilience correlated with biological subtypes but not asymmetry. This suggests a way for us to predict resilience to better personalize treatment and eventually find ways to increase resilience.

Food insecurity is a global problem with one out of every six people in the United States relying on charitable food assistance organizations to meet their needs in 2022. However, organizations like food banks are underfunded and overwhelmed, relying on donors to continue aid. One food bank in the Seattle area received 70% of their donations from individual donors as financial aid, and a significant contribution comes from grocery stores in the form of food surplus. Previous research has shown that tax deductions motivate grocery stores to donate and limit food waste, however little is known about the care involved in the decision of selecting food to be donated. The quality and quantity of donations varies depending on the store and department. I explore how managerial discretion explains why different grocery store departments choose to donate food or throw it away. I conduct one-on-one structured interviews with three managers in four grocery stores in a Seattle neighborhood to find what factors influence the amount and quality of donations. Additionally, I uncover the internal and external constraints of managers in different departments. Managerial discretion has been studied in various firms yet it has not has not been explored explicitly within the context of donations. Similarly to charitable donations, which are scarce in the context of individuals making philanthropic decisions within organizations. I predict managers’ values and opinions on charity, the grocery store’s philanthropic organizational structure, and the perceived strength of the food bank-to-manager relationship will influence the quantity and quality of donations. I also predict that managers in stores with a higher philanthropic public image will contribute donations of higher quantity and quality. The findings from this study will help identify factors to increase the quantity and quality of donations so that stores can donate higher quality food.

Plants recognize herbivore-associated molecular patterns (HAMPs) during herbivory that activates signaling to induce immune defenses. Caterpillar oral secretions contain Inceptin 11 (In11) which is a HAMP recognized by legumes such cowpea via Inceptin Receptor (INR). Thus, In11 and INR are a model system to study proteins involved in HAMP induced defenses, including Kunitz trypsin inhibitors (KTIs). It is known that KTIs are serine protease inhibitors with anti-herbivore activity; however, the precise role of In11 induced KTIs and the effect of cysteine content variation in cowpea KTIs remains unknown. Here, we show that selective removal of cysteines has a negative effect on KTI function in cowpea experiencing herbivory from the fall armyworm (Spodoptera frugiperda). We found that cowpea KTIs act as antiherbivore proteins against the fall armyworm when expressed in Nicotiana benthamiana, as we saw reduced weight gain on larvae feeding on leaves expressing wildtype KTI. Furthermore, we found that KTI function was negatively affected by the removal of cysteines, and larvae fed leaves expressing any of the mutant gained more weight than those feeding on wildtype. We hypothesize that these findings are due to reduced protein stability because we did not detect mutant KTIs in frass samples by westernblot. Understanding KTI protein structure and how it influences protein function is important for designing and selecting antiherbivore proteins to be used for plant defense in agriculture.

Plants are generally unable to move reasonable distances as single adult organisms. This lack of mobility necessitates robust pathways that allow for response to environmental stimuli which can cause changes in the plant’s morphology to adapt to their changing environment. One such trait is flowering, and it is crucial for plant survival because it allows for plants to reproduce. The vast majority of agricultural plants are flowering plants, and robust growth and reproduction of these plants is especially important for the growing human population. This experiment aims to understand the interaction between an Arabidopsis thaliana nitrogen response gene named NITRATE-INDUCIBLE GARP-TYPE TRANSCRIPTIONAL REPRESSOR 1 (NIGT1), and an Arabidopsis gene which controls flowering called FLOWERING LOCUS T (FT). NIGT1 is known to modulate plant flowering despite being a nitrogen response gene. Here, I used a Dual-Luciferase Reporter System to test whether NIGT1 proteins directly interact with the FT promoter to regulate FT gene expression. In this system, the FT promoter is used to drive expression of Firefly Luciferase rather than FT. Separately, the NIGT1 gene coding region will be highly expressed under a constitutive 35S promoter. It will also be fused to the transactivation domain of the viral protein VP16 which converts transcriptional repression into transcriptional activation, as NIGT1 is known to be a transcriptional repressor. The VP16 transactivation domain will be repeated as four tandem repeats, forming the construct called VP64. The gene constructs of interest will be inserted into Nicotiana benthamiana using agrobacterium infiltration, then the leaf material from N. benthamiana will be used for the Luciferase assay. I hypothesize that the binding of NIGT-VP64 to the FT promoter will cause increased in Firefly Luciferase production compared to the absence of NIGT1-VP64. Higher amounts of Firefly Luciferase will result in greater luminescence, which we will quantify with a luminometer.

While environmental writing is nothing novel, environmental journalism as a beat and research area is relatively recent. The available literature on the field provides insight into the challenges environmental reporters face and what their environmental backgrounds in the field are, but little research details how they respond to challenges and why they originally chose the beat. This research examines how environmental journalists respond to the challenges they face, what values they attribute to the beat, and why they chose to write about the environment. My hypotheses are that these reporters have had influential experiences in nature; they see their work as a form of social activism; and they rely on their social connections as a form of support. For my sample, I first used random systematic sampling, followed by purposive sampling to reach targeted demographics, such as gender and race. I conduct semi-structured, in-depth interviews to collect my data, with a goal of at least 15 interviews or until saturation. So far, I have conducted five interviews that average around 45 to 60 minutes each. Most of them grew up as outdoorsy people, all of them see their work as important, and most find that talking to someone about challenges they encounter is helpful. Through more interviews, I hope to begin recognizing strong trends in responses to compare to my hypotheses. In addition to filling the gaps in the literature on environmental journalism, this research provides these reporters with a chance to talk about their experiences and challenges. Additionally, building on this beat as a research area could help provide data to create a meaningful support network for environmental reporters. As climate change intensifies and more people feel its effects, it’s imperative that these journalists feel supported enough to continue highlighting climate solutions and inequities to advocate for climate action.

Oral contraceptives (OC), commonly known as the birth control pill, produce temporal hormonal infertility for women. While its effectiveness and reversibility are the major advantages of OC, users may experience varying side effects ranging from headache and nausea to blood clotting. Access to and perception of OC may also differ across cultures, further affecting the user’s experience with OC. For example, prior research has shown that OC usage is more common in the United States than in Japan. The purpose of the study is to examine how cultural factors such as accessibility and stigma as well as biological factors such as physical side effects influence American and Japanese college students' decisions and experiences surrounding OC usage. Between December 2023 to February 2024, I conducted an online Qualtrics survey and semi-structured phone interviews with female students at the University of Washington and Waseda University, Japan. In the survey, I included questions about why respondents used OC, perceptions of OC usage in their country, knowledge of how OC works, and experience with the side effects. I analyzed survey responses in R to assess differences in Japanese and American participants’ experiences and perceptions. For interviews, I thematically analyzed the transcription using ATLAS.ti. Through analyzing the responses of the two cultures quantitatively and qualitatively, I expect to highlight the difference in cultural perception of OC, as well as the common biological side effects that women may go through by taking OC. Through the real voices of women, this study can provide an understanding of how cultural and biological factors can potentially lead to positive and negative OC experiences. This can be especially useful for potential and experienced users of OC when making reproductive decisions.

The last few decades have marked an ongoing struggle between the highland Indigenous groups of Northern Thailand and the Thai government over the settlement and use of lands located inside of protected national parks. According to the Thai government, these native groups, including the Karen people, use practices that are harmful to the local environment and amplify climate change's effects. However, many others, including academics and Karen activists, argue that Indigenous knowledge and practices are in fact more beneficial to the environment than simply leaving nature alone. This paper examines the validity of these assertions by exploring how the Karen group interacts with the environment through their culture and what effect this has on environmental factors such as climate change and biodiversity. Findings from literary research, artifact observation at the Burke Museum, and interviews with various experts with experience involving these land rights issues suggest that Karen people’s culture is in fact what informs their respect for their local environment and the reason why their environmental practices, such as rotational agriculture, usage of natural materials for clothes and tools, and their general treatment of the natural world are mutually beneficial to both Karen people and the environment. My research also suggests that there is immense value in protecting Karen culture both within their communities and from the threats of the Thai government and its dominant culture. By better understanding and appreciating this knowledge that Karen and Indigenous peoples as a whole have based on years of experience on their native land, we will be able to understand the importance of protecting Native peoples’ right to their lands for both the good of the environment and those living within it.

In my research, I explore the role of children in Late Bronze Age Mycenaean society (ca. 1600-1100 BCE) by focusing on the relationships, socioeconomic and religious roles, gender dynamics, and burial practices of and for children, as well as ideas surrounding adolescence and the representation of children in Mycenaean art. Children hold a unique role in society, and it is important to understand the roles and perceptions of children. In doing so, we gain a better understanding of the experience of childhood as well as the culture as a whole. I begin by defining Mycenaean views on childhood, providing a framework to contextualize the discussion. This project is a synthesis of previously published data, incorporating the Linear B tablets, sites, excavations, and art from Late Bronze Age Mycenae, explored in this paper through comparative analysis. I utilize a similar approach when discussing the burial practices for children, focusing on burial architecture and style and archaeological reports on specific sites, such as Ayia Sotira and Mycenae. By studying the roles and lives of children, we gain insight into generational dynamics, family structure, gender roles, and domestic dynamics, which all contribute significantly to understanding the cultural structure of a society. Through the specifics of material culture, social spaces, and practices involving or surrounding children, I look to develop a larger picture of the experience and significance of children in Mycenaean society, in effort to give context and perspective for existing and future research surrounding Mycenaean culture.

Media coverage of the National Hockey League (NHL) has brought public attention to many accounts of physical and sexual violence, hazing, and illicit drug use by players and coaches over the past 20 years. My research investigates the institutional mechanisms the NHL, its teams, and the players union use in response to cases of criminal behavior by their athletes. I ask how consequences for player criminality vary by type of crime, status of player, and player network. I compiled a dataset of all incidents covered in the news or social media between 2009 and the present, and then identified the response type through qualitative coding and comparison. Given prior theorizations of men’s collegiate athletics as crime-facilitative environments based on low punishment risk and high temptation to participate in criminal deviance, I expect to find that fine-related punishments are levied more frequently against high-status NHL players and high level player networks than are playing-time related penalties, protecting their ability to continue contributing to a franchise’s game performance. I also predict violent crimes will more frequently include playing-time related penalties, with League Commissioner approval more consistently mandated. I anticipate the team sub-organization to most frequently levy punishments for player criminality. This research introduces a more comprehensive examination of NHL player criminality, extending beyond existing approaches of case-based analysis. Importantly, this allows for future comparison to leagues such as the National Football League and its handling of player criminality over time. More broadly, I clarify the consistency with which deterrence measures are employed by the organization, contributing to the body of literature analyzing private organizations and their governing power over employees.

In recent years, the Evangelical church in the United States has begun to see a decline in attendance which mirrors earlier declines in more mainstream denominations. Some who leave Evangelical congregations go on to host podcasts, write books, and speak at conferences about their experiences inside Evangelicalism and their journeys out of it. As they do so, these ‘Exvangelicals’ create meaning around their experiences, form new identities and communities around their newfound disbelief, and create spaces for, and inform the journeys of, other Evangelicals who are questioning their participation in the American Evangelical church. The intensity and frequency of the creation of new identities and communities around a former religious affiliation seems to be unique to Evangelicalism. This paper examines how and why these individuals come to leave Evangelicalism and create identities, media, and physical and psychological spaces oriented around that choice and considers whether the unique culture of American Evangelicalism may contribute to the likelihood of forming these communities. I conduct a qualitative analysis of 30 publicly available interviews of individuals who have left American Evangelicalism and now participate in public discussions of their exits. I expect to find common narratives around the strictness of family-of-origin religious experiences, feelings of betrayal or disillusionment, and similarly held political and social values among interviewees, as well as cultural and personality traits which may predict which individuals choose to become outspoken in the movement. It provides a better understanding of the newly formed and growing group of self-identified ‘Exvangelicals,’ their experiences, and the process of group identity they have undertaken, as well as describing the ways they connect and interact, and how they replicate aspects of the church they have left behind.

Existing studies show the importance of migration in ensuring both the livelihood and safety of migrants around the world, and that factors such as ethnicity and caste have a significant impact on an individual’s ability to migrate. My study aims to further explore the association between caste/ethnicity and the order of starting, intermediate, and most recent destinations a migrant passes through. To do so, I perform a sequence analysis on places traveled to by participants of the Chitwan Valley Family Study (CVFS) dataset. CVFS focuses on the Chitwan Valley region in Nepal, a country that hosts a variety of ethnic groups and has a wide range of destinations that its inhabitants migrate towards, with a much higher rate of human migration than other countries of similar population size or GDP. Using the dataset, I also search for common factors between caste groups, such as the kinds of intermediate locations they may travel to before heading to a more permanent destination, and how long certain castes take to migrate either due to legal issues, their own caution or the resources available to them. I expect people of lower caste to face more difficulty in migrating due to how caste profoundly influences social life in the region. This transforms how we view the impact of caste or ethnicity on migration, not just isolated to the Chitwan region but across any region involving the migration of multiple ethnic groups. It is also relevant to note that sequence analysis has little pre-existing use in migration sociology, especially when focusing on migration across multiple continents. Therefore, this study presents a new method of studying migration along with a better understanding of what goes on between the assumed starting and most recent point of a migrant’s journey.

This paper examines several ancient sources for instances in which figures exhibit disconnection, aversion, or repulsion towards the erotic sphere that is so valued in societies. The paper will mainly focus on two myths: those of Narcissus (as illustrated in Ovid’s Metamorphoses) and Hippolytus (as shown in Euripides’s Hippolytus, Seneca’s Phaedra, Ovid’s Metamorphoses, and Ovid’s Heroides). In this essay, I argue that these figures can be read as asexual and that their untimely ends were brought about by their refusal to conform to the societal expectations placed on men–often characterized by vigorous sexual appetite, which would eventually lead to a man passing down his bloodline and fulfilling his duty to his oikos. Since myth often reflects real life, and works as a tool people can use to think about themselves, it can be argued that the inclusion of asexual-coded figures supports the contention that the asexual identity has always existed, even if the words to describe it are a more recent development.Therefore, the Narcissus and Hippolytus episodes are valuable pieces of evidence for both ancient and modern discourses of sexuality stepping outside the bounds of allonormativity. From a methodical standpoint, I first analyze the ancient texts, including secondary sources on them, then look at studies and papers on modern-day asexuality and examine the myths through this lens.

Affordable housing has recently emerged as a central focus of many policymakers’ agendas, especially in big cities, and they have implemented a variety of reforms to address this. In Seattle, the Mandatory Housing Affordability (MHA) program was rolled out in 2017, and then implemented citywide in 2019. The MHA created zones within the city, with new developments in MHA zones required to either build a certain number of rent-restricted affordable housing units for low-income people, or pay into a fund for affordable housing. Studies have shown mixed impacts of programs like the MHA on the housing supply; research about the impacts of such programs on housing and especially rent prices, however, is limited. Additionally, there is evidence that the effects of such policies are heterogeneous, varying by location and other factors. In this paper, I test for changes in rent and housing prices in Seattle overall using a difference-in-differences analysis. I also test for spatial variation within Seattle in these rent and housing prices by comparing MHA zones with non-MHA zones. The implications of these results will be important in addressing the next steps in housing policy. Since the MHA was a novel program in the inclusionary housing policy space, analyzing its effectiveness is crucial to determining how future policies should be shaped.

Implicit bias, rooted in unconscious attitudes, fuels discrimination based on race, gender, or sexual orientation, disproportionately impacting marginalized groups. Despite the development of interventions addressing provider awareness of implicit bias, the advancement of clinical education through technology has been slow. In the UnBIASED research project, we investigated the usability of ConverSense, a personalized communication assessment tool to raise healthcare providers' awareness of bias in their communication with patients. This web-based tool measures social dimensions such as warmth, interactivity, engagement, and assertiveness from recorded patient-provider visits, and visualizes these patterns through graphs and embedded clips. In this study, we (PT, NE) examined whether ConverSense meets usability standards through heuristic evaluations conducted by design experts. Six healthcare technology experts participated in the evaluation of ConverSense using Nielsen's 10 usability heuristics. Experts documented usability issues for each heuristic and rated their severity on a scale from 0 (not a problem) to 4 (catastrophic problem). Through our analysis, we (PT, NE) identified three cross-cutting themes: 1) Poor design, where experts noted the absence of undo or delete buttons, making navigation challenging, and the distracting color scheme on graphs; 2) Data visualization issues, with experts expressing difficulty interpreting charts and uncertainty about what is considered ideal or good communication. One expert said “It's unclear what is considered ideal/good…for each gauge chart, high interactivity, engagement, and warmth I would assume are ideal/good. But how the charts are displayed in the system I cannot know for certain”; 3) Ambiguity in information presentation, where experts sought more definitions for measured social dimensions and recommended training links to help them identify personal actions they can take to improve their communication. This study underscores the value of incorporating expert feedback and addressing usability issues to improve tools like ConverSense to address implicit bias and promote equitable patient-provider interactions.

In the United States, education has historically been utilized as a vehicle of cultural assimilation. What that looks like, and its goals, has varied across time and region. Initially starting as the dominant white, anglo-saxon, protestant culture violently and brutally oppressing and subsuming the cultures of all others (including Black Americans, Indigenous peoples, and immigrants of a variety of backgrounds), and in current times moving towards a co-existence state in which multiculturalism and cultural pluralism is championed. It has yet to be determined as to whether the education system has caught up to societal shifts and what it means in terms of racially and culturally non-dominant student experiences. The primary question is how has assimilation-based educational practices impacted culturally non-dominant students in K-12 public schools? I conducted a qualitative synthesis across federated search databases evaluating common themes of resiliency and success, risk factors and failures, and avenues of potential change. I included qualitative peer-reviewed studies from the year 2000 or later, that focused on K-12 public schools in the United States, and that focused on culturally non-dominant students. Preliminary findings include language barriers being insufficiently addressed, culturally insensitive and irrelevant curriculum, lack of acknowledgement on the cultural perception of the value of education within teaching pedagogues, black/white racial paradigms of diversity and achievement gap, insufficient modeling of Americanness that reflects all students and an accurate history of the United States, and gaps with data aggregation and the resulting need for further research. Implications and future directions include key data disaggregation in research and school-based data collection, a reconsideration of what constitutes “standard” curriculum, and teacher education focused on humanistic and equitable principles.

The use of space to represent abstract concepts is integral to how humans understand and process the world around them. This study is interested in the Western tendency to organize abstract information on a left-to-right basis. This includes the organization of temporal information (the mental timeline) as well as numerical information (the mental number line). Researchers have examined this phenomenon in conjunction with Western left-to-right writing systems and have hypothesized that the development of the mental timeline and number line is strengthened as writing and reading experiences develop. The present study seeks to examine the effects of early literacy skills on the development of the mental timeline and number line, as well as compare how children organize temporal and numerical information. Preschool-aged children (N=24, aged 3.5-4 years, data collection is ongoing) were presented with three cards that either featured events in a story (temporal activity) or different quantities (numerical activity). Participants were then asked to arrange these cards in order of first to last for the temporal activity and smallest to biggest for the numerical activity. Participants also participated in a Concepts of Print assessment to assess their early literacy skills. The number of LTR lines created was compared to Concepts of Print score. Temporal sequences were most commonly represented through left-to-right lines and numerical sequences were most commonly arranged non-linearly, with neither suggesting an association with early literacy. These findings raise further questions on the origins of the mental timeline and number line, as well as imply that literacy skills are not the sole contributor to this development. In examining how literacy skills may contribute to enhancing spatial cognitive abilities related to spatial representation prominent in STEM subjects, we can reevaluate the traditional dichotomy between the arts and STEM fields.

This global change study examines the multiple-stressor impacts of heat and plastic leachates on a symbiotic clonal cnidarian, the aggregating anemone, Anthopleura elegantissima. Marine heatwaves and ocean plastics are two forms of anthropogenic pollution that are increasing and predicted to rise in future ocean conditions. In Puget Sound, intertidal marine organisms are most at risk of exposure to these combined stressors. In summer, low tides at noon leave intertidal organisms in stagnant warming water or fully exposed to desiccation. Marine heatwaves, like the one that occurred in June 2021, caused water temperatures to spike along Puget Sound coasts. Concurrently, road run-off and sewage likely expose intertidal organisms to higher concentrations of plastic leachates. Leachates are derived from machine-washed polyester clothing microplastics, polyvinyl chloride sewage pipes, and non-source point pollution that is swept through watersheds toward the coasts. Plastic pollution in the form of leachates is understudied in coastal ecosystems, compared to thermal stress. Plastic-derived leachates are the complex cocktail of chemicals that leach from plastics into the environment and are considered pollutants of emerging concern. We do not fully understand the impacts they have on the physiology of marine organisms, and even fewer studies address their impacts in the context of marine heatwaves. We will test physiological and photophysiological responses of aggregating anemones to thermal stress and plastic leachates, separately and combined. We will develop respirometry and light response curves for each of the treatment conditions and a control. We hypothesize that the cnidarian host will show increased metabolic activity indicating stress under both types of pollution, and that photosynthetic efficiency in the algal symbiont will increase with leachate exposure. We hope to use the results of this study to better understand how anemones and other cnidarians like corals are affected by the threats of plastic pollution and global warming.

 The Circumgalactic Medium (CGM) is an extended structure surrounding galaxies, populated with hot diffuse gas and cold dense clouds of gas. The CGM acts as an intermediary between gas within galaxies (Interstellar Medium, or ISM) and gas between galaxies (Intergalactic Medium, or IGM). The mass of a given galaxy's ISM at the time of its formation is much too small to fuel star formation over galactic timescales, so gas must be accreted from the CGM to continue fueling star formation, and insights into the properties of this gas could improve our understanding of galaxy evolution.  During galaxy formation, gas heats up as it falls into dark matter halos from the cosmic web, forming the hot CGM. This gas then cools radiatively (through metal lines and Bremsstrahlung emission in the UV and X-Ray). The time required for this gas to cool and collapse depends on the properties observed from emission—hotter gas has more energy to radiate, high density or metal-rich gas cools more efficiently. If the cooling time is short, heating is required to prevent the gas from collapsing onto the galaxy; one such mechanism is the galactic winds driven by supermassive black holes and supernovae. The details of these heating processes are still uncertain and are a topic of active research in the field. For example, recent observations from the X-Ray telescope eROSITA have been shown to conflict with numerical hydrodynamic simulations of galaxy formation.  In our work we construct analytical models of the CGM with parameters such as density and temperature distribution and spatial extent. We calculate the gas mass and luminosity of the CGM to constrain the reasonable values of the model parameters. We will then use detailed spectral models of gas emission to compare our calculated luminosities to those observed and relate them to heating mechanisms.

Massive stars place powerful constraints on stellar evolution and exhibit a wide range of exotic evolutionary phases. They play a crucial role in regulating their environments, driving the chemical evolution of host galaxies, and establishing energy equilibrium through feedback processes. Stellar variability, notably, acts as a profound probe into the poorly-constrained physics of massive star evolution, illuminating intrinsic properties such as surface gravity. Drawing upon the collective insights from past literature on the dynamics of stellar rotation and surface gravity, this project delves into the correlation between variability metrics from the Zwicky Transient Facility (ZTF) and surface gravity measurements from the Apache Point Observatory Galactic Evolution Experiment (APOGEE) data. By merging these datasets, our aim is to use modern statistical methods to decode the relationship between observed variability and surface gravity in massive stars, shedding light on their rotational behaviors and structural changes over time. This endeavor not only seeks to deepen our understanding of stellar mechanisms but also to improve the precision in classifying stellar masses by utilizing variability as a key diagnostic tool. We endeavor to connect empirical observations with theoretical frameworks, paving the way for future advancements in our comprehension of stellar dynamics and their cosmic significance. Our results will place key constraints on the study of massive stars.

The largest scale structure in the universe creates a cosmic web. Nodes of the web are connected by mega-parsec scale filaments of warm gas, galaxies, and cold dark matter. Cosmic filaments are typically assumed to form and cool uniformly in many cosmological models. Recent works suggest that the internal structure, particularly with regard to the cooling mechanism, may be more complex. Gas clouds larger than a characteristic length appear to shatter as they cool below 106 K, fragmenting into smaller cloudlets. Other factors may also contribute to a more turbulent and irregular structure. The behavior of these filaments has implications for many of the issues at the forefront of astronomy. Cold dense clouds could create Lyman limit systems in the early universe, restricting the the distance that ionizing photons can travel. Further, the structure of these filaments may influence the mass of dark matter and constraints on cosmological models. To study these systems, I am simulating the formation of a single cosmic filament using a cosmological hydrodynamics code. Together with my mentor, I derived functions for the initial displacement and velocities of the particles from theory to create the desired collapse of matter into a filament. I then run the simulations with these initial conditions on a supercomputer, enabling simulations with tens of millions of particles. The simulations have mass resolutions that have not been previously achieved, allowing us to better understand the internal behavior. Preliminary findings indicate that there is indeed a complicated structure within the filaments.

The lithium isotopic composition of metasomatized rocks preserves a history of fluid movement that may be related to large subduction zone earthquakes. High pressure and temperature conditions within subduction zones cause the dehydration of hydrous minerals, and the resulting fluid can raise the pore pressure and trigger slip along the plate interface. The role of fluid movement in subduction zone processes can be better understood by constraining the duration of these events. These relatively fast and periodic fluid increases are recorded by chemical diffusion between the fluid and rock. Fluid containing lithium within a subduction zone can drive the diffusion of lithium into the surrounding rock. Lithium has two stable isotopes that diffuse at different rates, ⁶Li diffusing faster than ⁷Li, creating spatial heterogeneity in the isotopic composition of the reacted rock. Metamorphic rocks from the Western Alps have a reacted rind structure where fluid interaction occurred within an extinct subduction zone. The period of this interaction was examined using lithium isotopes. I prepared these samples for isotopic analysis, first weighing rock powders, then digesting them in acid, and finally separating the lithium with cation exchange columns. Lithium isotopes were measured on a multi-collector inductively coupled mass spectrometer. The spatial distribution of lithium isotopes along the profile from the reaction rind to the unreacted core, together with a thermodynamic model of Li diffusion through the rock, constrain the duration of the fluid interaction and provide insight into the role of fluid in catalyzing slip along the plate interface. I expect the duration of this fluid contact to be short, consistent with pulsed fluid movement within subduction zones. Modern subduction zones, including the Cascadia Subduction Zone that underlies Seattle, pose seismic hazards that can be better understood by examining the relationship between fluid movement and slip in extinct subduction zones.

Zooplankton are important primary consumers in the marine food web and lead an important role in carbon cycling in the open ocean. Understanding what influences zooplankton community composition can help us understand the impacts of climate change on this delicate relationship. Current research shows that zooplankton communities change based on currents and chemical cues, but there is a lack of data about the community structure of zooplankton in the Western Equatorial Pacific, specifically between 5°S and 5°N along 167°W. Data was collected from 28 December 2023, through 10 January 2024, on the R/V Thomas G. Thompson near American Samoa. A closing zooplankton net with 200μm mesh was used for net tows from 200m to surface at stations between 5°S and 5°N along the 167°W longitudinal line. Net tows were processed by counting and identifying zooplankton groups from subsamples and data was converted to abundance utilizing standard equations in RStudio. Zooplankton abundance increased from 5°S to the equator and decreased after the equator to 5°N. Species diversity (Shannon-Weiner) was lowest at the equator and highest at 5°N and 1°S. Calanoid copepods had the highest abundance over all sites, and north of the equator, calanoid copepods and gelatinous zooplankton (larvaceans) dominated most of the species composition. There were no significant relationships between species community composition and temperature, salinity, nutrients, or currents. With many processes occurring with zooplankton in the open ocean, it may be that multiple variables are impacting the resulting diversity and abundance relationships. Additionally, data was collected during the 2023 – 2024 Strong El Niño, which could have impacted the abundances and species presence due to higher water temperature and stronger currents. Monitoring zooplankton composition over time is vital for monitoring the health of our oceans as it has implications for global fisheries and carbon cycling.

Eelgrass (Zostera marina) is a marine flowering plant that provides important ecological, environmental, and economic services in the Puget Sound. Z. marina reproduces both asexually and sexually; however, the cellular regulation behind each mode of reproduction is poorly understood. Establishing Z. marina’s regulation behind sexual reproduction will improve predictions of Z. marina’s resilience to anthropogenic pressures. FLOWERING LOCUS (FT) is a highly conserved gene that promotes floral development in flowering plants and is well-characterized in the model organism A. thaliana. As a marine flowering plant, we hypothesized that a homologous, functionall FT exists within Z. marina. We identified 13 candidate Z. marina FT homologs (ZmFT), and from analysis of relative flowering time, 5 homologs of interest emerged: 2 that promote flowering, 2 that inhibit flowering, and 1 that has striking sequence conservation to A. thaliana FT, yet has no observable impact on flowering. To determine how intracellular distinctions alter the function of these respective Z. marina FT homologs, I am currently investigating putative protein-protein interactions of the ZmFT homologs with components of the A. thaliana floral activating complex, such floral promoting transcription factors, and scaffold proteins . I can characterize these protein-protein interactions using Yeast Two-Hybrid assays (Y2H) and BiFC fluorescent microscopy In which protein-protein interactions between ZmFT and A.thaliana floral complex proteins yields either yeast strains with conferred prototrophy, in Y2H, or a fluorescent signal, in BiFC. From my Y2H and BiFC work, I will characterize interactions of the ZmFT homolog proteins, with known components of the A. thaliana flowering pathway. The results of this study will help define how Z. marina regulates flowering onset and sexual reproduction. 

Ocean acidification (OA), the lowering of the ocean’s pH due to human-driven increases in atmospheric CO2, threatens many coastal communities and industries, and is thus linked to high environmental, economic, and societal losses. Yet, OA and its local context remains under-discussed by educators, industry workers, and community members. To address this shortcoming, NOAA’s Ocean Acidification Program, the International Alliance to Combat Ocean Acidification, and the Aquarium Conservation Partnership collaboratively created the “Exploring Our Changing Ocean: Impacts and Response to Ocean Acidification in the U.S.” StoryMap collection. The six StoryMaps, which I researched for and storyboarded, support localized OA education, outreach, and calls to action by showcasing regional OA trends, impacts, and community responses taking place across each of NOAA’s six Coastal Acidification Network regions. The StoryMaps intend to 1) Increase understanding about impacts of carbon emissions on our oceans and contribute to aquariums' place-based storytelling efforts on addressing climate change in their communities, 2) Reach new audiences, specifically members of the public visiting U.S. aquariums and marine education centers, although traditional audiences, such as academics, government leaders, and seafood growers can engage as well, and 3) Accelerate calls to action by showcasing detailed personal calls to action which help move OA activities beyond science. Currently, I am working with the United Nations Foundation on effectively distributing the StoryMaps to partner aquariums and implementing the content into larger climate change narratives and outreach. The StoryMaps can be shared as interactive displays, virtual-learning materials, or hard-copy outreach materials in participating aquariums, as well as through networks like educators, non-profits, conservation organizations, or international climate leadership fora. Through regionalized storytelling, this project will increase awareness of local OA trends and responses, facilitating enhanced awareness and action in at-risk communities.

Although oft-cited, the effects of campaign finance are neither well-understood nor well-evidenced; in particular, the supposed vote-buying motive --- contributions given with the expectation the recipient legislator will change their vote on specific bills --- lacks systematic evidence, and literature is mixed both in findings and in methodological quality. Nevertheless, since 2000 (before which much of the literature of “vote-buying” developed), gross campaign expenditures have exponentially increased; prima facie, this suggests that contributors expect returns to their campaign contributions and that (if their expectations are not systematically irrational) campaign finance does affect political outcomes. Building on the public choice theory of regulation, I theorize agents pay the cost of campaign contribution to produce political pressure on a recipient politician, who must then trade-off between the support of competing interest groups. Thus, it is expected that increased campaign finance contributions from interest groups that favor a bill’s passage are associated with a ceteris paribus increased likelihood of the recipient politician supporting the bill. Moreover, because interest groups are heterogeneous in their ability to generate pressure, the estimated treatment effect should systematically vary by contributor type. Previous literature has generally attempted to find statistical associations between contributions from a select few contributing groups and votes on a handful of selected bills (often selected because they are ex ante expected to give significant results), while also failing to incorporate adequate regression controls. Hence, this paper attempts to reconcile the literature’s differing results by using logistic regression to examine a large dataset of US House members’ legislative votes from 1980-2022 and concurrent campaign finance data. This paper's primary contribution is to extend current methods into a large-n analysis, the statistical strengths and methodological limitations of which are informative for future research into interactive mechanisms in economic theories of regulation.

The most prominent international organizations have emphasized their commitment in aiding the global transition to a carbon neutral world, which is estimated to cost hundreds of trillions of dollars. Despite these global efforts to mitigate climate change, countries, even when faced with similar levels of threat to climate disasters, prioritize the immediate issue differently. This study explores the relationship between economic security and state climate initiatives as a way to explain countries' varying commitment to climate change mitigation. While many scholars focus on the ability democracies have to combat climate change, I build upon and synthesize theories of economic opportunity, welfare programs, and class solidarity to hypothesize that both individual and state economic security impact states' prioritization of climate initiatives. I argue that economic security is a likely influence on the magnitude of effort which states put into climate initiatives, as it reflects both individual ability to safely fulfill essential needs and state ability to adapt capital. To test my hypothesis, I run a multivariate regression analysis to examine whether key indicators of economic security correlate to state carbon emissions per capita, controlling for regime type, institutional trust, political unrest, and median age of the population. I expect to find a negative relationship between economic security and state carbon emissions per capita. These findings would suggest that improving economic security is essential for the development and success of future climate initiatives. Understanding the underlying roadblocks of long term climate mitigation is crucial in guiding climate policy and optimizing climate aid.

The goal of The Treaty on the Non-Proliferation of Nuclear Weapons (NPT) is to prevent nuclear weapons proliferation, promote peaceful nuclear energy cooperation, and pursue global disarmament. Signatory countries meet every five years for the NPT Review Conference to assess the implementation of the treaty and strengthen its measures. The Preparatory Committees of the NPT are held three years prior to the Review Conference so signatories can discuss relevant topics. Studies on the NPTs effectiveness focus on non-proliferation compliance and overlook cooperation for the peaceful uses of nuclear energy. In this study I map how the distribution of topics discussed in the Preparatory Committees change over time, how levels of state participation change over time, and if change in state participation in the Preparatory Committee correlates to increased transfers of nuclear reactor technologies. I employ a three-part approach in this study. First, I use topic modeling for categorizing the Preparatory Committee documents in seven categories. I assign the following labels to the categories by reading the most relevant documents in each one: Safeguards and Verifications, Non-Proliferation Policy and Disarmament, Conflict Resolution, Peaceful Nuclear Energy Cooperation, Implementation Assessments, New Initiative Proposals, and Logistics. Second, I generate “relevancy scores” between the names of the countries and the Preparatory Committee documents by using an embeddings model to quantify country’s participation. I then compare participation levels to nuclear technology transfers. Collectively, this research has shown that discourse on peaceful nuclear energy cooperation in the NPT forums often centers around the advancement of safeguards against proliferation. I also find that changes in participation do not correlate with changes in numbers of nuclear technology transfers. Thus, the NPT forums are platforms for dialogue on strengthening security against nuclear threats unrelated to tangible growth of peaceful nuclear technology usage.

Timely and accurate rental measures are needed by market participants to assess the state of the housing market and shelter costs, which are primarily based on rent and rent-derived measures, representing about 40% of the Consumer Price Index (CPI). In addition, subnational measures are needed for a range of business operations and policy implementations. Property transactions are recorded and often publicly available. This has allowed the development of well established house price indices that are available at high temporal and spatial resolutions. This is not the case with rental contracts. Rental measures based on survey data are key to the CPI and other private and public sources have also been used to create rent measures in recent years. In our research, we review existing rent measures showing substantial variations in estimates of rent levels and trends across measures. We then focus on the strengths and limits of listing based measures. Finally we use listing data collected from several online platforms over the 2017-2023 period to compare them to existing measures. Our findings do show the potential and limits for developing repeat rent measures based on asking rents and the adjustments needed to make such data usable.

Economic sanctions as a means of pressuring nations to improve human rights protections or end human rights abuses have become an increasingly common practice in recent years. Although the efficacy of sanctions for humanitarian ends remains under heavy scrutiny, the United States Department of State continues to both implement new sanctions and enforce existing policies of this kind. However, there is a notable discrepancy between regimes accused of human rights abuses by non-governmental organizations and those receiving these types of sanctions. This research aims to unveil potential factors that may explain this gap. I theorize that while investment and trade may protect a country from economic sanctions in an effort to keep certain markets open, past or present adherence to communist ideology increases the likelihood of receiving sanctions on the grounds that communism remains a perceived threat. To test my theories, I identify a set of countries that are currently verified by third party organizations as human rights violators. At this point, I conduct multivariate regression analysis to observe the relationship of both economic interest and conflicting ideology and the presence of sanctions citing human rights violations. While I expect to find that while both factors contribute to the presence of sanctions as outlined, I also theorize that economic interest will have a greater influence. This study serves to identify specific influences on sanctions that will enrich future discourse on their implementation.

Tuberculosis remains a global public health threat due to the rising number of multi- and extensively drug resistant strains of the causative pathogen Mycobacterium tuberculosis. Development of novel drugs and an understanding of their resistance mechanisms is urgently needed. Aminothiazoles (AmT) are potent molecules with killing activity against M. tuberculosis; these compounds act as copper ionophores and target a key enzyme (enolase) by displacing its Mg²⁺ co-factor, a substance required for its activity, with Cu²⁺ imported by the compounds. Spontaneous mutations in an essential protein export system (the Esx 3 Type VII secretion system) confers resistance to AmTs. My research focuses on understanding how mutations in the secretion system cause AmT resistance. We hypothesize that copper imported by AmTs could disrupt other metallo-proteins including EccA3, a key ATPase of the of the Esx-3 secretion system that hydrolyzes ATP into ADP and inorganic phosphate, and that resistance mutations (e.g. E237K) reduce Mg²⁺ co-factor displacement by Cu²⁺. To test this hypothesis, I expressed wild-type (WT) EccA3 and mutant EccA3 [E237K] proteins in Escherichia coli BL21(DE3) expression strain and purified the proteins via Ni-NTA His-tag chromatography. Subsequently, I measured the activity of the purified EccA3 (WT) and EccA3 [E237K] proteins via an ATPase assay based on colorimetric detection of free inorganic phosphate released by ATP hydrolysis. I aim to understand whether copper inhibits EccA3 activity through this assay, anticipating that copper reduces EccA3 (WT) ATPase activity while EccA3 [E237K] ATPase activity is unaffected. Thus, my work will provide an avenue for understanding AmT resistance in M. tuberculosis.

Major Histocompatibility Class I (MHC) Molecules serve as a window into the cell, whereby T-cells can use their T-cell receptor to recognize foreign peptides presented on the MHC of a cell and induce apoptosis. Unfortunately, for diseases such as cancer, cancerous mutations may only be a single residue different from the native protein, resulting in similar mutant and wild-type peptide-MHCs. This small difference often results in negative selection of T-cell receptors that recognize mutant peptide-MHCs, leading to an absence of T-cells that can target cancer cells. This issue motivated us to use de novo protein design to generate binders with a high level of specificity between mutant and native peptide-MHCs. Using deep learning based protein design methods such as RFdiffusion and ProteinMPNN, we have generated promising in silico designs against a range of target peptides and MHC alleles. After selecting our top designs, we tested them using yeast surface display against our target MHC molecules with peptides loaded from KRas, PIK3CA, MAGE, and TP53 mutants. We observed binding events to all targets; some designs also had specificity to their respective mutant peptide-MHC over the wild-type peptide-MHC. After further work we have shown that the same design scaffold can bind to multiple peptide-MHC targets after slight redesign, similar to native T-cell receptors, holding promise that we could easily and quickly repurpose these scaffolds for new targets. Following these results we will incorporate our binders into T-cells as chimeric receptors and test for the ability of our binders to activate T-cell signaling and cell killing. This method of targeting peptide-MHC molecules is promising as a novel and rapid way to target cancer.

Researchers consider autoimmune disorders and immunodeficiencies to be two completely unrelated conditions. However, it has been shown that patients can develop both diseases in spite of opposing etiologies. This has led researchers to examine the commonalities between autoimmunity and immunodeficiency in terms of pathogenesis, clinical manifestations, treatments and, more recently, genetics. With genome sequencing becoming more prevalent and easier to access, researchers have been able to look at disorders of the immune system from the genomic point of view, identifying genetic mutations of key genes. One such gene is cytotoxic T-lymphocyte associated protein 4 (CTLA-4) because of its known immunoregulatory role in terminating immune responses. CTLA-4 directly regulates the development of T-cells by activating T regulatory cells, but it can also affect the development of B-cells in downstream effects. In this literature review of a multitude of sources, we show that heterozygous mutations of CTLA-4 can lead to patients developing both autoimmune and immunodeficiency manifestations such as hypogammaglobulinemia, autoimmune cytopenia, lymphoproliferation, autoimmune enteropathy, etc. Such mutations point toward commonalities in both autoimmunity and immunodeficiency, leading researchers to believe that these two diseases are more alike than different. With this newfound discovery, treatments can be developed to target the CTLA-4 gene in hopes of treating both autoimmunity and immunodeficiency in patients.

People with diabetes have an increased risk of developing cardiovascular disease (CVD). Hyperglycemia is the hallmark of diabetes, but diabetic dyslipidemia with increased circulating lipid levels is also present, which is believed to contribute to the augmented CVD seen in diabetes. The Cluster of Differentiation 36 (Cd36) receptor mediates fatty acid and lipoprotein uptake in macrophages. Lipid-loaded macrophages are a key feature of atherosclerosis, the underlying CVD pathology. Preliminary data suggest that monocytes (macrophage precursors) are lipid-loaded via increased Cd36. However, it is unclear what drives the increased Cd36 expression in diabetes. Reduction in blood glucose, but not lipid levels, in diabetic mice reduced monocyte cell surface Cd36 expression. Based on these preliminary data, I hypothesize that hyperglycemia induces increased Cd36 mRna expression in monocytes in diabetes. To address whether glucose or lipids increase monocyte Cd36 mRna expression, I will isolate monocytes from the bone marrow of non-diabetic mice and stimulate them ex vivo. To address if hyperglycemia alters Cd36 expression, I will stimulate monocytes with 4 glucose conditions: 5.6 mM, 15 mM, 30.6 mM D-glucose, and an osmotic control of 5.6 mM D-glucose and 25 mM L-glucose. The 5.6 mM represents non-diabetic blood glucose conditions. To address if dyslipidemia alters monocyte Cd36 expression, I will use the same 4 glucose conditions in conjunction with 50 µg/mL of VLDL, a triglyceride-rich lipoprotein that is elevated in diabetic dyslipidemia. Following a 24-hour stimulation, I will isolate monocyte RNA and use qPCR to determine the amount of Cd36 mRna. If elevated glucose induces an increase in Cd36 expression, this suggests that hyperglycemia stimulates increased Cd36 expression in monocytes in diabetes. However, if the presence of VLDL is required to observe an increase in Cd36 mRna, this indicates that dyslipidemia is needed for increased monocyte Cd36 mRna expression. Results from this study will help us understand the relationship between lipids and hyperglycemia in the context of diabetes-induced monocyte lipid loading.

Over 500 million adults worldwide have type II diabetes (T2D), which has caused 6.7 million deaths in 2021 alone. Yet, accessible and sustainable treatments remain limited and many address the symptoms of the disease rather than the root causes, one of which is insulin sensitivity. Recent research shows that therapies targeting glucose-regulating mechanisms in the brain that do not directly affect insulin secretion can enhance diabetes remissions in T2D rodent models. Such treatments may revolutionize diabetes treatment and public health. The Scarlett Lab’s previous studies show that exogenous intracerebroventricular administration of hypothalamic fibroblast growth factor 1 (FGF1) resulted in sustained hyperglycemic remission and increased insulin sensitivity in the mediobasal hypothalamus (MBH), suggesting that FGF1 signaling in the brain impacts glucose metabolism. FGF1 is also endogenously expressed in the MBH, but the functional role of endogenous FGF1 is still unknown. Thus, I hypothesize that endogenous FGF1 signaling in the MBH regulates glucose and energy homeostasis. To test this, I utilize adenosine-associated virus as a vector for gene editing therapy to knock out the Fgf1 gene from the MBH in adult male Fgf1-floxed mice. I then monitor food intake, body weight, and blood glucose levels; perform insulin and glucose tolerance tests; and measure insulin and glucagon levels using enzyme-linked immunosorbent assay. Finally, I use immunohistochemical analysis at the end of the experiment to validate Fgf1 knockout. Based on pilot studies, I expect to observe that MBH Fgf1 knockout significantly increases body weight and impairs glucose tolerance and insulin sensitivity. Follow-up work can elaborate on investigating the effect of a high-fat diet on FGF1 signaling and function. This project illuminates the functional role of endogenous FGF1 signaling in regulating glucose and energy homeostasis and contributes to the development of novel therapeutic strategies to treat diabetes and obesity.

Over 38 million Americans have diabetes, with 90% of diabetic Americans having Type 2 Diabetes. Diabetes causes an increased risk of complications, including diabetic kidney disease (DKD), a disease that affects kidney filtration. This occurs in the glomerulus, a specialized capillary network of single-layered endothelial cells on one side and podocytes on the other, and their extracellular matrix (ECM). Injury, reduced function, or changes in the ECM of either of these cells cause abnormal filtration and kidney disease. Preliminary data from our group suggests that ECM remodeling driven by endothelial cells is a key feature in DKD in both our mouse model and in humans. Additional preliminary data from our lab indicates that two metalloproteases involved in ECM remodeling, ADAMTS6 and ADAMTS9, are increased in endothelial cells in diabetes via elevated lipids. My hypothesis is that very-low-density lipoprotein (VLDL), a lipid often increased in diabetes, induces increased endothelial cell ADAMTS6 and ADAMTS9 expression, contributing to remodeling the ECM and altering the filtration capacity of the glomeruli. To investigate this, isolated endothelial cells from non-diabetic mice were stimulated with varying VLDL concentrations, alone or with elevated glucose, to simulate diabetes. Following the stimulation, I isolated the RNA from these cells and measured the mRNA expression of Adamts9 and Adamts6 using real-time PCR. To test if endothelial cells isolated from mice with diabetes would respond differently, similar experiments are being carried out in cells isolated from diabetic mice. Western blots are used to verify the altered protein expression I observe. I am utilizing already acquired kidney sections from mice with diabetes with different lipid levels and determining the glomerular endothelial cell ADAMTS9 and ADAMTS6 expression by immunohistochemistry. Results from these experiments will help us understand the mechanisms through which endothelial cells respond to diabetes and thus contribute to DKD.

Biofilm is a community of bacteria enclosed in an extracellular polymeric substance (EPS) attached to a surface. Inside the biofilm, bacteria can collaborate to increase their survival. The EPS also protects bacteria from drug penetration, leading to increased antibiotic resistance. Therefore, biofilm formation is often linked with chronic bacterial infections. Pseudomonas aeruginosa is an opportunistic pathogen that often causes chronic lung infections in cystic fibrosis patients. It is also a common model for studying biofilm formation. The initial step for biofilm formation is bacteria attaching to and sensing a surface. Upon surface contact, P. aeruginosa may produce cyclic adenosine monophosphate (cAMP), which is a universal second messenger that regulates cellular functions in both eukaryotes and prokaryotes. In P. aeruginosa, cAMP is synthesized by two adenylate cyclases, CyaA & CyaB, and degraded by a cAMP phosphodiesterase, CpdA. cAMP is a key regulator for P. aeruginosa virulence by upregulating the production of the type III secretion system, the type II secretion system, and the type IV pili. However, recent observations in our lab suggest that cAMP may also contribute to the homeostasis of the cell envelope. To investigate this phenomenon, I used microscopy to characterize the cell morphology of strains with different cAMP levels and found that increased cAMP levels lead to longer cells. I also found that high cAMP strains are more sensitive to ꞵ-lactam antibiotics specifically, while low cAMP strains become more resistant. Ongoing work includes characterizing the genetic factors that connect cAMP and ꞵ-lactam sensitivity, as well as using microscopy to determine changes in cell envelop induced by cAMP. Overall, this work reveals an important role of cAMP in bacterial physiology and provides insight into the complex relationship between virulence and antimicrobial resistance.

The retina is a unique neuronal structure in the eye that facilitates vision. Many diseases cause the death of retinal cells and this can lead to blindness. Frogs and fish have retinal stem cells that can repair the retina after retinal cell death; these stem cells are concentrated in a region called the ciliary marginal zone (CMZ). It was thought that humans lack these cells; however, we have discovered a region of the retina that has some features of the CMZ. We call this the Late Proliferative Zone (LPZ). One of my research goals was to determine whether the LPZ in humans also contains retinal stem cells that could be harnessed to repair the injured retinae. To start, I measured the area of small cuttings of fetal retinal tissue grown in culture, called retinospheres (RSs), over time and identified a window from 250-325 days gestation in which the LPZ of the human retina continues to grow after the rest of the retina is quiescent. This result shows that the cells of the LPZ can make new retinal cells much later than we thought, supporting the idea that these are retinal stem cells. My second goal was to find factors that can stimulate the growth of these cells. I tested several factors known to be important for the stem cells in frogs and fish. I found the effects of these factors on the types of neurons made by the LPZ. In sum, investigating different ways to manipulate the LPZ provides the field with insight into what is needed to regenerate cell types lost in blinding diseases.

Pseudomonas aeruginosa is a ubiquitous environmental bacterium and an opportunistic pathogen of wounds, cornea, and the Cystic Fibrosis lung. P. aeruginosa is also a model organism for the study of bacterial biofilm formation. Biofilms are multicellular communities that form from bacterial growth concomitant with the production of extracellular polymeric substances (EPS). EPS includes polymers such as polysaccharides, DNA, and proteins; these polymers provide structure and protection to the biofilm cells. Proteomics experiments by the Parsek Lab and others have demonstrated that a notable component of the biofilm matrix are the secreted proteases. Secreted proteases have defined roles in virulence and nutrient acquisition, but their role in the biofilm matrix of P. aeruginosa has not been explored. I hypothesize that these secreted proteases recycle nutrients, remove cell waste, and protect cells from host immunity. To test my hypothesis, I generated a mutant strain of P. aeruginosa that lacks the six major secreted proteases. While we see that loss of the proteases does not impact planktonic growth, preliminary data suggests that loss of proteolytic activity results in moderately increased biofilm formation. Using a general proteolysis assay relying on casein hydrolysis, I have determined the relative contribution of each of the six proteases to the total proteolytic capacity of P. aeruginosa in planktonic growth. I will further test the impact of the proteases on biofilm growth in different growth environments, including under flow conditions and in artificial sputum medium. I will also assess which proteases contribute the most to proteolysis during biofilm growth. My work fits into a growing body of literature that suggests that the biofilm matrix is not an inert scaffold, but is instead a dynamic and active network.

Sambucus newtoni is a type of elderberry (Adoxaceae, Angiospermae) from the late Eocene epoch. It has not received much research attention since its discovery in the early 20th century. Researchers identify it based on its leaflets (parts of compound leaves), with other traits inferred from more modern Sambucus species. As a result, its flower morphology has remained unknown. Here, I contribute to our understanding of S. newtoni by describing an excellently preserved compression fossil of a floret (small flower) from a S. newtoni cluster of florets, a so-called cyme. The fossil was collected from lake deposits in the Florissant Formation, Colorado. I described and analyzed the specimen with a microscope and observed extremely well-preserved petals and anthers (male flower parts). The flower sits next to a fossilized fly specimen on the adjacent sedimentary rock layer. The close similarities of the flower of S. newtoni to that of the extant species S. javanica (Chinese elder) may help confirm recent common ancestry as originally suggested through comparisons of the leaflets. Further, improved understanding of S. newtoni through study of the new specimen may open the door to greater understanding of Sambucus evolutionary history as a whole.

In place of an adaptive immune system, pattern recognition receptors (PRRs) that perceive host-derived herbivore-associated molecular patterns (HAMPs) induce immune signaling cascades in plants. Inceptins are a class of proteolytic peptides that originate from chloroplastic ATP synthase that are produced in the oral secretions of all studied species of caterpillars in the order Lepidoptera. Upon introduction to the plant, inceptin-11 (+ICDINGVCVDA−) binds to Inceptin Receptor (INR), triggering a signaling cascade to up-regulate defensive hormones, specialized metabolite toxins, and other direct resistance mechanisms against herbivores. However, the full ecological extent of the effects of the INR signaling cascade is poorly understood. Here, we show that inceptin-11 signaling in common bean (Phaseolus vulgaris) can mediate the attraction of predatory wasps (Polybia sp., Polistes sp.) as an added, indirect line of defense against herbivore threats. Using a near-isogenic line of P. vulgaris containing a 103 base pair deletion in the INR locus, we found that Polybia and Polistes preferentially forage on sibling lines with wild-type INR that respond to herbivore threats. Siblings with fully functioning INR produce nearly three-fold more (E)-4,8-Dimethyl-1,3,7-nonatriene (DMNT), an established volatile wasp attractant, than the deletion line. Our results demonstrate that INR can leverage cross-kingdom predator-prey relationships to aid in defense of the plant. Moreover, our near-isogenic line of P. vulgaris provides a genetic resource for studying the role of pattern-triggered immunity in indirect defenses.

In the face of a global crisis, the poaching of pangolins (Manis spp) has emerged as a dire threat, with Africa at the epicenter of this illicit trade; these enigmatic creatures, sought after for their scales and meat, face exploitation driven by illegal activities that endanger their populations and disrupt the balance of the African ecosystems. Studying the lineage distributions of pangolins could prove crucial for their protection as it provides valuable insights into their evolutionary history, genetic diversity, and ecological adaptations. This information may be essential for formulating effective conservation strategies, understanding their vulnerabilities to diseases, and identifying key habitats for preservation. We aim to address these gaps, by exploring whether pangolin (Manis temminckii) lineages from Mozambique can be genetically distinguished from those from Southern Africa and by investigating the region of origin for two scale samples seized by law enforcement in Singapore in 2019. We conducted genetic and phylogenetic species delineation by sequencing two mitochondrial genes, Cytochrome b oxidase (cytb) and the Control Region (D-loop), from pangolin skin tissues from 10 villages in 3 provinces in Mozambique to complement existing data from Southern Africa. We will then use RAxML (Randomized Axelerated Maximum Likelihood) for phylogenetic analysis and to estimate phylogenetic trees based on our collected data. This multi-locus approach ensures robust species delineation and provides essential support for species recognition. Our focus on mitochondrial DNA (mtDNA) in Mozambique aims to make a reference map of the distributions of genotypes throughout Southern Africa, and our research contributes valuable genetic insights, offering a nuanced perspective on the spatial dynamics of pangolin populations. Ultimately, these findings play a pivotal role in addressing the complex issue of illegal wildlife poaching in Africa, providing essential resources for conservation efforts and informed management strategies as well as contributing to the field of forensic science for counter wildlife trafficking.

Angiogenesis, or the formation of new blood vessels, is crucial for normal bodily function but is especially important in diseases that cause blood vessel breakdown such as diabetic vasculopathy. Angiogenesis is regulated by activation of the Tie2 receptors in endothelial cells, which have two main ligands: angiopoietin-1 (Ang1) and angiopoietin-2 (Ang2). Ang1 binding has been shown to stabilize blood vessels and inhibit vascular leakage, while Ang2 antagonizes these effects. We have previously shown that a computationally designed Tie2 super-agonist which presents eight copies of the Ang1 F-domain strongly activates Ang1-like signaling in human umbilical vascular endothelial cells (HUVECs). In this project, we hope to assess the Tie2 super-agonist’s ability to rescue diabetes induced blood vessel defects in a diabetic blood vessel organoid (BVO) model. To model diabetic conditions, a three-dimensional blood vessel organoid model has been cultured in a high glucose media along with inflammatory cytokines associated with the diabetic phenotype. Western blotting and immunofluorescence staining will be used to assess the relative quantities and localization of proteins involved in vascular stability and inflammations upon treatment with the Tie2 super-agonist. Vascular degeneration is a very harmful condition associated with many prevalent diseases including diabetes, so the Tie2 super-agonist could potentially be a new therapeutic drug candidate for treating blood vessel dysfunction in patients with these conditions in the future.

Climate change and global population growth are driving the need for more sustainable methods for growing crops used in agriculture and the production of biofuels. To address these challenges we are exploring the role of the plant microbiome in host plant tolerance to environmental stresses related to climate change. In particular, we are investigating whether endophytes, microorganisms that colonize the internal tissues of plants, make the host plants more tolerant to drought or low-nitrogen conditions. We are currently optimizing the process of DNA extractions of fruit and poplar trees which were inoculated with beneficial nitrogen-fixing bacteria, and grown in either water or nitrogen-limited conditions. We then purify high quality microbial DNA and use polymerase-chain reaction (PCR) to optimize strain-specific primers (SSP), which target specific DNA sequences in the genomes of our endophytes in the presence of competing DNA. This allows us to gain an understanding of where they colonize, and to demonstrate that the trees were successfully colonized by our endophytes to support growth and drought tolerance data collected from inoculated and uninoculated controls. By ensuring the SSPs only target our strains of interest, we differentiate our endophytes from other members of the plant microbiome. These primers are then used in Droplet Digital PCR (ddPCR) to quantify their relative abundance. Using this information we hope to demonstrate that beneficial endophytes can be used as a sustainable method for improving drought and low nitrogen tolerance in plants, both in agricultural and biofuel applications, reducing the consumption of nitrogen fertilizers and water for irrigation in these sectors.

Dilated cardiomyopathy (DCM) is a leading cause of heart failure around the world. Inherited mutations cause the left ventricle of the heart to enlarge, thinning the heart muscle wall and decreasing the overall function of the heart. In my research project, I will determine if disrupting fibroblast function by knocking out a key developmental signaling factor, p38, can improve, or even reverse, DCM disease characteristics. Specific Aim 1 will be to determine the effects of p38 knockout-induced fibroblast dysfunction on cardiomyocyte function and structural remodeling in late-stage DCM. The rationale is that myocytes in DCM have poor contraction and structurally remodel to longer, thinner morphologies, which occurs in our DCM mouse model around 4 months of age. I expect to see less of these characteristics with the p38 knockout. Specific Aim 2 will assess cardiac fibroblast proliferation and fibrosis in response to disabling cardiac fibroblast function late into the DCM disease process. The rationale is that studying and observing the dynamics of the fibroblast population is critical when understanding the effects of fibroblasts and the p38 knockout model on DCM. In previous studies, the Davis lab identified that cardiac fibroblasts maladaptively respond to inherited DCM mutations in cardiac myocytes, worsening the whole heart. I expect to see less fibroblast proliferation in the p38 model. P38 is essential for fibroblast signaling pathways and functionality, so by knocking it out I will be able to test if fibroblasts are a viable therapeutic target for patients with DCM.

Test anxiety is a prevalent issue among K-16 students, with approximately 20-40% reporting its adverse effects. This pilot study assessed the preliminary efficacy of a Short Mindfulness Activity to Reduce Test Stress (SMARTS) recording in mitigating test anxiety, indicated by heart rate (HR) physiological markers. Our research team’s undergraduate students assembled a student advisory panel to receive feedback on existing mindfulness recordings and narrow down the most effective traits of each one. From the feedback, we designed a 7-minute mindfulness recording to be administered to University of Washington Bothell Introductory Biology students before an exam. Using a randomized controlled trial design, 29 students were randomly assigned to the mindfulness recording group (n = 15) and to the control group (n = 14). The intervention group was exposed to the SMARTS recording before an exam while the control group studied in the hallway. We found that listening to the SMARTS recording reduced the increase in heart rate due to the exam by an average of 8 beats per minute. Additionally, 56% of participants expressed intentions to continue using the recording before future exams, suggesting its perceived efficacy in alleviating test anxiety. This research suggests promising implications for integrating mindfulness interventions into educational settings to support students in managing test anxiety. Further study is warranted to evaluate the intervention’s efficacy in a fully powered trial. In addition, we will explore heart rate variability to provide insight into the role of parasympathetic activity in stress management.

Sporadic inclusion body myositis (sIBM) is an acquired progressive inflammatory muscle disease. It is most commonly seen in individuals over 50 years old and affects more men than women. Symptom onset is generally gradual and characterized by progressive muscle weakness and atrophy. Weakness often starts in the quadriceps and finger flexors but can affect other muscles in the arms and legs as the disease progresses. Difficulty swallowing also can be present. The disease remains challenging to diagnose due to its non-uniform presentation. There is currently no cure or standard treatment for sIBM as it is unresponsive to corticosteroids and immunosuppressive drugs. In this study, we investigated the prevalence of and associations between different features considered in the diagnosis of sIBM. We reviewed the electronic medical records of adult patients diagnosed with sIBM using ICD-10 codes at the University of Washington Medical Center from 2003 to 2023. Data was collected including creatine kinase (CK) levels, presence of the anti-cytosolic 5′-nucleotidase 1A (NT5c1A) antibody, pulmonary function testing, presence of dysphagia, muscle strength testing, muscle biopsy findings, electromyography (EMG)/nerve conduction studies, and magnetic resonance imaging (MRI). Statistical analyses were performed to identify the presence of sIBM phenotypes and correlations between them. This study confirms the heterogenous presentation of sIBM and highlights the associated diagnostic challenges this presents. Understanding both typical and atypical presentations is key to preventing delayed diagnosis and misdiagnosis commonly seen in this patient population. Timely diagnosis allows for more tailored management of disease-related symptoms and can help to eliminate the unnecessary administration of ineffective medication and invasive testing. In addition, further characterization of sIBM phenotypes may lead to improvements in both current diagnostic criteria and considerations for clinical trial outcome measures.

With this project, researchers for the Harry Bridges Center for Labor Studies from Seattle, Tacoma, and Bothell campuses seek to gain a better understanding of the intricate relationship that exists between the UW undergraduate experience and employment, as well as inform students on existing resources and support services both on and off campus. To sufficiently measure unique student experiences, we have sought survey responses through canvassing, contacting departmental and Registered Student Organization (RSO) listervs, and visiting in-person classes. Over the course of the past 6 weeks, we have made 73 points of outreach at the Seattle campus, 62 at Bothell, and 44 at Tacoma, resulting in 430 survey responses as of February 7th, 2024. Our Qualtrics survey will run through the end of winter quarter. We will then also offer paid interviews for 15-20 students to supplement survey data through further examination of survey themes and trends. Modeling UCLA Labor Center’s Unseen Costs: The Experiences of Workers and Learners in Los Angeles County, we will then compile our survey results and interview stories into a cumulative research report about why UW students work while pursuing their undergraduate degrees, what fields they have worked in, and any associated workers’ rights violations they may have faced while in these positions. With this research, we aim to create a more supportive environment for our student workers, and to help educate our respective campuses to meet their student body needs.

Autism spectrum disorder (ASD) is defined as a complex developmental condition involving persistent challenges with social communication, restricted interests, and repetitive behavior. The development of autism spectrum disorder (ASD) has been associated with altered brain development in infants with high familial likelihood for developing the disorder. Some of the same subcortical regions associated with altered brain development in ASD are also associated with sleep problems, including the hippocampus. Healthy sleep is an integral contributor to brain development during childhood and there is increasing evidence that chronic sleep loss can lead to neuronal loss and impaired functioning. Additionally, the multi-site, NIH-funded Infant Brain Imaging Study (IBIS) has previously reported sleep problems are associated with hippocampal alterations in infants who developed ASD. However, the relationship between sleep problems and altered brain development in autistic school-age children is not well understood, even though clinically significant sleep problems affect 75-80% of this population. Based on prior findings from the IBIS network, this project will examine the relationship between hippocampal volume and sleep problems in 137 school-age children with high familial likelihood of ASD (HL-ASD), no diagnosis of ASD (HL- noASD) and low familial likelihood of developing ASD (LL). Our prior findings suggest there will be differences across the three school aged groups in hippocampal volume, sleep duration, and sleep quality. We also expect that hippocampal volume in the HL-ASD children is differentially affected by sleep quality and sleep duration compared to the HL-noASD and LL children. Understanding the connections between sleep patterns and autism will help steer future research endeavors examining a largely understudied but significant issue. Additionally, this project will provide new insights into the negative consequences associated with chronic sleep loss in autistic children, affecting both their daily functioning and their quality of life.

The T3 Watershed Experiment Upland Silviculture Study explores innovative approaches to managing forests in the Olympic Experimental State Forest (OESF) managed by Washington state DNR. This model study is an addendum to the T3 study exploring different treatments to enhance ecological heterogeneity through the implementation of variable-density plantings in the context of marigold growth. Variable-density planting increases heterogeneity by adding a variety of clumped areas across a unit which has the potential to enhance biodiversity and habitat, improve ecological resiliency, increase carbon sequestration, and optimize tree growth. We chose marigolds to act as a model specie for conifers due to its short life cycle, ease of use, and ability to growth tall and straight. Our team planted marigolds using three treatment groupings including small clump, large hex clump, and regular spacing (control)— implemented in six boxes within a controlled greenhouse environment. In Autumn 2024, volunteers and I collected comprehensive data on various growth metrics. Later, I measured the biomass of leaves, stems, reproductive shoots, and total biomass to determine biomass variations. I analyzed the effects of treatment on the stem straightness/sway, mortality, biomass of different tissues, and social class proportions. This research contributes to the broader understanding of how variable-density plantings influence vegetation dynamics, providing valuable insights for silviculture practices. The findings aim to inform future land management strategies by providing data that supports the hypothesis that variable density planting may provide numerous benefits without decreasing timber harvests.

The robotic exploration of Mars has found that the early atmosphere was similar to current day Earth, suggesting that life could have existed on Mars in its past. The former atmospheric conditions and potential ancient rivers and lakes are preserved in the sedimentary rocks found across the surface. Interpretation of the Martian sedimentary record in Gale Crater, a possible ancient lake, requires differentiating between a variety of processes that alter sediment chemistry. Our study will contribute to the reconstruction of source rock composition based on the sedimentary records in ancient river systems on Mars. Iceland is a useful analog to ancient Mars as it has a similar climate and geologic makeup as well as similar environmental features, such as glaciers, volcanoes, rivers, and lakes. Here we characterize the chemical composition of source rocks in a cold, basaltic Mars analog source-to-sink system in Iceland and compare them to adjacent sediments. If the source rocks are primarily composed of palagonite, a glassy product of hydrothermal alteration of volcanic glass that weathers easily, we hypothesize that palagonite is the dominant component of the sediments. We analyzed the samples using Micro X-ray Fluorescence (μXRF) to examine thin sections of rock for changes in the elemental composition, and X-ray Fluorescence (XRF) to measure the bulk chemical composition of rocks and sediments. We used thin section classification to quantify the percent proportion of altered rock (palagonite). In source rocks with relatively high amounts of palagonite (greater than 10%), there was no significant chemical difference. The sediment samples are higher in Al, Si, and Fe and have less Mg and Ca. The difference in sediment and source rock chemistry indicates that another process is occurring, such as chemical weathering, sediment sorting, or that palagonite is a major portion of the sediments.

Methicillin resistant Staphylococcus aureus (MRSA) is an antibiotic resistant pathogen that causes severe illness and thousands of deaths each year in the US. It spreads within the community through improper hand hygiene and is often found in hospitals and on public transport surfaces. This poses a danger to the public, specifically to vulnerable populations such as the elderly and immunocompromised. This study in Seattle, Washington compares the prevalence of the antibiotic-resistant MRSA bacterium on public transport surfaces both proximate and distant to Harborview Hospital. Swabs from bus stops and pedestrian call buttons were collected outside of Harborview Hospital and urban areas of Capitol Hill, a Seattle neighborhood approximately one mile away from any major hospitals. The swabs were streaked onto Tryptic Soy Agar plates, gram stained, and streaked on Mannitol Salt agar plates;catalase and coagulase tests were run to help further confirm the presence of Staphylococcus aureus. The resulting colonies were then screened for antibiotic resistance using the Kirby Bauer Disk Diffusion method. One instance of potential MRSA was isolated from a crosswalk button in Capitol Hill. A higher MRSA prevalence on surfaces close to hospitals could establish a link between the spread of pathogenic bacteria from hospitals to Seattle's city surfaces but was not found in this study. These results suggest that the spread of MRSA in Seattle may have more to do with foot traffic and public transportation usage. However, the presence of MRSA on urban surfaces puts sensitive populations at risk regardless of its source. Practicing good hand hygiene can help curb the spread of MRSA in the community.

 Fragranced products such as cleaners, air fresheners, and colognes are pervasive in the United States. The fragrance in these products release volatile organic compounds (VOC), some of which are hazardous and have been shown to cause environmental and health impacts including headaches, liver and kidney dysfunction, neurologic impairment, cancers, asthma and other respiratory diseases. In this study, I investigated the VOC levels of fragrance products in retail environments to determine if they exceed levels recommended by Leadership in Energy and Environmental Design (LEED) guidelines. I measured concentrations of total VOCs (TVOC) in eight retail stores selling fragranced products. I conducted continuous sampling while moving within three feet of fragranced products. In department stores, I also collected a second sample in an adjacent department, at least 40 feet away from fragranced products. To establish a baseline, the final collection was taken outdoors away from other VOC sources (i.e. car exhaust) at each site. To measure TVOCs, I used a high-sensitivity, photoionization detector (PID). Because there is no U.S. standard for safe levels of VOCs in an indoor environment, I used the LEED guideline equivalent of 500μg/m^3 with a conversion factor of 3.767 to compare my ppb findings. My research showed the TVOC levels in fragranced areas of stores were at levels 10 times higher than that of outdoor environments, reaching levels of 1000 ppb. The VOC levels found in these retail environments are likely to impact the health of employees and possibly customers. More research is needed to determine the exact composition of VOCs emitted from these consumer products and the hazard they present to the health of customers and employees.

The measurement problem is the challenge of reconciling the probabilistic and deterministic aspects of Quantum Mechanics. In this study, our primary aim is to unravel the measurement problem by investigating the possibility that quantum collapse occurs according to the probabilities presented by Born's Rule due to a perturbation on the system, appearing during the measurement process. Our group employs a multifaceted approach, where we examine the interplay between time dynamics and classical limits, alongside the influence of time-(in)dependent perturbations. Computational simulations serve as our primary tool in this exploration. One part of our group worked with a 3-well system with a time-independent perturbation, another part looked at a 2-well system with a time-dependent perturbation, and the last part saw what a time-independent perturbation does to a 2D-well system. We anticipate that our investigations will uncover critical parameters that are expected to yield probabilities consistent with Born's Rule, a foundational principle of Quantum Mechancis. This research points towards a potential reconsideration of quantum collapse as a dynamical, affected by the perturbation introduced by measurement. While preliminary, these findings contribute to the ongoing discourse in Quantum Mechanics and may offer insights for future theoretical developments and applications.

The large-scale production of fossil-based plastics and its lack of biodegradability pose a significant threat to the environment, calling for the fabrication of sustainable polymer alternatives. Recent developments have been made in designing bio-based materials derived from plant and algae cells and tissues, transforming biological matter into compostable engineering materials. Despite these advancements, the effects of macromolecular composition and chemical binding patterns in shaping the mechanical properties of these bioplastics are still not fully understood. Therefore, this project aims to explore how the removal of macromolecules, specifically proteins, from various algae alters the microstructure and macroscopic properties of the resulting bioplastics. To explore this process, we performed two extraction procedures, extracting larger groups of molecules or targeting proteins specifically, on six different algae species: Ulva species, Gracilaria parvispora, Halymenia hawaiiana, Caulerpa lentillifera, Spirulina, and Ulva expansa. The extraction residues were mechanically tested and further characterized through FTIR, BCA assay, and SDS-PAGE. The water-extracted residues produced bioplastics with decreased strength, indicating the extracted material is essential to structural binding of the bioplastics. The BCA assay of the residues after protein extraction show that protein is successfully removed from the studied species, and the FTIR spectra confirm the expectation that the extraction is incomplete. Future experiments include optimizing the protein extraction and performing a large-scale extraction for mechanical testing. These findings open the door to a deeper understanding of the processes governing bioplastic properties and allow for the optimization of these materials based on their chemical composition.

Staphylococcus aureus is an important human pathogen that has increasingly developed resistance to antibiotics and antimicrobials. It is important to understand how genetic mutations influence antibiotic resistance to anticipate how the organism is able to evolve and combat emerging resistance. My project aims to fully characterize the impact of variants in the norA gene, which is associated with efflux-mediated resistance. The Salipante lab has developed a system for precision genome engineering in S. aureus by constructing a vector that is able to conduct recombineering while suppressing DNA repair, and a separate vector for killing unmodified bacteria using programmed CRISPR/Cas9. Using this system, I will construct a library of genetic mutations in the S. aureus genome that will encompass all possible single mutations in norA and to subsequently test the fitness costs of those mutations in the presence and absence of antibiotics.We will recombineer into S. aureus randomly mutagenized oligonucleotides that encode a silent mutation that renders transgenic bacteria immune to targeted CRISPR/Cas9 cutting. After killing off unmodified bacteria, the transformed population will be composed entirely of different mutants, so that with a large population size, we can statistically ensure that all possible random mutations are represented. We will analyze the gene sequence from the initial population, after the population has expanded in the absence of drugs, and after exposure to different concentrations of drugs, to compare the relative representation of each specific mutation. Changes that are detrimental under a condition will be underrepresented relative to the starting population, while those that are beneficial will be overrepresented. Mutations that are lethal will not be recovered in the first place. Our project has the potential to fully and comprehensively address clinically important questions regarding antibiotic resistance evolution in this gene.

Snow is an essential resource, especially in the Pacific Northwest, where it provides clean drinking water, irrigation water, and helps prevent wildfires. Despite the importance of the snowpacks in the Pacific Northwest, the composition of snow is not well documented. The ion concentrations in snow can indicate where the snow came from. I have created a dataset of the concentrations of anions found in snow throughout Washington State. Snow samples were collected primarily throughout the Cascade Mountain Range by volunteers using sterilized plastic containers. Samples were melted and then analyzed using ion chromatography to quantify the anions in each sample. Samples were found to contain concentrations of fluoride, chloride, nitrate, bromide, phosphate, nitrite, and sulfate. These anions have previously been found to be present in glaciers, clouds, and freshly fallen snow. Cataloging these anions provides an important baseline from which to observe how human and natural impacts, including climate change, are affecting the snow in the Pacific Northwest.

Artificial intelligence has made significant progress in image classification, an essential task for machine perception to achieve high-level image understanding. Despite recent advances in vision-language fields, image classification is still challenging because classes are inherently multimodal. Traditional approaches provide solely images or descriptions of a given class, but this does not properly encapsulate the entire concept. I aim to understand if utilizing multiple modalities when classifying images will allow for more robust and generalizable image classification. Here, I leverage Ensemble Learning to form a weighted sum of the embeddings produced by my classifiers. The ensemble consists of an image-to-text classifier that utilizes generated class descriptions and an image-to-image classifier that uses generated images for prediction. My results show that this improved conceptual representation from the multimodal approach directly results in higher classification accuracy. Additionally, this approach allows for these improvements without any additional context, training, or fine-tuning, meaning this method is completely zero-shot. Improved image classification directly impacts a myriad of fields, and enhances how humans use computing technology. Whether it be more accurate diagnoses in medical imaging or robust crop monitoring in agriculture, improvements in image classification extend across disciplines and aid the quality of life for all.

Alzheimer's Disease (AD) impacts over 6 million people in the U.S, but there are currently no fully effective treatments. Ageing is the biggest risk factor for AD and is associated with cellular changes called senescence. Cellular senescence describes a natural process in cells, leading to cell cycle arrest and metabolic changes due to insults from aging/disease processes. Factors contributing to senescence include DNA damage, and others. A risk factor in neurodegeneration is the ageing of microglia- our brain's immune cells that maintain a healthy brain. Senescent microglia express a senescence associated secretory phenotype- a combination of inflammatory proteins released into their environment- that enhances neurodegenerative processes. My project investigates the relationship between microglial senescence and AD by comparing the levels of senescence markers in AD brains, healthy young brains, and aged brains. I hypothesized that AD brains will contain the greatest amount of senescence markers, followed by aged brains, then healthy young brains. I performed immunohistochemistry for p16Ink4a and gammaH2AX (two robust senescence markers) on 10 human individuals (5 male/5 female per cohort) who donated their brain post-mortem. p16Ink4a is involved in cell cycle regulation and gammaH2AX signals DNA damage. The brain samples were also stained with Iba-1 to identify microglia. A confocal microscope imaged the samples and data was analyzed using the IMARIS software and ImageJ. Senescence markers were quantified in each cohort and localized in microglia or non-microglia cells. I expect to see the greatest amount of p16Ink4a and gammaH2AX in AD brains (specifically AD microglia), with the least amount in healthy young brains. I also expect co-localization of gammaH2AX and p16Ink4a in my samples. Understanding the relationship between microglial senescence and AD pathology could aid in finding methods to target cellular senescence. Slowing down this process could be a usefull tool in decreasing the progression of AD.

NASA is currently developing communication infrastructure for the lunar landscape in preparation for its Artemis missions to the moon. When rovers explore remote areas on the moon, where radio signals may not reach, there is a need for methods to facilitate both communication from base camps and help the rover reconnect to the network. The goal is to develop a deep learning model to predict radio signal quality and maximize communication by autonomously relocating lunar rovers to areas with optimal signal strength. Channel State Information (CSI) data provides insight into how a signal propagates from transmitter to receiver, including data for the signal magnitude, phase, and ray interactions with the environment. I investigate feature selection methods with different combinations of simulated CSI data to train our Recurrent Neural Network (RNN) deep learning model and analyze the resulting performance. Previous research demonstrates one way to improve the prediction of a model is by utilizing information at the hidden layers, the internal layers between input and output data. I explore this method and aim to capture patterns over time with our CSI input data and RNN architecture for predicting the magnitude of the next ray hit. We expect that using additional information at hidden layers will help us understand the relationships between input data and help optimize the model. We anticipate validating the model through the use of real CSI data using physical experiments to replicate signal interaction in a lunar environment. Our work contributes to the development of communication technologies for upcoming lunar explorations. It also provides insight into the role deep learning can play in radio frequency propagation, paving the path for further research in this area.

Achieving high performance in fixed-wing, unmanned aerial systems necessitates efficient wing assemblies which often entail significant design and production costs. Balancing measures associated with performance, production, reliability, and maintainability adds further complexity to wing design. I present here my current work on the use of Cellular Compressive Wing (CCW) architecture as a viable solution for achieving low structural mass and high flight efficiency while simultaneously enhancing production, maintainability, and reducing costs. To confirm the approach, a wing planform utilizing CCW has been developed based on specific aircraft performance requirements. Computational Fluid Dynamics and Finite Element Analysis have been leveraged to generate estimates of dynamic planform load distributions and CCW interface load characteristics. These simulation methods have in turn been used to guide the design of wing cell interfaces optimized for additive manufacturing techniques employing photopolymers and composite thermopolymers. Application-specific bench-test and in-flight hardware are currently being constructed and tested for direct experimental validation of dynamic planform and CCW interface loads.

Microdissected tissues from tumors have shown promise in preserving tumor microenvironment, which is critical for effective preclinical screening evaluation for drug development and cancer precision therapy. The tumor microenvironment’s microvasculature has an essential role beyond nutrient delivery and has been challenging to maintain ex vivo. Microdissection of cancer tissue into thousands of uniformly sized “cuboids” permits simple organization and perfusion of tissue using microfluidics to preserve the microvasculature. Here, we present a promising flow protocol in a 24-well microfluidic platform. The cuboids were cultured in our platform for different periods with different flow rates and without flow. The tissues were then fixed, stained with CD31, and cleared for fluorescence confocal microscopy. We demonstrated that this flow preserves the microvasculature of the tumor microenvironment of microdissected cancer tissues. The development of such a tool can enable medium-high throughput drug screeing and lead to improved cancer diagnostics and therapies for humans.

Saccades are rapid eye movements that are essential for tasks like reading, and their accuracy is maintained through motor adaptation across life stages and in response to neural injuries of diseases. Our long-range goal is to identify the neural mechanisms of such saccade adaptation. While saccade adaptation has been intensively studied before, our understanding of the neurophysiological basis of this phenomenon is based largely on amplitude-decrease adaptation. However, the clinical relevance of amplitude-decrease adaptation is marginal at best, as overshooting saccades rarely occur in real life. Most erroneous saccades fall short of their target, necessitating an amplitude-increase adaptation. This is what we propose to investigate in this application. The challenge in studying amplitude-increase adaptation lies in the requirement of numerous saccades over extended periods to observe significant amplitude alterations. Our preliminary studies, conducted over daily sessions spanning more than a month, discovered a bimodal distribution of adapted saccades characterized by low and high gains, with gain defined as the ratio of saccade amplitude to the target step. Low-gain saccades surface initially, reaching peak velocity saturation leading to a gain increase saturation. Conversely, high-gain saccades emerged after extensive trials, characterized by their reduced velocities and prolonged durations, suggesting a novel adaptation mechanism through the fusion of consecutive saccade pairs. Our project aims to (1) characterize this newly found adaptation mechanism and (2) investigate the superior colliculus's (SC) role, which is pivotal in commanding saccade size, in generating both low and high-gain adapted saccades and the integration of saccade pairs. The overarching impact of this research lies in its potential to enhance our understanding of motor adaptation's role in recovering from motor deficits caused by neural damage. Understanding these changes may help refine the strategy of rehabilitation for patients with saccade dysmetrias, and perhaps motor hypometrias in general.

The effective treatment of individuals with HIV relies on maintaining therapeutic drug concentrations, necessitating accurate measurement of antiretroviral (ARV) drug levels. Current methods, such as liquid chromatography tandem mass spectrometry (LC-MS/MS), are limited by cost and accessibility. Our research addresses this gap by developing the INTEGRase activITY (INTEGRITY) assay for measuring integrase strand transfer inhibitors (INSTIs), a leading class of ARV drugs. This 2-step assay quantifies INSTIs using a DNA strand transfer reaction and quantitative polymerase chain reaction (qPCR). The presence of INSTI drugs disrupts the strand transfer reaction, inhibiting full-length target DNA formation, which is then measured through real-time qPCR. My work focused on optimizing the limit of detection of INTEGRITY by altering the strand transfer reaction conditions and protocol. Specifically, I conducted experiments altering INSTI drug concentrations and optimizing pre-incubation times of integrase with the drug to enhance the LOD. I observed that preliminary incubation of integrase and INSTI drugs for 5 minutes at 37 degrees Celsius improved the LOD of INTEGRITY by an order of magnitude. The simplicity of the INTEGRITY assay, utilizing standard laboratory equipment, holds immense promise for broadening access to routine clinic-based ARV drug level monitoring. This advancement has the potential to significantly enhance HIV care on a global scale by offering a cost-effective and accessible solution for monitoring therapeutic drug concentrations.

TAR DNA binding protein 43 (TDP-43) is an RNA/DNA binding protein that forms pathological aggregates in most amyotrophic lateral sclerosis (ALS) and half of frontotemporal lobar degeneration (FTLD) cases. Knockout of TDP-43 in animal models leads to neurodegeneration and motor deficits, but overexpression of wildtype TDP43 leads to the same events; therefore, TDP43 protein homeostasis is critical to prevent ALS/FTLD. To achieve this homeostasis, TDP-43 autoregulates its own mRNA splicing, resulting in multiple TDP-43 isoforms, some of which go through non-sense mediated decay to regulate overall TDP43 levels. However, other isoforms encode unique proteins with differing C-termini, leading to variable cellular localization. It is unknown if these alternative, protein-coding isoforms are predominantly associated with ALS/FTLD or if aging changes the frequency of these isoforms. To determine how TDP43 overexpression yields these different isoforms and interacts with aging and ALS-like symptoms, we created a novel approach to overexpress human TDP43 via Adeno-Associated Virus (AAV) delivered through retro-orbital injection, leading to ALS-like motor deficits. Surprisingly, when tested in older and younger mice, we found the older mice were paradoxically protected from severe motor deficits and mortality. To determine if tardbp alternative splicing is linked to ALS-like symptoms and aging, I designed and validated primers and protocols to measure the nine tardbp mRNA isoforms in mice via quantitative real-time polymerase chain reaction (qRT-PCR). I have started to determine if hTDP43 overexpression leads to differential splicing compared to mice injected with a sham AAV in these old and young mice. Once this is done, we will clone the most interesting differentially spliced isoform in an AAV and inject that AAV and a full-length TDP43 AAV into mice to see if the spliceform causes increased toxicity, manifesting in worsening motor deficits and mortality.

Matrescence, the transformative journey into motherhood, encompasses profound physical, emotional, and psychological changes with lifelong implications. Postpartum mothers face various challenges including recovery from childbirth, sleep deprivation, hormonal and cognitive changes, insufficient support, and social isolation, often leading to depression and anxiety. While traditional medical interventions address some issues, alternative modalities of care such as horticultural therapy (HT) are gaining traction in postpartum care due to their holistic approach. This paper investigates the potential benefits of incorporating HT into postpartum care, aiming to explore its effectiveness in treating stress and related conditions, addressing psycho-social challenges, and integrating insights from other alternative therapies like occupational therapy (OT) to enhance postpartum wellness. Through an exploration of existing literature and a proposed pilot study, this research aims to fill the gap in understanding HT's efficacy specifically for postpartum mothers. The study proposes a comprehensive approach targeting physical and psycho-social aspects of maternal well-being through 12-week HT sessions with 24 postpartum mothers aged 18-35, incorporating health screenings, socialization, and horticultural activities. The study anticipates improvements in mental health, reduced postpartum depression symptoms, enhanced social support, and positive mother-infant interactions. Statistical analysis and qualitative assessments will evaluate intervention effectiveness, engagement, and feasibility. Understanding the transformative potential of HT for postpartum mothers advocates for a paradigm shift in maternal healthcare towards holistic and patient-centered approaches. Integration of HT into postpartum care models aligns with the evolving healthcare landscape and addresses the multifaceted challenges of matrescence, particularly in marginalized populations, emphasizing early intervention and culturally sensitive practices. This research underscores the importance of innovative interventions like HT in addressing the complex needs of postpartum mothers and calls for further exploration and integration into healthcare practices.

Translation seeks to convey meaning between one form of communication and another. The ways in which translation is completed can concentrate power and influence in ways that favor those whose languages and communication methods are dominant. This distribution of power and influence has important historical and modern consequences. This social science research project explores how translation has impacted nation to nation relationships between Indigenous people and colonial governments in North America. The treaty making history of the Blackfoot Confederacy will be considered as a case study, with a focus on the making of Treaty 7 between the British Crown in Canada and the Blackfoot Nations of Siksika, Kainai, and Piikani as well as other First Nations. Treaty 7 was signed in 1877 in a geographical, social, ecological, and cultural context that was heavily influenced by treaties, colonial westward expansion, and diminished bison populations. This case study on translation is being conducted through a review of firsthand accounts of the making and signing of Treaty 7 and a literature review of documented Indigenous oral histories of the event. These reviews will be conducted with consideration for the shifting power dynamics at play during that time. Finally, current academic work on language reclamation will be considered for its potential to support Truth and Reconciliation efforts, and a greater respect for Indigenous sovereignty. Language is a tool for carrying out translation, and carries significant elements of culture. Indigenous languages have a connection to the lands where they come from that are important for understanding current social and ecological challenges. Modern language reclamation efforts may have potential for restoring a more balanced distribution of power and offering solutions to these challenges.

When recalling an event, remembering when things happened is difficult for both children and adults. Across many cultures, we get around this problem by representing time spatially. One ubiquitous way adults and older children use space to represent time is by using a mental timeline: a linear, mental model of time. However, we do not know to what extent the mental timeline might be a product of our biology versus a cultural invention. This study examines whether 6-month-old infants spontaneously map temporal order to a mental timeline. In this task, infants are first habituated to a triplet of images that are either presented sequentially from left-to-right (linearly) or non-linearly across the screen. During the testing phase, infants see the same set of triplets presented centrally in a congruent (where objects are presented in the same order as shown during habituation) and an incongruent (where objects are not presented in the same order) pattern. If infants spontaneously represent temporal order using a mental timeline, they should learn the sequence more efficiently when presented from left-to-right relative to nonlinearly and, consequently, discriminate between the congruent and incongruent sequences at test. In Experiment 1 (N=41, data collected in France), the sequences involved puppet faces. There was no effect of habituation condition on looking times at the test trials. In Experiment 2, we replicated the study using simplified stimuli. Preliminary data (N=22) suggests that again, the proposed hypothesis is not supported. Data are now being collected for an updated version of the task. These results suggest that infants do not spontaneously map time to a mental timeline, suggesting that the development of a mental timeline may be a product of cultural experiences such as exposure to written language.

Prior research has shown that parent spatial language use predicts child spatial language use, and as a result, a child’s spatial ability. In our current study, we seek to identify factors that influence differences in both parent and child spatial language use. Using a picture book called “Flashlight”, which contains no words, parents were instructed to read aloud to their children (N=75 dyads, range: 2-4 years). Parents created their own story based on the pictures. After the storybook task, the parent and child were given blocks to play with for 5 minutes. Both these tasks seek to examine what types of language are naturally produced between parents and children, such as spatial words, questions, and number words (e.g., “at”, “What animal is that?”, and “one”). The videos of these sessions were manually transcribed. Parents also completed a series of questionnaires on their beliefs about the utility of play and their child’s language, motor, and communication skills. We hypothesize that the more questions a parent poses, the more spatial language a child will produce. This study is multifaceted in that it examines total language used overall (tokens), different types of language, free play, and spatial abilities. Studying these aspects of communication and underlying factors that impact the type of language parents use can lead to a better understanding of children’s spatial development and the factors influencing it. For example, parents who believe that play offers children unique opportunities to learn spatial concepts might use more spatial language with their children during play, which could scaffold their child’s spatial development. By identifying these factors, we can develop ways to increase spatial language use between parents and children, thus facilitating spatial development.

For my Capstone Project, I initially considered crafting a 20-minute monologue reflecting on my journey as a non-traditional undergraduate student at UW. My experiences, ranging from humorous misunderstandings with faculty members to instances of mistaken identity at school events, offered a rich tapestry of anecdotes. However, as I delved deeper into my narrative, I realized that my story echoed the archetypal "Hero's Journey" found in both mythological tales and contemporary dramas like "The Matrix," "The Hunger Games," and the "Harry Potter" series. Motivated by this insight, I expanded my project to encompass broader themes, weaving together elements of my cultural heritage with universal motifs. After extensive research, I discovered the perfect vehicle for my vision: the Turkish myth of "Wild Dumrul." This ancient tale follows the exploits of a young wild man who imposes tolls on travelers crossing a dry riverbed until a transformative encounter with death alters his fate. My Capstone Project will take the form of a dramatic production enhanced by traditional folk dance and music. Alongside a captivating solo folk dance performance, I will provide a brief exposition on the myth's resonance with Greek mythology, particularly the parallels with the story of Alcestis. I am thrilled to collaborate with a diverse ensemble of nearly 20 individuals, including members of the Turkish community, the Seattle theater scene, and talented actors from UW's School of Drama. Together, we aim to bring "Wild Dumrul" to life on stage, inviting audiences to embark on a journey of discovery and reflection. I am deeply excited and honored to spearhead this ambitious endeavor, and I look forward to sharing the magic of "Wild Dumrul" with our community.

Old (> 4 million years ago) ice drilled at the Allan Hills, Antarctica, can help us understand how Earth’s atmosphere has changed in the past. The bubbles trap bits of the atmosphere when they form, which can be analyzed to see what the climate was like. However, the preservation of this old ice depends on ice flow dynamics, possibly including localized shearing (one side of the ice is getting pulled faster than the other), that are difficult to observe. Bubbles in the ice become elongated when the ice around them deforms from strain. Over time, surface tension processes tend to restore bubbles to spherical. Thus, they can indicate the directions of recent or ongoing strain in the ice. We analyze thin/thick section images taken from four samples of Allan Hills ice. The images include information on grain size (size of individual ice crystals) and bubble size, shape, and distribution. We use the Segment Every Grain (SEG) model, a Python package based on the Segment Anything Model developed by Meta, to automatically calculate the sizes and shapes of bubbles in an image. We validate this method by comparing the values it returns with those obtained using another segmenting software, ImageJ, and manually calculated measurements. We can see if automated calculations are reliable enough to use regularly. So far, the SEG model has analyzed one image and it has made mostly correct bubble identification. The data shows that most bubbles are either elongated and small area, or round and large area. It is expected that the SEG and ImageJ models are close to humans in accuracy. The bubble orientations that we measure show the predominant directions of strain in the ice. Future work will use these data along with models of bubble elongation to estimate the strain rates at the Allan Hills.

The Allan Hill region of Antarctica has produced the oldest ice core samples ever recovered, which provide insights into Earth’s climate history prior to the existing 800,000 year ice core record. However, the highly disturbed nature of this ice complicates straightforward dating and interpretation. Understanding the scales of preserved climate records in this old ice will enable deeper insights into the variability of climate over the last 2 million years. Here I study a section of ice from ALHIC1901, an ice core recovered from the Allan Hills in 2019. This section has three parallel sets of water isotope measurements, and they all show a small but significant dip. However, the cause of this dip remains unclear. The goal of this study is to test whether this isotope change could be a glacial-interglacial transition preserved for 1.3 million years, or whether diffusion should have eliminated any climate signal. To investigate this question, I apply a simple water isotope diffusion model that takes as inputs temperature, an initial water isotope profile, and a thinning history, and provides as an output the resulting water isotope profile after a given number of years. I identify the range of possible temperature, thinning, and initial water isotope signals for this ice. I use these as inputs for the diffusion model, and compare the results to the ice core record to evaluate if the observed water isotope signal can be climatically driven. Constraining the cause of this water isotope signal will improve our understanding of fine scale paleoclimate proxy changes in the extremely old Allan Hills ice cores, enabling new insights into past climate variability.

Anthropogenic CO₂ emissions are rapidly warming the Earth, pushing it towards climate states without societal precedent. Past warm periods in geologic history can offer important insight into how the Earth responds to increases in atmospheric CO₂. One such period is the early Eocene (EECO; ~50 million years ago), which was characterized by global-mean temperatures ~13 °C higher than during the late 20th century, and is often offered as a potential analog for future unmitigated warming. Atmospheric CO₂ concentrations during the EECO are highly uncertain, but estimates range from 500-1400 ppm. Understanding EECO climate may also help us interpret past changes in biological systems; there is evidence, for example, that significant mammalian reorganization occurred in North America in response to high temperature and precipitation during the EECO. Because we do not have direct observations of the EECO, climate models are useful tools for understanding temperature and precipitation during this period. I use eight climate models from the Deep-Time Model Intercomparison Project to characterize zonal-mean temperature and precipitation under different atmospheric CO₂ concentrations, ranging from present-day CO₂ concentrations to 9x present-day CO₂ concentrations. I analyze temperature and precipitation in the annual-mean and across seasons. In the present-day climate, atmospheric heat transport and its efficiency (“diffusivity”) have been shown to govern the meridional temperature gradient and influence the shape of the hydrological cycle. To better understand the modeled Eocene hydroclimate, I calculate diffusivity across space and seasons. Ultimately, we seek to understand how diffusivity changes under different climatic conditions, and the physical mechanisms driving zonal-mean climate under elevated CO₂ levels. We expect that diffusivity will be relatively invariant with CO₂. This analysis can provide insights for how temperature and precipitation patterns may change under high CO₂ levels in the future.

Rhabdomyosarcoma (RMS) is a devastating soft tissue sarcoma most commonly affecting children. The components in the RAS-PI3K-MAPK signaling pathway are frequently mutated in RMS and are being studied as a critical target for treatment. A MEK1/2 inhibitor, trametinib, has shown clinical efficacy singly or in combination with other drugs in improving survival outcomes of patients with cancer types harboring mutations in the MAPK pathway. In addition, vincristine is a common chemotherapeutic drug used to treat RMS, but its efficacy has been limited due to RMS cells developing resistance. The study aims to determine whether combined treatment of vincristine and trametinib could more significantly inhibit RMS cancer growth compared to treatment with each drug alone. For this study, a clonal zebrafish line bearing RMS tumors labeled with a fluorescent protein was first treated with a range of trametinib concentrations to find the appropriate dosage for optimal effectivity yet minimal toxicity. Then, RMS-bearing zebrafish received two doses of trametinib, vincristine, or a combination of both through intraperitoneal injections (IP). Finally, we collected tumors from each treatment group to assess treatment-induced changes in gene expression by quantitative RT-PCR and changes in tumor cell proliferation and cell death by immunohistochemical stains. The changes in tumor volumes were determined by quantifying the intensity of tumor fluorescence in the ImageJ software. Preliminary findings showed that zebrafish tumors treated with the combination of vincristine and trametinib significantly inhibited tumor growth compared to the other with each agent alone. Thus, the results of this in vivo study on zebrafish may introduce a drug dosage combination with translational potential in creating a new treatment composed of lower doses of each drug while increasing potency against chemotherapy-resistant tumors and lessening side effects in patients. The end results will translate into improved survival outcomes and well-being of RMS patients.

Alzheimer’s disease and related dementias (ADRD) are neurological disorders that affect cognitive function, including impaired memory, thinking, and reasoning, primarily in older adults (OA). Cultural and linguistic differences can pose additional challenges. The experience of ADRD differs between patients from diverse backgrounds, and language barriers can create additional challenges when communicating with healthcare professionals. Our overall objective is to improve healthcare processes for persons with ADRD and limited English proficiency (LEP) in the United States. In this phase of a larger qualitative study, we are seeking to understand the experiences of older adults experiencing dementia whose primary language is Chinese and their family members. To understand the experiences of this population, we are conducting semi-structured interviews with OA with LEP and ADRD and their family members. I am performing qualitative data analysis along with three other team members. Data gathered from these interviews is transcribed and coded using a codebook developed using the inductive method. In this poster, I present key themes that are emerging in our preliminary analysis. There are significant barriers in patient-provider interactions between healthcare providers, OA with ADRD, and their family members, in the context of LEP. A complex dynamic is formed as providers seek to convey information to the OA and family members who play different roles in an OA’s life, including but not limited to acting as interpreters. OAs struggle to adjust to their progressive symptoms as they lose autonomy over time while navigating life with ADRD. Language barriers, lack of understanding of cognitive decline, and denial can also exacerbate problems. By identifying challenges experienced in patient-provider communication in the context of ADRD and LEP, we hope to work with healthcare providers to equip them with a better understanding of how to serve this demographic.

In response to acute genotoxic stress, such as chemoradiation therapy, stem cells undergo temporary cell cycle arrest at the G1/S phase transition. This state, called quiescence, is reversible once stress-free conditions allow reentry into the cell cycle. We have previously identified the underlying mechanism behind quiescence in Drosophila Germline Stem Cells (GSCs) and human-induced pluripotent stem cells (hiPSCs). Mitophagy, or autophagy of the mitochondria, is required to enter quiescence. Surprisingly, we have observed a reserve of cyclin E (CycE) associated with the outer mitochondrial membrane that’s present in normal GSCs and hiPSCs but is reduced in quiescent stem cells. The role of CycE in quiescence remains unclear. Previously we have shown that reduced levels of CycE via inhibition of mTOR have driven cells toward mitophagy-dependent quiescence. This reveals that mitophagy serves as an alternative mechanism of CycE inhibition in contrast to the typical p21-mediated inhibition. Additionally, Parkin, a ubiquitin ligase activated by a serine/threonine kinase PINK1, is a key protein involved in mitophagy required for quiescence, and it has been found that CycE is a degradation target of this protein complex. Our hypothesis is that CycE degradation is necessary for entry into quiescence. To test this we upregulated CycE with a deleted portion of its PEST domain, which is a target for ubiquitination, under UAS-GAL4 control and used the GSC spectrosome morphology to observe quiescence. We observed a six-fold reduction of quiescent GSCs with overexpressed CycE, and hence concluded that CycE degradation is necessary for entry into quiescence. Determining the mechanism of CycE in stem cell quiescence is critical to understanding how cancer stem cells can avoid chemoradiation therapy. This project allows us to characterize the role of CycE within mitophagy and strengthen our understanding of the mechanisms that govern the cell cycle and quiescence.

Mind-wandering, characterized by the inward direction of attention towards thoughts, represents a widespread cognitive phenomenon that can disrupt task-related attention. Evidence indicates that the frequency of mind-wandering increases during sustained attention tasks, and previous research has proposed several hypotheses, such as resource-depletion, control-failure, and resource-control hypothesis, to explain the increases as well as the declines in task performance from their perspective. This study first aims to investigate the impact of incorporating a strategy of break on the frequency of mind-wandering during a Sustained Attention to Response Task (SART). It seeks to determine whether a break strategy can mitigate the frequency of mind-wandering and to identify which of the aforementioned hypotheses best fit the experiment outcome. Specifically, it examines whether a rest break or a task-switching intervention can reduce the frequency of mind-wandering, comparing these effects to a no-break condition. Forty students from the University of Washington will participate in a four-part experiment that includes two 480-trial SART tasks, an intervening break condition (rest break, task-switch, or no break), and a concluding questionnaire. Mind-wandering will be measured through thought-probe methodology, and task performance will be assessed using error rates and response times. Additionally, subjective workload and fatigue level will be assessed using NASA Task Load Index (NASA-TLX) questionnaire. Based on the three existing hypotheses and behavioral data, the second aim of the study is to construct three cognitive architecture ACT-R models, and by comparing different models to reflect potential mechanisms of the role of breaks in mind-wandering.

Maladaptive aggression characterizes - or is comorbid with - many neuropsychiatric illnesses, and can have devastating effects on individuals, their caretakers, and healthcare professionals. Human aggression is typically demarcated as exhibiting either reactive (defensive) or appetitive (rewarding) components. Despite a significant clinical awareness of the differences between these aggression presentations, preclinical characterization of their relative circuitry and associated neuronal mechanisms are absent. Using recently established protocols within our lab, we are able to study and compare these aggression phenotypes in outbred male mice in a high throughput manner. Briefly, for appetitive aggression, we train mice to self-administer a novel subordinate intruder over 7 days using a trial design. In the reactive condition, we non-contingently administered intruders with the same frequency distribution as the appetitive mice. In the current experiment, we used CD1xVgat-Cre mice injected with pGP-AAV-syn-FLEX-jGCaMP7s in the rostral lateral septum (LSr) to examine cell-type specific activity via fiber photometry. GABAergic activity in the lateral septum has historically been implicated in the control of reactive aggression, but its role in appetitive aggression is unknown. My roles in this project include behavioral testing and filming of the mice, as well as scoring these videos for first attacks following intruder presentation. Using these timestamps, I will next analyze the changes in population level dynamics across different time points of aggression motivation, seeking, and consumption using the open source photometry analysis program GuPPy. We expect that the photometry results for mice in reactive and appetitive environments will show different patterns of activity, with more GABAergic activity during the consumption of reactive aggression. Interestingly, our preliminary results also show an increase in GABAergic activity when mice press the lever for a trial on which they subsequently attack, indicating that GABAergic activity may drive appetitive aggression seeking.

Continuous gravitational waves (CW) from sources such as a non-axisymmetric spinning neutron star have not yet been detected. If CW from neutron stars exist, the weak signals could be hidden within noise. The search for CW signals is impeded by the presence of narrow spectral artifacts (lines) caused by instrumentation or the environment at the Laser Interferometer Gravitational Wave Observatory (LIGO). Better identification of line noise would make a detection more likely, and a detection has the potential to expand our current knowledge of neutron stars. A non-machine learning (non-ML) line finding algorithm is currently used on a daily basis at the LIGO Hanford and Livingston Observatories. A different approach to line finding utilizes a machine learning (ML) algorithm. My research uses both the ML and non-ML line finding methods, comparing the accuracy and efficiency of these two methods applied to various data sets. Expected results are that the non-ML method is currently more accurate and efficient from a computing resources perspective, but that the ML approach has the potential for high accuracy and adaptability, and will eventually be more efficient in human hours. The impact of my research is to implement more accurate and efficient line finding algorithms. This work is important because automated line finders at LIGO Hanford and LIGO Livingston save researchers hundreds of hours of work and could be set up to alert researchers on changes in noise behaviors.

Repetitive transcranial magnetic stimulation (rTMS) is a psychiatric treatment which has shown promise for experimental treatment of memory loss in Alzheimer’s Disease. rTMS uses a coil and electric current which is able to create a magnetic field that can depolarize neurons noninvasively and induce synchronized activity of large populations of neurons, ultimately inducing, lasting changes through synaptic plasticity. Alzheimer’s disease patients show disruptions in the Default Mode Network (DMN), a network of brain regions which is typically active at rest. The DMN has an important role in memory consolidation and is disrupted in Alzheimer's Disease. We hypothesize that strengthening the default mode network through rTMS applied to area left 8AV of the frontal cortex will create improvements in patient memory. To answer this question, we are performing a single-blind, single-arm, randomized cross-over trial of rTMS on early-stage Alzheimer's disease patients. Region 8AV is located by using MRI scans obtained before patients receive either the sham or experimental procedure. This region was chosen due to its connection to the default mode network and previous promising TMS research. Our primary outcome measure is the speed of forgetting, a new, reliable index of memory function obtained by fitting a computational model of episodic memory to behavioral data from an adaptive memory test. Due to the frequent use of rTMS in mood disorder treatment, we are using depression and anxiety scales to track possible mood improvements as a secondary outcome measure. MRI scans will also be analyzed to see if the experimental treatment caused any structural differences in patient brains. Should our hypothesis be correct, we expect to see improvements in memory or cessation of memory decline in patients. Successful treatment would provide a novel target for Alzheimer’s Disease treatment using rTMS, and additional evidence for the continued investigation of rTMS for Alzheimer’s Disease.

Neuropsychiatric disorders, such as major depressive disorder, pose a difficult challenge for healthcare providers. Treatments for such disorders vary in efficacy and come with detrimental costs. Historically, preclinical animal models have failed to incorporate the nuances of volitional human social behavior. This project used chronic social defeat stress in which mice experienced bouts of antagonistic encounters to induce depression-like behaviors in male and female mice, this was followed by self-administered social interactions within an experimental chamber in which lever presses were reinforced by social contact. The goal is to develop preclinical animal models that can be assessed to identify neural mechanisms responsible for stress-induced social motivation. Male and female mice will train to self-administer social interaction with a sex and age-matched housing partner over the course of ten 12-trial sessions. Next, experimental male and female mice will be subjected to both social and witness defeat (observation of social defeat) sessions followed by social self-administration. Before and after the 10-day social stress sessions, we will test social reward seeking via non-reinforced self-administration of social reward followed by a progressive ratio test. Brain tissue will be collected and prepared for immunohistochemistry and whole-brain clearing. Social defeat decreased social reward seeking behaviors in male mice. Witness defeat did not alter social reward seeking in males but increased seeking behavior in female mice. Social stress can be used to discern differences in social motivation in male and female mice as a result of stress-induced factors. There is potential in using whole-brain activity mapping to identify brain structures activated during social reward following social stress. We hope to build a technical tool for the field that can encompass whole-brain activity responsible for social stress responses by utilizing nuclear localization and retrograde tracing.

The potential cognitive benefits of mindfulness meditation, such as its effects on memory processes, are gaining increasing attention. The growing interest in mindfulness meditation as a technique to improve cognitive functions, particularly long-term memory, motivates this investigation. The purpose of this study is to determine how mindfulness meditation affects the individual’s rate of forgetting in long-term memory as practice days increase. Using a computational model of episodic memory fitted to data obtained from an adaptive memory test, the study is able to monitor and predict the rate of forgetting for each individual and acquire a deeper understanding of how mindfulness meditation influences individual differences in memory retention and retrieval. The study will recruit thirty healthy participants through the Psychology Subject Pool of the University of Washington. The participants will be randomly assigned to either a group that practices mindfulness meditation or a control group that engages in podcast listening. For six days, participants will complete online memory tasks before and after each mindfulness meditation session or podcast listening session, while their performance data will be analyzed to model the rate of forgetting. To assess the principal effects of mindfulness meditation on the rate of forgetting, statistical analyses, such as linear mixed models, will be employed. We hypothesized that participants who participate in mindfulness meditation will have a lower rate of forgetting than those in the control group who engage in podcast listening after six days. Moreover, the study also hypothesizes that participants engaging in mindfulness meditation will demonstrate a decreasing rate of forgetting as the number of practice days increases. The finding of this study may inform interventions using mindfulness to enhance memory retention, benefiting those with age-related cognitive decline or stress-related memory challenges. It contributes to understanding the potential of mindfulness in cognitive function and memory-related disorders.

Vestibular disorders are prevalent, particularly in aged people, and can cause significant impairment to quality of life including spatial disorientation. The sensory hair cells (HCs) in vestibular end organs sense head motions. Vestibular HCs degenerate as we age. Interestingly, vestibular end organs generate neural signals that terminate in the hippocampus. These connections contribute to the regulation of spatial memory and learning, although the mechanism is not completely understood. We are exploring whether loss of vestibular inputs to the hippocampus affects neurogenesis of granule cells in the dentate gyrus. A prior study showed a gradual increase in proliferative activity of granule cell progenitors after bilateral labyrinthectomy. We wondered if the same effect would occur when vestibular HCs are destroyed and if regeneration of HCs reverses the effect. We used a mouse that enables near complete killing of vestibular HCs upon injection of diphtheria toxin (DT). We administered the cell division tracer, BrdU, to experimental (HC-depleted) and control mice, all of which were adults, using 1) a pulse-fix paradigm to capture cells in the cell cycle or shortly thereafter and 2) a pulse-chase paradigm to study cells that differentiate into neurons. I am counting BrdU-labeled cells in the hippocampal dentate gyrus at different times after HC damage and in undamaged controls. In my poster, I will be presenting preliminary data that tests the hypothesis that we will see increased neurogenesis once HC inputs are lost. Once the study is completed, we hope to determine whether and how vestibular HC destruction and regeneration impact hippocampal neurogenesis. By exploring the relationship between the vestibular system and the hippocampus, we will build our understanding of how spatial orientation is regulated and whether vestibular regeneration can improve spatial orientation, which may inform on how to treat people with vestibular disorders in the future.

 The Amazon rainforest is one of the most biodiverse regions on this planet and home to thousands of species of butterflies. Our pilot study aimed to investigate correlations between vegetation diversity and temperature and butterfly species richness in the Madre de Dios region of southeastern Peru. This study was conducted across three sites of differing plant diversity, including a monoculture orchard, a mixed orchard, and a rainforest. We hypothesized that there would be a strong positive correlation between species richness and vegetation diversity, resulting in high species richness in the rainforest. We also predicted that each species would only be found within one site, as most butterflies rely on specific plant species for reproductive purposes. Finally, we hypothesized that the hottest times of day would have the least amount of butterfly activity. By attracting butterflies with baited banana leaves, we were able to record an array of species during multiple, daily 45 minute observational sessions. After classifying photographs of butterflies during our sessions to morphospecies and averaging species richness, we found that the sites with higher vegetation diversity had a wider range of species. Most species demonstrated specialist behavior, found only in one site. Out of 26 morphological species, only three were observed at more than one site. Limitations in our sample size made any data surrounding behavior across a temperature gradient inconclusive. By critiquing this pilot study and analyzing its small set of data, we suggest protocols for future research on butterfly species richness and activity.

Primary care providers are responsible for utilizing well-child-check-ups to screen toddlers for autism spectrum disorder and immediately refer children with increased likelihood to diagnostic and intervention services. Participating in intervention services at young ages is extremely impactful as it often allows children to develop their social, cognitive, and language skills. However, it is estimated that only 40% of children who screen at an increased likelihood for autism are being referred to recommended services by their providers. It is especially important to look at autism referral rates as autism is half as likely to receive a referral from a provider than other developmental concerns. This mixed-methods study used a video vignette and focus groups to investigate factors influencing primary care providers’ decision- making regarding autism referral in a well-child-check-up setting. We examine the role of provider attitudes, autism knowledge, and distribution of attention (information providers attend to during a visit). We anticipate that providers will exhibit low knowledge of autism and intervention, and believe information about autism that has empirically been proven to be false. Additionally, we expect that providers pay less attention to clinically significant behavioral information than physical information during a well-child-check-up. Overall, we aim to understand what influences provider’s referral decisions to provide insight on a development of a future primary care provider intervention that supports the increase of autism referral rates. Ultimately, improving the developmental prognosis of children with autism.

Glutamine synthetase (GS) is a highly regulated enzyme critical for converting glutamate to glutamine and associated with ammonia assimilation. Dysregulation in the GS interconversion process can lead to hyperammonemia, potentially resulting in death or brain damage. GS is conserved across prokaryotes and eukaryotes. Among enzymes, glutamine synthetase has the ability to polymerize but the functional characteristics of its self-assembling filaments remain unknown. This study aims to elucidate the occurrence of filament formation in GS and its effects on enzyme activity. We hypothesized that filaments might influence the association of GS substrates or allosterically regulate the enzyme. I purified GS from Pseudomonas aeruginosa, Mycobacterium tuberculosis, and Helicobacter pylori using Ni-column and size exclusion chromatography (SEC). The focus was primarily on Pseudomonas GS, examining it under various buffer conditions (Mg²⁺, Co²⁺) through negative staining. Under magnesium conditions (10 mM), dodecamer strcture of GS was observed and filaments was induced under cobalt conditions (10 mM). To investigate the structural mechanism of filament formation further, we utilized cryogenic electron microscopy (Cryo-EM) to create a model of the GS filament interface and identifying involved residues. Additionally, I am conducting mutagenesis on key residues of Pseudomonas GS to disrupt filament formation. This research holds significant implications for metabolic engineering, as understanding the structure and role of filament formation in GS could lead to new therapeutic targets in metabolism.

The ventral tegmental area (VTA) is a key region in the brain’s mesolimbic circuitry playing a role in regulating reward processing, aversion, motivation, and stress-related behaviors, housing both dopamine (DA) and GABA-expressing neurons. GABA neurons in the VTA form direct connections with DA neurons, modulating dopamine and influencing reward-related behaviors. This study aims to characterize two distinct VTA GABA populations marked by expression of the nociceptin gene (Pnoc) and corticotropin-releasing hormone binding protein (Crhbp). Using double transgenic mice and viral targeting, we aim to map the projection patterns of these two populations. We anticipate that Crhbp-GABA expressing populations will innervate the Ventral Pallidum and Lateral Habenula brain areas as these neurons also coexpress Vglut2, a marker for glutamate neurons which are known to project to these regions, whereas, Pnoc-GABA expressing populations might represent the local GABA population that synapses onto DA neurons within the VTA. To assess their functional role, we will optically activate these populations during a real-time place preference task (RTPT) using channelrhodopsin-2 (ChR2). We hypothesize that activation of Pnoc-GABA neurons will result in a negative valence response and support real time place aversion. On the other hand, Crhbp-GABA neurons may have a more varied effect on valence response, based on their coexpression of Vglut2, during the RTPP task. By understanding the roles of VTA GABAergic populations marked by Pnoc and Crhbp expression, we can gain insights into the neural mechanisms involved in reward processing, motivation, and stress, often dysregulated in psychiatric disorders. This understanding could ultimately inform the development of targeted therapeutic interventions for psychiatric conditions characterized by maladaptive behavioral responses to stress and other stimuli.

Meningiomas, the most common type of primary human brain tumor, arise from the thin fibrous membrane that covers the brain and spinal cord. Most grow slowly and are diagnosed when they disrupt brain function or lead to persistent headaches. While many meningiomas can be cured by surgery, ~20% of them cannot be fully resected or display increased growth, invasion and destruction of adjacent brain and skull. Effective control or eradication of these ‘High Grade II/III’ meningiomas is clinically challenging. To identify new agents and treatment measures, our project uses both computational and experimental approaches in concert to identify new and potentially better therapies. As part of this effort, we are using PISCES, a machine learning model, together with augmented drug and radiation combination datasets to predict potential new therapy synergies. The best predictions from PISCES will then be tested experimentally in our cell line model versus standard-of-care treatments.  My presentation summarizes work to characterize genomic, drug and ionizing radiation sensitivity data on IOMM-Lee, a Grade III human meningioma cell line disease model. We detail how A.I.-driven analyses of IOMM-Lee and related meningioma datasets led us to test new drug pairs and drug-radiation combinations predicted by PISCES to be more effective in killing IOMM-Lee tumor cells. This translational cellular disease model and project are part of a long-term effort to develop better ways to rapidly and efficiently identify and validate new treatment options for brain tumors and other human cancers that can be taken directly to clinical trial.

PARP inhibitors are a revolutionary precision cancer therapy that inhibits PARP function and causes synthetic lethality in cells with homologous recombination deficiency (HRD) such as cells with BRCA1/2 loss. As such, BRCA1 presence plays a key role in PARP inhibitor sensitivity, providing a basis to predict treatment response based on BRCA1 expression in patient tumors. In addition to protein loss mutations, hypermethylation of BRCA1 gene may also result in the loss of BRCA1 expression, and subsequent HRD and potential PARP inhibitor sensitivity. The NRG-GY005 clinical trial focuses on the development of a clinically useful predictor of PARP inhibitor sensitivity/resistance to spare toxicities for patients unlikely to derive benefit. I will first characterize BRCA1 expression in randomized and blinded tumor samples as a biomarker for response to PARP inhibitors in patients. I will then relate BRCA1 protein expression to methylation status of BRCA1. Combining these two aims will describe BRCA1 function/presence in tumors, and better define overall homologous recombination deficiency to optimize patient-specific treatment between PARP inhibitors or other precision therapies. I conducted immunohistochemistry (IHC), a targeted staining with antibody MS110, towards BRCA1 to identify protein presence in tumor tissue and have compiled and analyzed droplet digital PCR (ddPCR), IHC, and clinical treatment reports. I am currently working through Aim 1 by performing IHC staining on tissues from the first phase of the NRG-GY005 clinical trial and will continue investigating whether a lack of BRCA1 expression is associated with cancer cells that are responsive to therapies. In combining IHC and ddPCR assays, we can compare BRCA1 presence with methylation to analyze tumor cases without BRCA1 expression and contribute toward identifying biomarkers to inform patient-specific treatment for individuals with recurrent ovarian cancer. I anticipate that cases lacking BRCA1 expression have an increased likelihood of hypermethylation in the tumor, causing loss of BRCA1.

This project evaluates the flight mechanics of a class of Autonomous Underwater Vehicles (AUVs) known as gliders using digital simulation via Python code. When used in the real world, these gliders perform vertical profiling of important marine quantities like temperature and salinity. These are then used by oceanographers and others to help them gain a deeper understanding of the ocean environment. The overall goal of the project is to optimize the trajectories of a fleet of vehicles to minimize energy consumption while maximizing uniformity of ocean coverage. Using specific engineering data on real world glider flight as well as public domain ocean environmental models, I have coded a custom Python application with guidance from my mentor at the Applied Physics Lab. We chose a computer simulation of glider flight so that multiple variables could be easily manipulated without the risk of losing a valuable glider if certain parameters are not favorable. This simulation produces data that describes the vehicles’ locations and energy states over time. The software is structured such that important parameters are specified in an easily-modified configuration file. The parameters I alter include geographic area, the number of gliders, the maximum flight depth, the vehicle’s available buoyancy range, and the glide angle. Then, using the data that the simulation produces, I analyze variations in energy consumption, uniformity of coverage, and the time required for each glider to reach their destination. The oceans, which cover about 70% of the planet’s surface, have a huge impact on the climate and health of the Earth as a whole. The result of this analysis is useful to real-world AUV operations by helping determine how to program them to fly more efficiently and maximize their utility as scientific instruments.

Challenges with social-emotional reciprocity, such as initiation of and response to social interactions, are a core diagnostic criteria of autism spectrum disorder (ASD), and many children with ASD experience challenges with expressive language. Prior research demonstrates parents are accurate reporters of language and fine motor skills development, but may under report social emotional reciprocity. The goal of my study is to discern how well parent concerns around child social-emotional reciprocity and language correspond to observed virtual interactions with their child. In a larger study investigating interventions for children with social communication concerns, parent participants of toddlers aged 25-41 months (n=41) completed the Toddler Autism Symptom Interview (TASI), a structured interview about core autism symptoms, over Zoom. Parent-child interactions were recorded in the same session while they completed the TELE-ASD-PEDS (TAP), a telehealth ASD assessment for toddlers. As part of the larger study, I am assisting with coding child expressive language, including use of words and multi-word combinations. For my independent research, I am coding child behavior on two tasks from the TAP, ‘calling the child’s name’ and ‘approaching others to play’. ‘Calling the child’s name’ is coded as the number of times the child stopped what they were doing to look at their caregiver when their name was called, while ‘approaching others to play’ is the number of times the child approached their caregiver during a 2-minute period where their caregiver was instructed to sit back and ignore their child. I will compare coded behaviors with corresponding items on the TASI. I hypothesize that parent reports will be more consistent with coded observations for language concerns than for social-emotional reciprocity behaviors. Understanding whether social-emotional reciprocity is being accurately reported by parents can help clinicians provide accurate diagnoses for ASD, especially given the critical role of parents in the diagnostic process.

Fifth Harmony’s BOSS music video, which showcases female empowering choreography, has been viewed 267 million times. The choreographer behind it, Sean Bankhead, said in an interview the higher you go the bigger the budget and the bigger the ego of the artist that you are working with. He went on to say about how when he was working with Missy Elliot, she would challenge him to pull things out of him that he didn’t know he had and he was told "start fresh tomorrow you can do better.” As hard as it is to break into this industry, choreographers continuously mention being extremely proud of sharing their work with millions of people. From the outside, it seems like music video choreographers have so much freedom in artistic choices like style of the piece and movement influences, but how much do choreographers truly get to create the exact choreography that they want? The exploration of music video choreography is really important because music videos are a way for the dance artform to be shared widely, but in scenarios like the one above, commercial popularity influences the creation of a choreography. Bankhead also mentioned that in music videos, he incorporates a snippet of choreography that even non-dancers can do, and these are usually the pieces that go viral. How do Los Angeles dance industry choreographers strike a balance between their own artistic vision and commercial pressures to create viral videos to become successful in the LA music video scene? My research seeks to answer this question through analysis of YouTube interviews, close reading of popular dance music videos, and interviews with choreographers.

All plants have symbiotic relationships with microbes known as endophytes. These endophytes, often bacterial, live within plant tissues and help improve resiliency in the face of environmental challenges. In our changing climate, we have become increasingly dependent on the unsustainable practice of using nitrogen-based fertilizers to promote plant growth. With the hypothesis that wild plants growing in nutrient-limited sites would have endophytes particularly strong in providing nutrients to the plant host, our team collected plant tissue samples from several sites along the Bogachiel River Trail located in the Bogachiel Rainforest near Forks, WA. We successfully isolated endophytes from these samples, characterizing each strain based on their ability to grow on nitrogen-limited growth media. We used acetylene reduction assays (ARA) as a proxy to quantify nitrogenase activity levels of fast-growing microbial strains. The strains we quantified with the most nitrogen-fixing activity were collected from the primary successional zone of the river, consistent with previous findings of the UW Plant Microbiology Laboratory. We later identified these promising strains through 16S DNA sequencing, and using bioinformatics platforms, we searched for genes involved in nitrogen fixation. The properties of nitrogen-fixing endophytes can be investigated as an alternative to nitrogen-based fertilizers, sustainably increasing plant productivity in environments where nitrogen is limited. Future plant experiments can now be performed using our strong nitrogen-fixing strains to determine if these microbes have plant growth-promoting properties. The isolation and further study of endophytes like these will be crucial in the future of sustainable agriculture.

The effects of the fungal pathogen Swiss needle cast (SNC) on its host species, Douglas-fir, and its timber harvest repercussions have been researched. However, more research is needed regarding the effects of SNC on understory species and, consequently, browsing ungulate species. This study analyzes the effects of various thinning methods on SNC and SNC’s influence on understory species richness, diversity, and cover. The goal is to articulate a clear dynamic of SNC in Sitka spruce and Douglas-fir - western hemlock zones to provide insights for guiding forest management. I will analyze the Olympic Natural Resource Center’s pre and post-treatment data from Siuslaw National Forest with statistical analyses to articulate patterns in SNC presence, thinning, and understory change. The early, mid, and late seral/thinned treatments vary in density and by species replanted. I expect that thinning will decrease SNC abundance and increase understory species richness. Additionally, I predict that the stands replanted with red alder and conifers will see a higher abundance of understory due to red alder’s nitrogen-fixing ability. The enhanced understanding of SNC and thinning’s interplay aims to educate current and future forest managers about ecologically responsible management.

 Forest edges impact a wide range of factors, including species composition, tree growth, and tree density. Understanding the nature of edge effects on forests is critical to guiding forest management practices and forest yield modeling. The Long-Term Ecosystem Productivity study (LTEP) was established to study the effects of silvicultural treatments on long-term forest productivity. In this project, I aim to understand the effects of forest stand edges in different forest management regimes in the Sappho LTEP site. I am using data collected in two treatment units : one that was planted with a 50/50 mix of Pseudotsuga menziesii and Alnus rubra, and one that was planted with only P. menziesii. Field data was collected on the edge of units within the buffer and within the mensuration plot, and included tree species composition, spatial position, and diameter at breast height (DBH). These ground data are paired with LiDAR remote sensing to verify tree locations and delineate the “edge” of experimental units. I used ArcGIS to calculate Euclidian distance to the edge (DTE). I am performing exploratory statistical analysis to determine relationships between DTE and tree species composition, DBH, trees per acre (TPA), and will also compare effects of DTE between experimental units. I predict that tree in-growth will demonstrate codominance between shade tolerant and intolerant species in edge plots, whereas shade tolerant species will be favored in mensuration plots. I expect that in both units, TPA will be higher, and as a result, DBH will be lower in the edge plots than in the mensuration plots. Results from this project will inform if and how forest managers should manage stand edges. The impacts of edge effects on tree species composition, DBH, and TPA will have direct implications for timber yield and carbon sequestration, which may not be accounted for in current models.

Lipotoxicity in cells occurs when lipids in biological tissue accrete to a toxic level. This toxic buildup is associated with obesity and type 2 diabetes, two of the leading causes of death around the world. Various studies have induced obesity in mice through a diet consisting of high levels of saturated fats. Some of these studies also investigated the effects Adefovir Dipivoxil, an antiviral and inhibitor of mitochondrial DNA replication, had on these obese mice. My research investigates fibroblasts that have been grown in a high-lipid environment and how Adefovir Dipivoxil affects these cells. We hypothesized that administering Adefovir to these cells would halt apoptosis and prevent further progression of lipotoxicity by stabilizing levels of triacylglycerol synthesis. To test this, I induced lipotoxicity in the cells using palmitic acid, a saturated fatty acid. Next, I treated the cells with Adefovir Dipivoxil. To measure the efficacy of the drug, I used a staining kit to measure the ratio of live to dead cells before and after administering the drug. Furthermore, I used a dye-based assay to measure the intracellular triglyceride levels before and after treatment. In addition to treating fibroblasts with palmitic acid, I treated a new set of fibroblasts with oleic acid, an unsaturated fatty acid, to determine how Adefovir acts on these cells. Data from this research will contribute to further understanding the mechanism of lipotoxicity on various cells, as well as the role that mediators of mitochondrial function like Adefovir Dipivoxil could play in treating lipotoxicity. On a broader scale, we hope that this research will provide insight into future treatments for obesity and type 2 diabetes.

General anesthesia (GA) is administered as a sedative in nearly 60,000 surgeries daily in the United States. Yet, there is a very limited understanding about how GA impacts brain activity, leading to induced loss of consciousness and pain sensation. Preliminary work in the Heshmati lab has highlighted key subcortical structures that are engaged during anesthesia, but it remains unclear how activity in these regions and across the brain regulates awareness or pain sensation as anesthesia is induced (“induction”), maintained at a steady state (“maintenance”) and removed (“emergence”), as is done during surgeries. My work aims to identify the neural circuits that regulate the loss of consciousness and pain sensation during GA by recording local field potentials (LFP) from mice as they undergo volatile anesthetic isoflurane (ISO). During LFP recordings, I will insert small electrodes into highlighted regions of interest, to capture low-frequency extracellular voltage signals generated by the synchronized activity of nearby neural populations during the three periods of interest: induction, maintenance, and emergence from isoflurane GA. I will analyze the amplitude fluctuations and frequency patterns to identify synchronized oscillations within subregions and assess the level of synchrony, or coherence, across different regions. Given previous findings on the shared and opposed involvement of subcortical regions in pain and anesthesia, I expect to observe coherence among some of the regions, such as the amygdala and hypothalamus, but potentially anti-correlation within specific subsections, such as central vs. basolateral amygdala. Through these experiments, I will be able to monitor the effects of isoflurane anesthesia through a temporally-defined electrophysiological lens, capturing real-time activation dynamics of large neural populations across induction and recovery from anesthesia. Thus, my research aims to further develop our understanding of the brain under GA, by providing novel insight into the neural circuits regulating wakefulness and pain during surgical procedures.

Mice missing NADH: Ubiquinone Oxidoreductase Fe-S Protein 4 (NDUFS4) are a model of Leigh Syndrome, a mitochondrial disease. NDUFS4 KO mice show reduced lifespan and high levels of acetylation of mitochondrial proteins. The anti-diabetic acarbose increases survival in NDUFS4 knockout mice and reduces mitochondrial protein acetylation, as deduced by reduced acetylation of SOD2 on Lysine 68. SIRT3 is a NAD+/- dependent deacetylase that deacetylates mitochondrial proteins. We sought to determine whether acarbose reduces acetylation in NDUFS4 mice via SIRT3. NDUFS4 and SIRT3 double KO mice were fed acarbose and dissected at 35 days of age. Brain samples were collected and analyzed via western blot. We determined the levels of acetyl K68 SOD2, total SOD2, and actin for each sample. Our results show that acarbose reduces acetylation in knockout NDUFS4 regardless of the presence of SIRT3. Acarbose reshapes the intestinal microbiome and the production of short-chain fatty acids in NDUFS4 KO mice. We are currently devising in vitro experiments to determine whether short-chain fatty acids can reduce mitochondrial protein acetylation in NDUFS4 knockout cells. This research investigates if short-chain fatty acids are responsible for acetylation of mitochondrial proteins. The results can be significant because the acetylation of mitochondrial proteins can prevent diseases from being as deadly by extending lifespan. The work completed during this research is hopeful to be applied to a human model, in which this model can be used to treat mitochondrial diseases.

Dopamine (DA) producing neurons of the ventral tegmental area (VTA) in the midbrain regulate reward association learning and motivation. These DA neurons are modulated by neuropeptides and can be separated into distinct subpopulations based on differential gene expression, regulation of activity, and projection patterns. But how these different patterns are established and contribute to distinct functions of DA subpopulations remain poorly understood. One potential key component for these neuropeptides is the transient receptor potential canonical (TRPC) channels. Specifically, we identified the gene encoding TRPC type 6 channel (Trpc6) as having enriched expression in the VTA DA neurons. To determine whether Trpc6 is differentially expressed in VTA DA subpopulations, I utilized the quantitative, multiplexed in situ hybridization methods. Using wild-type mice, I probed for the expression of tyrosine hydroxylase (Th), a marker of all DA neurons, and Trpc6 as well as markers of two subpopulations, corticotropin releasing hormone receptor 1 (Crhr1) and cholecystokinin (Cck). The analysis showed that Trpc6 expression is significantly higher in the Crhr1 subpopulation, of 81%, than in the Cck subpopulation, of 66%. Because neuropeptides like neurotensin increase calcium concentration in DA neurons, we hypothesized that TRPC6 contributes to these neuropeptide-evoked calcium signals. To investigate the role of TRPC6 in DA signaling, I used a viral-based CRISPR/Cas9 approach to induce selective mutagenesis of TRPC6 in specific DA subpopulations. Then, I assessed the calcium responses of subpopulations to neurotensin by measuring the amplitude and neurotensin-evoked oscillations using acute brain slices. We expect the calcium responses to decrease more in the Crhr1 subpopulation than in the Cck subpopulation compared to the control as the Crhr1 population has higher Trpc6 expression. By elucidating the role of TRPC6, we hope to contribute to discovering pharmacological interventions for diseases caused by dopaminergic system dysfunctions such as Parkinson’s disease and substance use disorders.

Social interactions in humans have shown to improve pain outcomes and diminish the development of mechanical hypersensitivity (allodynia) following injury. This effect is known as social buffering of pain; however, the underlying mechanisms are not well understood. Prior preclinical studies focused on forced social interactions between unfamiliar mice, lacking translational value to patients. To fill this gap, our research explores how volitional social behavior shifts pain sensitivity and affect following a neuropathic injury. Volitional interaction is key to socialization as individuals usually socialize because they want to, not due to force, which makes studying how mice voluntarily interact with each other important. To determine how volitional social interaction impacts both sensory and affective (emotional) components of pain, we use male and female mice who have received a spared nerve injury (SNI). Trained in social self-administration, mice learn to lever-press to engage with a familiar conspecific. Mice are then tested in von Frey where thin plastic filaments of increasing weights are applied to the mouse hind paw before and after SNI. These filaments do not cause pain, rather elicit a pain response of withdrawing the paw. To determine sensory sensitivity, the weight when the animal's paw is withdrawn is recorded as percent change from baseline. To determine changes in affective pain, the amount of time the animals hold their paw up, following withdrawal, is recorded as percent change from baseline. We found that male and female mice show significant attenuation in their mechanical hypersensitivity following volitional social interaction compared to mice deprived of volitional social interaction. Males show even less mechanical sensitivity, indicating that males may be more impacted by social analgesia than females. Understanding the divergent responses between male and female mice and the role of volitional social interaction in pain modulation, offers potential avenues for developing novel therapeutic strategies.

Data visualizations are critical to understanding large, complex datasets. For example, visualizations help us detect missing and erroneous data values, identify potential relationships between data variables, and review the output of machine learning models. That being said, customized visualizations can be difficult to create, because they require the use of specialized toolkits such as ggplot2, Vega-Lite, or D3. In this research, we study how people use a browser-based toolkit called D3 to program custom visualizations, with the long-term goal of creating AI-assistants to help people code in D3. However, to train rigorous AI models, we need a large input corpus of D3 examples that is accurate and reliable. In this presentation, we share our progress towards building this training corpus. First, we mined hundreds of real-world examples from the web. Then, we analyzed these examples to understand how visualization users take complex D3 programs and break them down into easily understandable parts, which we call components. Currently, we are investigating how these different components can be remixed and reused to create D3 visualizations for datasets that a model may not have encountered before. Future research will involve collecting more examples to grow our corpus, and training AI to use the corpus to generate documentation and relevant examples to help new visualization users better understand D3.

Attention-deficit/hyperactivity disorder (ADHD) is a neurodevelopmental condition characterized by differences in attention, focus, and emotional regulation. We investigate the relationship between parent temperament, including negative affect, and emotional socialization in ADHD parent-child dyads (parents and their children with ADHD).  Temperament refers to innate behavioral traits shaping one’s personality. Individuals with ADHD are known to experience higher rates of temperamental negative affect. Negative affect is a temperament factor that includes significant aversion from feelings like sadness. Emotional socialization refers to response methods of emotion regulation and can be categorized as supportive and unsupportive. Our study is impactful because ADHD has high heritability rates, yet there is little research on parental ADHD symptoms and associated temperament differences’ effect on ADHD families. We hypothesize that parents who report more ADHD symptoms and/or higher levels of temperamental negative affect use less supportive emotional socialization strategies and more unsupportive strategies when interacting with their children with ADHD. To test this hypothesis, I assist with administration of parent self-report measures and assess negative talk during laboratory-based, video-recorded parent-child interactions. Parents complete the self-report Adult Temperament Questionnaire (ATQ), which evaluates negative affect frequency. We measure negative talk by coding frequency of verbal disapproval of the child’s behavior/ attributes during parent-child interactions, where I instruct parents to perform standardized tasks with their children and code parental verbalizations into categories like negative talk using the Dyadic Parent-Child Interaction Coding System (DPICS). I then use bivariate correlation analysis to determine the likelihood that the two variables occur together and are linked. Through our anticipated findings, we hope to better inform care for children with ADHD and provide more resources for ADHD parent-child dyads. We want to identify emotional state-based targets to use in parental interventions to better support emotional regulation strategies in ADHD families.

Traumatic spinal cord injury (tSCI) is a devastating condition that causes sensory and motor dysfunction and permanently impairs normal life. Spasticity is one of the most common complications associated with tSCI that limits independent functional abilities. Spasticity is defined as a velocity-dependent increase in muscle tone, in response to passive movement and it is accompanied by pain and stiffness. Unfortunately, current treatments provide only transient and often incomplete relief of spasticity and individuals often experience long-term adverse effects. Through a collaborative project between three labs, we aim to develop a durable non-invasive electrical stimulation treatment to alleviate spasticity. I participated in preparing the model of spasticity by performing spinal surgeries on the cervical spine of rats. To evaluate spasticity, we studied the loss of Rate-Dependent Depression (RDD) of the H-reflex which is considered the electrophysiological hallmark of spasticity. To do so, I fabricated an electrode nerve cuff that was implanted on the median nerve of the rodent’s forearm to study the H-reflex of the affected muscle in the rat’s forelimb. I then recorded and analyzed the temporal development and change of spasticity. H-reflex results validated the spasticity model by showing RDD reduction in injured rats compared to the uninjured rats. The developed treatment shows promising modulation of the H-reflex and recovery of RDD in injured animals. Additionally, to measure velocity-dependent muscle tone, we developed a robotic device that passively moves the rodent’s forearm at different velocities. Employing this robotic behavioral assessment allows me to objectively assess the effect of stimulation on spasticity in the rodent forelimb. Obtained data reveals the muscle resistance to be three times higher in the injured rodent. This novel therapeutic stimulation protocol could potentially be used for clinical use as a non-invasive therapy, to help patients with spasticity in the hand after suffering from cervical tSCI.

Syphilis remains a serious global health concern, underscoring the need for better control strategies. If left untreated, the syphilis agent, Treponema pallidum subsp. pallidum (T. pallidum), can persist for decades due to its ability to evade the host immune response. Antigenic variation of the surface-exposed outer membrane protein TprK is believed to mediate persistence. TprK contains seven discrete variable (V) regions. TprK antigenic variation occurs via non-reciprocal gene conversion between the variable regions in the tprK expression site and 53 donor cassettes (DCs). We previously engineered a T. pallidum strain impaired of antigenic variation (SS14-DC^KO) by eliminating 51 of the 53 DCs. Rabbits infected with the DC^KO strain developed an attenuated infection phenotype with a reduced burden of T. pallidum cells compared to wild type (WT). Therefore, we hypothesized that if immunosuppressed rabbits are infected with SS14-DC^KO, the disease would undergo similar progression to rabbits infected with the WT strain. In this study, two rabbit groups (n=8) were either immunosuppressed with Depomedrol or untreated. Four rabbits in each group were infected intradermally with SS14-WT or SS14-DC^KO strain on clipped backs. Sera was isolated weekly to measure antibody titers using VDRL and TPPA. In addition, DNA was isolated from lesion biopsies to perform TprK profiling. Based on TprK profiling data, synthetic peptides for the most abundant V5, V6, and V7 variants were used to measure antibody reactivity by ELISA. Antibody titers measured by TPPA and VDRL were similar between immunosuppressed rabbits regardless of infecting strain. TprK profiling shows that the most abundant V sequence at the inoculum decreases over time in lesions as humoral reactivity to these peptides increases. Overall, this research demonstrates the role that TprK plays in the persistence of T. pallidum during syphilis infection and the need for novel control strategies.

Parental emotion socialization is how parents can use positive strategies such as modeling and discussion of emotions with their children. This skill is especially valuable for families with attention-deficit/hyperactivity disorder (ADHD), a highly heritable condition that affects one in eleven children and one in fifteen adults in the United States. Up to 50% of parents with ADHD are affected by emotional dysregulation. However, there is a lack of knowledge of how parental emotion socialization is impacted by parental emotional regulation abilities in ADHD families. The current study investigates this relationship through laboratory-based recordings of parent-child interactions to measure parental emotion-focused reactions as well as EEG recordings to detect biomarkers related to emotional regulation abilities in parents. I lead these video-recorded parent-child interactions and am trained to code them using the dyadic parent-child interaction coding system (DPICS), which systematically scores these dyadic conversations. For the EEG portion of the study, data analysis includes examining event-related potentials of interest, including the late positive potential (LPP). The LPP relates to emotional expression in response to a visual stimulus and is the variable that relates to parental emotion socialization. We measure the relationship between frequency of observed parental emotion-focused reactions and EEG-based measures of parental emotion regulation. In addition to laboratory-based measures of parental emotion regulation and parental emotion socialization, self-report measures are additionally collected. We expect parent participants with greater neural responses associated with emotion regulation to demonstrate greater frequency of parental emotion-focused reactions when interacting with their child during observed parent-child interaction. Because emotional socialization is linked to predictors of mental health, academic success, and peer relationships, understanding this relationship is critical. Establishing a better understanding of how parental emotion regulation affects parenting allows for individualization of behavioral treatments for families with ADHD.

Prosocial behavior is important to many species and its disruption is a hallmark symptom of many diseases and disorders such as autism. Previous research has shown that the mesostriatal network, which consists in part of neurons located in the midbrain that make the neurotransmitter dopamine and release it downstream in the nucleus accumbens, plays an important role in prosocial behavior. Dopamine neurons express the potassium ion channel Kv7.3 and mutant variants of Kv7.3 have been identified in individuals with autism spectrum disorder (ASD). Here we investigate how the Kv7.3-R2C variant found in ASD patients impacts social behavior and dopamine release in mice. We use viral strategies to express human Kv7.3-R2C in mouse dopamine neurons and a genetically encoded dopamine sensor (dLight1.3) in the nucleus accumbens during a social operant task. We found that while Kv7.3-R2C expression decreased social reward in an operant task rewarded with social behavior, dopamine release was similar between groups. Findings from this study help improve our understanding of how deficits in the mesostriatal pathway may lead to decreased social interaction in patients with ASD.

Autism Spectrum Disorder (ASD) is a prevalent neurodevelopmental condition affecting approximately 1% of the world population. Characterized by impaired social interactions, the precise etiology of ASD remains elusive, but emerging research suggests deficits in the mesostriatal pathway may contribute to deficits in social reward processing. In order to better understand the role of the mesostriatal pathway in social reward processing, I performed in vivo microscopy coupled with viral delivery of genetically encoded fluorescent sensors to record nucleus accumbens (NAc) neuronal ensemble activity during an operant task in which mice were trained to press a lever in order to gain access to a social partner mouse. I found distinct populations of neurons that were more active or less active during the social reward task at discrete time points. Understanding the neurobiological mechanisms involved in social reward may pave the way for novel medications and treatments for ASD.

The Pacific Northwest (PNW) saw an unprecedented heatwave between June 25 to July 3 of 2021, with temperatures reaching up to 15℃ above the climatological mean. Previous research has examined the impact of this event on plants in Western Washington and Oregon through observational studies, and has focused on the economic implications for poor crop turnout. We used remote sensing data to take a top-down approach and examined how all plants throughout the PNW fared during and after this historical heatwave. Solar induced fluorescence (SIF) and Near-Infrared Reflectance of vegetation (NIRv) are two remotely sensed products that have been used to estimate plant health and gross primary productivity (GPP). SIF is more closely connected to plant processes like photosynthesis but has a short record (2018-2021) compared to VIIRS NIRv (2012-2021). We compared the responses of SIF to NIRv and found that both vegetation indices increased in trees and woody savannas, but decreased in grasslands and crops. However, SIF showed more intense and geographically larger increases in areas covered by trees. We then compared these vegetation indices to in-situ flux tower measurements of carbon fluxes, which did not always agree with SIF during the heatwave in woody areas. This study shows how remote sensing can further our understanding of how extreme events impact plant health, which is increasingly important as heatwaves become more intense and frequent in the future.

Skin cancer is the most common cancer in the United States, with 5.5 million annual cases. It is strongly associated with ultraviolet (UV) radiation that generates many mutations. There is thus a need to investigate what mutations drive skin cancer. The CDKN2A gene encodes the p16 tumor suppressor protein, and silencing of the CDKN2A gene plays a key role in cancer progression. Our previous mouse study showed that chronic UV irradiation to mouse skin induced mutations at the mouse Cdkn2a/p16 promoter. However, the impact of the mutations at this promoter on cancer progression remained unclear. We hypothesized that these UV-induced mutations at the Cdkn2a/p16 promoter may inhibit the binding of transcription factors, thereby suppressing the expression of the p16 tumor suppressor and leading to cancer development. The mutations at the Cdkn2a/p16 promoter were found in the DNA sequence that was similar to the ETS transcription factor binding motif. Also, it has been shown that ETS2 is expressed in the skin. Thus, we tested whether mouse ETS2 binds to the mouse Cdkn2a/p16 promoter by measuring light signals from luciferase that is expressed under the control of the promoter. Overexpression of mouse ETS2 resulted in higher light signals than no overexpression control, indicating that ETS2 binds to the mouse Cdkn2a/p16 promoter. We also tested mutated promoters that carry the UV-induced mutations. With overexpression of mouse ETS2, the mutated Cdkn2a/p16 promoter showed lower light signals than wild-type promoter, implying that the mutation at the promoter inhibits the binding of ETS2 transcription factor to the promoter. Although further investigations are needed, these results suggest that UV-induced mutations affect the binding of transcription factors. This study highlights the importance of investigating promoter mutations that may contribute to cancer progression.

The antibiotic penicillin is highly effective at treating the STI syphilis, caused by the bacterium T. pallidum. However, the United States has seen increases in syphilis cases every year for the past 20 years; congenital syphilis cases have risen more than 219% from 2017 to 2021 and overall syphilis cases have risen 32% from 2020 to 2021. This situation demonstrates the need for an effective vaccine as current approaches are not working. The aim of this project is to utilize phage immunoprecipitation sequencing (PhIP-Seq) techniques to assist in the development of an effective vaccine in rabbits and eventually humans. To this end I have been using PhIP-Seq techniques to systematically profile the immune responses to vaccine candidates and T. pallidum infections in rabbits. When rabbits are immunized with a cocktail of three strains of the protein TprC we saw a protective immune response against treponemes (resulting in no viable treponemes) whereas an immunization with TprD saw reduced immune protection. I used PhIP-Seq methods - informed by next-generation sequencing (NGS) and differential expression analysis - to determine the epitope-specificity of antibodies in polyclonal serum samples from rabbits immunized with these vaccine candidates. Epitope-specificity comparisons between the resulting antibodies of the two immunogens can shed light on regions of these proteins critical for protection against treponemes. In the next few months I plan to integrate alanine scanning mutagenesis into the project to assess amino acid binding specificity and accurately identify crucial residues for antibody-binding. The fusion of scanning mutagenesis with PhIP-Seq will allow me and the other research scientists assisting with the project to refine of the effectiveness of our existing vaccine candidates.

Infants perceive speech and acquire language amidst noisy and complex auditory environments. Thus, elucidation of the cognitive mechanisms governing speech perception under noisy conditions is crucial. Cortical encoding of the speech envelope has been one approach used to study speech-in-noise perception in adults. For infants, research shows that Infant Directed Speech (IDS) facilitates cortical encoding of the speech envelope in quiet conditions more than adult direct speech. However, it is unclear whether infants are able to track the IDS speech envelope amidst competing speech. To investigate this, we recorded the neural responses from 40 typically-hearing infants (20 seven-month-olds, 20 eleven-month-olds) to continuous IDS using electroencephalography (EEG) in three conditions: Quiet, Co-located Noise, and Separated Noise. The target stimuli consisted of naturally recorded IDS produced by two female English speakers. The noise stimuli consisted of a four-person babble constructed from audiobooks read by 2 male and 2 female English speakers. We presented stimuli at an overall level of 70 dB SPL via speakers placed at 0°, +90°, and -90° azimuth to infants sitting on a caregiver’s lap in a sound-attenuated booth. Our team analyzed EEG signals using the Multivariate Temporal Response Function (mTRF) toolbox in MATLAB. This backward modeling approach assesses whether the stimulus envelope can be reconstructed based on the recorded neural responses. Reconstruction accuracies greater than chance were observed in all three conditions for the majority of infants, suggesting that we were able to decode the speech envelope in both quiet and noise. Participants demonstrated the capacity to process speech, even amidst competing auditory stimuli, emphasizing speech perception competencies from an early developmental stage. These results support using the envelope model and mTRF method as a feasible method for investigating the development of speech-in-noise perception in infants and young children.

As the world’s reliance on Lithium-ion batteries increases for technologies like electric vehicles, we need to improve battery performance. Traditional Lithium-ion batteries are composed of planar electrodes whose thickness can be optimized for energy capacity or charge rate (power). Thinner electrodes have a faster charge/discharge rate but low energy capacity while thicker electrodes have slow charge rates but higher energy capacity. Three-dimensional (3D) electrode structures that deviate from traditional planar electrodes can mitigate these trade-offs by allowing for fast ion transport while still maintaining a high ion quantity. One structure of interest due to its theoretical performance improvements shown in literature is a line patterned electrode. Line patterned electrodes have material and structural design features that greatly impact battery performance; it is therefore important to have methods to characterize and quantify features prior to battery testing. To rapidly and accurately analyze 3D line electrode feature sizes, we have developed an image processing code that analyzes cross-sectioned images of 3D line electrodes made from battery materials, enabling automated quantification of features such as line width, spacing and height. Cross-sectioned images are converted to black and white, which can then be processed by a function to detect the line edges and calculate the feature sizes. The results of our image processing code were compared to manual measurements to quantify accuracy. We draw connections between 3D line patterned electrode features and Lithium-ion battery performance to demonstrate how 3D electrode structures can be tuned to improve performance. This material is based upon work supported by the U.S. Department of Energy’s Office of Energy Efficiency and Renewable Energy (EERE) under the Advanced Manufacturing Office (AMO) Award Number DE-EE0009112. The views expressed herein do not necessarily represent the views of the U.S. Department of Energy or the United States Government.

Lithium-ion batteries (LIBs) are vital energy storage devices for electric vehicles (EVs). Conventionally, LIBs have planar electrodes that present trade-offs between energy and power (charge/discharge speed) due to ion diffusion limitations. EVs require a high energy battery to enable long mileage ranges while also being able to charge quickly (< 15 minutes). 3D battery electrodes can potentially overcome this trade-off, achieving both high energy and power by leveraging 3D structures that create fast ion transport pathways. However, a scalable manufacturing process for 3D electrodes is needed. We are investigating processes for this, and we need a method to characterize our 3D electrodes. There is no method to automatically quantify the features within these 3D structures, which is required for rapid, high quality analysis. By accurately measuring 3D electrode feature sizes, correlations between features and optimal battery performance can be determined. We hypothesize that fabricating fine 3D features (order of 10s of microns) will improve battery performance. To address this need, I have developed an image processing script that characterizes 3D electrode samples. I investigate how threshold values improve accuracy in comparison to manual measurements and am able to achieve < 10% error. I also connect the code’s feature size measurements to our manufacturing process operating conditions to inform how manufacturing conditions can be altered to precisely control feature sizes, which impact battery performance. We expect that higher operating frequencies for our manufacturing process will result in our target fine feature 3D electrodes, achieving high-performance Lithium-ion batteries. This material is based upon work supported by the U.S. Department of Energy’s Office of Energy Efficiency and Renewable Energy (EERE) under the Advanced Manufacturing Office (AMO) Award Number DE-EE0010226. The views expressed herein do not necessarily represent the views of the U.S. Department of Energy or the United States Government.
 

Many transgender and gender diverse (TGD) people desire gender-affirming hormone treatment (GAHT) to alleviate discomfort due to the misalignment of one’s gender identity with their secondary sex characteristics, though little is known about its effects on the body. One area of interest that may be affected by GAHT is the gut microbiota (GM). GM and the sex hormones (estrogen, testosterone, and progesterone) have been shown to interact bidirectionally, referred to as the “gut microsexome.” At puberty, the commensal microbiota of males and females diverge due to circulating sex hormones. This difference is hypothesized to contribute to sexual dimorphism of disease prevalence between cisgender males and females, although little research on the effect of GAHT in the TGD population exists. Alteration of the gut microbiota, or dysbiosis, also has many adverse effects that overlap those of testosterone GAHT (THT) such as acne, weight gain, and hypercholesterolemia. Dysbiosis has also been shown to lead to intestinal and systemic inflammation by disrupting immune function. More information on the effects of THT on the gut microbiota is necessary to counsel transmasculine clients effectively. In this study, we injected mice born of the female sex biweekly with testosterone enanthate dissolved in sesame oil versus sesame oil alone. We profiled GM of mice throughout treatment using 16S rRNA sequencing and measured markers of inflammation in serum to assess the effect of THT on both the population of GM and intestinal and systemic inflammation. We expect that the mice receiving THT will have differentially abundant gut microbiota and increased concentration of inflammatory markers compared to controls. The findings of this study will serve as a basis for further studies exploring additional analysis of the gut microbiota and inflammation in both transmasculine and transfeminine people receiving GAHT.

Atherosclerosis is characterized by the accumulation of lipids, inflammatory cells, and fibrous tissue in arterial walls, forming plaques. Plaque accumulation can lead to stenosis and potentially severe outcomes such as myocardial infarction or stroke. The gut microbiome, including Collinsella aerofaciens, is believed to play a role in the prevention or development of atherosclerosis. Gut bacteria can directly influence systemic inflammation, a factor correlated with the pathogenesis of atherosclerosis, and produce metabolites that alter the disease course. This study explores the potential link between C. aerofaciens and atherosclerosis by investigating the abundance of C. aerofaciens in the gut microbiome of individuals with and without atherosclerosis. We collected 179 stool samples from participants at the Kisumu District Hospital HIV Clinic in Kenya and conducted a comprehensive analysis of their gut microbiomes. 100 participants had carotid ultrasonography, categorized as showing atherosclerosis with visible plaque or intima medial thickness ≥ 0.7 mm. We employed bacterial 16S ribosomal RNA gene sequencing to characterize the stool microbial composition and noted that the relative abundance of C. aerofaciens was 2.6-fold less in participants with atherosclerosis (p=0.006). To validate these findings, I employed a Quantitative Polymerase Chain Reaction with a cloned plasmid control for targeted quantification of C. aerofaciens. We found 6.9-fold more C. aerofaciens copies per total 16S in Kenyan adults without atherosclerosis versus with (p=0.020). This suggests a potential protective or mitigating role for this bacterium in cardiovascular health. Future work could include assessing changes in C. aerofaciens abundance over time and its association with cardiovascular disease progression. Additionally, in vitro or preclinical studies could reveal the specific mechanisms by which C. aerofaciens influences atherosclerosis development and progression. This research contributes to our understanding of the intricate interplay between the gut microbiome and atherosclerosis, offering insights that may inform future therapeutic strategies and personalized interventions for cardiovascular diseases.

In the rapidly evolving landscape of remote work, the challenges associated with recalling important information from meetings and missing meetings have increased. One potential solution is to use large language models (LLMs) to summarize meetings to help participants catch up after meetings are over. To have a better understanding of this topic, we systematically reviewed 17 existing commercial tools and research prototypes for LLM-generated meeting summaries. The results show that existing solutions fell short of supporting users to verify and validate the comprehensiveness and accuracy of the generated summary, hindering users from trusting the summary. To address this, the project aims to design and build a more trustworthy LLM-generated meeting summary tool. Specifically, we propose that LLM-generated summary should progressively display relevant meeting information based on the importance of the information and the user’s goals, and include trustworthiness cues to aid users in making accurate trust judgments of the summary. Our preliminary interviews and a literature review showed that users are more hesitant to trust the AI summary when the information is consequential, such as when they missed the meeting or specific action items. While trustworthiness cues such as quotes or links to raw transcripts could increase users’ trust, irrelevant and redundant information erodes people’s trust. To further validate these observations, we will conduct a formative interview study. We will show participants mid-fidelity prototypes exemplifying the key design decisions and elicit their feedback on appropriate trustworthiness cues, desired ways to indicate their goals and intentions, and expectations on the importance of different portions of a summary. These empirically supported insights will inform the final design of a trustworthy LLM-generated meeting summary tool, which we plan to implement and evaluate in the next step.

The Poolos Lab has made significant progress in elucidating the regulation of tau phosphorylation in the brain and its relationship to both Alzheimer’s disease (AD) and temporal lobe epilepsy (TLE). However, conflicting data exists in the field regarding whether patients with epilepsy exhibit increased or decreased tau phosphorylation and expression compared to health controls. From our preliminary findings using mass spectrometry, we hypothesize that tau undergoes dephosphorylation at several amino acid sites in TLE, as opposed to the hyperphosphorylation observed in AD. To validate this hypothesis, I conducted western blots to separate sample proteins based on their molecular weight via gel electrophoresis. Western blotting has increased sensitivity compared to mass spectrometry in measuring protein phosphorylation levels. I assessed changes in tau expression and phosphorylation using phospho specific antibodies that quantify site specific tau phosphorylation levels. These samples are derived from hippocampal tissues obtained from a chemo-convulsant rat model of TLE, which mimics the chronic seizures experienced by human patients, and compared to age-matched naive controls. Densitometry is employed to quantify the relative amount of phosphorylated tau, and a two-tailed t-test statistical analysis confirms significant changes in tau phosphorylation and expression between tissues from chronically epileptic animals and control subjects. Additionally, I plan to conduct western blots on human tissue from TLE patients to generalize our findings from animal models, deepening our understanding of tau dysregulation in epilepsy. Given the increased risk of premature death and adverse effects on physical and mental health experienced by epilepsy patients, our research holds significant implications for the well-being of epileptic patients and their loved ones. By identifying alterations in tau phosphorylation, we aim to develop a biomarker of epilepsy in cerebral spinal fluid and the blood, thereby advancing diagnostic tests and potential treatments for epilepsy.

The drug resistance in tuberculosis (TB) is a rising concern for the diagnosis and treatment of the disease. Being able to detect the presence of drug resistance accurately and rapidly in the patient strain is essential for improving individual treatment outcomes and reducing further transmission of resistant strains, which are more costly and difficult to treat than drug-susceptible strains. However, the current methods come in short in point-of-care (POC) settings, due to problems such as long processing time, high complexity, and necessity for specialized personnel/equipment. Oligonucleotide ligation assay (OLA) provides a high sensitivity and specificity against TB drug resistance, and here, I have developed a novel lateral flow test (LFT) device that incorporates OLA into it, which have shown comparable specificity and sensitivity against traditional protocol of OLA in lab setting followed by LFT. Moreover, the simplicity of the design enables further incorporation of other techniques such as isothermal DNA amplification, for a compact, one-step TB drug resistance diagnostic device for low-resource environment. 

The Campanian stage of the Cretaceous (~84–72 million years [Ma]) was the zenith of dinosaur diversity. Western North America is highly fossiliferous and preserves Campanian-age rock units throughout the Western Interior Basin. Most studies that investigated dinosaur diversity from this interval used data obtained from macrosites (e.g., skeletons), whereas few have investigated vertebrate microfossil sites. Vertebrate microfossil sites are a rich source of data on biodiversity (e.g., taxon richness, relative abundance) and how it changes through time. The Judith River Formation of north-central Montana is rich in vertebrate microfossil sites, preserving 4 million years of the Campanian (~79–74 Ma). Here we aim to observe patterns of dinosaur diversity in the Judith River Formation by quantifying dinosaur taxon richness and relative abundances based on dinosaur teeth from two stratigraphically and temporally separated microfossil sites. These sites are the lower Makela-French 1 (~77 Ma) and the upper Clamfetti (~75 Ma). Presently, we have 300 specimens out of a planned 400. We hypothesize that changes in diversity and abudance occurred between these two sites. Our preliminary results reveal a change in dinosaur diversity between Makela-French 1 and Clamfetti. Hadrosaurs and ceratopsians are present and relatively abundance at both sites, whereas ankylosaurs decrease in abundance from Makela-French 1 to Clamfetti. Small herbivores like pachycephalosaurs and hypsilophodonts are rare at both sites. Theropods show similar patterns to the herbivore’s trends. Tyrannosaurs and dromaeosaurs are common at both sites, whereas troodontids are absent from Makela-French 1. These preliminary findings reflect diversity patterns that are not easily observable solely through the collection of dinosaur macrofossils. Our continued collection of fossils from Makela-French 1, Clamfetti, and additional sites will increase our sample size and provide better fine-scale resolution of dinosaur diversity patterns during this crucial interval in their evolution.

The Cretaceous-Paleogene (K-Pg) mass extinction (66.052 Ma) is one of the most important events in mammalian evolution as it was the catalyst for mammals to diversify and fill the ecological holes left by the extinction of the non-avian dinosaurs. This extinction event impacted all groups of mammals, including the multituberculates, one of the longest-lived and most successful clades of Mesozoic and early Cenozoic mammals. The Constenius vertebrate fossil locality is in the lowermost Tullock Member of the Fort Union Formation in Garfield County, northeastern Montana, deposited within the first 28,000 years after the K-Pg mass extinction (66.052-66.028 Ma). Constenius is a very rich but understudied fossil locality that provides a snapshot of the immediate aftermath of the mass extinction. In this study, we used qualitative descriptions partnered with linear measurements to identify 44 lower fourth premolars (p4s) to the lowest possible multituberculate taxon. We recognize three genera of multituberculates from Constenius: Cimexomys, Mesodma, and Stygimys. The presence of these multituberculates supports the previous assignment of Constenius to the Pu1 interval zone of the Puercan North American Land Mammal Age (early Paleocene, 66.052-65.820 Ma). Further work on this project will include expanding the dataset to include other multituberculate dental specimens, such as upper premolars, and conducting a geometric morphometric analysis with the lower fourth premolar specimens to further confirm taxonomic identifications.

Vaccines have successfully reduced global infectious disease burden, but there is room to improve vaccination technologies. Because many pathogens infect at mucosal sites, a goal of new vaccines is to promote strong mucosal and systemic antibody and T-cell responses. Integrated fiber microneedle devices (iFMN) are a novel oral vaccination method that may achieve this goal. These devices are patches with a polymer backfill matrix and multiple >1 mm pyramidal needles that penetrate immune cell-rich mucosal tissue in the mouth, inducing immune responses at draining lymph nodes. To test the hypothesis that priming with iFMN delivery of a DNA vaccine increases mucosal and systemic antibody responses after systemic booster immunization with the same vaccine, male rhesus macaques (n=6) were primed with an iFMN delivery of a DNA vaccine encoding Influenza A Virus (IAV) Nucleoprotein (NP) at weeks (0) and (6). The macaques then received a single boost of the same NP DNA vaccine at week (12) using the proven delivery modality of Gene Gun epidermal delivery (GG). Mucosal secretions (including bronchoalveolar lavage, saliva, and nasal/tracheal swabs) and serum were collected 2-4 weeks before and after each immunization. I conducted enzyme-linked immunosorbent assays (ELISAs) to quantify antigen-specific IgG and IgA binding antibody at each timepoint. To characterize the priming effect of iFMN oral delivery on systemic and mucosal antibody responses, I compared these animals’ responses to macaques (n=8) previously immunized with a single GG dose of the same NP DNA vaccine. The iFMN-primed animals had robust post-GG boost NP-specific IgG responses in serum but these responses were not significantly higher than for macaques boosted solely with GG DNA. These results demonstrate that iFMN delivery did not effectively prime for robust systemic and mucosal antibody responses. Additional experiments will be done to confirm these findings.

 The human influenza A Virus (IAV) is a contagious respiratory pathogen that can lead to severe maternal illness during pregnancy. IAV infections during pregnancy impart a higher risk of preterm birth, but the mechanism is unknown. We used a nonhuman primate (NHP; pigtail macaque, Macaca nemestrina) model to investigate the immunopathogenesis of IAV infection during pregnancy with a focus on the link between lung infection and placental injury. We hypothesized that a greater IAV infection in the lungs of a pregnant macaque would result in immune injury of the placenta. Pregnant macaques (n=8) were inoculated with 1 x 10^7 plaque forming units (PFU) of the IAV strain A/California/07/2009(H1N1) or media. Blood and bronchoalveolar lavage fluid (BAL) was collected at baseline and at several time points until Day 5, the study endpoint, when Cesarean section and necropsy was performed. I used a tissue culture infectious dose (TCID50) assay to quantify IAV in maternal lung tissues. The TCID50 titer, determined at the dilution where 50% of wells exhibit cytopathic effects, provides a quantitative assessment of actively replicating virus. Further, the laboratory quantified cytokine and chemokine concentrations from the placenta providing an opportunity to correlate viral loads in the lung with placental immune response. All lungs had evidence of replicating virus by TCID50 with a median PFU of 8 x 10^3. Higher viral loads were significantly correlated with higher concentrations of placental interleukin-1 beta (R= 0.94, p<0.0016) and interleukin-4 (R= 0.97, p<0.005). These results are the first to link the IAV viral load in the lungs with the cytokine response in the placenta. In summary, the data suggests that a higher load of replicating virus in the lungs correlates with a placental inflammatory response that may increase preterm birth risk.

Human Immunodeficiency Virus 1 (HIV-1) is a lentivirus and the causative agent of Acquired Immunodeficiency Syndrome (AIDS). HIV encodes a viral protease, the function of which is required for viral replication. The host innate immune sensor CARD8 detects HIV protease activity, leading to inflammasome activation during HIV infection. Inflammasomes are cytosolic innate immune complexes that recruit Caspase-1 and lead to secretion of pro-inflammatory cytokines and lytic cell death. Humans encode a single CARD8 gene; however Old World Monkeys (OWMs), the hosts of Simian Immunodeficiency Viruses (SIVs) encode two copies of CARD8. The function of the OWM CARD8 is unknown. To characterize the function of OWM CARD8s, I cloned CARD8 from representative OWMs and tested their responses to HIV and SIV protease in two ways. First, I determined if OWM CARD8s are capable of forming an inflammasome in response to HIV-1, a panel of SIVs, or the broad CARD8 activator VbP. I found that most, but not all, OWM CARD8s are functional but not responsive to HIV-1/SIVs. Human CARD8 senses HIV-1 through viral protease cleavage of its N-terminus. To determine if this lack of response of OWM CARD8s is due to the absence of viral protease cleavage, I will next perform western blots comparing human and OWM CARD8 proteolysis in the presence of absence of HIV/SIVs. My data suggests that the species-specific differences in CARD8 alters its capacity to detect viral proteases. We speculate that the absence of HIV-like pathogenesis in OWMs with endemic SIV may in part be due to the absence of CARD8 inflammasome activation to SIV protease.

Through the production of a 150-page policy report, our team analyzes international activity in the Arctic related to the loss of summertime sea ice that opens new shipping routes, creates new opportunities for research extraction, and dramatically impacts the way of life for Indigenous Arctic communities, notably the Inuit. In the past decades, the Inuit and other Arctic Indigenous peoples have organized and created for themselves significant political power, leading to self-determination and self-governance initiatives and involvement in the Arctic Council, which is the leading forum for decision-making on Arctic issues. However, the growing economic value of the region also attracts non-Arctic states like China, who want to become a part of decision-making discussions on the region and benefit from the seasonal open ocean in the future. Given the growing power of Indigenous peoples and growing interest in the region, our report addresses two questions: what impact and influence do the policies of Arctic Indigenous Peoples, particularly Inuit, have on the roles of nation-states including non-Arctic nations such as China? And what are the implications of China’s Arctic policy on Arctic Indigenous Peoples, Canada, the United States, and beyond? We use qualitative research methods, mainly the review of primary and secondary source documents that situate our analysis. Our report also draws on meetings with government agencies and indigenous organizations in Ottawa. We will produce a series of recommended policies and areas for further attention that are meant to inform and guide nation states and organizations as they engage in the Arctic. Our analysis of Indigenous and international relations in the Arctic is important because the Arctic represents a unique and unusually successful arena for cooperation between nation states, Indigenous representation, and other non-state actors. This cooperation will only become more vital as global activity in the region increases.

Mentoring is an effective way to improve diversity and retention in STEM fields. The UW Psychology Undergraduate Mentoring Program (PUMP) matches psychology students from underrepresented groups with mentors, including undergraduates, graduate students, faculty, and alumni. Matches are made based on a combination of career interests and personal identities. In this study, we aim to understand predictors of mentees’ success in the program, defined by feelings of belonging, academic preparedness, and satisfaction with the mentoring relationship. These predictors include meeting characteristics and match characteristics. We hypothesized that in-person meetings, more frequent meetings, and greater similarities between mentors and mentees would be linked with greater success. 32 out of 168 mentees completed our survey on Qualtrics in Spring 2023. We measured meeting frequency (“How often did you meet with your mentor during [quarter]?”), format (“In what modality did you meet with your mentor? In-person, video/phone calls, texting, email”), match characteristics (e.g., whether the mentor and mentee shared similar demographics such as BIPOC, first-generation, or gender; how far apart they were in their class levels), belonging (e.g., “I know other people in the psychology major”), preparedness (e.g., “I feel prepared to pursue the psychology major at UW”), and satisfaction (e.g., “My mentor and I have had a successful mentoring relationship”). We will conduct correlations and multiple regression analyses to test the prediction that in-person meetings, more frequent meetings, and greater similarity between mentors and mentees will be linked with higher feelings of belonging, preparedness for the major, and satisfaction with the mentoring relationship. We hope to demonstrate that controllable variables such as these can improve program outcomes. Identifying the most effective program strategies, like in-person meetings or demographics-based matching, can aid mentees in our program and may also guide the development of new mentoring programs in other departments.

Mycobacterium abscessus are non-motile bacilli that cause soft-tissue and pulmonary infections, commonly in healthcare settings or patients with cystic fibrosis. Though it is considered an opportunistic pathogen, its many virulence factors signal its potential for evolution into a true pathogen. Upon infection, the bacilli are internalized by macrophages, forming granulomas to contain the infection. Macrophages can harbor bacilli during infection stages and induce drug resistance by expelling toxins through ABC transporters. Treatment is often challenging as M. abscessus is intrinsically resistant to many antibiotics. Current treatment uses a combination of two or more intravenous drugs and one or more oral antibiotics over several months. Treatment success is challenged by patient adherence and may also be impacted by drug efflux by macrophage ABC transporters. Transporters identify certain drugs as toxic to the body and try to flush them out of the cell. Since Mycobacteria infect macrophages, these channels pose a significant disadvantage to treatment since the cell will actively efflux the drug, preventing the drug's intracellular concentration from increasing to an effective level against the bacilli inside. Certain drugs are known to inhibit ABC transporters, and the addition of these inhibitors in treatment could increase bacteriocidal activity and reduce the development of drug tolerance. First, we will determine the drug's minimum inhibitory concentrations to each inhibitor to see if there is an intrinsic activity on M. abscessus. Next, we will use the Human THP-1 cell line infect with Mycobacterium abscessus and treat with known ABC transport inhibitors in concert with a current therapeutic drug, Clarithromycin. They will then be plated at various time points to determine the colony-forming units. If efflux by macrophage transporters reduces the efficacy of Clarithromycin, bacteriocidal activity will increase between the combination therapy and the clarithromycin-only treatment. These results may improve the current treatment regimens for M. abscessus.

When a cell undergoes stress conditions, such as oxidation or aging, an increase in protein instability can occur and prevent proper cell functions. Small Heat Shock Proteins (sHSPs) are molecular chaperones that work to maintain a healthy proteome by associating with misfolded “client” proteins to delay aggregation under such conditions. HSPB5, a human sHSP, is ubiquitously expressed throughout the body. HSPB5’s disease mutant, R120G, is a defective chaperone associated with cataracts and desmin-related myopathy. It is still unknown how this mutation is detrimental despite many years of research. My research aims to understand how this mutation retunes the electrostatic properties of HSPB5, affecting its chaperone activity. Residue R120 is part of an electrostatic network that helps create an important structural feature in the folded region of HSPB5, the alpha-crystallin domain (ACD). In the unmutated (WT) protein, the ACD surface is overall positively charged. Substitution of the positive R120 to glycine alters both ACD’s structure and electrostatics. I generated two mutants, R120K (retaining positive charge) and R120D (switching to negative charge) to investigate how R120 plays a role in ACD’s conformation. Using a negatively-charged molecule, ATP, as an “electrostatic” probe in 2D NMR, I observed differences between its binding affinity to my R120 variants. I found that only R120K ACD behaves similar to WT ACD, suggesting a possible correlation between charge potential and ACD’s interactions with ATP. Currently, I am investigating if charge potential affects chaperone activity through aggregation assays with a client protein, human γD-crystallin, found in the lens and implicated in cataracts. I predict that WT and R120K, with similar electrostatic properties, will have similar chaperone activity. R120G and R120D, prevalently in an “active” state, will have higher chaperone activity. Understanding how such mutations affect HSPB5’s conformations and chaperone activity is a step forward in understanding sHSPs’ chaperone mechanism.

Small heat shock proteins (sHSP) are a family of molecular chaperones whose function is to delay the harmful aggregation of other proteins. Protein aggregation is associated with neurological disorders such as Alzheimer's disease and Parkinson's disease. In many tissues, multiple sHSPs are coexpressed and tend to assemble into hetero-oligomers. Hetero-oligomers are complexes of two or more different protein species. The extent and mechanism by which these hetero-oligomeric complexes form is yet to be fully understood. The goal of my discovery-driven research is to assess how the properties of sHSP hetero-oligomers differ from the properties of homo-oligomers. In my project, I focus on three sHSPs that are highly expressed in muscle: HSPB1, HSPB5, and HSPB6. Each of these proteins exhibit different behavior when on their own. HSPB1 and HSPB5 form a distribution of large homo-oligomers, whereas HSPB6 forms a small homo-dimer. One of the most characteristic properties of the small heat shock proteins is formation of oligomers that span different sizes. Thus I am primarily determining the sizes and composition of the sHSP hetero-oligomers. I performed a comprehensive study to characterize the sizes of the hetero-oligomers using three complementary methods: analytical size exclusion chromatography, mass photometry, and native gel electrophoresis. I have found that HSPB6 is able to readily incorporate into hetero-oligomers as the concentration of the other sHSP is increased, and that the complexes are formed in a distribution of intermediate sizes. I am currently working on assessing the ability of the hetero-oligomers to act as molecular chaperones by aggregation assays. I predict the hetero-oligomers will delay protein aggregation more efficiently than HSPB6 on its own. The findings of my project give insight into why sHSPs are coexpressed and form hetero-oligomers in cells. Understanding these hetero-oligomers sheds light into the complex pathways of sHSP function. 

Phosphoribosyl Pyrophosphate Synthetase (PRPS1) is an enzyme in the nucleotide biosynthesis pathway that makes a molecule necessary for de novo nucleotide synthesis. It is known that PRPS1 protein hexamers can stack into linear filaments in the presence of ADP and phosphate. When these filaments are broken, catalytic activity is lost, and it is hypothesized that enzyme inhibition is lost as well. Mutations in PRPS1 lead to a wide spectrum of diseases in humans. In addition, changes in cell regulation of the enzyme have been linked to cancer. Motivated by research that connects PRPS1 phosphorylation to increased cancer proliferation, my project investigates the effects of phosphorylation on PRPS1 structure, enzyme activity, and inhibition properties. I have transformed plasmid DNA containing the PRPS1 phosphomimetic mutations S47E, S103D, and S308E into E. coli strains BL21 and pLysS. I then grew overnight bacterial cultures and induced protein expression using IPTG. After verifying protein expression with gel electrophoresis, I purified the protein from bacteria using nickel resin affinity and size exclusion chromatography. Having made and purified protein mutations that mimic phosphorylation, I conducted a negative stain screen to analyze filament formation trends. This has yielded preliminary findings that S47E and S103D phosphorylation mutations of PRPS1 break enzyme filament formation. Variation in filament formation between mutations points to the importance of phosphorylation location and its potential impact on enzyme activity and inhibition. To assess the catalysis of the phosphomimetic mutations in PRPS1, I will conduct biochemical assays which measure the activity and inhibition of the enzyme. Through these ongoing experiments we will learn how phosphorylation modifies PRPS assembly and activity and the implications of PRPS1 dysregulation in cancer proliferation.

Elementary school educators are primary influences on children’s development by helping shape their behavior with peers and in the classroom. Employees in education are deferred to by their newer coworkers regarding student interactions and set precedents for students’ school environment. New and seasoned educators’ disciplinary practices greatly vary when addressing students exhibiting problematic behaviors which may differently affect student-instructor relationships. I use an existing dataset from a larger study testing the impact of a novel implementation strategy on the implementation of a universal social, emotional, and behavioral program in nine elementary schools from one district. Educator participants in this study completed a survey battery when starting and ending the school year and the district provided quarterly discipline records via a universal prevention program. The findings identify trends in student disciplinary actions between new (<5 yrs) and experienced (>5 yrs) instructors. Understanding the relationship between teacher tenure and the treatment of student misbehavior is crucial to demystifying the manifestation of poor student enrichment and hostile student-instructor relationships. These findings could suggest how differential disciplinary practices by educators based on their years of experience influence student learning.

Seriously ill hospitalized patients and their families experience elevated stress, anxiety, and depression, resulting from factors like the complexity of serious illness, fragmented medical care, and transitions across settings. They struggle to navigate treatment options, make difficult decisions, and express their values and goals to their clinicians. In addition to communication challenges between patients and clinicians, poor communication among healthcare team members exacerbates the stress associated with serious illness. The aim of this study was to continue work from previous Facilitating Communication Studies to gain knowledge from bedside nurses' experiences with collaborative communication (CC) between patients and families and their healthcare team, and to elicit ideas from nurses about what might improve CC within the hospital system. We defined CC as patients and families being active participants with their healthcare team working towards shared goals of care. We recruited and interviewed 15 bedside nurses from ICU and acute care units at Harborview Medical Center. Following each interview, we input each participant's audio-recorded content into a matrix table, created summaries of each transcript, and synthesized content across transcripts to distill major themes. Nurses reported that breakdowns in CC most often occur when multiple clinicians are involved, during transitions of care, and during the discharge process, resulting in confusion and distress for the patient, family, and bedside nurse. Suggested solutions to overcome these barriers include (1) promoting in-person interactions by rounding as an interprofessional team, (2) inviting bedside nurses to goals of care meetings, (3) implementing a CC training program for clinicians to standardize the CC approach among team members, and (4) empowering patients and families to initiate CC by educating them about their role in their healthcare. These suggestions should be implemented in a future Facilitating Communication Study to reduce distress for seriously ill patients, their families, and their healthcare team. 

The climate crisis is predicted to increase the occurrence of extreme weather events such as heat waves and polar fronts, which will disproportionately affect vulnerable populations such as people experiencing homelessness (PEH). As climate change increases social and economic instability, policymakers need to be informed on how to effectively protect the interests of our most vulnerable populations. Building off of studies that have analyzed the impact of heat and cold on hospitalizations among PEH, I examine the health effect of extreme weather events in King County, San Francisco County, and Los Angeles County. Temporary emergency shelters are one of the main strategies used to protect the wellbeing of the PEH population during extreme weather events. Moreover, I intend to determine the effectiveness of emergency shelters in reducing the impact of extreme weather on the death toll and/or health complications of PEH. Utilizing data on health impacts among PEH, daily maximum/minimum temperatures, and cooling shelters, I will use regression models to determine the relationship between maximum/minimum daily temperature and health outcomes and if cooling shelters reduce negative health impacts during extreme weather events. I expect to find a positive relationship between extreme temperatures and hospitalizations, as prior studies have observed, but that this relationship will be weaker on days on which cities open emergency shelters. However, it is plausible that because emergency shelters have restrictions on access, limited bed capacity, or inadequate public outreach, the impact of emergency shelters on hospitalizations may be insubstantial. My results will contribute to a growing body of literature that points to a relationship between extreme weather and poor health outcomes for PEH. This evidence will have implications for the importance of allocating funding towards the shelter system and other social service efforts, while clarifying the effectiveness of the emergency shelter system.

While much has been done to understand rape as an act of genocide and as a war tactic, little scholarly work has focused on how the intentional wartime spread of HIV/AIDS in the Rwanda genocide led to a slow genocide of the Batutsi. This work argues that the spread of HIV/AIDS resulting from the rapes that occurred during the Rwandan Genocide has led to the continuation of the slow genocide against the Batutsi people. I analyzed seventeen semi structured interviews of rape survivors that were conducted between 2007 and 2008. These interviews were published in Sandra Chu et. al. The Men Who Killed Me Rwandan: Survivors of Sexual Violence. By using thematic coding, my analysis of the interviews concluded that Tutsi women were intentionally infected with HIV by the Interahamwe so that the women themselves would be turned into biological weapons of genocide that could be used to inflict a delayed death-sentence on another Tutsi. Roughly 80% of the women who survived the Rwanda Genocide were raped, and of those women, 70% contracted HIV. With the lack of medical treatment in Rwanda for HIV/AIDS, those who test positive for the disease have 7-15 years to live before they will die a slow death. The intentional viral spread of HIV and the subsequent deaths from AIDS is a direct result of sexual violence committed as a tactic in the 1994 genocide. Despite being purposefully killed due to their ethnicity, women who died of HIV-related illness in Rwanda are not calculated in the death toll for the genocide against the Batutsi in 1994. This work expands our conception of the long-term effects of the rape campaign led by Hutu militias that intended to inflict death upon survivors of the initial violence from April to July 1994 in Rwanda.

Why does human rights law fall short in protecting people from human rights abuses made by governments acting in the name of self-sovereignty? In this project, I analyze these questions in the context of human rights abuses, such as extrajudicial killings, false imprisonments, rapes committed by law enforcement officers, and forced disappearances in Panjab, India. To do so, I use official reports from non-governmental organizations, an original interview I conducted with a survivor of Operation Blue Star, which was an attack on one of the holiest shrines for Sikhs, process-tracing, and human rights legal analysis. After developing the political and legal framework, I present my research in the form of three narrative case studies. I argue that the acts orchestrated by the Indian government and the tactics they employed to eradicate Sikhs constitute genocide under human rights law. Nonetheless, Sikhs struggle in gaining recognition due to extreme efforts by the Indian Government to cover-up and malign the names of Sikhs through their own governmental groups, and an international political environment that discourages foreign governments from condemning Indian government actions. With the research conducted in this project, links between other human rights violation in India can be found. The lack of accountability of the Indian government in multiple cases can be uncovered, which will prove how even with the existence of human rights law, governments have the most power.

Despite the extensive literature on Canada and European countries, there is relatively little research on paid maternity policies in U.S. states due to its recency. Economic theory would suggest that fertility rates would increase with a paid maternity leave policy as the cost of having a child would be effectively subsidized. A strong family leave plan could be the deciding factor for many families to have a child. However, current research is inconclusive with there being no studies on U.S. fertility rates specifically in mind. The goal of my research is to provide insight on the way in which the U.S. fertility rate may be affected by paid leave programs. To accomplish these goals, I will be studying the implementation of paid maternity leave in Washington state specifically. I will be using a difference in difference method to measure the effect of this policy. I will be using data from the National Center for Health Statistics and the American Community survey for my fertility rate estimates. My research is in progress; however, I would expect either slight or no change in fertility rates in Washington based on previous research in the field. The goal of this research is further insight to nationwide implementation of this program by analyzing a specific possible implication of this policy and to contribute to the literature of paid maternity leave on fertility rates.

Nepal, despite its heavy reliance on agriculture, is witnessing a decline in agricultural productivity and an increasing dependence on food imports. This phenomenon, coupled with factors such as land fragmentation and labor shortages, has led to a masked "Silent Green Revolution" reminiscent of past agricultural movements. Agrochemical overuse and over-intensification has raised concerns about the long-term sustainability and health implications of such practices. In this paper I explore interplay between agricultural practices, farmer health, and environmental sustainability in Nepali farming communities. With the help of a translator, I orally conducted a Farm and Farmworker Health Survey in Phoolbari and Baluwa, two villages in the Kavrepalanchok District with both commercial and subsistence farmers. I found that while commercial farmers favored agrochemicals for their cost-effectiveness, subsistence farmers preferred organic alternatives for their safety and sustainability. However, there was a general lack of awareness among farmers regarding the symptoms of pesticide exposure and proper safety measures like Post Harvest Intervals and Personal Protection Equipment, indicating a need for targeted health and procedure awareness programs. Furthermore, the study highlighted the potential health risks associated with agrochemical use, including exposure to carcinogenic heavy metals like cadmium and glyphosate residues. These findings underscore the urgent need for sustainable agricultural practices that prioritize the health of both farmers and consumers. By centering traditional farming methods and indigenous knowledge and implementing initiatives that address occupational safety and provide proper training and infrastructure, Nepal can pave the way towards a more sustainable and equitable agricultural future.

The World Health Organization reports that drowning is the third leading cause of unintentional injury deaths worldwide. Intervention efforts are crucial to prevent drownings from occurring and maximize survival in people who do experience drowning. While multiple factors contribute to the risk of drowning, age affects drowning settings, patterns, and rates. Therefore, drowning prevention efforts should be tailored based on age. However, most drowning prevention interventions focus on pediatric populations, which fails to address the increased injury burden in older adults that disproportionately affects aging societies. To address this knowledge gap, our research team conducted a formal rapid review of published drowning prevention efforts in adults aged 50 years and older. We will analyze our results using Haddon's Matrix, a systemic tool that provides a comprehensive view of different types of prevention acting on discrete injury components. Our search strategy yielded 1914 titles and abstracts, of which we eliminated 1850 studies as irrelevant to our research question. We then conducted full-text screening on 64 studies, retaining a total of 7 studies for data extraction, which is currently ongoing. Preliminary results show discrepancies in the age definitions of older adults and a failure to address equity issues related to drowning prevention in this population. Many studies included older adults as part of the study population but did not stratify outcomes by age. Hence, no determination could be made regarding the intervention’s efficacy in older adults. Finally, most older adult drowning prevention efforts targeted a behavior change in the host rather than modifying the environment, which is often a more efficient method of injury prevention. Our preliminary results illustrate the need for a global collective effort to diversify drowning prevention and research efforts to alleviate the drowning injury burden in older adults.

As our relationship with public space continues to grow, pedestrianization remains an important tool for reclaiming our space for public social function. Pedestrianization of The Ave is both academically and statistically supported, yet there remains little movement in its favor. Through the lens of placemaking, this project examines the best practices for campaigning the pedestrianization of The Ave in a way that fosters community ownership. Incorporating insight from historical reviews and stakeholder surveys, this research identifies preferred engagement strategies for community stakeholders. Utilizing these findings this research conducts a pilot study and creative visioning that centers community culture. This will result in concepts of a pedestrianization that will best gain public support and feelings of community ownership. These strategies and materials will be compiled as a “campaign” that has the potential to be implemented in the near future. As community planning in the University District seems to be very disconnected from its stakeholders, this project is rooted in community empowerment, seeking to connect people to systems.

Exclusionary zoning laws — which limit population densities and land uses in specific neighborhoods — are a typical feature of American municipal land use regulation. An extensive body of evidence links traffic-related air pollutant (TRAP) exposure to adverse health effects. Using zoning data and a model of TRAP levels in cities across the Seattle metropolitan area, I hypothesize that TRAP exposure will be greater on average in zones where higher-density housing is an allowed use, and lower on average in zones reserved for lower-density housing. I used the software package QGIS to spatially join zoning and air pollution data and used the software package R to perform correlation analyses between zone types (classified by maximum population density) and three common TRAPs (NO₂, black carbon, and ultra-fine particles.) This research highlights the public health implications of normative policy regimes like exclusionary zoning. These results can assist elected officials and planners in pursuing a more geographically distributive approach to increasing housing supply in the Seattle area, in order to minimize the TRAP exposure burden – and associated adverse health effects – faced by residents.

Cytomegalovirus (CMV) is a nuclear-replicating DNA herpesvirus that rearranges the nucleus to form a kidney bean shape during infection. CMV-induced cellular rearrangement is disrupted by knockout of a histone variant, macroH2A1. This disruption leads to significantly decreased infectious progeny for CMV. We examined how different transcriptional profiles during CMV infection of macroH2A1 knockout cells and found several host genes were misregulated in the absence of macroH2A1. One such gene is KIF1A, a kinesin-3 motor protein found in neurons. I previously found that KIF1A is induced in primary human foreskin fibroblasts (HFFs) during infection, which is unusual since KIF1A is not normally expressed in fibroblasts. Interestingly, KIF1A is not induced during CMV infection in the macroH2A1 knockout cell line. This led me to hypothesize that macroH2A1 is required for induction of KIF1A expression. To test this hypothesis, I overexpressed KIF1A in HFFs to establish if I can rescue the defect in infectious progeny. Wild-type and macroH2A1 knockout HFFs transduced with a plasmid containing the KIF1A gene are analyzed using Western blotting and plaque assays to determine KIF1A expression and viral titers. I anticipate that the defect is rescued by overexpressing KIF1A in macroH2A1 knockout cells. CMV is the leading infectious cause of birth defects in the United States, making its mechanisms of infection a key area of study for development of antiviral therapies.

Skin is a densely innervated sensory organ that protects us every day from environmental trauma. As a barrier organ, skin is susceptible to frequent damage that must be promptly and properly healed to prevent infection and restore sensory function. Our lab uses adult zebrafish as a model to study skin injury and repair. Adult zebrafish skin is similar in composition to human skin and transparent, lending itself to high-resolution microscopy. Previous experiments in our lab revealed that dynamic, skin-resident immune cells known as Langerhans cells (LCs) rapidly engulf cellular and axonal debris after injury in the zebrafish skin. Calcium signaling regulates phagocytosis and cell motility in other immune cells, but the role of calcium signaling in LCs is unstudied. Through skin explant assays, various injury paradigms, and confocal fluorescence microscopy, I have established a model for monitoring calcium signaling in LCs. I found that LCs exhibit rapid, transient calcium flashes under homeostatic conditions. However, upon engulfment of large cellular debris generated by precise laser-ablation of skin cells, LCs exhibit an atypical sustained calcium signal lasting an hour on average. To test the requirement of calcium during engulfment by LCs, I treated skin with the drug Thapsigargin to perturb calcium flux. I confirmed that Thapsigargin increases intracellular calcium in LCs and keeps intracellular calcium concentrations elevated for hours after drug addition. During Thapsigargin treatment, I showed that LCs formed phagocytic cups around cellular debris but engulfed fewer laser-ablated corpses compared to controls. Thapsigargin-treated LCs also experienced normal migration to a wound site. My results indicate that calcium flux regulates LC engulfment of large debris, but not through migration. Identifying the molecular mechanisms underlying LC motility and debris removal is ultimately relevant to understanding skin repair and disease states in which the wound healing response is attenuated, such as in chronic wounds.

The synthesis of key organic molecules often requires toxic, expensive, non-reusable reduction agents and extreme conditions. In recent years, electrochemistry has emerged as a sustainable alternative to standard methods, but this approach is often hindered by high energy barriers for electron transfer to the substrate. Electrocatalysts address this challenge by shuttling charge between the electrode and dissolved substrates, accessing lower transfer barriers, and reducing the overall energy needed. Current electrocatalysts, however, cannot stabilize reactive intermediates, which often leads to harmful side reactions and degradation of the electrode. We hypothesize that redox-active supramolecular cages can address this limitation by both shuttling charge and providing unique microenvironments capable of stabilizing intermediates. Previously, we synthesized two tetrahedral supramolecular cages that incorporate redox-active perylene diimide (PDI) and pyromellitic diimide (PMDI) motifs. Using cyclic voltammetry, we then showed that both cages can lower the voltages required for the electroreduction of vicinal dihalides to alkenes, indicating electrocatalysis. To better understand these results, I used density-functional theory (DFT) calculations to obtain computer models of the PDI and PMDI cages. These DFT-optimized structures revealed significant differences in charge density between redox centers due to electron-donating functional groups, which may explain why the PMDI cage lowered the substrate reduction voltages more than the PDI cage. With these models, I have also studied the shape and volume of the cages’ internal cavities, thereby providing information about substrate compatibility. I am conducting additional DFT analysis to understand how modifications to the ligand motifs may alter the electrocatalytic behavior. By continuing to investigate supramolecular cages for reductive electrocatalysis, I aim to contribute to the development of low-waste synthetic strategies for the production of alkenes and other commercially significant organic compounds.

Vulcanized rubber, the main component of tires, is prized for its chemical durability and thermal stability. These properties, however, make disposal difficult and contribute to the increasing problem of polymer waste. To broaden the applications of end-of-life tires, we developed a method to chemically upcycle polybutadiene, a primary component of vulcanized rubber, via selenium-mediated allylic amination. We hypothesized that functionalizing the backbone of crosslinked polybutadiene with sulfonamide groups—without breaking their double bonds—would result in favorable thermal properties, creating a new life for the crosslinked polybutadiene. We used infrared spectroscopy and scanning electron microscopy with energy dispersive X-ray spectroscopy to confirm the aminated crosslinked polybutadiene’s molecular structure and differential scanning calorimetry and thermogravimetric analysis to measure its changes in thermal properties. Our research has future implications for the reduction of tire waste and reprocessing of other end-of-life crosslinked polymers.

Hydrostatic pressure can be used to tune the electronic properties of atomically thin layered materials by decreasing the interlayer spacing, thereby enhancing the strength of interlayer interactions. In moiré systems, pressure can be used to create flat bands in samples with twist angles away from the usual ‘magic angle’. Twisted trilayer graphene (tTLG) has a Dirac band superimposed atop a flat band. These two bands can hybridize in a finite displacement field making it possible to further tune the flat band. The flat band is host to a variety of flavor-polarized correlated states, which may be an important ingredient in generating the exotic superconducting phases seen in tTLG. Although pressure could provide a new avenue for tuning these correlated states, a high-pressure study has not previously been performed on tTLG owing to the challenges of applying pressure to layered 2D materials: limited sample space, difficulty mounting the sample, and challenges in establishing electrical contacts. In this talk, I will discuss advances we have made in addressing these issues via a custom printed circuit board (PCB). The PCB provides a sturdy platform for mounting samples, and has gold pads to enable wire bonding. I will also discuss ongoing high-pressure electrical transport measurements of tTLG nanodevices. This work could elucidate further the origin of the unusual superconducting phase seen in tTLG, and provide a blueprint for future high-pressure studies of 2D materials.

Solar energy is a promising form of renewable energy that will play a major role in reducing carbon emissions. Perovskite-based solar cells have attracted significant attention due to their high power conversion efficiency (PCE), which reached 26.1% this year, surpassing commercial silicon’s (23.3%). High PCE, low cost of materials, and ability to be solution processed make perovskite solar cells a prime candidate to replace silicon. However, efficiencies are still below the theoretical limit and these materials suffer from limited operational stability. To tackle these problems, scientists have focused on minimizing active layer and interfacial defects which act as barriers for charge extraction in a solar cell, lowering the device efficiencies. Defects also electronically dope the perovskite layer, changing the recombination kinetics in the sample. The goal of this project is to quantify how the electronic doping and defect concentration of the perovskite sample is affected by surface passivation treatements via fluence dependent photoluminescence (PL) and time resolved photoluminescence (TRPL) spectroscopy. By solving the kinetic equations at the basis of charge recombination, we can extract the rate constants that correspond to different charge recombination pathways. We pioneer a global fitting analysis to simultaneously fit TRPL and PL measurements for robust determination of these kinetic constants that are subsequently used to determine the doping density of films before and after passivation. We show that the electronic doping density is higher than previously reported in literature, and that this doping is reduced with a surface passivation treatment. We collaborate with the University of Arizona to correlate our measured electronic doping density to electrochemically measured defect densities on the same samples. This work will provide an implementable tool to quantitatively assess electronic doping and defect density values for various perovskite compositions which will be useful for optimizing future solar cell devices.

To form Human Immunodeficiency Virus type 1 (HIV-1) virions, the HIV-1 Gag polyprotein multimerizes, traffics to the cell membrane, assembles into virions, and buds as viral particles. HIV uses the infected cell proteins to support viral assembly and export from the cell. Although some cellular proteins have been identified that participate in viral assembly and budding, the spatial and temporal "cellular proteome" is not known. Insights as to the order of proteins involved in facilitating assembly and budding provide information on viral infection and potential therapy targets. I am utilizing a split-APEX2-mediated proximity labeling method to identify which host factors interact with Gag during assembly. The basis of the method is splitting APEX2 into two segments encoded into separate Gag sequences so that the APEX2 segments rejoin for complete enzymatic function when Gag dimerizes and multimerizes. Once the APEX2 enzyme reconstitutes, we activate it to biotinylate cellular host proteins within ~20 nm of Gag during the processes of virion formation. I aim to deliver this system to primary human CD4+ T-cells and macrophages via two lentiviral vectors containing transgenes for either AP-Gag-P2A-EGFP or EX-Gag-P2A-mCherry. Cells express the dual fluorescence markers EGFP and mCherry if both Split-APEX2 domains are present, and we enrich these cells by cell sorting. APEX2 is restored and enzymatically active, allowing proximity-dependent biotinylation of Gag-host cell proteins for SILAC-based quantitative proteomic mapping during virion assembly. We are using this method to quantitatively map the Gag-host protein "interactome" in the infected cellular microenvironment in subtypes of human primary CD4+ T-cells and macrophages. Therefore, this research studies Gag interactions of potential HIV-1 host dependency factors in CD4+ T-cells and macrophages, the natural cell populations for HIV-1 infection. Previously, we conducted a similar study in HEK293Ts, and we anticipate elucidating a similar Gag-host cell protein analysis with this study.

Human immunodeficiency virus type 2 (HIV-2) is endemic in West Africa and is also found in areas with socioeconomic ties to the region. Historically, advancements in the treatment of HIV-2 infection have been slow compared to HIV-1, and there is an urgent need to identify effective antiretrovirals (ARVs) for people with HIV-2 (PWH2) who harbor drug-resistant virus. Lenacapavir (SUNLENCA®) may help fill this gap in the HIV-2 treatment landscape. Lenacapavir is a long-acting, injectable ARV that targets the HIV-1 capsid protein and is approved for use in adults with multidrug-resistant HIV-1 infection. We recently showed that lenacapavir is highly active against HIV-2 isolates in culture, albeit with a 11- to 14-fold lower potency relative to HIV-1. These data suggest that lenacapavir binds to HIV-2 capsid cores in a manner similar to that observed in HIV-1. To further test this hypothesis, I am using site-directed mutagenesis (SDM) to introduce specific amino acid changes into the putative lenacapavir-binding region of the HIV-2 capsid protein. These changes fall into two categories: 1) mutations that convert the targeted residue in HIV-2 to the corresponding amino acid residue found in HIV-1; and 2) mutations that are associated with the development of lenacapavir resistance in patients with HIV-1 infection. To date, I have constructed a 5.5-kilobase plasmid clone that contains the capsid-encoding region of HIV-2ST (plus downstream flanking sequences) in a pBluescript plasmid vector. I introduced the desired mutations by SDM, and verified that the plasmids are correct via automated Sanger sequencing. My next step is to clone the mutated capsid sequences into a full-length clone of the HIV-2 ST genome for virus production and culture-based drug susceptibility testing. These studies have direct implications for improving HIV-related care and public health in West Africa and other areas where significant numbers of PWH2 reside.

Spatial skills are crucial for understanding and navigating the physical world around us. Previous research indicates spatial development begins in infancy, with sex differences present early on. These skills are predictors of later STEM achievement, highlighting the importance of nurturing them from a young age. Our study explores the impact of spatial language and play style on infants’ spatial development. We replicated a change detection mental rotation task for infants using an eye-tracker (N = 115, 47 girls, M age = 11.21 months). As infants preferentially look at novel stimuli, we recorded looking time as a measure of recognition of a change in stimulus. Additionally, we observed parent-infant dyads during play, and parents completed a survey on their infant's play, language, and motor skills. We hypothesized that parents would use more spatial language during spatial play than non-spatial play. We were also interested in whether play and language use varied by child sex. Infants showed no preference for the novel stimulus in the mental rotation task, suggesting it might not effectively measure mental rotation in infancy. Parent-infant dyads varied in engagement with the spatial toy, and no significant sex differences emerged from these measures. Due to the null findings in our change detection task, we will be designing and testing a new task for measuring mental rotation ability in infancy. Furthermore, we will also conduct analyses on the relationship between individual differences in spatial language and play to explore how these factors impact the development of spatial abilities. These findings provide insights to better support spatial development in early life.

Cannabis use has dramatically increased in response to legalization in the U.S., with U.S. sales jumping 46% from 2019 to 2020. ᐃ9-tetrahydrocannabinol (THC) is the primary psychoactive compound in Cannabis, and it has been shown to modify learning and motivation amongst regular users. Learning and motivation are key central processes primarily organized by the prefrontal cortex (PFC) brain region. I sought to test effects of THC on PFC activity during appetitive Pavlovian conditioning in mice- a behavior in which a subject learns to pair two stimuli together over time. Doing so provided much needed insight into learning and motivation under the effect of THC. THC acts on the endocannabinoid CB1 receptor, a presynaptic signaling protein responsible for modulating neural activity throughout the brain, with robust expression in the PFC. To monitor neural activity during behavioral trials, we implanted optic fibers into the PFC and virally expressed biological sensors. We used VGLUT1-Cre mice with a Cre-dependent GCaMP6f sensor to selectively target pyramidal glutamatergic activity during conditioning. We also utilized machine learning tracking software, SLEAP, to analyze behavior through video recordings. In our conditioning paradigm, animals were presented with a houselight and a sucrose reward, which they consolidated an association between after many trials. The mice experienced 5 days of Pavlovian conditioning, and I injected a moderate i.p. dose of THC (5 mg/kg) to one cohort, while another was given a vehicle before undergoing further trials. Our preliminary results showed that glutamatergic activity correlated with learning and association to the cue over time. We expected and observed that THC decreased the signals across the animals and reduced motivation. We categorized THC-induced behavior using SLEAP, a program tracking the mouse’s body parts to capture real-time movement, and found that locomotion decreased and resting behaviors increased in the THC cohort.

The peptide neurotensin (NTS) has been known as a regulator of dopamine neuron activity and its system, which modulates numerous functions in the brain. Although ample research has now demonstrated that NTS in Ventral Tegmental Area (VTA) increases dopamine release in some regions, much remains unknown about the endogenous sources of NTS in the VTA and the impact of physiological NTS release. Recent NTS mapping data from the Soden lab demonstrated that there is a NTS projection from Periaqueductal Gray (PAG) to the VTA and also to hindbrain regions including the ventral medulla. This project investigates the effect of this interconnection between these three regions on the dopamine system. Utilizing advanced techniques in circuit mapping, mice will be injected in the VTA and the ventral medulla with fluorescent Retrobeads or a retrograde virus (rAAV2 anti mcherry or GFP), which are taken up by synaptic terminals and migrate up the axon retrogradely to label cell bodies, one color assigned for each region. Following euthanasia, mice brains will undergo immunohistochemistry such as histology and in-situ RNA staining. Then, data will be collected using imaging microscopes for results and further analysis. If NTS neurons in the PAG have green and red expressions, this will indicate that the same population of neurons send axons to both downstream regions, compared to PAG NTS neurons with only one color, indicating the presence of two separate neuron populations. Experiments on retrograde mapping of NTS inputs will contribute to building onto our current knowledge about VTA-PAG-ventral medulla circuit and effects on dopamine neurons following their interplay. In the end, our goal is to establish a novel understanding of endogenous NTS signaling mechanisms, mediation of complex reward processes, and treatment targets with experimental outcomes, giving rise to the development of therapeutic interventions towards addiction and related psychiatric disorders.

A key neuromodulatory system involved in anxiety disorders is the locus coeruleus noradrenergic system (LC-NE), which projects broadly throughout the central nervous system. The LC is stress responsive and tonic activation of the LC and its projections to the BLA is anxiogenic. Previously, the Bruchas Lab has used two-photon calcium imaging to show that a powerful stressor (predator odor) increased synchronous activity of LC neurons. They also found that mimicking this predator odor evoked activity with optogenetics altered the activity of individual neurons downstream in the BLA in a β-adrenergic receptor (β-AR) dependent manner. Although these data support the LC's involvement in promoting aversion and increasing anxiety-like behavior, the specific neurotransmitter, neuronal cell types, and receptors responsible for these effects remain unidentified. Therefore in hopes of identifying these specific signaling molecules and neuronal cell types and receptors, I first used fiber photometry and a novel biosensor (GRABNE2m) to detect norepinephrine (NE) release in the BLA while mice were exposed to a predator odor. I found that predator odor produced robust increases in NE release in the BLA compared to control odor (n=5, 3 male, 2 female) Further, we found that optogenetic activation of terminals from the LC to the BLA produced very similar levels of NE release compared to what was evoked by predator odor. To determine the cell type and receptor that is sensing this stress-induced NE release, I used a CRISPR/SaCas9 virus, developed in collaboration with Dr. Larry Zweifel’s lab, to knock-down β2-adrenergic receptors (β2-ARs) in glutamatergic BLA neurons to test their causal role in stress-induced anxiety-like behavior. CRISPR knockdown of β2-ARs in the BLA blocked several stress-induced anxiety-like behaviors (n=4, 4 female). By understanding the circuit-based mechanisms of how stress-induced anxiety is regulated, researchers could identify potential targets for therapeutic treatments of anxiety disorders.

2023 was the Year of Open Science, part of a national push requiring public accessibility to the products of federally funded research. The Infant Brain Imaging Study (IBIS) Network is one such federally funded multi-site research network collecting neuroimaging data to study early brain development in infants with Down Syndrome. Data from the IBIS will be part of the INCLUDE Data Coordinating Center (DCC), a National Institutes of Health (NIH) managed open database intended to accelerate Down Syndrome research through sharable data. Understanding attitudes towards data sharing is important to ensure public trust in scientific research, and though much research exists assessing attitudes towards public sharing of genomic data, there is a lack of knowledge regarding attitudes towards public sharing of neuroimaging data. To combat this gap, I helped develop the CARDS-DS (Capturing Attitudes towards Research and Data Sharing in Down Syndrome) questionnaire, a novel parent-report measure to understand the perspectives of families participating in longitudinal Down Syndrome neuroimaging research. Currently, I am interviewing families enrolled in Down Syndrome neuroimaging research to get their feedback on the effectiveness of the questions asked in the CARDS-DS, a method called cognitive interviewing. Following this pilot study, CARDS-DS will be implemented throughout the IBIS Network. As such, CARDS-DS queries parent attitudes towards aspects of research participation that are particularly relevant to the INCLUDE DCC goals, namely: attitudes towards sharing of a child’s neuroimaging, genomic, and behavioral data, perspectives on data privacy, preferences for return of research results, and attitudes towards ongoing engagement with research teams throughout a longitudinal project. The anticipated data will reveal how attitudes may differ between parents from different sociodemographic or educational backgrounds, how attitudes may differ between the sharing of neuroimaging versus genomic data, whether attitudes are associated with study retention, and how attitudes may change over the course of longitudinal participation.

The mental health of family caregivers who care for elders who have passed is often dismissed by healthcare providers and society because of the age of their family members. When an older adult dies, their death is seen as expected and as the natural ending to having lived their life. However, there is still a tremendous grieving process that family members, friends, and caregivers experience with the death of a loved one. After losing a loved one, family caregivers report experiencing higher rates of depression, anxiety, and ambiguous grief. Additionally, many family caregivers find that medical help decreases drastically after the death, thus creating a sense of isolation. Current studies tend to explore one aspect of the experience of caregiver loss. Furthermore, much of these findings are based on data that is homogeneous in race and social status with an overwhelming majority being white and middle class. No existing studies have attempted to untangle the complexity of emotions that come with family caregiving. This systematic literature review starts to fill these gaps by answering the question: What are the psychological impacts experienced by family caregivers following the loss of an elderly loved one they provided care for? The databases used to identify the articles include PubMed, Web of Science, and PsycINFO. Approximately 15 articles have been identified that meet the inclusion criteria. The preliminary findings are themes of support changing before and after death, relationship with caregiving, relationship with family members cared for, psychological adversities (anxiety, depression, and loneliness), and religion as a way to find peace. These findings will help family caregivers see trends in their experience, helping with the loneliness that comes while grieving. It will also be a starting point to show healthcare workers what to look for when determining whether the grieving process is chronic.

In this study, Interpretive Phenomenological Analysis (IPA) is utilized as a framework to examine the experiences of hospital social workers who care for unhoused patients. Past research suggests that hospital social workers feel unprepared to appropriately support unhoused patients; this study aims to expand upon that research by introducing a type of intervention that supports the care of this population (based on Relational-Cultural Intervention or RCT) as a way of equipping social workers with additional tools to address this perception. Existing literature indicates that RCT can be a meaningful, reparative intervention to help unhoused individuals find affirmation and support through relational re-engagement. Through the provision of information about relational-cultural techniques, as well as advice on how to utilize these techniques, this research seeks to consider hospital social workers’ impressions of RCT’s potential usefulness as well as increase respondents’ perceptions that interventions performed with unhoused patients can offer meaning and value to both the patient and the social worker. Existing research suggests that meaningful relational exchanges have value for increasing an individual’s confidence in others' support and willingness to engage with services, programs, and other individuals, even in the absence of immediate options for accessing housing.