Light sheet imaging differs from other forms of microscopy by illuminating the entire plane of an ex vivo extracted brain sample and computationally compiling these planes post-hoc into a 3-dimensional image. This technique allows users to preserve the structural integrity and map biological units such as cells, axons, and genes onto a standardized brain atlas. Using the SHIELD clearing, preservation, and staining protocol, we prepared fluorescently labelled ex vivo whole, intact brains for light sheet imaging and used the ClearMap open source pipeline on the Hyak cluster operated by University of Washington-IT to overcome computational limitations. While ClearMap registers images and identifies cell counts per brain region, it lacks quality assurance metrics as well as figures for visual confirmation of preprocessing performance. To overcome these deficits, we have built a quality assurance package for ClearMap that includes registration validation and group-level quantification figures. In testing this package we analyzed multiple mouse brain samples exhibiting Fos expression patterns under varied conditions. Using this dataset we found that registration performance between expected and actual slices measured in F1 score averaged 0.7 in medial sections of the brain and we were able to visually inspect the alignment of subregions identified in a standardized brain atlas. Additionally, we created an interactive script to visualize heatmaps that indicate expression patterns for user-defined brain regions on an animal level. This work will enable rapid validation of collected data and third-party package performance while streamlining the analysis, interpretation, and visualization of results. In the future we intend to include useful statistics (one-way ANOVA, classification models, etc.) to provide additional functionality and further increase the speed and quality of data analysis.

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.

Loudness is a major component of how comfortable someone is with the sounds they hear, and looking at how people perceive the loudness of sounds around them is a crucial part of understanding how people process sound. Previous research by Dr. Yi Shen's laboratory has created an application that allows rapid measurement of loudness across frequencies and levels. The purpose of my project is to expand upon what was found using the previous application. To do so, I am measuring people’s perceptions of loudness across multiple bandwidths. In this project, I guide participants to listen to a sound stimulus in a specific condition, and then select what they perceive the loudness of said sound stimulus to be on an application. Multiple bandwidths of sound stimuli are presented (examples of bandwidths being presented include "Puretone", "1/2 Octave", etc.) and participants then rate how they perceive the loudness of each sound on a scale from "Not Heard" to "Too Loud". By utilizing the application to look at said stimulus bandwidths and performing computer modeling of the results, we gain a greater understanding of how a person’s loudness preferences from one ear can differ across frequency, level, and stimulus bandwidth. We can then analyze these interactions and create a more comprehensive model of the auditory system. I anticipate that results will show participants' bandwidths are consistent with the proposed bandwidths of prior models. This project has future implications regarding rapid identification of issues within a person’s auditory system, how comfortable they are with the loudness of what they hear, and the ability to deliver personalized interventions in assistive hearing devices accordingly.

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.

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.

Marine debris, classified as solid, man-made litter and material that has been lost or discarded in the ocean, is a persistent pollution issue in coastal regions around the world, and Puget Sound is not an exception. This research aims to investigate the distribution and abundance of marine debris throughout various regions of Puget Sound over time. Since 1987, the Washington Department of Fish and Wildlife (WDFW) has conducted annual trawls to assess bottom fish populations in Puget Sound. The contents of these trawls provide valuable representation of the soft-bottom habitat, including the organisms and debris that inhabit the seafloor. The WDFW’s extensive records of these surveys include the region, depth, type, and abundance of debris collected in each trawl. Using this data, I am exploring patterns in how different types of debris are spatially distributed, examining trends in abundance over the last two decades, and identifying hotspots for debris accumulation in Puget Sound. Through the use of R programming, I conduct analyses and fit statistical models to better communicate my findings. The results of this study provide a better understanding of the dispersal of aluminum, plastic, glass, fishing gear, and other debris that lies at the bottom of Puget Sound. Awareness of these patterns is vital to informing effective clean up and guiding prevention efforts to make community waters cleaner and safer for both humans and marine life.

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.

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.

This interdisciplinary project has the goal of investigating the interplay between behavior and biology using the context of thermoregulation in an invertebrate animal model system. Specifically, our approach involves the design, construction, and implementation of an ethoscope apparatus that examines behavioral responses associated with exposure to ethanol vapor (EtOHvap) in Drosophila. The methodology involves integrating a customized EtOHvap delivery and removal system into an ethoscope apparatus into which we incorporate a thermal gradient within the chamber. This customized ethoscope allows for the assessment of Drosophila behavioral responses prior to, during, and after EtOHvap exposure. Analysis of these data will focus on a comparison of the behavioral responses and how they reflect the effects of acute EtOHvap exposure in the form of temperature-preference-based behavior. Equipped with a Raspberry Pi and HD camera, the ethoscope apparatus captures and records Drosophila movements with software using OpenCV-Python to analyze the resting positions along the thermal gradient, and then compares them with the post-exposure resting positions. By working with Drosophila, a well-studied animal model, our design allows us to rapidly identify and examine behavioral responses as potential indicators of thermoregulatory responses to EtOHvap and forms the basis of future directions of research that utilize behavioral responses to understand underlying body temperature physiology and homeostasis. This research intersects with other works that have reported on the effects of ethanol exposure in other contexts and different model systems, including vertebrates. Thus, findings from this research not only inform this specific investigation but also have implications for understanding differences in how different animals regulate body temperature through behavior.

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 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.

Ethnic/racial minorities often face challenges associated with adjusting to a dominant or new host country. These challenges, known as acculturative stress, include difficulties with behavioral, emotional, and social adaptations, and are linked to adverse outcomes. Understanding the degree to which, for whom, and in what contexts acculturative stress may affect mental health is important in designing culturally-informed interventions. Yet, there are several limitations in the current literature on acculturative stress. First, most studies focus on Latino or Asian ethnic groups or international students/cross-national migrants; it is unclear how acculturative stress is differentially associated with health across populations. Second, most studies have focused on mental health outcomes, whereas other important outcomes (academic, relationship) are neglected. Using records from a large-scale meta-analysis project, we conducted a scoping review of research on acculturative stress to characterize the heterogeneity across studies. We performed literature searches using keywords (e.g., “acculturative stress,” “bicultural stress”) in databases including PsycINFO, and identified 3746 relevant studies. Abstract and full-text screening yielded 681 published and unpublished articles eligible for quantitative analysis. Primary studies were included if they measured acculturation in the context of intercultural adaptation and migration, acculturative stress, and health, academic, and relationship outcomes. We coded and summarized sample characteristics of all articles (e.g., % immigrants, % female/woman, mean age). We will randomly select 50 articles that examine acculturative stress and its associations with mental health, academic, and/or relationship outcomes. We will review and present common outcome measures, instruments assessing acculturative stress, and conclusions. We expect the scoping review to indicate patterns, variability, and gaps in the acculturative stress literature. Results will inform future research on overlooked outcomes and understudied populations, and shed light on necessary basic scientific information (e.g., mediators, modifying conditions) that will support the development of culturally-informed interventions and policies.

Parkinson's Disease (PD) is a progressive neurodegenerative disease characterized by slowness or stiffness of movement and cognitive impairment. PD is characterized neuropathologically by Lewy Body (LB) aggregates that include lipids, proteins and oligomerized alpha-synuclein. Mutations in the gene glucosidase, beta acid 1 (GBA), are not only the most common genetic risk factor for PD but also accelerate the progression of the disease. We hypothesize that mutations in GBA may mediate faster spread of pathogenic protein aggregation from neuron to neuron. Our previous work has implicated GBA in extracellular vesicle (EV) regulation, suggesting a non-cell autonomous mechanism for GBA accelerating propagation of LBs. To test this hypothesis, we are first exploring how GBA influences EV biogenesis in neurons and astrocytes by examining endolysosomal trafficking in GBA mutated neurons and astrocytes, as well as controls, differentiated from human induced pluripotent stem cells (iPSCs). Our initial results indicate that neurons heterozygous for a GBA null mutation have impaired endolysosomal trafficking with enlarged early endosomes and lysosomes, while astrocytes heterozygous for GBA null do not have impaired early trafficking. These results suggest that GBA mutations differently affect different cell types in the brain and improve our understanding of how GBA influences the spread of LB pathology. I image the iPSC derived neurons and astrocytes using a confocal microscope, for endolysosomal markers and distributions. The goal of this work is to identify novel therapeutic targets for slowing PD progression. 

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.

Glomeruli are the basic filtration unit of the kidney. The current understanding of its physiology is limited by the partial or 2D analysis of its structural components. The Vaughan Group uses optical super-resolution microscopy in combination with advanced chemical labeling techniques and powerful data analysis approaches to perform high-resolution 3D reconstruction of the whole mouse glomeruli. Overall, the work has the potential to provide a novel understanding of the glomerular structures and how they are altered in aged and diseased conditions. The labeling of the overall morphology of the glomeruli is achieved by chemically labeling the distribution of abundant macromolecules (carbohydrates, amine, and DNA) using Fluorescence Labeling of Abundant Reactive Entities (FLARE). Though we could visualize the general physiology of the sample with FLARE, incorporating specific targeting of molecules with FLARE is still challenging. My role is to optimize the FLARE protocol to add the capability of labeling the distribution of specific molecules using immunolabeling. The most challenging part is that all the fluorophores labeled prior to FLARE will be bleached out by the strong oxidation step while labeling carbohydrates. I am focusing on exploring possible workarounds to incorporate immunostaining with FLARE. The only way to bypass the bleaching fluorophores is to label dyes after the FLARE. However, the FLARE involves the gelation part, and the gel makes antibodies which are linked to fluorophores hard to get into the sample. So, instead of using regular secondary antibodies, I use biotin and then link to the streptavidin dye, which is smaller in size and easier to enter the sample. With this optimization working, we could incorporate whatever target of interest with high resolution on top of three general stains provided by FLARE, giving us an extra degree of information for our 3D reconstructions of glomeruli.

Epigenetic factors, including histone marks, change the patterns of gene expression without altering the DNA sequence. Variations in such marks are known to account for the ability of stem cells to differentiate into various cell types, but a preliminary experiment conducted by a former member of the Vaughan Group, Dr. Marcus Woodworth, has revealed that even in a phenotypically homogeneous, terminally differentiated cell population, the presence of H3K27me3, a repressive histone mark, varies on the HOXC gene of human retinal pigment epithelium (RPE1) cells at single-cell, single-loci level. My role is to evaluate the two possible origins of such heterogeneity: inheritance (histone mark varies due to events that happened during differentiation, or the random drift after differentiation, and the variations are kept within each lineage of cells), and multiple mark co-repression (one histone mark varies, but summing its effect with another histone mark that perform a similar function lead to the observed functional homogeneity), and to validate that such pattern exists among a broader range of genes. To achieve these ends, I profile selected histone marks (H3K27me3 and H3K9me3) on genes that experience different types of regulations during differentiation (HOXC, GAPDH and SIX6), using imaging-based methods, including the time-lapse imaging of live cells to map out lineages, and expansion microscopy (ExM) to capture fluorescently labeled histone marks at single-loci level. If the hypothesized origins are true, a significant difference in the number of histone mark clusters around the genes of interest would be observed between cells of different lineages, and complementary variation patterns would be observed between H3K27me3 and H3K9me3. The study reveals the nuances of histone mark dynamics on the single-cell, single-loci level, and optimizes an imaging-based method that has the potential for multiplexing at high spatial resolution, thereby providing a powerful tool for further studies on epigenetics.

The kidney is composed of thousands of filtration units called nephrons. Within each nephron lies a tuft of capillaries, the glomerulus, that filters from the blood through a filtration barrier. Over time this filtration barrier thickens, ultimately causing decreased blood filtration. A main marker of this age-related degradation are telomeres which comprise the ends of chromosomes and protect the coding DNA from degradation. If telomeres become too short, the coding region of DNA will begin to degrade. To combat this, telomere shortening signals for cells to enter a state of permanent cell cycle arrest, senescence, which prevents replication of cells with degraded DNA. Accurately quantifying telomere length will enable the development of correlations between cell lineage and structural changes within the kidney. I hypothesize that Expansion Microscopy (ExM) and quantitative-Fluorescent In-Situ Hybridization (Q-FISH) will allow me to determine the relationship between physiological changes in the filtration barrier and single-cell telomere length. ExM enables a superresolution cellular view by embedding a tissue sample in a swellable hydrogel, achieving four-fold isotropic expansion. This technique confers greater resolution of Q-FISH signal versus traditional confocal microscopy. To determine telomere length, I developed custom analysis scripts to quantify Q-FISH signal brightness. Preliminary results indicate an increased brightness of younger mice compared to their aged counterparts. Additionally, to receive a base-pair output I compared the Q-FISH signal to the signal of a DNA region of known length, Major Satellites, determining young telomeres to have an average base-pair length of 30 kb. I am validating these results in collaboration with the Miller Lab using next-generation sequencing techniques. Future work includes concurrent application of general physiology stains to identify and measure the glomerular filtration barrier physiology. Results from this method will allow for a wealth of information regarding the relationship of single-cell telomere length and glomerular structural health.

Alcohol and other substance use are associated with a range of negative physical, mental, and social consequences including blackouts, unintended injuries, involvement in interpersonal violence, and other health problems. Young adults in college are particularly at risk for alcohol and other substance use. While existing research on reduction interventions often emphasized individual responsibilities of those that engage in alcohol and substance use, peer interventions remain an overlooked approach to harm reduction. Bystander interventions have been shown to be effective in preventing and reducing the harm of sexual assaults among college students. However, there is less empirical attention on applying bystander interventions to reduce the negative consequences of substance use. An important first step of understanding how to incorporate bystander interventions to address substance use is to have a reliable and valid measure assessing these behaviors. This study aims to develop and examine the psychometric properties of a new bystander behavior scale specific to substance use. Data came from a large multi-site survey study with 930 college students (Mage = 19.39, 69.6% female, 58.1% White). I will conduct exploratory factor analysis to extract underlying factors that best explain the observed correlations between the items. Next, I will examine convergent and discriminant validity of the scale scores by correlating them with established instruments such as Drinking Refusal Self-Efficacy Questionnaire and Personal Assessment of Responsible Drinker Identity. Individuals who score higher on the bystander behaviors measure are expected to score higher on self-efficacy in resisting alcohol and be more likely to identify as responsible drinkers. Results will provide initial validation data for the bystander behaviors measure and can be used in future research and intervention implementation that address substance use among college students.

Parkinson’s disease (PD), the second most common neurodegenerative disorder, is characterized by Lewy bodies, pathogenic protein aggregates that include alpha-synuclein oligomers. The missense mutation p.G192R in the RAB39B gene was recently found to cause X-linked dominant PD. Loss of function mutations in RAB39B are associated with X-linked intellectual disability and autism spectrum disorder. RAB39B is a member of the human Rab GTPase family which plays a role in early autophagosome formation and is implicated in intracellular vesicular trafficking. This project investigates how defects in endolysosomal trafficking caused by the p.G192R mutation in RAB39B gene leads to parkinsonism and neurodegeneration. Because RAB39B is highly conserved, we developed a Drosophila model as human RAB39B and Drosophila RAB39 share 75% similarity in amino acid sequence, including 100% identity at p.G192 and flanking amino acids. Using CRISPR/Cas9 genome editing, we created a RAB39G196R Drosophila model that we are currently characterizing for possible neurodegenerative phenotypes. We are examining locomotor deficits and lifespan in RAB39G196R mutant flies compared to isogenic controls, as well as protein aggregation by Western blot. Complementary to the Drosophila model, we developed a human neuronal model by generating induced pluripotent stem cells (iPSCs) from peripheral blood mononuclear cells (PBMC) of an affected male and similar age unaffected male family member kindred with X-linked PD due to the p.G192R mutation. We are investigating endolysosomal trafficking defects in neurons differentiated from iPSCs using antibodies specific for early and late endosomes and lysosomes. We are also examining whether insoluble ubiquitinated protein aggregates and oligomerizes alpha-synuclein are present in RAB39BG192R neurons compared to control neurons. Understanding mechanisms underlying the pathogenesis of X-linked Parkinson’s disease will elucidate the development of PD and potential novel therapeutic targets.

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.

Marine ecosystems are currently experiencing changes at unprecedented rates. Implementing camera technologies to visually monitor vulnerable marine ecosystems is becoming increasingly common. Images and video can capture a variety of biological indicators of ecosystem health including species abundances, ecological interactions, and individual health and behavior which can be challenging to assess in other ways. As massive amounts of high-quality photographic data are accumulating, the tools to analyze them are often not readily available to scientists. Therefore, collected images may not be analyzed at the timescales required for scientists to assess and respond to these rapid ecological changes. To address this, we have developed a series of machine learning models that can analyze this surplus of image and video data into meaningful insights for researchers. We built the Subaquatic Ecosystem Analysis and Population Estimation Network (SEAPEN) by meticulously labeling millions of images from oceanographic datasets using a model-assisted labeling protocol. Our original models began with human annotated models, but as accuracy and precision progressed, we were able to rapidly iterate and expand SEAPEN’s capabilities. The processed images are then put through our custom training framework and are run until they output trained TensorFlow and TFLite models. These models are then sent through a series of statistical tests and further quantized until they reach our high set of QA/QC standards. SEAPEN has been tested using a variety of image data from Ocean Observatories Initiative to accurately classify organisms at deep sea ecosystems. SEAPEN is also capable of assessing coral bleaching rates, estimating fisheries stock population size, and quantifying the presence of marine debris. By utilizing large amounts of previously unused public oceanographic data, SEAPEN helps put the tools necessary to process old and new ocean data quickly into scientists’ hands.

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.

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.

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 mechanistic target of rapamycin, mTOR, functions in the mTORC1 complex with another protein called raptor as a master regulator of eukaryotic cellular metabolism thereby regulating cell growth including from cancer, cell death including after stroke, inflammation, and aging. In a forward genetic screen for hypoxia resistant mutants, the Crowder lab recently identified a missense mutation in the daf-15 gene, which encodes C. elegans raptor. The mutation produces a heat-sensitive reduction of raptor function, hereafter referred to as daf-15(rf). At 20°C, daf-15(rf) is normally hypoxic sensitive, at 22°C very hypoxia resistant, and at 25°C incapable of normal development. Raptor negatively regulates autophagy, a mechanism for breakdown and recycling of proteins and organelles. Activation of autophagy has been found to promote hypoxic survival in C. elegans and higher organisms. Thus, we hypothesized that activation of autophagy was responsible for the hypoxia resistance of our daf-15(rf) mutant. To test this hypothesis, we first asked whether we could detect increased autophagy using fluorescently-tagged autophagy proteins at 22°C in daf-15(rf) but saw no effect compared to wild type. Next, we asked whether a C. elegans transcription factor, HLH-30, that promotes expression of autophagy proteins was activated by daf-15(rf) and found activation at 25°C but not at 22°C. Finally, we tested whether proteins essential for autophagy were also necessary for the hypoxia resistance of daf-15(rf). By generating double mutant strains, we showed that animals with daf-15(rf) but without essential autophagy proteins were still hypoxia resistant. Thus, we conclude that C. elegans raptor regulates hypoxic sensitivity by an autophagy-independent mechanism. These findings demonstrate that raptor can control hypoxic cellular injury by mechanisms distinct from autophagy. Such mechanisms, if identified, could be targeted for treatment of cancer, stroke, and other diseases where hypoxia plays a role.

Early-Stage Alzheimer’s Disease (ESAD) is characterized by the development of beta-amyloid aggregates (Aβ42) and phosphorylated tau (pTau) leading to mild cognitive decline and variable personality changes. Because specific diagnostic criteria have not yet been established for ESAD at middle age, there is no way of knowing who might be susceptible and who might be resilient to more severe neuropathology and dementia in later years. The geroscience concept assumes pathways associated with aging are also associated with age-related diseases including ESAD. Therefore, a simple skin biopsy procedure shown to predict resilience to aging in middle-aged mice should be able to predict resilience to ESAD in middle-aged mice. An adeno-associated-viral (AAV) vector system carrying pathogenic components of AD, Aβ42, and pTau, was used to induce ESAD in 23-month-old C57BL/6 mice. Before receiving the AAV vector, 2 mm ear punch biopsies were performed, and the rate of closure was measured over 3 weeks. The study ended when mice were 26 months of age, and the closure rate for each mouse was calculated and correlated with behavioral and neuropathological features of EASD. Preliminary observations will help address the question of whether the healing rate of a simple skin wound can predict susceptibility to the burden of AAV-mediated ESAD. It is expected increases in physical resilience will be associated with increased wound closure, and thus, mice with increased wound closure will have greater resilience to the onset of ESAD neuropathology. This could have highly impactful implications for the early treatment of ESAD in human patients thus preventing the irreversible and fatal progression of dementia associated with late-stage AD. In addition, DNA from skin biopsy cores could be used to obtain DNA methylation signatures for determining biological age thus providing an enriched, translationally relevant data set.
 

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.

Alzheimer's Disease (AD) is incredibly complex such that development of neuropathology and cognitive impairement is driven by multiple pathways. Therefore, targeting these pathways simultaneously, could provide a more effective treatment for AD compared to any single drug. Rapamycin, acarbose, and phenylbutyrate each have independent but overlapping effects on multiple pathways involved in cellular respones to pathogenic beta amyloid such as inflammation, glucose homeostasis, synaptic integrity, autophagy, and DNA damage. To test the safety and effectiveness of a cocktail of these three drugs, a proof of concept experiment was undertaken in transgenic mice carrying mutations for genes associated with early onset AD (5xFAD). These mice express neuronal amyloid plaques, a major feature of AD neuropathlogy. Transgenic and wild type mice were given either a control feed or feed containing the drug cocktail starting at 4 months of age and continued until 12 months of age. Medicated transgenic mice showed significantly less cognitive impairement in a spatial navigation learning task and reduced amyloid plaque levels in the hippocampal brain region compared to untreated transgenic mice. Immunohistochemistry will be used to identify specific biomarkers for inflammation, synaptic integrity, glucose homeostasis, autophagy, and DNA damage in the hippocampus of treated and untreated transgenic mice. Observation from this study will suggest the need to conduct additional preclincial experiments testing this specfic drug combination for a successful approach to treat Alzheimer's Disease. 

Alzheimer's disease (AD) is a neurodegenerative age-related disease characterized by the presence of amyloid-beta aggregates and hyperphosphorylated tau tangles. It has been well documented that cognitive decline and changes in age-related pathways are associated with disease progression. Mitochondria play an important role in degradation of amyloid protein through a mitochondrial protein-mediated quality control system. This pathway can break down with increasing age and lead to the overwhelming presence of amyloid, disrupting normal mitochondrial activity. This damage leads to the formation of more Aβ plaques and neuroinflammation, contributing to the pathogenesis of AD. Mitochondrial regulators may be potential therapeutic drug targets but models are needed to help identify and characterize them. In this regard, an Adeno-Associated-Viral (AAV) vector was used to induce AD protein expression in the brains of old mice. 40 Male and 40 Females mice aged 24 months were infected with either the AAV-AD or AAV-SHAM vector and given 3 months for expression of the proteins to build. Mice were euthanized and brain tissue collected into formalin, with the hippocampus cut into slides for immunohistochemistry (IHC). Data generated from these mice has shown trends in decreased synaptic integrity, increased inflammation and DNA damage associated with expression of the vector proteins. Utilizing the same model, this experiment aims to understand how expression of the AAV-AD proteins may be associated with known roles of mitochondria and characterized pathways in the early stages of AD. IHC was performed using antibodies specific for PITRM1, a mitochondria protein degradation regulator, and PINK1, responsible for mitochondrial-mediated cell death (mitophagy). Imaging software “ImageJ” will be used for quantitative analysis of the stains. This study will help clarify an association between varying levels of AD protein expression and mitochondrial regulation, providing valuable information for enhancing therapies aimed at preventing the progression of early stage AD.

Alzheimer’s Disease (AD) is a progressive brain disorder that debilitates memory, learning, and decision-making. Early-stage AD represents the initial phase where individuals are still able to function independently, but with increasing age, their condition steadily progresses to dementia and loss of independence. Because a significant number of the aging population is affected by AD, understanding the neuroinflammatory processes would help develop more effective strategies for treatment. Examining markers such as MCP-1 and TNF-alpha, known to be associated with inflammatory response, will help identify the modulatory processes that lead to mild cognitive impairment associated with early-stage AD. Subsequently, higher levels of inflammation markers within the brain leads to mild cognitive impairment. This research study involved 40 C57BL/6 mice, 20 males and 20 females (21 months old), retro-orbitally infected with 80 µL of neurotrophic AAV-AD vector or AAV-Sham for a duration of 2 months before humane euthanasia. Brains were collected, and specific regions were examined by immunohistochemistry (IHC) and digital imaging to assess the expression levels and distribution of the inflammation markers. Preliminary observations showed that hippocampal regions of the brain from mice with early-stage AD had higher staining intensity for MCP-1and TNF-alpha compared to respective areas in Sham mice, suggesting increased inflammation is a very early lesion that develops in the presence of AD pathogenic components that might be controlled by anti-inflammatory drugs. The preliminary data suggests that the characteristics of AD manifest in part due to the neuroinflammatory response of brain factors that change with onset AD.

It is well documented that pet cats develop age-related diseases similar to humans with chronic age-related diseases, including Alzheimer’s disease (AD). Since pet cats live in the same environment as their owners and by extension are subjected to the same environmental stressors, older pet cats are an excellent mammalian model to study therapeutic targets to slow or reverse brain aging. However, aging within the brains of pet cats is not well characterized, partly because valid reagents have not been identified. This study was designed to test several human-specific antibody reagents that identify non-neuronal cells, aging pathways, and Aβ amyloid and phosphorylated tau (pTau) seen at autopsy in brains from patients with AD. Archived brain samples, collected from pet cats at autopsy, were graciously provided by the veterinary pathology departments at University of California Davis campus and University of Pennsylvania. Immunohistochemistry staining was done to detect: 1) Microglia, a non-neuronal inflammatory reactive cell type, using an IBA1-specific marker; 2) An inflammatory pathway using an MCP-specific marker; 3) Amyloid plaques using E610, an Aβ42-specific marker; and 4) pTau fibrillary tangles using AT8, a pTau-specific marker. A digital imaging software program was used to generate a heat map to visualize staining and quantify results. It was found that brain samples from older pet cats had increased inflammation as determined by high staining intensity of microglia and MCP1. Brains from several cats showed evidence of amyloid plaques and pTau tangles. These observations suggest that the human-based reagents tested can identify analogous cell types, pathways, and pathogenic components of AD in brains from pet cats. These prototype reagents can now be used to begin the task of characterizing neuropathology in deceased pet cats donated to the Cat Alzheimer’s disease Program at the University of Washington.

The causative agent of malaria, Plasmodium spp., generated 608,000 deaths worldwide in 2022 according to the World Health Organization and disproportionately threatens endemic areas of Africa. Plasmodium sporozoites infect the host by entering the bloodstream through the skin following bites by female Anopheles mosquitoes. From there, sporozoites migrate to the liver and infect hepatocytes. A single sporozoite-infected hepatocyte is capable of producing thousands of merozoites, which go on to enter the bloodstream. Complete elimination of infected hepatocytes is necessary to achieve sterile protection. In order to observe adaptive and innate immune cell localization towards infected hepatocytes, we applied fluorescence microscopy on livers in a BALB/c rodent model of malaria. Naïve unvaccinated mice were infected with sporozoites of Plasmodium yoelii, a rodent malaria parasite. Two important cell populations are recruited to infected hepatocytes. The first are tissue resident memory CD8+ T cells (Trm), which are crucial in pre-erythrocytic protection. The second are Kupffer cells, which are specialized liver macrophages. To measure these adaptive and innate cell populations, respectively, we applied fluorescently-labeled antibodies to mark the parasite as well as Trms and Kupffer cells. After staining the collected liver tissue and imaging with a widefield fluorescent microscope, we visualized recruitment and measured immune cell proximity quantitatively within a region of interest of the area surrounding infected hepatocytes using microscopy imaging analysis software. This method will be used to test the hypothesis that Trms and Kupffer cells are induced following sporozoite challenge in the rodent malaria model.

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.

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.

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.

Mitochondria are the main oxygen consumers in eukaryotic cells and as such are the primary organelles affected by oxygen deprivation, hypoxia. Hypoxia alters the size and shape of mitochondria, called the mitochondrial dynamics, but their role in hypoxic cell death is unknown. The Crowder lab has recently discovered that a mutation in the Mechanistic Target of Rapamycin Complex One (mTORC1) protein Raptor confers hypoxia resistance in the nematode C. elegans. mTORC1 is a master regulator of metabolism and is known to affect certain aspects of mitochondrial biology. Given these two facts, we tested the hypothesis that the hypoxia resistance of the C. elegans Raptor mutant is from alterations of mitochondrial dynamics. First, I showed that hypoxia induces small, rounded mitochondria in C. elegans caused from mitochondrial fission. Second consistent with the hypothesis, I showed that the mitochondria appear to have more normal morphology before and after hypoxia in the Raptor mutant. However, not consistent with the hypothesis, a C. elegans mutant with excess mitochondrial fission was not hypersensitive to hypoxia. Then combining the hyper fission mutant with the Raptor mutant did not diminish the hypoxia resistance produced by reduced Raptor function. Thus, our data demonstrates abrogating mitochondrial fission is not necessary for the hypoxia resistance produced by the Raptor mutant and leads us to reject our hypothesis. By exploring the interaction of mitochondrial fusion and fission with Raptor, we are beginning to understand how these important organelle and metabolic regulators combine to control hypoxic cell death.

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.

The recognition of individuals as sufficiently "Darwinian" entities for natural selection has led to the exploration of clades as alternative units of selection to species in the field of philosophy of biology. Previous research has demonstrated that clades meet the criteria of "Darwinian" individuals, intriguing philosophers to explain natural selection at the clade level. This research employs an Abstract Data Type (ADT) approach from computer science, constructing a data structure (DS), and attempts to implement the clades ADT both with and without species. The objective is to scrutinize the inner structure of clade and unearth the interplay between clades and species, a relationship not explicitly implied in their definitions. Preliminary findings affirm the technical feasibility of implementing clades ADT as a clade-tree data structure with species, while the feature of each level of clades remains unclear: (1) Issues of trait identification emerge when assigning properties to organisms within clades; (2) The empirical nature of the data used to construct the data structure representing clades limits holistic demonstration of evolution. On the other hand, a clade-tree data structure without species encounters challenges: (1) Historical organisms within inner stages cannot be distinctly separated without defined species-like boundaries (2) If properties are specified to species-level, the species-free intention of the data structure becomes questionable. The emergence of a dependency on species introduces a compelling reason for reconsidering the ability of clades in explaining natural selection, particularly for those philosophers who oppose species as a unit of selection.

The Climate-Ready Landscape Plants Project (CRLP) evaluates landscape plant performance under different irrigation treatments across six different geographic and climatic regions. Under the CRLP, four overlapping taxa Physocarpus ‘Diabolo’, Physocarpus ‘Little Devil’, Cercis occidentalis, and Cercis canadensis were chosen to investigate whether xylem anatomy changes in response to water deficits across three different planting sites: University of Washington, Oregon State University, and Utah State University. At each site, 24 replicates per taxa and 8 plants per irrigation treatment were tested under a two-year trial period. In year one, plants were established using the same water treatment. During year two, plants were introduced to irrigation treatments of 20%, 50%, and 80% based on the Water Use Classification of Landscape Species (WUCOLS). Plants were randomly assigned a position in the plot 2m away from each other under one of the three irrigation treatments. Stem cross sections were stained and imaged with vessel sizes analyzed using ImageJ. I expect both Cercis canadensis and Cercis occidentalis to display smaller vessels per xylem area reducing vulnerability to drought induced cavitation across the three treatments. I expect Cercis canadensis to display stronger acclimation to drought conditions because of its high tolerance for cold temperatures across the three sites. I expect Physocarpus ‘Diabolo’ and Physocarpus ‘Little Devil’ vessels to display lesser plasticity in xylem size and density across the three treatments and sites. Compared to the Cercis genus, Physocarpus is ornamentally selected and hybrids of the Physocarpus genus experience reduced water use efficiency with a lesser ability to acclimate to drought conditions. Most plants used for horticulture have not been tested under different irrigation treatments. This research can promote the production and utilization of low water-use plants to create well adapted landscapes under a changing climate for horticulture efforts on a local and industrial scale.

Brain-derived neurotrophic factor (BDNF) plays a critical role in neuronal function with potential implications for cognitive health, including involvement in adult neurogenesis. A decline in BDNF levels is associated with mild impairments in learning and memory. The hippocampus, known for its involvement in learning and memory processes, serves as a focal point for investigation in the brain due to its responsiveness to environmental stimuli, including exercise. There is an existing knowledge gap concerning whether running promotes an increase in BDNF levels within the hippocampus at very old ages, despite BDNF's importance in neuronal function and its potential implications for cognitive health. This study was designed to investigate whether physical exercise influences BDNF levels in the hippocampus of aged mice. Aged C57BL/6 mice were allowed access to running wheels, or locked running wheels, for three days, after which their brains were collected, post-euthanasia for neuropathology assessment. Immunohistochemistry (IHC) was performed with an anti-BDNF antibody by measuring BDNF presence, since lack of BDNF levels signifies lost neurons. QuPath digital imaging techniques were employed to provide a quantitative measure of the potential impact of running on hippocampal BDNF expression. Both the average and the variance of total distance run during voluntary wheel running decreased with age. Elevated BDNF levels were observed in the hippocampus of running mice compared to sedentary counterparts. The study provides insight into the potential impact of exercise on neurotrophic support in the aging brain. Such findings suggest a beneficial effect of exercise on neurotrophic support in the aging brain, and indicates the need for further investigations into lifestyle stratergies for promoting resilience to brain aging and cognitive decline in older adults.

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.

mTOR, the mechanistic target of rapamycin, is a serine/threonine kinase that regulates protein synthesis, cell growth, and metabolism in response to nutrients and energy in most eukaryotes. mTOR consists of two distinct complexes, mTORC1 and mTORC2. These complexes can be further divided into three components: mTORC1 consists of mTOR, Raptor, and mLST8, and mTORC2 consists of mTOR, Rictor, and mLST8. mTORC1 is critical in metazoan development and has been implicated in aging, cancer, diabetes, cardiovascular disease, and hypoxia. Previously, the Crowder lab conducted a mutant screen in C. elegans for hypoxia resistant mutations, and identified a missense reduction of function mutation in the daf-15 gene, the C.elegan ortholog of Raptor. A unique feature of this mutation is that the function of Raptor can be turned on and off by varying temperature. It has normal hypoxia resistance at 20 degrees, increased resistance between 21-22, and developmentally arrests at 25 degrees. I and the other authors made use of this conditional developmental arrest phenotype to screen for genetic suppressors. Using genetic mapping, sequencing, and complementation testing, we have identified multiple mutations in three different genes responsible for restoring Raptor function. One of the genes identified in this manner was rnf-126. Results show mutations in rnf-126 suppress the Raptor mutation. A null mutation in rnf-126 similarly suppressed the Raptor mutation. Previous work has implicated mammalian rnf-126 in degradation of the mTORC1 complex in cancer cells, suggesting that reduced levels of daf-15 may produce hypoxia resistance. We tested this hypothesis using auxin-mediated degradation of daf-15, finding that auxin-treated animals are hypoxia resistant. Current work by myself and others will further investigate how rnf-126 controls Raptor function and hypoxia sensitivity. Elaborating the function of this gene will define novel mechanisms whereby Raptor and mTORC1 controls metabolism, hypoxic injury, and development.

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.

This study explores the adaptive dynamics of Pseudomonas aeruginosa (P. aeruginosa), a bacterium that often plagues cystic fibrosis patients. In P. aeruginosa, many genes are regulated through quorum sensing (QS), a form of cell-to-cell communication. In wild-type P. aeruginosa, strain PAO1, LasR acts as a QS regulator, controlling several genes, including those encoding proteases, as well as two other QS regulators, RhlR and PqsR. QS-regulated proteases are secreted by P. aeruginosa into the environment, benefiting the whole population– a cooperative process. When grown in conditions requiring proteases for growth, such as on casein, mutated P. aeruginosa emerge that are capable of using proteases without producing any themselves. While disadvantageous to the community, these “cheaters” experience a fitness advantage. Over time, the proportion of cheaters increase until the population cannot support itself and collapses. Related to QS is the transcription factor MexT, that, when active, down-regulates RhlR. Another transcription factor, MexS, down-regulates MexT expression. In PAO1, MexS is inactive, resulting in constitutively expressed MexT. I was interested in how variants of MexT and MexS that alter expression of QS regulators might influence cooperative behaviors in P. aeruginosa. I compared survival and function of wild type PAO1 to two engineered strains, one with restored MexS function (MexS+) and one with MexT deleted (ΔMexT). I passaged these strains in casein broth for 30 days, evaluating every five days for the emergence of protease-negative mutations. I am also analyzing the genetic composition of each population, focusing on categorizing lasR and a gene involved in peptide transport, psdR. Preliminary results show fewer mutations in psdR than expected in all strains, and fewer mutations in lasR in the ∆mexT strain. Continued exploration of the complex QS and gene regulatory pathways in P. aeruginosa may point to mechanisms of virulence and to potential targets for antibiotic therapies.

Music interventions are gaining attention due to the evidence they improve health and cognition in elders. In the US, there is an increasing number of Mandarin-speaking elders whose cultural background affects their aging experience. After English and Spanish, Chinese is the third most spoken language in the US. Three and a half million households in the U.S speak Mandarin or Cantonese. However, there is little information regarding specific music choices or culturally-tailored music interventions for Mandarin-speaking elders to improve their health and aging experience. The purpose of this study was to gain insights into Mandarin-speaking elders’ views of aging, ageism, and the relationship between music and health. We conducted a qualitative study that involved interviewing Mandarin-speaking elders in Mandarin. The interview consisted of questions related to their cultural perspectives on aging, experience with ageism in the US, and the impact of music on their health. The participants selected music to play during the interview and discussed what the music meant to them and their thoughts on how music affects their health. Four interviews in Mandarin have been conducted to date. Initial findings reveal that filial piety and respect are recurring cultural themes that influence participants’ perspective and experience of aging. Internal ageism was identified in one of the interviews while the participant discussed their experience with ageism. Participants expressed that music benefits their health by providing emotional support and outlet, motivation, reminiscence of the past, brain activity, and stimulation through physical movement such as dance. The findings increase our understanding about the role of culture in the aging experience and music’s role in enhancing health for Mandarin-speaking elders. Future studies can use this information to develop music-based interventions tailored for Mandarin-speaking elders to improve their health.

 Music is considered a therapeutic tool to improve physical, cognitive, and emotional health. The nature of music varies across cultures and backgrounds around the world. This research project explored the connections between music, aging, and ageism in older adults from Kenya and the therapeutic role of music in improving physical, cognitive, and emotional well-being. This project considers cultural similarities and differences between Kenya and the United States and examines the unique nature shaped by different cultures. In this study, we explore how music affects the perception of age in older adults from Kenya and their perceptions of ageism. This research project focuses on using interpretive research method where data is collected through interviews to gain a deeper understanding of the topic. The study involves three older adults from Kenya between the ages of 60 and 65. The interview questions focus on understanding the cultural influences of Swahili speakers in the context of music and ageism. Through research on the differences in aging between Kenyan and American older adults, this project seeks to identify cultural factors and explore ways that music can enhance well-being among older adults. This project provides valuable insight into cultural aspects that influence aging and the potential benefits of music-based interventions. The findings have the potential to improve our understanding of how cultural differences influence attitudes toward aging and the impact of music on the health and well-being of older adults.

Nearly 70% of older adults hospitalized in an intensive care unit (ICU) experience delirium, a risk factor for long-term cognitive impairment that persists beyond discharge. The severity of critical illness, coupled with the 24-hour care provided in the ICU, is associated with significant disruptions to sleep and the circadian rhythm. These circadian rhythm disturbances, which affect up to 80% of ICU patients, may decrease the efficacy and benefits of interventions to improve cognitive function. Few intervention studies have been conducted testing circadian-based approaches to optimize timing of interventions to prevent cognitive decline in older ICU survivors. The study aims are: 1) to assess the feasibility, adherence, tolerability of morning or afternoon sessions of a computerized cognitive training intervention, and 2) to explore the role of individual chronotype on intervention usability and acceptability in older ICU survivors. Participants are randomized to one of three arms: morning computerized cognitive training sessions, afternoon computerized cognitive training sessions, or usual inpatient care. Participants assigned to the intervention groups complete daily 30-minute cognitive training sessions for up to 7 days or until hospital discharge. Wearable sensors monitor circadian rhythm patterns (via continuous body temperature and activity/sleep), and participants answer a questionnaire to determine their individual chronotype (“morning” versus “evening” circadian preference). Upon study completion, participants provide quantitative and/or open-ended feedback via surveys. We hypothesize that participants whose assigned intervention timing aligns most closely with their individual chronotype will demonstrate higher intervention adherence and will report higher tolerability and acceptability. Data collection is ongoing; results will investigate the potential of circadian-based and chronotherapeutic interventions to mitigate cognitive impairment in older ICU survivors. Additional research is needed to develop personalized interventions that integrate individual circadian rhythm and chronotherapy as targets to accelerate cognitive recovery throughout critical illness.

Cardiopulmonary bypass (CPB) is required for most cardiac surgeries. More importantly, CPB is used during heart surgeries to circulate blood out of the patient's body for surgeons to operate on the heart under optimal conditions without blood obstructing their view. However, CPB has been shown to induce systemic inflammation which can lead to complications including multiorgan dysfunction. Lack of understanding, specifically not enough knowledge of the molecular mechanisms of post-CPB inflammation has been a major obstacle to improve treatment methods. To better understand these mechanisms, we performed mRNA and Assay for Transposase-Accessible Chromatin (ATAC) sequencing on total circulating leukocytes from neonatal patients undergoing CPB. Prior data identified myeloid cells, particularly monocytes, as the major cell type driving transcriptional responses to CPB. Furthermore, Interleukin-8 (IL-8) and Tuman Necrosis Factor alpha (TNF-α) were inflammatory cytokines found to be robustly upregulated in leukocytes in patients. To further explore these findings, we performed in-vitro experiments of running THP-1 human monocytic cells to CPB-like conditions, including artificial surfaces, high shear stress, and cooling/rewarming. These experiments were collected and studied at times post shear allowing us to study and contrast how the blood is affected and recovers from CPB. Additionally, we performed drug treatment experiments with egtazic acid (EGTA) to see how calcium signaling may benefit post-CPB inflammatory response. Observing the opening and closing of chromatin of the sheared and the static blood has shown positive correlation that IL-8 and potentially IL-2 contribute to patient inflammation. We hypothesize that specifically these regions, IL-8 and IL-2, are the areas to determine where the root cause and solution are to post-CPB complications. Finally, further analysis with Topologically Associating Domains (TAD) was shown to increase chromatin density and activation in post-CPB samples in IL-8 and IL-2 genomic regions. We hope this research can reduce complications with patients going through CPB.

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.

The COVID-19 pandemic spotlighted Asian American (AA) mental health and their foreign position, underscored by anti-Chinese rhetoric and policies and the concurrent surge in hate incidents, crimes and xenophobia against AAs. However, framing COVID-19 as the sudden cause of anti-Asian hate facilitates misconceptions about AAs: it undermines a storied history in which AAs have long survived and challenged their treatment as “forever foreigners.” To understand the extent to which anti-Asian hate and mental health challenges among AAs during COVID-19 was a unique event in AA racialization, it is paramount to understand AA marginalization and its implications for mental health prior to COVID-19. To do so, I use the 2016 Collaborative Multiracial Postelection Survey (CMPS). The survey gathers data on respondent attitudes across a diverse set of political, psychological and socio-demographic measures and is ideal for Asian ethnic subgroups and intersectional analyses of gender, age and immigration generation. My analysis includes bivariate and multivariate regression analyses using the sample of AAs (N= 3006). Thus far, I have found that a substantial proportion of AAs indicate low feelings of belonging in the U.S., personal experiences of racial discrimination, and depressive symptoms. For the project’s next step, I will examine the extent to which AAs share a common foreign experience prior to COVID-19 and how this contributes to depressive symptoms. I expect that there will be significant subgroup differences at the intersections of ethnicity, gender, and immigration generation in the relationship between experiences of racial discrimination, feelings of belonging, and depressive symptoms when community involvement and local contextual factors are considered. This work helps situate anti-Asian hate during the pandemic as part of a more extensive history of racism in the U.S. and broadly, contributes to more informed mental health services by highlighting the implications of racism for mental health.

I am interested in producing Human Papillomavirus (HPV) type specific monoclonal antibodies (mAbs) from B cells derived from adolescent females and young women vaccinated with the licensed human papillomavirus vaccine, Gardasil 9, which provides protection against HPV types 6, 11, 16, 18, 31, 33, 45, 52, and 58. Fluorescently labeled pseudoviruses for these nine HPV types, as well as phenotypic markers of memory B cells are used to isolate HPV specific memory B cells via fluorescence-activated cell sorting (FACs). Gene transcripts for Immunoglobulin (Ig) heavy and corresponding light chain variable regions are amplified through Reverse Transcription Polymerase Chain Reaction (RT-PCR), cloned into their respective IgG1 backbone vectors, and subsequently transfected into eukaryotic cells to produce antibodies. HPV types [6, 11, 16, and 18] have been previously produced by our lab. I aim to expand on this work by developing antibodies against types 31, 33, 45, 52, and 58. I will characterize these mAbs for HPV binding and neutralization. Production of these well-characterized, type specific antibodies are useful for HPV studies, because they provide standards in HPV serologic assays. 

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.

In recent years, organizations have debated whether elements of “white supremacy culture”, including a sense of urgency, may create inequitable environments for people of color. We aimed to empirically investigate whether cultures of urgency in professional settings undermine the recruitment of racially minoritized groups. Undergraduate participants (N = 219) read job advertisements for two jobs: one where urgency culture is highly valued (e.g., “swiftly reset priorities at any given time”) and one with less sense of urgency (e.g., “adjust priorities based on capability”; order counterbalanced). Participants then reported how likely they would be to apply to each job. We hypothesized that individuals from racially minoritized groups will be more likely to apply to lower-urgency jobs over high-urgency jobs. The results show that most participants preferred lower-urgency jobs. However, countering our hypothesis, individuals from racially minoritized groups held more positive attitudes towards high-urgency jobs (e.g., how well they think they'd perform in the job) when compared to white participants. This finding casts doubt on the proposed elements of "white supremacy culture". It urges for more empirical research on how different racially minoritized groups may perceive these elements in work settings. Furthermore, our sample consists mainly of Asian Americans, which does not speak for the experiences of other racially minoritized groups. Our future research will focus on diversifying samples collected.

DNA-binding proteins (DBPs) capable of targeting specific DNA sequences play key roles in genetic regulation and manipulation in both natural and synthetic contexts. Aided by advances in machine learning and protein engineering, the design of entirely new DBPs is now possible. However, early attempts yielded small, single-chain proteins that were unable to induce changes in genetic expression despite successfully binding to their target DNA sequence.  It is thought that contacts made by these early de novo DBPs are not sufficient to maintain binding in the presence of other proteins involved in DNA transcription, preventing the DBPs from altering gene expression. In contrast, many natural DBPs consist of protein complexes composed of two identical subunits called homodimers. These complexes have increased intermolecular contacts with DNA, enhancing their DNA-binding affinity. In hopes of improving the affinity of current de novo DBPs, machine learning tools like RFdiffusion, ProteinMPNN, and AlphaFold2, were invoked to engineer homodimerization domains onto these monomers. Promising designs were filtered, ordered, synthesized, and tested for efficacy as genetic inhibitors in E. coli. Homodimers that consistently and specifically bind to their target DNA sequences and induce genetic repression in E. coli have promise as tools for genetic engineering and manipulation. In future research, successful designs could also have applications in synthetic gene circuits and serve as foundations for de novo allosteric transcription factors for use as biosensors.

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.

Family medicine requires effective dissemination of its growing research base to inform practice, education, and policy. The new Consensus Reporting Items for Studies in Primary Care (CRISP) guidelines may contribute to success. We will describe the pathways primary care (PC) research follows from presentation to publication and test if encouragement to use the CRISP guidelines is associated with increased acceptance and publication rates. We are conducting a confidential online survey in two phases of everyone who presented original research at the November 2023 meeting of NAPCRG (North American Primary Care Research Group), using the Qualtrics platform. Currently in progress, Phase 1 collects data on presenters, studies, research reports, author teams, submission processes, acceptance rates, and publication outcomes. Bivariate and multivariate analyses will identify factors associated with submission, acceptance, and publication. In Phase 2, a randomized controlled trial (RCT) will assign participants to either an observation-only control group or an intervention group receiving the CRISP guidelines. A follow-up survey at 6-9 months will assess presenters’ experiences with acceptance and publication of their written reports. The ongoing Phase 1 survey of 659 presenters worldwide includes diverse professions, specialties, scientific disciplines, and research roles. The presented studies include a broad range of research methods, study designs, problems, populations, and research questions. We will describe presenter experience with the submission, acceptance, and publication of these studies and examine associations with characteristics of the researchers, studies, and reports. The later Phase 2 RCT and follow-up survey will test for differences between the CRISP guideline group and the observation-only control group in success with acceptance and publication. Study results will describe the current practices and patterns of submitting and publishing reports of PC research to guide the dissemination and implementation of research findings to help improve patient care and population health.

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.

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.

Déjà Vu is an emotional experience that occurs less as we age and is most frequent when traveling, having vivid dreams, or brain injury. Research of déjà vu may prove vital for advancing interdisciplinary research, our interpreting of the world, and answering, arguably, many of life's greatest mysteries: human consciousness and even comprehension of life & death. Societal views of déjà vu have often labeled it abnormal, mysterious, and magical. As déjà vu transitioned from the realms of literature to scientific inquiry in the late 1800s, it captured the interest of the rapidly developing psychological and neurological disciplines. This project seeks to help clarify this issue by exploring how the explanations of déjà vu shifted from literary and artistic interpretations to an empirical research question. In this project, a literature review of historical and contemporary studies of déjà vu will be conducted to help map the conceptual overlaps, theoretical shifts, and methodological progression of research on déjà vu. Understanding how déjà vu's reconfiguration is an empirical question will help us better understand the entangled relationships between the body and experience (i.e., embodied experiences). Additionally, advancing interdisciplinary research about the experience of déjà vu may be vital to advancing phenomenological and scientific (e.g., neuroscience) understanding of many of life's greatest mysteries, from differences in human perception to diverse experiences of lived reality.

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.

There is currently a lack of tractable data on what was published in England in the early 20th century. However, this information exists in printed volumes of The English Catalogue of Books, which have been digitized through the HathiTrust digital library. The English Catalogue of Books, released in the UK by the trade publication Publishers’ Circular, provides a yearly record of books published from the mid-19th to the early 20th century. Each catalog has been converted to plain text automatically through optical character recognition (OCR). Our aim was to parse this plain text into user-friendly data on books published each year in the UK. However, the OCR-generated text often contained errors and inconsistencies that prevented the effective extraction of data on books listed in the catalogs. Thus, we aimed to gauge the accuracy of existing methods for parsing the catalogs and to tailor processes for gathering data from catalogs published between 1908 to 1922. By writing regular expressions to capture, split, and match the patterns of bibliographic entries, we were able to improve the accuracy of processes for extracting data. Our solutions increased the number of publications for which information was accurately being captured by on average 28%, and we were able to record information on over 21,500 books that previously had not been captured. We also created a summary dataset from the catalogs with information on the overall output of the publishing industry during these years. By analyzing and visualizing this publishing data, we were able to show that fiction was the most frequently published genre during the period. We anticipate this project to be the foundation of more work towards efficiently parsing The English Catalogue of Books in order to offer insights into the British publishing industry in the 19th and 20th centuries.

Over the past few years, patients have been identified with debilitating phenotypes due to mutations in MSTO1, a nuclear gene. These patients often have distal muscle loss and weakness leaving patients incapable of walking but to date there is no known treatment. One barrier to progress is that virtually nothing is known about MSTO1 function, making the development of therapeutics for these patients extremely challenging. The goal of this project is to use an unbiased approach to discover functions of MSTO1. To do this, I will find genetic interactors utilizing yeast to perform an unbiased screen. Yeast DML1 is the homolog to MSTO1 and is required to keep the yeast cells alive. This screen will identify genes in the yeast genome that support survival of cells lacking DML1 when the gene is overexpressed. We utilize an auxin-degron system that targets DML1 for degradation when the yeast are grown with auxin. To find genes from the yeast genome that keep the cells alive when DML1 is degraded, I express random fragments of genomic DNA. Those genes must, therefore, be linked to DML1 function in some way, thus providing insight into what MSTO1 does, how it works, and how to help MSTO1 defective patients. I have obtained hundreds of yeast colonies that survive without DML1 when other genes are overexpressed. Currently, I am extracting these overexpressed DNAs to determine the gene(s). This work is an essential step toward fully understanding MSTO1 function in cells and we plan to characterize these connections in yeast and human cells.

We use genes to survive and reproduce, but this means that genes can hold our survival hostage to ensure their own propagation, without providing any benefit to us. Selfish genes are a brutal, and poorly-characterized, demonstration of this concept. Instead of producing beneficial proteins, they produce nonessential proteins that prevent individuals who carry the selfish gene from successfully reproducing with non-carriers. One such example, the PEEL-1/ZEEL-1 system, is natively found in C. elegans. In this system, PEEL-1 is a toxin protein that kills cells when it is expressed without the antitoxin protein ZEEL-1. My aim is to determine the mechanism of toxicity employed by PEEL-1. AlphaFold predictions suggest that PEEL-1 contains an amphipathic helix. The amphipathic property of this region is hypothesized to play a critical role in PEEL-1 toxicity. To test this hypothesis, I am conducting a Deep Mutational Scanning (DMS) on the amphipathic helix of PEEL-1, with the goal of identifying key polar or nonpolar residues in this region that are essential to PEEL-1 toxicity. First, I generate a library of single-residue PEEL-1 mutants. Second, I transfect these constructs into HEK293T cells. I sample this initial pool of cells for sequencing, to identify which PEEL-1 mutants the pool carries, and in what proportions. Third, I induce expression, exposing each cell to the effects of the PEEL-1 mutant they carry. Only the cells expressing loss-of-function mutants survive. Now, I sequence this final surviving pool of cells, similarly to the initial pool. Mutations that drastically alter the polarity of the residue and thus break the overall amphipathic structure of the region are expected to be overrepresented in the surviving pool. This result would provide a broader understanding of the various methods of cell death in nature and provide novel insight into how animal-derived selfish genetic systems function.

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.

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.

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.

Reward-seeking behavior, crucial for survival, is hijacked by various neuropsychiatric disorders, notably substance use disorder. Prior work implicated reward-seeking with the dorsomedial striatum (DMS), a brain structure expressing the endogenous opioid dynorphin (dyn) in 50% of its neurons. Previous studies also linked dyn, signaling through the kappa opioid receptor (KOR), with escalation and reinstatement in drug-seeking, however, the exact mechanisms of dyn-KOR signaling remain unknown. Recent experiments from the Bruchas lab show that supraphysiological release of dyn in the DMS with excitatory optogenetics elicited reward-seeking behavior, demonstrating that dyn-KOR is sufficient for reward-seeking behavior, but not that it is necessary. To provide evidence of necessity, I plan to inhibit dyn-KOR signaling in the DMS by expressing the novel inhibitory optogenetic tool Platynereis dumerilii (PdCO) that we recently showed to be useful for such studies (Wietek et al., Nat Methods, 2024). PdCO is expressed in a target neuronal population and begins inhibiting under 473 nm light. Hence, in this study, PdCO will be injected into the DMS and expressed selectively in dyn neurons. Following recovery, mice will learn a self-administration procedure to study reward-seeking behavior, where sucrose is delivered when mice poke their nose in the correct 1 of 2 holes, with a 5s light cue before reward delivery. Following learning, I will inhibit dyn release in a counterbalanced fashion by activating PdCO using 20 Hz pulsed 473 nm light. I anticipate that inhibiting dyn release would result in reduced sucrose consumption. Finally, I plan to use an extinction learning procedure, where dyn release is inhibited during perceived reward delivery. Here, I hypothesize that inhibition will accelerate extinction learning. By leveraging the specificity of optogenetics on various procedures, this study begins to isolate the exact mechanisms by which dyn-KOR signaling impacts reward-seeking behavior with potential insights for substance use disorder interventions.

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.

Acorn barnacles (Balanus glandula) are among the most common species found in the Northern Pacific Ocean, inhabiting rocky surfaces across intertidal zones. These barnacles’ distribution is attributed to various factors, including surface exposure, spatial competition, and predation. The size and abundance of barnacles may serve as an indicator of the primary abiotic and biotic processes that influence barnacle populations in highly dynamic intertidal systems. This study tests our hypothesis that higher abundance and smaller-sized barnacles will be observed at higher tide elevations, while lower abundance and larger-sized barnacles will be observed at lower intertidal elevations on the shore. In September 2023, at Friday Harbor, (San Juan Island, WA) we photographed 10 quadrats (24x24cm) at low (0m), mid (1m), and high (2m) elevations. We observed barnacle distributions at two sites along the San Juan Channel. Barnacle abundance and size were measured with ImageJ. We calculated the mean and median barnacle size at each elevation and location. Small barnacles (0-1.5mm) were predominant in the high elevations, whereas large barnacles (>3mm) dominated the lower elevations. Our results are associated with a combination of abiotic and biotic factors. Abiotic factors include greater desiccation risks in the high intertidal elevation. Biotic factors involve food availability, with larger barnacles having greater access to the nutrients coming from the ocean, along with spatial competition, where higher abundance may lead to increased competition, thereby reducing available resources for each individual. Differences between sites may be attributed to diverse characteristics unique to each location.

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.

The use of opioid drugs for pain management in postoperative settings has been widespread since the 1860s. However, the risk factor for developing an opioid use disorder (OUD) has increased substantially with its continued use, with addiction rates of more than 10% for those taking opioids. An obstacle to abstinence in opioid addiction are the adverse side effects that occur during cessation of drug use once dependence has formed, including nausea, anxiety, vomiting, and depression. The Nucleus Accumbens (NAc) is part of the mesocorticolimbic reward pathway. Decades of pharmacological studies demonstrate that nearly all abused drugs evoke dopamine release within the NAc, thus altering innate systems for how reward is processed. The activity of NAc neurons is strongly regulated by efferent excitatory input from numerous brain regions. The paraventricular thalamus (PVT) a relatively understudied brain region, regulates behavioral responses to reward and aversive stimuli as well as to drugs of abuse such as morphine. Our preliminary data demonstrate that the activity of these projections is highly regulated by the Cannabinoid 1 Receptor (CB1), which mediates the primary psychoactive effect of cannabis. This is particularly relevant on account of recent clinical findings demonstrating that activation of CB1 can ameliorate the aversive effects of opiate withdrawal. Using fiber photometry (which uses fluorescence emission of the calcium sensitive fluorophore, GCaMP, as a proxy measurement for neural activity), I have shown that this circuit is activated by aversive stimuli and inhibited by rewarding stimuli. Furthermore, treatment with morphine can attenuate the pain-induced activation of this circuit. However, whether cannabinoids can influence this circuit's activity to reduce withdrawal symptoms remains untested. Our research will contribute to our understanding of the neurophysiological basis for opiate withdrawal and how cannabinoids could represent a novel class of therapeutics for the treatment of opiate use disorder.

Addiction is characterized by the compulsive use of substances despite adverse consequences, a process closely linked to dopamine-induced changes in the Nucleus Accumbens (NAc) and its role as the brain's "reward center." The NAc integrates information from various brain regions, including the Paraventricular Thalamus (PVT), to produce motivated behaviors. Recent studies have identified the PVT, especially its anterior segment (aPVT), as a critical hub in addiction neurocircuitry, but findings have been inconsistent, likely due to the PVT's heterogeneity and the specific neurochemical and anatomical properties of its connections to the NAc. Prior research has shown that aPVT neurons, identifiable by neurotensin expression, send excitatory projections to the NAc, which are modulated by endogenous cannabinoids (eCBs). These interactions suggest a complex regulatory mechanism. Preliminary experiments used techniques including transsynaptic viral tracing and in vivo calcium imaging, to study the activity dynamics of NAc neurons, particularly those expressing Proenkephalin (PENK) and receiving aPVT inputs, during reward-seeking tasks. I propose to extend these findings by employing a multidisciplinary approach that combines experimental neuroscience with sophisticated computational analysis. By applying dimensionality reduction techniques, clustering algorithms, and machine learning models to neural and behavioral data, I aim to map the functional connectivity within the NAc and elucidate the roles of specific neuronal ensembles in reward-seeking behavior. This comprehensive analysis will not only clarify the neurobiological underpinnings of addiction but also contribute to the development of targeted therapies for addiction and related disorders, leveraging the unique intersection of computational neuroscience and behavioral analysis.

 The locus coeruleus (LC), a nucleus in the pons of the brainstem, is the primary source of norepinephrine (NE) in the brain and is responsible for regulating arousal and stress responses. The LC also has roles in pain, avoidance and reinforcement learning, exploration, and feeding. How LC neurons are mediated to generate these distinct responses has yet to be defined. One area of interest is the peri-LC, which sends local GABAergic projections to the LC and is thought to modulate physiological arousal through direct inhibition of the LC. The peri-LC is composed of various populations of GABAergic neurons that may contribute to the inhibition of the LC. We were interested in exploring the role of neurokinin1/substance P, nociception, and enkephalin GABAergic neurons. Using vGAT-cre, Tac1-cre, Noci-cre, and Penk-cre mice strains, we injected an excitatory DREADD into the peri-LC to be able to activate genetically defined neural subpopulations within the peri-LC. I ran the mice through open field and elevated zero maze testing to assess anxiety levels. The activation of all peri-LC^GABA neurons led to a decrease in anxiety-like behavior, which is consistent with an inhibition of the LC and a decrease in LC^NE release. In contrast, the activation of peri-LC^Tac1 neurons increased anxiety-like behavior, while the activation of peri-LC^Noci and peri-LC^Penk neurons had no behavioral effect in these assays. Together, these results suggest that the different peri-LC^GABA subpopulations likely represent distinct nodes that differentially integrate novel stimuli to influence distinct behaviors. Understanding how the peri-LC regulates LC function can give insight into the neurobiological basis of arousal and stress-related disorders, which informs our understanding of neuropsychiatric pathologies.

Climate change is increasing glacial melt worldwide, causing freshening events in marine ecosystems that rapidly decrease salinity. In the Salish Sea, summer low salinity events occur regularly from the Fraser River meltwater. This negatively impacts many organisms including sea urchins, which are resilient to other climate change impacts like marine heatwaves and reduced food availability. Previous studies found that low salinity impacts tube foot mobility and function in green sea urchins (Strongylocentrotus droebachiensis). We replicated the Salish Sea freshening events with a treatment of 20 PSU saltwater (control 30 PSU) to investigate the impacts on red sea urchin (Mesocentrotus fransiscanus) coordination and spine movement. This study implemented new techniques to quantify urchin motility to determine the negative effects that low salinity had on sea urchins over 96 hours. We quantified spine movement using image analysis to track individual spines of urchins in low and regular salinities. We also measured righting time, which is an indicator of urchin coordination. We found that urchins exposed to low salinity had significantly longer righting times and less spine movement overall. The low salinity treatment spine movement and righting time were highly correlated (p < 0.01), but not in the ambient salinity tanks (p > 0.01). We found that lower salinity waters have the potential to reduce urchin coordination and movement, which may impact urchin populations and kelp forests in the changing climate. Thus, there may be a lessened impact on the kelp forests needed to maintain ocean health.

Zooplankton are vital to the marine food web, supplying nutrients and energy from primary producers to secondary consumers. During Diel Vertical Migration (DVM), zooplankton travel between depth and the surface during day and night to capitalize on food and avoid predation. This study investigated diel differences in zooplankton community composition at two locations, one exposed and one protected, in the San Juan Channel, WA over four days in September 2023. Zooplankton were collected using net tows from surface waters at both sites during day and night times. Samples were analyzed using a stereoscope and different taxonomic groups were counted. Copepods were the most abundant zooplankton taxa at both locations, with mean abundances up to 1000 individuals per cubic meter. At the exposed site, there was a significantly higher (p<0.05) abundance of zooplankton at night versus during the day. The exposed site had significantly higher diversity than the protected site at night (p<0.05). At both locations, species richness was significantly higher (p<0.05) at night compared to day. The exposed location also had significantly higher richness (p<0.05) compared to the protected location during the day. Our results indicate that zooplankton abundance and diversity in surface waters of the San Juan Channel are controlled by DVM, and differences in locations perhaps due to exposure to different flow regimes. This study reinforces the flexibility of zooplankton community composition and emphasizes the importance of understanding factors that influence changes in the base of the marine food web.

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.

Investigating the mechanisms behind asymmetric cell division (ACD) not only provides insight into an integral aspect of development in organisms such as C. elegans, D. melanogaster, and in human stem cells, but it also has the potential to elucidate the cellular functions that result in disease processes like cancer. ACD describes the cellular activity that results in a singular parent cell dividing into two daughter cells with two distinctive molecular compositions, and, in turn, fates. Drosophila melanogaster neural stem cells, called neuroblasts, are intrinsically polarized and divide asymmetrically. This process involves an uneven distribution of cell-fate-determining proteins that eventually designate the function of the daughter cell. Because of this polarity establishment and orientation, these daughter cells can have highly specialized functions. My work uses Drosophila melanogaster neuroblasts to screen for novel mutations responsible for compromising ACD. I'm specifically interested in finding genes that are involved in the generation of cell size asymmetry. Finding these mutations is imperative because, by doing so, we can identify critical genes involved in this process of interest. To that end, I knocked-down candidate genes with RNAi in neuroblasts and assayed the localization dynamics of myosin and microtubules– two critical components of the ACD and symmetric division process. Through confocal, live-cell imaging, I then examined the dynamics of these cellular proteins as they relate to wild-type ACD to see if my chosen candidates were responsible for this process. I anticipate finding novel cytoskeleton regulators that are important for establishing cell size asymmetry. In the future, finding the genes behind ACD will provide great insights into the mechanisms behind both development and disease in addition to the field of stem cell biology overall.

As our world progresses through technological advancements, much of our planet regresses as an effect of climate change, highlighting a need for underutilized resources to be brought to the forefront of industry. One avenue for transforming abundant resources into useful chemicals for generating sources like fuel is the catalytic upgrading of biomass derived molecules. However, catalysts traditionally used for these reactions are not stable to contaminants in biomass mixtures, such as water or organic acids. For biomass derived molecules to serve as precursors for biofuel and other related energy sources, more stable and efficient catalysts are needed. Our group has recently shown that a bifunctional acid–base MOF with co-localized acidic and basic sites outperforms a MOF with randomly dispersed acid–base sites for the aldol condensation reaction. To further demonstrate the importance of having the acid and base groups co-localized, I synthesized and tested three control frameworks for comparison: (1) a framework with no functionality, (2) a framework with only acidic sites, and (3) a framework with only basic sites. I then tested stability and recyclability of the bifunctional acid–base frameworks by conducting recycling experiments. I resubjected the same sample to reaction conditions for a total of 5 cycles. After each cycle, I used 1H NMR to quantify the conversion of starting material to ensure that there were no changes in catalytic activity. Lastly, I used powder X-ray diffraction (PXRD) to ensure that the catalysts maintained their crystalline structure after 5 cycles. Here I show that metal–organic frameworks (MOFs), a class of porous crystalline solids, can be used as efficient and recyclable catalysts for the aldol condensation, an important reaction for biomass conversion. Overall, this work illustrates the stability and reusability of metal organic frameworks as catalysts and thus their potential for utility in biomass upgrading reactions.

 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.

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 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.

In the US, our primary healthcare is mostly delivered via the fee-for-service model. Interactions between providers and patients under this model are mediated by insurance companies. In order to bypass health insurance companies, some primary care physicians have opened direct primary care clinics which charge a monthly fee for unlimited care. As part of my undergraduate thesis for the anthropology honors program, I investigated two clinics operating under this model in the Pacific Northwest using participant observation and semi-structured interviews with physicians. My aim was to determine how the direct primary care model affects the agency of physicians and influences the therapeutic process via the clinical encounter. I found that physicians in the US face a unique dilemma of role conflict between the competing identities of business owner and doctor. Additionally, I explored how the expectations of patients differ for male and female physicians and how direct primary care can play into this. My findings highlighted the inadequacies of our existing healthcare system in the United States. Further, I concluded that although direct primary care is a solution to this broken system for some patients and some physicians, systemic changes must be made in order to make primary care a more desirable field for physicians and to provide equitable and quality care for all patients.

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.

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.

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.

The Disability Inclusive Development Initiative (DIDI) is a project that brings together students and faculty from various fields to promote the human rights of persons with disabilities. Our current project builds upon the findings of a 2020 Memorandum of Understanding (MoU) between the International Disability Alliance (IDA) and the United Nations High Commissioner for Refugees (UNHCR). We are conducting a comparative analysis of asylum procedures in Uganda and Colombia, specifically examining how these procedures consider the rights of persons with disabilities. Our goal is to ensure that refugees and asylum-seekers with disabilities have access to fair and accommodating processes. We are also looking into the challenges faced by persons with disabilities during the asylum process and how governments are addressing these challenges. To achieve these aims, we are conducting a comprehensive literature review, including primary legal research, using resources from the University of Washington libraries. Additionally, we are consulting with experts working in this field. Our team is dedicated to producing a report for the United Nations High Commissioner for Refugees, which will outline our findings, highlight effective practices, and offer recommendations for future actions.

University of Washington Food Pantry (referred to as "food pantry" in the rest of the article) has been providing free food and supplies to faculties and students for years. Visitors to the food pantry have been increasing since 2021, and even quadrupled from winter 2022 to winter 2023. I observed that the visitors to the food pantry are predominantly people of color since my start of volunteering at the food pantry in April 2022. Other observations include most visitors studying science or engineering majors. I designed a google form survey where visitors fill out during each visit which includes information including race, gender identity, major of study, age, and hometown. I conduct analysis to compare the demographic and academic profiles of food pantry visitors as collected through a Google Form survey during each visit, against the corresponding parameters of the entire student population at the University of Washington. I anticipate that there will be a higher percentage of STEM (Science, technology, engineering, and mathematics) majors visiting the food pantry, and a higher percentage of people of color visiting the food pantry compared to the entire student population at the University of Washington. Findings from this research aim to highlight the inequalities in higher education and provide evidence for underlying financial or social disparities in order to inform and raise awareness among university policies and support systems on resource distribution. The findings can also serve as the basis of long term future study on impact of food pantry use on academic performance, graduation rates, and post-graduation outcomes.

During the rise of the BLM movement of 2020, confederate monuments became a focal point of national debate, with heightened awareness of their historical context and implications for racial equality. My research seeks to leverage this moment of increased attention to explore how the presence of symbols of marginalization near voting stations may impact voting intentions and overall experience. In this study, Black and White Americans are presented with a simulated map along with a set of instructions that lead them to a nearby polling station. In the experimental group, participants received a map featuring Confederate monuments while participants in the control group received a neutral map with directions that avoided any mention of Confederate monuments. Once the participant correctly identifies the voting station on the map, they are asked a series of questions not only on the usefulness of the map but also on factors such as safety, comfort, the value of the vote, personal belonging, and likelihood of voting. Findings suggest Black participants exposed to Confederate monuments expressed a diminished value of their vote and reduced intention to vote, whereas White participants indicated no changes in voting intentions, perception of vote value, or a desire to change voting locations. As an extension to this study, I am currently conducting a replication study where participants are explicitly informed that the map represents a neighborhood in the South. After navigating the map, participants are asked not only about the impact of factors such as comfort, safety, value, personal belonging, and the likelihood of voting but also regarding their relative importance and influence on voting behaviors and intentions. Collectively, these studies have important implications for racial disparities in voting intentions and for advancing a more inclusive and equitable electoral process within the United States.

Within the mainstream misogynistic and homophobic narrative of Chinese society, radical feminists and lesbians face significant challenges. But the issues each of these groups face and their communities are separate and seldom intersect. This project addresses this gap between lesbian and feminist communities while aiming to comprehend how individuals within them perceive themselves and each other. It also explores the possibilities for solidarity and greater communication between the lesbian community and the radical feminist community in China. Through interviews with Chinese radical feminists, lesbians, and lesbian feminists, this project aims to bridge the divides between Chinese lesbians and radical feminists and to facilitate discussions about negotiating different aspects of one’s identity.Both lesbians and radical feminists are engaged in reconstructing intimate relationships. With the current rise of feminism in China, lesbians are reevaluating gender roles in intimate relationships. At the same time, some radical feminists are exploring lesbian feminist ideas, such as gender separatism. This project draws on theories from the second wave of the feminist movement in the U.S and queer theories to highlight important conversations in contemporary China with the goal of stimulating discussions on envisioning intimate relations beyond the framework of a heteropatriarchal society. Preliminary analyses of interviews conducted in winter 2023 show that many radical feminists and lesbians have struggled to assert their identities and find belonging in a community. Additionally, the lack of platforms and offline spaces for gender and sexuality communities leads to the isolation of individuals. The project to be presented will offer a more detailed interpretation on the reasons behind such isolation and miscommunication between the two groups. On a broader level, this project aims to offer a perspective on how we can understand the relations between people and power dynamics in this current world.

Gender identity has become a central issue in ongoing political debates regarding cultural shifts in America. For many, nonbinary and other gender-variant identities seem new and confusing, but there is evidence of these identities stretching back to classical antiquity. The god of wine, madness, and theater, Dionysus, emerges as a consistently gender-variant figure in ancient literature and art. For this project, I examine the construction and performance of Dionysus' gender in antiquity by performing a close nonbinary reading of Euripides' tragic play, the Bacchae, in conversation with current scholarship on gender and sexuality in Ancient Greece and Rome. My research seeks to interrogate the appropriateness of applying modern gender-variant labels to antiquity, investigate the ways in which Dionysus both resists and subverts binary gender categorization, and consider what meaning modern nonbinary and gender-variant people can extract from ancient representation. I argue that applying modern gender labels to antiquity aligns with current practices in scholarship regarding binary gender identities. Through a nonbinary lens, I reveal the image of a clearly nonbinary Dionysus who cannot be contained by either ancient or modern gender binaries -- a force of gender chaos. With my critical analysis of the Bacchae, I shed light on new considerations of the motivations behind Dionysus' display of divine wrath and the cultural implications of gender construction and performance in the play. My project finds that nonbinary and gender-variant identities have long existed even in societies that seemingly embrace a gender binary.

Evaluating women in Ancient History could be difficult as patriarchal societies limited the representation of women, which resulted in little or biased documentation. Another obstacle was evaluating the accuracy of evaluating the authenticity and accuracy of reported events when there is such limited evidence of certain figures and events in Ancient History . However, regarding the feminist martyr narrative, I was able to evaluate the effects of the representation in martyrdom and how they could be applied to modern feminist narratives. To investigate the significance of the texts of women and martyrdom to values in the Patristic Era of Christianity, I analyzed women in the context of the Roman Empire compared to the Early Christian Church using primary sources. Introducing the consequences of martyrdom, I addressed the etymology of the word “martyr” and the connotations associated with the term. As my primary sources, I used the accounts of Perpetua and Blandina because their narratives were among the most well-known. Studying the divergence of feminine virtues of the Roman Empire and Early Christianity, I explained how the differences influenced the divisions and spread of the Early Christian Church. I also studied the retelling of their stories throughout history, which illustrated society’s expectations of women as opposed to the possible motivations of the martyred women. I discovered that the stories of martyred women were altered throughout history to perpetuate different ideals of femininity but at a closer look, the original narratives depicted a progressive depiction of women. This research exposed how the representation of women could be altered to fit an agenda and revealed the values of the authors of the narratives as well as the surrounding culture. This could be used to evaluate the representation of influential women figures today as well as the consequences of altering feminist narratives.

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.

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.

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.

Changing climate has resulted in cascading consequences to processes within the natural world, influencing physical, chemical, and biological components of our ecosystems. To properly prepare for changes within these natural systems and mitigate impacts, it is important that we understand underlying mechanisms that are being influenced by this change. One such mechanism that has a known correlation with warming temperatures—a factor of our shifting climate—is increased metabolism of cold blooded organisms. As their metabolism increases, they must consume additional food to offset the increased energy demands. To investigate this issue, I worked with the University of Washington’s Applied Ecology Lab and the National Oceanic and Atmospheric Administration’s (NOAA) Northeast Fisheries Science Center to examine whether predation by predatory fishes in the Northwest Atlantic has increased over the past 50 years. Specifically, I examined diet information for 17 commercially important fish species from 1973-2021, enabling us to capture any shifts over time. To determine if the observed changes were related to environmental conditions, I tested various climate indices as possible forcing factors. Additionally, I examined changes in the body condition of these fishes. To do so, I fit time series models to the data to estimate annual changes in consumption and body condition, and determine the degree to which the climate indices were correlated with these changes. I further hypothesized that the life history, habitat, and range of the given species may also be related to shifts in diets. These results will help inform us about potential “winners and losers” with respect to climate change within the Northwest Atlantic ecosystem, enabling management agencies to target groups that are negatively impacted by these environmental trends. Additionally, our analyses can provide insight on potential shifts in ecosystem dynamics that we would expect to see in relation to future climate change.

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 recent decline in the body size of sockeye salmon (Oncorhynchus nerka) returning to Bristol Bay, Alaska has been associated with increased competition within the marine environment as these populations have increased in abundance. As an anadromous species, different populations of sockeye salmon return to freshwater environments, occupying streams of varying sizes to which they have evolved habitat-specific adaptations to local habitat conditions. In particular, fish spawning in small streams are substantially smaller than ecotypes that spawn in the deep water of rivers and lakes where sexual selection promotes large body sizes. We hypothesized that density-dependent marine growth in sockeye salmon would be most intense for large-bodied spawning ecotypes, compared to small-bodied populations where there is less evolutionary pressure to achieve large body size. Using general linear mixed-effects models, I compared the effect of run size on growth rates in sockeye salmon from a range of streams of different sizes. My preliminary results suggest that ecotypes spawning in large water bodies (stream mouth >5m wide, rivers, and beach spawners) show stronger density-dependent marine growth than ecotypes spawning in small streams (stream mouth <5m wide). These results demonstrate that evolutionary selection for spawning success as adults affects the development programs of juvenile salmon while in the ocean.

Marine heatwaves have altered ecosystems globally, including changing community composition and facilitating the spread of invasive species. In south Puget Sound, Washington (USA), non-native Pacific oysters (Magallana gigas) have been farmed extensively for almost a century and grown in enhancement sites, however, they have only recently recruited in the wild. This study explores how the appearance of Pacific oysters was related to spatially (eight sites) and temporally (decade) warmer summer water temperatures in south Puget Sound and compares oyster persistence across five sites where recruitment occurred. The largest recruitment event from 2012-2020 was in the summer of 2015, in the middle of the east Pacific Blob marine heatwave which led to warm water temperatures off the west coast of North America. Throughout the study period, the number of oyster recruits each year was positively correlated with warmer water temperatures. Oyster population densities differed across the five sites where recruitment occurred and generally declined after 2015, but showed no site by year interactions, which is consistent with spatially-variable recruitment and similar post-recrutiment survival. Mean oyster shell heights also differed among sites, which could reflect different growth trajectories or recreational harvest patterns. This study supports the claim that warming sea surface temperatures may interact with species introductions to change modern biogeography. 

One outcome of strong intraspecific interactions is the top-down regulation of juveniles by older individuals in a population, resulting in cohort dominance. Due to their distinct spawning patterns and high variability in juvenile recruitment, many species of fish experience these interactions. Arctic char (Salvelinus alpinus) is a commercially and ecologically important species native to Alaska whose intraspecific interactions are only vaguely understood. The goal of this study was to assess the evidence for suppression of recruitment by older individuals that both compete with and cannibalize younger individuals in a population. I used Arctic char fork length data from Little Togiak River, Alaska, between 1972-2023 to construct annual size distributions. I conducted a wavelet analysis for periodicity to determine whether there was distinct cyclicity in the size distribution of individuals in the population, as would be expected by a population regulated by a dominant cohort. The analysis suggests periods of 10-15 years, the natural life span of Arctic char, where a new cohort arises from a successful recruitment event and suppresses subsequent generations through competition and cannibalism. Improving the understanding of top-down intraspecific regulation in Arctic char can help inform fisheries policy and provide additional insight into Alaskan ecosystem functions.

Modern astronomical survey telescopes, like the Vera C. Rubin Observatory and Extremely Large Telescope, are projected to produce terabytes of data each observing night, raising the need for efficient machine learning algorithms to flag astronomical data for further study. One such algorithm is the Random Forest (RF) algorithm, which has previously been demonstrated to process some 2.36 million different galaxy spectra data (in 12 hours over 128 CPUs) for any potentially unique or even undiscovered phenomena. However, this previous demonstration used spectra taken from the combined light of a given galaxy. RF algorithms are ensembles of structures called “decision trees,” which categorize data points with value-comparison questions. This method can be extended to create a metric to calculate how unusual a data point is compared to other points in the dataset. Our project will extend this RF algorithm for the Mapping of Nearby GAlaxies survey (MaNGA), which accounts for the fact that spectra vary with regions of a galaxy. We explore the behavior of the RF algorithms when accounting for these spatially-varying features. Our methods include the generation of synthetic data to train random forest algorithms, RF model hyperparameter searches, and comparison of models. Furthermore, our project compares the similarity of our results to the results from Baron and Poznanski (2016), which previously applied the RF algorithms to the 2.36 million spectra data. We present the conclusions from the RF algorithms and whether prevalent emission lines are flagged by the algorithms, such as hydrogen-alpha and O-III lines. Our project also discusses the features characteristic of outlier galactic region spectra data. Successful implementation of the RF algorithms to process data pipelines from upcoming large surveys has the potential to accelerate the rate of astronomical discoveries to unprecedented levels.

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.

Dark matter is a theoretical form of matter that doesn’t interact with light or conventional matter despite its large expected abundance in our universe. One potential candidate for dark matter, predicted in models, is weakly-interacting long-lived particles (LLP). The ForwArd Search ExpeRiment (FASER), located in the Large Hadron Collider (LHC) at CERN, uses detectors to search for LLP produced in proton-proton collisions. Upon hitting the detectors, particles generate electronic hit signals that are used to reconstruct the decay products of LLPs. Analyzing these tracks may offer insights into the properties and characteristics of LLP. The FASER detector is made of four tracking stations: 1, 2, 3, and interface tracker, each comprising of three layers with eight Semiconductor Tracker modules in each layer. Due to the imprecise installation of these tracking stations, misalignment occurs preventing accurate track reconstruction. To address this issue, I aim to execute an iterative local Chi-square alignment test to determine alignment parameters for each station individually and collectively using previously collected FASER data. I hypothesize that modules will have improved residual values and sensitivity after alignment.

Ocean salinity, a measure of salt concentration in seawater, is a key variable influencing water density and controls important processes like mixing and stratification. In dynamic coastal systems like Puget Sound, temporal and spatial salinity variability can present a challenge for making high resolution salinity measurements that can characterize these patterns. Salinity is typically measured using widely used conductivity cells, but they can be costly, provide spatially limited observations and can have technical limitations including drift and biofouling. This project therefore developed and validated a cost-efficient buoyancy-driven profiling float for the indirect determination of salinity for use in these dynamic coastal areas. The float is built from stock hardware and electronics, and 3D printed components. The float measures in-situ temperature and pressure through electronic sensors and determines water density at a given depth by achieving neutral buoyancy (e.g., float density equals water density). Neutral buoyancy is achieved by using a stepper motor to precisely move a piston to displace up to 6% of the float’s volume in water. Salinity is then calculated using the TEOS-10 equation of state using these three known parameters. This approach could enable the large-scale production of floats to obtain high-resolution data to better quantify patterns and change in dynamic coastal systems.

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.

Cybersecurity is one of the fastest-growing areas of study within the social sciences, as the internet is becoming an increasingly important piece of the international security system. Russia, along with the cybercriminal gangs that operate there, is one of the key global actors in cyberspace - making it crucial to research in order to better understand the global security landscape. My research centers on the governance structures that have emerged within cybercriminal organizations due to the anarchic, ungoverned nature of the internet. I am seeking to understand what structural factors prevent members of cybercriminal organizations from defecting, whether that be joining another organization or going to law enforcement. Traditional criminal organizations can rely on the threat of violence to prevent this type of defection, but due to their geographically dispersed nature and the anonymity the internet affords their members, cybercriminal organizations often have no way to credibly threaten violence. To understand defection and cybercrime organizations, I analyze the history of traditional criminal norms and traditions in Russia’s vorovskoi mir (thieves’ world) along with in-depth textual analysis of a trove of over 60,000 leaked internal chats from a prolific Russian ransomware gang with the use of natural language processing (NLP). I expect that my findings will show that cybercriminal gangs, without the threat of violence available to them, are forced to resort to positive incentives encouraging members to remain loyal. However, when these incentives fail, I expect to find that defections have very little impact on the overall functioning of the organizations. My research is aimed at contributing to a better understanding of the cybercriminal actors at large in our world today, and provides insightful observations about cybercriminal organizations that not only contribute to theoretical discussions but also have real-world policy implications.

How do organs have such consistent and reproducible shape, form, and volume? One factor of this complex phenomena is cell-cell adhesion. Cell-cell adhesion plays a vital role in organ formation, as it is an essential driver of cell shape, cell arrangements, and tissue structure. To determine the role of adhesion in organ formation, I define the role of E-Cadherin, a cell-cell junction projection that adheres neighboring cells. The developing renal system of Drosophila, Malpighian tubules, are an excellent system because I can selectively manipulate expression of E-Cadherin in the organ and can utilize fluorescence microscopy to observe how these changes affect tubule morphogenesis. I observe where the adhesion protein is located during organ growth, and what happens to organ growth when expression of the adhesion protein is reduced. To track the dynamic localization of E-Cadherin, I take measurements of specific location of E-Cadherin between cells and concentration of E-Cadherin throughout organ development. I expect the concentration of E-Cadherin to increase during elongation, and that it will be enriched in more looped parts of the organ. To define the requirement of E-cadherin during organ formation, I use RNA interference to reduce E-Cadherin expression. Because of how vital E-Cadherin is in other developmental morphogenetic processes, I expect a decrease of expression to have profound impacts, leading to severe organ developmental defects. I measure these defects by comparing cell shape change and organ shape in control and E-Cadherin reduced organs. The results of this study will not only help us understand Malpighian tubule morphogenesis, but it will also help us understand organogenesis more generally. Elucidating the precise mechanisms behind cell behavior, shape, and cell-cell interaction has important human health implications and will enable work in many other fields such as cancer, regenerative treatments, tissue growth, and organ synthesis.

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.

Although the relationship between the structure, function, and physiology of organs is well documented, the mechanisms by which cells collectively coordinate into three-dimensional tissues and organ components remains unknown.The countless factors that inform the morphogenesis of mammalian organs poses a challenge to understand organogenesis from first principles. However, the Malpighian tubules of the fruit fly Drosophila offer an excellent model system for investigating this question due to their rapid development, relative simplicity, and the degree to which scientists can manipulate variables that affect their development. These tubules are the renal equivalent of the fruit fly excretory system; further, many of the genes involved in sculpting these tubules are conserved from flies to humans. One conserved gene is the fly homolog of β-catenin, which is known to play an essential role in cell-cell adhesion. The goal of my research is to define how β-catenin impacts organ morphogenesis. To do this, I use fluorescence microscopy and live imaging to compare wildtype Drosophila to those with decreased β-catenin expression. Using tissue-specific fluorescent protein tagging, I can differentiate Malpighian tubule cells from other embryonic cells under the microscope so that their shapes can be analyzed, and I control the level of β-catenin expression specifically in Malpighian tubule cells using RNAi. Due to β-catenin’s integral role in cell-cell adhesion, I expect to find localization of β-catenin to the cell membranes of the tubules, with high concentration along membranes undergoing the greatest adhesion or motion, and interrupted tubule morphogenesis in reduced-expression lines. I also suspect that cells may completely fail to adhere and will be unable to transmit tension effectively along the tissue. The results of this experiment will contribute to our understanding not only of Malpighian tubule morphogenesis, but of one of the components of morphogenesis in general.

Traditional vaccines use inactivated or live attenuated pathogens to elicit an effective adaptive immune response, but these vaccines can lack safety for immunocompromised individuals. Subunit vaccines–which display characteristic components of pathogens–are safe, stable, and readily engineered, but struggle to elicit a strong immune response. These next-generation vaccines require adjuvants (substances that stimulate the immune system) to increase efficacy. However, many currently used adjuvants lack well-understood mechanisms or wide applicability across vaccines. There is a need for new adjuvant platforms, and protein-based adjuvants are appealing because they are stable, readily engineered, and can be co-delivered with antigens on subunit vaccines. Toll-like Receptor (TLR) proteins are promising adjuvant targets that bind pathogen-associated molecules to activate the innate immune system. Of this family, TLR5 binds the bacterial protein flagellin, but flagellin is not a suitable adjuvant candidate because it is degradation- and aggregation-prone. Therefore, this project aims to design, test, and characterize novel protein-based adjuvants that can bind TLR5 and activate the immune system. Here, I test and characterize de novo mini-proteins that I have computationally designed to bind mouse TLR5 (mTLR5). I use cell-based binding and signaling assays to identify and characterize successful binders. Preliminary results show de novo mini-proteins bind and activate mTLR5 and key knockout mutations at the binder interface decrease signaling. Ultimately, this work hopes to provide a mouse model for these novel protein-based vaccine adjuvants with clinical aims. This project has wide-reaching public health implications, as vaccines offer the potential to improve the health and lives of countless individuals.

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.

Toll-like receptor 4 (TLR4) is an immune protein which binds lipopolysaccharide (LPS) present on the outer membrane of Gram-negative bacteria and activates the innate immune response. In mice, an mRNA splice variant composed of only the extracellular domain of TLR4 was shown to encode a soluble product (sTLR4) capable of inhibiting inflammatory response to LPS. sTLR4 has been recovered from human saliva and demonstrated to dampen the production of pro-inflammatory cytokines by macrophages. Prior work showed that TLR4 was also present in endometrial glands, uterine tube epithelia and endocervical glands. However, there are no published studies exploring the presence or role of sTLR4 in lower genital tract secretions. We tested primary female genital epithelial cells’ supernatants as well as human endocervical cytobrush and vaginal swab samples for the presence of sTLR4 by using a chemiluminescent immunoassay. We found sTLR4 in cervicovaginal secretions, with increased concentration of sTLR4 present in participants with endocervical ectopy and in those sampled during the proliferative phase of the menstrual cycle. Supernatants from endocervical cell lines possessed higher levels of sTLR4 than those derived from ectocervical or vaginal cells. sTLR4 concentration was not correlated with the presence of bacterial vaginosis, age, the concentration of common vaginal Gram-negative bacteria or with genetic variation in the TLR4 locus. By western blotting, we demonstrated that sTLR4 is composed of a ~100 kDa polypeptide, corresponding to the entire TLR4 ectodomain. In a reporter monocytic cell line, we showed dose-dependent inhibition of the LPS/Interferon-regulatory factor (IRF) pathway when LPS was preincubated with endocervical cells’ supernatants. These results point to an unappreciated form of innate immune regulation in the cervicovaginal niche, which could potentially open new avenues for understanding inflammatory disorders such as cervicitis and pelvic inflammatory disease.

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.

Coccidioidomycosis, also known as Valley Fever (VF) is caused by the fungus Coccidioides. Pigtail macaques (PTMs) bred at the Washington National Primate Research Center (WaNPRC) in Mesa, AZ are naturally infected with Coccidioides and are similar to humans in their physiology, symptoms, and immune responses. Populations with a weakened immune system, notably older individuals, are at risk for severe complications from infection. Additionally, there is evidence that males have a higher incidence of VF than females in endemic areas. I characterized the immune responses in a PTM model across age and sex to better understand how VF affects the immune response of these populations. Forty-two PTMs (2.25-19.24 years, 3.66-18.29 kg, 37 female, 5 male) at the WaNPRC were sampled for blood. The frequencies of immune cell subsets in whole blood were characterized by flow cytometry and compared for significant differences based on age and sex. I analyzed sex-based differences with Brown-Forsythe and Welch ANOVA t-tests and found no statistically significant differences. For age-based differences, we used a simple linear regression to analyze differences by age in immune cell subsets. We found that old PTMs (10.07-19.24 years) have higher activation of CD8+ T cells, myeloid dendritic cells, intermediate monocytes, and higher frequency of γΔ T cells and CD4+ γΔ T cells than young PTMs (2.25-9.69 years). Young PTMs have a higher frequency of CD45+ granulocytes, PD-1 High CD8+ T cells, plasmacytoid dendritic cells, and NK cells. By correlating older PTMs with higher immune cell activation, and younger PTMs with higher immune cell frequency, we have a better understanding of how a vaccine or treatment could be developed to support older individuals, who are at greater risk of severe infection.

Damage to the spinal cord causes one of the most debilitating injuries to the human body. The challenge of promoting the regeneration of this dense network of neurons and glia after spinal cord injury has been seen as insurmountable. However, new techniques emerging from the field of regenerative medicine have illustrated the possibility of encouraging the body to repair these injuries on its own. In the Wills Lab, we study the model organism Xenopus tropicalis, or the Western clawed frog, which has the ability to regenerate its spinal cord and associated tissue following amputation. My project focuses on how X. tropicalis uses the developmental morphogen Sonic Hedgehog (Shh) to re-establish the dorsal-ventral (DV) patterning of the spinal cord during regeneration. I have used cyclopamine, a Shh inhibitor, and SAG, an agonist, in order to perturb Shh signaling during regeneration. I then monitored the effect on DV patterning via immunohistochemical labeling of dorsal and ventral markers. Work so far has shown that Shh signaling is in fact necessary to the establishment of proper DV domains in the regenerate spinal cord. However, my research has also hinted that this specification is complex. Shh appears to have a more proliferative role early on, with patterning effects coming later. In addition, there appears to be an interaction between Shh and other signals that specify anterior-posterior polarity. Overall, my research so far has generated new evidence for how developmental signals are repurposed in the context of regeneration. 

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.

As anthropogenic causes of species declines (i.e. habitat loss, climate change) worsen, many carnivoran species are threatened with extinction. These extinctions could also decrease the overall diversity of carnivorans, which have evolved diets as disparate as herbivorous pandas and carnivorous lions, and body shapes as different as elongate mustelids and robust bears. Because phylogeny and dietary ecology influence the evolution of skull shape, we used skull shape to quantify diversity in carnivorans. In this study, I will investigate whether carnivoran species at high risk of extinction have similar skull shapes to one another and whether their average skull shapes are distinct from the average skull shapes of species with other conservation statuses (i.e., least concern). If this is true, specific skull shapes could eventually cease to exist, decreasing skull diversity in carnivorans. I used 3D geometric morphometrics and phylogenetic principal component analysis to quantify skull shape diversity, and I used the International Union for Conservation of Nature (IUCN) Red List of Threatened Species to determine each species’ conservation status. In preliminary analyses, a phylogenetic ANOVA and pairwise comparisons of skull shape against conservation status indicated that there are significant differences in average skull shape among conservation statuses, specifically between vulnerable species and both least concern and near threatened species. This study will reveal new insights into the conservation of carnivorans and carnivoran morphological diversity.

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.

Corepressors are proteins recruited by partner proteins to negatively influence the transcription of genes. TPL is a corepressor from the model plant Arabidopsis thaliana, and while we understand a lot about how TPL works, many mysteries still remain. My project aims to identify other proteins that work with TPL to form a transcriptional repression complex at a single-engineered promoter site. First, we created a synthetic repressor called dCas9-TPL that binds and represses the transcription of the RUBY reporter. The RUBY reporter is a visual marker designed to express throughout the entire plant, turning the green plant a bright purple. Our engineered RUBY line also carries two guide RNA binding sites in its promoter with sequences not found anywhere else in the Arabidopsis genome. This allows dCas9-TPL to bind to and repress this synthetic gene and not affect the transcription of other genes. Many of these plants have morphological phenotypes, and visual screening of the repressed RUBY line showed the plants turn a faint whitish-pink instead of bright purple, signifying that the repression by TPL is working. I have screened mutagenized populations of 40,000 individuals from the validated repressed RUBY plant strains using the Ethyl methanesulfonate (EMS) protocol, which creates new point mutations. I identified 257 individuals from 129 mutagenized families with bright purple organs, which signifies that the RUBY reporter is no longer repressing due to a putative TPL interactor being mutated. I will then proceed to form complementation groups and subsequent DNA sequencing to map the mutations. By identifying regulators of corepressor function in plant biology through downstream whole genome sequencing, I hope to learn principles that can inform cellular engineering across many organisms and better understand why certain mutations associated with transcriptional repression cause developmental defects or diseases like cancer in humans.

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.

The flow Z pinch is an innovative concept of magnetically confining a high-temperature, high-density plasma. It has a linear configuration, where the magnetic field generated from the axial current confines the plasma. Plasma impurities are produced via interactions between high-temperature ions and the electrodes or chamber walls. Bound electrons in these impurity ions are excited into higher energy levels and then de-excited, causing the release of photons whose wavelength can indicate the ion species. Typical impurity ions present in the Z pinch formed on the ZaP-HD device include C-III or C-V. Monitoring this line radiation can allow measurement of velocity, ion temperature, and impurity content. This research focuses on diagnostic instruments used to monitor these impurity emission lines. A Fastie-Ebert spectrometer is coupled to two light-sensing devices: a charged couple device (CCD) camera and a photomultiplier tube (PMT). The CCD captures an emission spectrum at a single time and the PMT measures the time evolution of a narrow-wavelength-band emission. However, due to the modification and backlash in the wavelength adjustment dial mechanism in the spectrometer, a new wavelength calibration is required. Wavelength calibration is conducted by viewing calibration lamps using the CCD and capturing spectra at various dial settings. Using known wavelengths from the calibration lamp, wavelength is related to dial setting by a second-degree polynomial interpolation function, and a chart of the dial settings of common ions monitored used for plasma parameters is created. PMT calibration is conducted by capturing CCD spectra at small incremental dial settings near one known wavelength. An electrometer records the real-time PMT output signal and allows tuning to the maximum of the signal which determines the calibration relative to the CCD spectrum. The diagnostic operation, calibration method, and resulting calibration curves will be presented.

Phytophthora belongs to a group of plant pathogens found in the Kingdom Protista and is known to be highly destructive to plants. The water mold has been found all over the U.S. but just in Washington and Oregon, there are over 30 different species. This water mold is a significant problem in the Pacific Northwest because it infects Pacific Rhododendron leaves. Comprehending the factors that contribute most to the diversification of the Phytophthora species is essential in sourcing and minimizing its spread. We anticipate that the presence of Phytophthora species in residential areas, influenced by human activities, will procure a higher diversity rate of at least 3 or more species. This expectation is rooted in the understanding that the human influence on these environments contribute to an increase in landscaping that works for the proliferation of diverse Phytophthora species. Our location sampling of choice for the bacteria was Chambers Creek located in Pierce County, WA. Chambers Creek has mostly residentially areas surrounding. We gathered four leaves from a rhododendron plant on the Pierce College Fort Steilacoom. We placed the collected rhododendron leaves into bags which were then submerged in the water and left undisturbed for a duration of approximately nine days for observation and analysis. We plan to inoculate the necrotic tissue into V8 agar. We plan to isolate DNA of the Cox-1 subunit protein and sequence it to then use bioinformatics to identify species of Phytophthora. We will use class data to determine if residential areas have an effect on Phytophthora diversity.

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.

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.

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 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.

Our undergraduate research project is on Phytophthora. Phytophthora translates to “plant destroyer”. The purpose of our research is to use watershed studies and bioinformatics to find patterns in the diversity of Phytophthora species found in local streams in Washington State. We assessed prior studies to estimate the probability of discovering Phytophthora in Chambers Creek, a local stream. Our hypothesis is, in comparison to freshwater, water sources closer to the ocean will have a higher diversity of Phytophthora species because they have more sources of contamination and higher saltwater concentrations. We chose the stream location based on previous watershed information we had. Additionally, we baited Chambers Creek with Rhododendron leaves in an attempt to grow Phytophthora. We then cultured the lesions on the leaves onto a V8 agar to grow Phytophthora. After that we isolated the DNA and “cleaned” it by using reagents to remove unused primers and deoxynucleoside triphosphates, then sent it off for sequencing. At this time, we used gel electrophoresis to determine the quality and quantity of our DNA. Lastly, we used bioinformatics to become familiar with sequence analysis using the National Center for Biotechnology Information Basic Local Alignment Search Tool. Three out of the four samples we obtained had no sequence matches but one sample was 100% positive for Phytophthora bilorbang. In conclusion our hypothesis was correct. Since our water source was subject to higher saltwater concentrations, it was more contaminated and had a higher diversity of Phytophthora species. Doing this research and discovering this information will help add to the knowledge of Phytophthora in Washington State. This can lead to where Phytophthora treatment is needed to protect our environment.

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.

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.

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.

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.

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.

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.

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.

 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.

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.

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.

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.

Earth’s sedimentary rock record provides us with a unique window into the co-evolution of life and the environment. One potential indicator of “paleoenvironment” in the stratigraphic record are stromatolites, which are laminated buildups constructed by microbes. The individual laminations of stromatolites (known as “laminae”) are typically treated as recordings of microbial responses to various environmental factors. Yet, the processes that drive the morphogenesis of laminae, as well as their evolution and variability through geologic history, remain poorly understood. Laminae often are described as having millimeter-scale thicknesses, but this statement has not been quantitatively characterized. The first step in linking laminae thickness to broader questions in Earth’s history is to describe the variation of such thicknesses through time and space. Here, we present a multi-scale analysis of stromatolite morphology from the 1.8 billion year old Pethei Group in Northwest Territories, Canada. We describe our analytical workflow, from collecting imagery through photography of polished hand samples and thin sections, to measuring thicknesses in images using a combination of manual and semi-automated methods. We contextualize laminae thickness measurements and larger stromatolite morphological features using three-dimensional reconstructions of bedding planes derived from photogrammetry. We complement this spatially and temporally constrained study with a database of published stromatolite laminae measurements that span 3.5 billion years of Earth’s history. We detail the development of this database, from compiling imagery in the literature to providing metadata for each measured sample, including information on depositional environment and mineral composition. We further describe the development of image and signal processing techniques for analyzing morphological features and assessing laminae periodicity. Our study provides a novel approach to testing historical trends and variations in stromatolite laminae thickness throughout geologic time and space, thereby enabling researchers to gain new insights into the evolution of life on Earth.

Quantum computers represent data through qubits, as opposed to bits in classical computers. These qubits can be implemented using various physical systems, including trapped ions where they are represented by the internal energy levels of individual ions confined within electromagnetic fields. Trapped ions are an attractive choice for qubit implementation since this system has the potential to meet all the DiVincenzo criteria, which are a set of requirements needed to build a mainstream quantum computer. To facilitate the development of mega-qubit (MQb) trapped-ion quantum technologies, the Scalable Quantum Research Lab is conducting extensive research on the persistent issue of collisions with background gas molecules, an error immune to the standard quantum error correcting codes. My research focused on answering the question, 'Can error rates be controlled through trap design?'. To answer this question, there are 3 parameters to determine: (1) trap height: vertical location of the ions from the surface trap; (2) trap depth: how strong the trap is (i.e., how stable is the trapping potential); and (3) trap anharmonicities: the coefficients associated with polynomial potential. These anharmonic potentials can accelerate ions after collisions, thereby increasing collision errors. These results were found using Particle-in-Cells simulations and computational analysis for error minimization. Optimizing the trap design allowed greater control over the collision error rate for a long ion-chain trap. In short, finding a way to control anharmonicity and trap depth using trap geometric optimization can reduce the additional measured errors in the bigger experiments. The results presented are a model of trapped ion and graphs showing relationship between different trap paramters and the three variables mentioned above. 

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.

Phagocytosis is a primary function of macrophages that facilitates the elimination of pathogens and the clearance of endogenous and exogenous debris from tissues. This function can become impaired by aging or exposure to environmental toxicants, which may contribute to impaired immune responses and wound healing. To determine the involvement of macrophage phagocytosis in different pathophysiological conditions, we developed an in vitro macrophage phagocytosis assay with lung macrophages and brain microglia from CD-1 mice (three male, three female). The tested groups included an untreated control group, a lipopolysaccharide (LPS)-treated positive control group to increase phagocytosis, a cytochalasin D-treated negative control group to inhibit phagocytosis, and an experimental pioglitazone-treated group to stimulate PPAR-g (nuclear hormone receptor and transcription factor that has been shown to stimulate phagocytosis). We used green fluorescent protein (GFP)-labeled dextran beads opsonized with serum to assess the phagocytotic abilities of the macrophages in each treatment group. We are currently analyzing data that will determine the percentage of macrophages that phagocytosed the beads (GFP+ macrophages) and the average number of beads phagocytosed among GFP+ macrophages per treatment. We predict that there will be fewer GFP+ macrophages in the cytochalasin D-treated group vs. controls, indicating that phagocytosis is properly inhibited. We predict that both LPS and the pioglitazone-treated macrophages will present more GFP uptake and GFP+ cells than the control. A comparison of pioglitazone and LPS uptake will indicate the relative efficacy of pioglitazone in enhancing phagocytosis. Because pioglitazone is thought to stimulate phagocytosis without causing a pro-inflammatory response like LPS does, pioglitazone treatment may be a way to improve the helpful functions of macrophages with impaired phagocytic functions, which can occur with aging or following exposures to environmental toxicants.

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.

Pain is the number one reason why patients seek medical treatment, yet current pain therapeutics such as opioids have limited efficacy and produce harmful side effects. This has produced a critical need for the development of novel therapeutics for the treatment of acute and chronic pain. Cannabidiol (CBD) shows promise as an analgesic, but the mechanism of action is not well understood as it interacts with several receptors such as cannabinoid receptors CB1 and CB2 and noxious nociceptors TRPA1 and TRPV1. I am investigating how CBD acts on the nervous system to disrupt nociception (pain perception) utilizing the Danio rerio model system and human embryonic kidney cell line 293T (HEK 293T). I use behavioral assays with genetic knockout models to interrogate the molecular mechanism of CBD-mediated analgesia and ratiometric Fura-2 calcium imaging of HEK 293T cells that express TRPA1 or TRPV1 to further elucidate their responses to combinations of CBD, heat, and allyl isothiocynate (AITC, a TRPA1 agonist). My preliminary results indicate that CBD is pronociceptive at low concentrations (10uM) and analgesic at high concentrations (20uM). Recent experiments suggest that CBD’s pronociceptive properties occur via TRPA1 activation, and that this sensitization attenuates CBD-mediated analgesia. I anticipate each CBD receptor knockout will alter CBD-mediated analgesia, with CB1 and CB2 null animals experiencing deficits, while CBD-evoked analgesia may be potentiated in TRPA1 and TRPV1 null animals. I anticipate observing heightened intracellular calcium concentrations when HEK293T cells expressing TRPA1 are perfused with CBD, and increased responses to AITC when cells are perfused with CBD. Importantly, this project creates a platform for the investigation and characterization of minor cannabinoids and other potential therapeutics, using behavioral phenotype based screening to aid in the development of novel, non-opioid analgesics that can revolutionize pain treatment.

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 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.

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.

Drug use has been stigmatized for more than 140 years. This stigma has influenced public policy, school curriculum, and treatment for drug addiction. Additionally, the US has a historical pattern of shaming and excluding people who use drugs, which is especially concerning as opioid overdose death rates have risen every year. This research aims to analyze this historical, morally centered stigma and its effects on people who use drugs, while exploring the role of epigenetics (i.e., gene expression changes) in opioid addiction and opioid addiction treatment. This Literature Review examined textbooks, journal articles, blog posts, book sections, and government websites. This review analyzes historical timelines relating to societal stigma, public policy, and addiction treatment, as well as how the epigenetic mechanisms of opioids relate to those of popular opioid addiction treatments. This research indicates that how popular treatments alter the gene expression changes seen in opioid addiction is unclear due to a lack of research in this area. My analysis of historical timelines shows that the “maintenance” model of treatment, seen with methadone and buprenorphine today, was created approximately 50 years before discoveries that shaped the science of modern epigenetics. Thus, today’s leading treatments were incapable of considering the role of epigenetics in opioid addiction during their development because they were introduced before the science was fully developed. This research suggests a possibility for future use of precision medicine methods for gene editing, like CRISPR-Cas9, as a treatment modality for drug addiction. This analysis also points towards a role of epigenetics in psychiatric conditions like Borderline Personality Disorder and altering gene expression changes associated with successful psychotherapy treatment, suggesting a possibility of precision medicine in a mental health setting as well.

Teachers play an integral part in a child's development as they are responsible for educating youth. Since COVID-19, schools are reporting rising rates of teacher burnout (52%) and students exhibiting problematic behaviors (e.g., disruptions [59%], disrespectful [52%], rowdiness [49%]). Disruptive classroom behaviors hinder the ability of teachers to adequately implement curriculum which results in a variety of disciplinary actions administered. My research analyzes the correlation between elementary school teachers' burnout rate and disciplinary actions taken. My data comes from a larger study that examined the implementation of a preventative social-emotional behavioral program in nine different schools. Teachers completed an online survey distributed in the middle of the academic year, including a measure on burnout, and the district provided systematically obtained disciplinary records (i.e., September to December) upon completion of the study. Preliminary analysis is ongoing to examine the relationship between teacher burnout and disciplinary actions recorded. This can be a first step towards improving teachers' professional well-being and creating a more prosocial classroom environment for student growth.

American military Veterans with a history of trauma exposure have a high prevalence of comorbid mental health diagnoses (e.g., traumatic brain injury (TBI), posttraumatic stress disorder (PTSD), substance use disorder (SUD)) with many overlapping symptoms, resulting in difficulty in diagnosis and treatment matching. Increasing attention has focused on understanding how the direct interactions between these medical and psychiatric diagnoses vary across military subgroups. Critically, current research lacks the usage of larger datasets encompassing wide arrays of interactions between these comorbidities and their unique relationships to disproportionately prevalent diagnoses such as TBI and PTSD. This study aims to investigate the interaction between TBI severity and psychiatric outcomes as well as explore these interactions using a comprehensive network analysis of prevalent psychiatric comorbidities. Data was obtained from Veterans serving in Iraq and Afghanistan who received VA healthcare in clinical visits between 2005-2019 and the United States Veterans Eligibility Trends and Statistics database (USVETS). Veterans will be stratified by TBI severity ranging from 1 (low severity) to 5 (high severity) using Comprehensive TBI Evaluation (CTBIE) and Neurobehavioral Symptom Inventory (NSI) scores. Incidence of all prevalent psychiatric comorbidities will then be calculated for each TBI category to determine the individual relationships between TBI and its comorbidities. This analysis will be followed by a network analysis of co-occurring psychiatric diagnoses to determine their associations in prevalence with one another. In doing so, we hope to create a more nuanced understanding of the mechanisms behind TBI and its psychiatric comorbidities to improve the physical and mental wellbeing of US Servicemembers.

Research shows that approximately 100 million people in the United States (US) have medical debt. However, there is a lack of qualitative research on how individuals experience and navigate the complexities of medical debt. To examine the broad implications of medical debt in the US, we interviewed 12 people from across the country about their experiences affording healthcare. Conducting virtual interviews, we asked people about the resources they use to pay medical bills, the impact of their financial struggles on their lives, and how they accrued medical debt. We analyzed our findings using qualitative coding software, Dedoose, where we identified two major themes. Our participants often lacked an awareness of secondary medical debt resources and instead primarily relied on close personal connections for aid. Second, more than half of our participants experienced feelings of stress. Our research findings show that medical debt exceeds the monetary costs owed to a hospital or doctor and include the severe emotional distress people experience mitigating medical debt, affecting their mental and physical health. Our findings are important because instead of blaming individuals for healthcare costs, they draw out the systemic barriers that prohibit access to affordable, quality healthcare in the US. Our findings also challenge cultural stigmas surrounding medical debt. We anticipate our research to be a starting point for further analysis of medical debt's broad financial and emotional ramifications and to encourage advocacy for a health system that no longer pushes millions of people into massive debt.

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.

Beavers are widely considered to be ecosystem engineers because of the way they construct, modify, and maintain their dams, which in turn, alters the surrounding ecological system. Beaver dams can act as a natural filter and help clean pollution from streams and rivers. The dams also slow down the water flow in a creek or river and form ponds, which means less erosion and, in some cases, less flooding. In spring of 2023, a family of beavers began constructing a new dam along Piper’s Creek, an urban stream in Carkeek Park, Seattle. This recent arrival of beavers provides an opportunity to investigate the impact of beaver dams on water quality in Piper’s Creek. This study investigates the levels of phosphate and nitrate above and below the beaver dams as both ions are commonly implicated in eutrophication events. We took three water samples at each of nine sites along the creek, six upstream from the dam, as a baseline, and three downstream. Ion chromatography was used to measure the concentrations of ions, including phosphates and nitrates. Initial results suggest that nitrate levels may vary seasonally, perhaps due to the presence of salmon carcasses and decaying vegetation in late fall. Longitudinal studies of water quality in Piper's Creek can help elucidate both seasonal variation as well as the impact of the beaver population in this important urban stream.

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.

Elevated concentrations of CO2 in classrooms are a persistent issue facing colleges because of both its ability to reduce academic performance by impairing cognitive function and as an indicator of the increased risk of airborne disease transmission in the classroom. The COVID-19 pandemic applied pressure on most colleges to upgrade ventilation systems including North Seattle College (NSC). Preliminary data collected pre-pandemic found that NSC classrooms’ CO2 concentrations exceeded public health recommendations. We resumed our study post-pandemic now that classrooms have returned to regular occupancy levels. We created updated ventilation efficiency curves by placing Aranet4 carbon dioxide detectors in classrooms with multiple classes under varying occupancy levels. We collected daily attendance of classrooms during each of the courses and their corresponding carbon dioxide levels. We were then able to produce a ventilation efficiency curve to contrast our preliminary data and offer a basis for further study. Initial findings suggest that NSC classrooms exceeding an occupancy of 30 people will surpass 1000 ppm, thereby reaching CO2 levels that impair cognitive functions. This is a sufficient improvement from preliminary studies which found that under previous ventilation conditions, classrooms’ occupancy was limited to 22 people. Continued monitoring of CO2 levels in academic spaces is crucial. With the growing popularity of remote teaching, and online learning becoming more accessible for students, one must wonder if our homes can meet the same standards we hold our classrooms to.

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.

T cells rely on cytokines for cell communication and regulating effector function to respond to infections and cancer. Cytokines are sensed by T cells when bound to surface receptors, activating common second messengers that modulate cell differentiation and distinct cell activity. Second messengers allow T cells to mount controlled and effective immune responses; extracellular signal-regulated kinase (ERK) is phosphorylated (pERK) primarily downstream of antigen-T cell receptor engagement to drive proliferation, differentiation, and survival; signal transducer and activator of transcription (STAT) proteins consist of polyfunctional transcription factors phosphorylated (pSTAT) downstream of cytokine signaling, inducing gene expression to regulate cell activity. Different cytokines give rise to distinct responses by activating the same set of second messengers, raising an unresolved question: how can T cells distinguish between different cytokines and mount distinct functional responses? My project aims to investigate the hypothesis that T cells encode information about cytokine identity and magnitude by activating and translating unique combinations of second messengers into distinct gene expression programs. I will stimulate splenocytes with interleukin (IL)-2, IL-7, IL-15, and IL-21 and measure pSTAT and pERK levels using flow cytometry. To analyze differential gene expression, I will analyze an unpublished single-cell RNA sequencing dataset to determine the effects of different cytokines on CD8+ T cell gene expression programs using single-cell sequencing analysis packages. Previously, I found T cells use combinatorial second messenger activity through differential activation of STAT3 and STAT5 in response to IL-2 and IL-21, and expect cells to upregulate effector and memory genes respectively. I expect IL-7 and IL-15 to differentially activate STAT5, respectively upregulating cell survival and memory response genes while both promoting memory cell proliferation. By elucidating relationships between cytokines, second messengers, and gene expression, we can further our understanding of T cell differentiation mechanisms to inform potential therapeutic targets to combat disease.

 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.

Fluoride is an important element for dental health as it helps to strengthen tooth enamel. Insufficient fluoride intake can lead to weakened teeth that are more susceptible to cavities, leading to further dental issues. On the other hand, dental fluorosis is a condition that causes white or brown speckling of the teeth due to overexposure to fluoride during the development of permanent teeth. For most people, a large portion of their fluoride intake is through water, but water from different sources is expected to contain different amounts of fluoride. Many Americans consume tap water, but bottled water is a popular choice. In fact, US consumers buy about 50 billion water bottles each year which equates to 13 bottles per person monthly. This mass consumption of bottled water raises important questions: 1) Is bottled water a sufficient source of fluoride? 2) How does it compare to tap water? To answer these questions, we collected tap water samples from six cities in the Puget Sound area, along with samples from six popular brands of bottled water. We then used ion chromatography to determine fluoride concentrations in each sample. We found the levels of fluoride in our six municipal sources averaged 0.702 +/- 0.031 mg/L as advertised, however, the levels within the different bottled water brands averaged +/- 0.282 0.307 mg/L, and none met the CDC recommended levels of 0.7 mg/L. The findings of our research are important for keeping consumers well informed about the possible risks or advantages associated with their preferred source of water and for upholding public trust.
 

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 Field of Cultural Mediation, along with Linguistic Mediation, has gained popularity in Italy as the country is increasingly a destination for immigration. As EU migration policies have become more selective, solidifying dangerous paths for “irregular migration” such as the Mediterranean and Balkan routes, reception systems become more complicated, borders become more violent, and public opinion polarizes (Lo Bianco, personal communication, 2023). In this context, it is increasingly important to ensure migrants have someone to facilitate communication between them and the various actors they encounter in an increasingly complicated and dispersed reception system built to dispel them. Thus, the Cultural Mediator, often a former migrant themselves, is increasingly employed by public institutions and social cooperatives to facilitate communication, integration, and to inform newcomers of their rights (Cuiban, 2019). There is inherent precariousness in being placed between these two often conflicting sides, requiring a high degree of social, emotional, and institutional expertise in addition to mere linguistic and cultural knowledge. Due to a lack of national regulation and decentralization, Cultural Mediators in Italy face social and emotional difficulties as well a lack of respect, proper regulation, payment, and support in their jobs. In this paper, I investigate the struggles Cultural Mediators in Italy face, which I argue is a product of the broader decentralization of the “Migration Industry”. By conducting surveys and Interviews regarding the Struggles Cultural Mediators in Italy face in comparison to existing primary and secondary resources, I aim to identify these struggles as well as their contexts.

 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.

 Nitrogen (N) has an important role in aquatic ecosystems, both as an essential nutrient for plant growth and a limiting factor maintaining the overall health and balance of an ecosystem. N concentrations exceeding 0.3mg/L in streams and rivers and 0.1mg/L in lakes and resevoirs pose risk for eutrophication events and ecosystem destabilization. This study tracks the presence of dissolved inorganic nitrogen (DIN) throughout Puget Sound watersheds to determine if concentrations pose a threat to these ecosystems. Our team collected water samples along streams located in the greater Cedar-Sammamish watershed. Ion chromatography was then used to determine the concentrations of nitrate (NO₃^-), nitrite (NO₂^-), and ammonia (NH₄⁺) in the samples. The DIN concentrations varied throughout samples, allowing us to map N levels throughout the watershed and pinpoint regions that are at-risk of eutrophication, as well as identify trends in the fluctuations of N levels. The results of this study contribute to efforts to monitor stream health in the greater Seattle metropolitan area in order to prevent the ecological cascades caused by direct and indirect effects of nitrogen pollution. 

Carnivorans display a wide array of adaptations in their mandibular morphology, potentially influenced by various evolutionary factors like sexual selection and niche divergence. The Mustelidae family encompasses a diverse range of carnivorous mammals such as otters, badgers, and ferrets. Although mustelids are predominantly carnivorous, there is significant variability in dietary habits, with weasels exhibiting strictly carnivorous behaviors, badgers tending towards omnivory, and otters adopting piscivorous diets. This study explores the relationship between ecomorphological differences in mandibular shape and different dietary groups within the Mustelidae family. We will investigate variations in the breadth and length of the coronoid process, a mandibular element associated with biting performance. We hypothesize that carnivorous mustelids will exhibit shorter mandibles and larger coronoid processes compared to their omnivorous and insectivorous counterparts within the family. We predict these morphological adaptations aid carnivorous species by enhancing bite force and optimizing prey processing efficiency. We will use geometric morphometrics on mandible scans from various mustelid species with different dietary habits using 3D Slicer. Additionally, shape variation and allometry will be assessed using phylogenetic regression and phylogenetic ANOVAs to determine the effectiveness of diet as a predictor of mandibular feature variation within the Mustelidae family. This study will reveal new insights on how diet influences mandibular shape and size in mustelids, which can be applied to all carnivoran families. 

The mandible is a functionally important part of the mammalian skull. Various selective pressures shape mandibular morphology, with the impact of diet on mandibular evolution across mammals intriguing evolutionary biologists. In this study, we investigate whether diet influences mandibular evolution across a variety of primate species. The diets of primates vary greatly (e.g., omnivory, insectivory, frugivory, folivory, and gummivory) that are characterized by a range of different physical properties that include toughness, nutrient accessibility, and size. Therefore, we hypothesize that primates with different diets will have distinct mandibular morphologies, particularly traits associated with the relative length and width of the whole primate mandible as they may impact the speed and force transmission during biting. To test these hypotheses, we utilized a database of 3D scans of primate mandibles from natural history museums. We quantified the mandibular shape and size by placing landmarks and semilandmarks on digitized 3D models of primate mandibles. We then quantified primate diets by ranking the relative importance of food items found in primate species based on a literature review. We then used phylogenetic comparative methods to test the effect of dietary ranks on primate mandibular size and shape. We predict that different dietary categories and ranking of food items will influence mandibular evolution. For instance, primates feeding on gums by gouging or scraping would require a higher mechanical advantage, resulting in a relatively shorter and more robust mandible. Our research could be helpful for future primate and mammal studies focusing on the selective pressures on the evolution of the mandible.

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.

Body shape provides useful insight into the diversity and evolution of vertebrate body plans. Previous research revealed that body shape scales with body size in carnivoran mammals, but whether this trend occurs in other mammals remains unknown. Our goal is to examine the relationship between body shape and body size in Leporidae, which consists of rabbits and hares. Leporids exhibit a unique gradient of locomotion types between the smallest rabbits that are primarily saltatorial and the larger hares that are primarily cursorial. We quantified body shape using the head-body elongation ratio and body size using the geometric mean of all measured traits from osteological specimens held at the Burke Museum and other natural history museums. We tested the allometric relationship between body shape and size using a phylogenetic regression and also tested if these allometric relationships differed between saltatorial rabbits and cursorial hares using phylogenetic ANCOVAs. We predict that the differences in locomotory modes within the family could influence the correlation between body shape and allometry, possibly leading to different amounts of correlation in rabbits and hares. Specifically, we predict that larger body size corresponds with more elongate body shape. Elongate body shape would facilitate cursorial locomotion more common in larger leporids. More elongate and flexible bodies would allow the forelimbs to reach farther forward, enabling longer strides while running. This study informs allometric relationships of body shape and size of saltatorial and cursorial members of leporids and can lead to future research into the relationship between locomotion and body shape in other mammalian clades.

Sexual dimorphism describes the distinct characteristics between males and females of the same species. These different traits can be traced back to many external and internal factors. Here, we investigated how temperature and habitat influenced the evolution of sexual dimorphism in the mandible of mustelids. Mustelids are a great clade to study sexual dimorphism because of their mandibular diversity that is associated with their diverse diets, behavior, and ecological niches. We test two hypotheses: first, we examined if mustelids in high temperature environments exhibit higher degrees of dimorphism in their mandibles compared to those in lower temperature environments. Second, we test how the degree in dimorphism varies among mustelids inhabiting different habitats. To test our hypotheses, we quantified sexual dimorphism of mandibular size and shape using 3D geometric morphometrics. We then used phylogenetic ANOVAs to test how habitats influenced the degree of mandibular dimorphism and phylogenetic regressions to test how temperature influences the degree of mandibular dimorphism. We predict that mustelids in high temperature environments will exhibit a higher degree of sexual dimorphism than those in low temperature environments. Higher temperatures could correspond to a limited amount of resources which could lead to interspecies competition for food resources. As males and females use different strategies to survive, sexual dimorphism in mandibles size may occur. In our habitat analyses, we predict that mustelids in deserts will exhibit a higher degree of sexual dimorphism compared to those found in other habitats due to the same factors of resource availability and competition. Deserts tend to have scarce resources compared to forests and we believe that sexual dimorphism will reduce this competition. This research is impactful because as climate change is increasing temperatures all over the world, we should be aware of how that is directly affecting species.

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.

Deep brain stimulation (DBS) is currently being studied as a potential treatment for Alzheimer’s Disease (AD). At this time, little is known about how prospective users of such devices view this kind of surgical intervention. User-centered design recommends early input from potential users of devices to ensure that their needs and values are recognized in the design process. From February 2021 to January 2022, 34 qualitative interviews were conducted with individuals who are at risk for dementia due to factors including family history, genetic biomarkers, or mild cognitive impairment. Subjects were asked for their perspectives on the hypothetical use of DBS devices to assist individuals living with dementia. One focus for such a hypothetical device is the improvement of short-term memory. My research examines interviewees’ perspectives on the value of improved memory as well as the value of forgetting, and how their stance informs their perspective on and personal interests in the hypothetical use of DBS to treat AD. Using Atlas.ti for coding and qualitative analysis, I found that a small number of interviewees expressed complete enthusiasm for, or complete rejection of, the device. The majority of participants were intrigued while concerned. I show how these interviewees answered questions such as, “Do you think there are any downsides to having a better memory?” and/or, “Do you think there is value in forgetting” in ways that suggest nuanced perspectives on the value of memory and forgetting. They voice concerns regarding potential drawbacks of the hypothetical memory device in relation to risk, trauma, and caring relations. As clinical trials for DBS in AD continue, understanding the values and concerns of prospective users will be vital information in both the design process and successful clinical trials.

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.

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.

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.

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.

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.

CD-1 outbred mice are a preferred mouse model used in behavioral research due to their genetic diversity and overall capacity for acquiring learned behavior. Psychiatric traits involve cognitive brain function, and psychiatric disorders always result in cognitive dysfunction. Therefore, behavioral research aimed to model psychiatric disease states can maximize translational value to human brain function and behavior by incorporating the study of effects on cognition in CD-1 mice, specifically. However, behavioral tests that target cognition sensitivity have been only made in other mouse strains. Here, we present a study establishing a human-compatible touchscreen-testing environment where we conduct the 5-choice Serial Reaction Time (5-CSRT) task, a mouse-friendly adaptation of the human continuous performance task on CD-1 mice from both sexes to phenotype baseline attention, impulsivity, and motivation. The 5-CSRT task is administered within automated Bussey touchscreen chambers and requires extensive behavioral shaping, habituation, tone, and screen touch training sessions designed to activate underlying neural circuitry involved in cognitive function. Before moving on to the next session, specific criteria must be met in order to promote acquired learning. These sessions become progressively more difficult, with an ultimate goal of priming the mice for the 5-CSRT touchscreen task, whereby animals need to find and touch a visual target being displayed from 5 available options on a touchscreen within a given amount of time to receive a food reward. To analyze and phenotype cognition, we combined the use of camera recordings to observe high-resolution behavioral dynamics in addition to summary task metrics that include accuracy, omissions, premature responses, and food-retrieval latency. Our results establish the use of CD-1 mice with human-compatible touchscreen-based approaches to facilitate an entry point in the effective modeling of psychiatric disorders that can map to human cognitive dysfunction.

Chemical communication is the oldest and most widespread form of communication across the tree of life, and markedly present among lizards. However, the drivers of chemical profile variations in this group remain for the most part uninvestigated. In South American lizards of the Tropiduridae family, semiochemicals are produced by epidermal gland organs called α-glands, exclusively found on the ventral side of male individuals of at least 40 species from four genera. The chemicals produced by these glands are hypothesized to interact with their environments in different ways since chemical species are naturally reactive and tend towards their lowest energetic state. Thus, the intrinsic properties of a semiochemical impact its survival and efficacy for communication. Given the diverse ecology and broad geographical distribution of tropidurids, we investigated whether variation in the chemical composition of α-gland secretions correlates with temperature, humidity, and habitat openness. We performed liquid chromatography-mass spectrometry (LCMS) to obtain the metabolomes of three different sample types. We sampled male skin containing the α-glands, undifferentiated male skin, and female skin. Environmental and chemical property data were extracted from online databases, literature, and field observations. Preliminary tests were done by making Venn diagrams comparing the metabolomes of each sample type. These revealed differences in metabolite compositions, notably between males and females as well as between glandular and undifferentiated skin. From the metabolomes of α-glands, we expect to see chemical species with properties that confer greater survival given the specificities of the environment. For example, given a lizard from a hot and humid environment, we expect the metabolome of the α-glands to contain higher molecular weight species with less functional group complexity. Understanding how environmental parameters drive the chemical composition of α-glands is expected to provide a deeper understanding of the evolutionary history of chemical signaling in terrestrial vertebrates.

Sickle cell disease (SCD) is an autosomal recessive disorder caused by the inheritance of two abnormal hemoglobin genes and can present numerous health complications due to the sickling of red blood cells. It can lead to multiple comorbidities, and can have adverse effects on pregnancy. SCD is very common in Sub Saharan Africa in mainly developing countries, but it can pose risks to pregnant women even in developed countries. There has been conflicting information about whether or not the presence of SCD poses a significant risk to pregnancy outcomes. To analyze this, I systematically reviewed previously published evidence on the effects of SCD on maternal pregnancy outcomes in women from multiple hospitals in different countries from 1973 to 2021. I did a matched comparison of the outcomes between mothers with SCD (HbSS genotype) and mothers without SCD (HbAA genotype) using data from 11 studies. For binary variables, I used log risk ratio as the effect size metric, and for continuous variables, I used standardized mean difference. I hypothesized that the presence of SCD would pose a significant risk to pregnancy and lead to higher occurrences of adverse pregnancy outcomes. I found that there was a significantly higher risk of adverse pregnancy outcomes in mothers with SCD and their offspring, compared to mothers without SCD and their offspring. Perinatal mortality and stillbirths increased significantly with SCD, as well as intrauterine growth restriction, urinary tract infection, eclampsia, and occurrences of preterm delivery and c-section. I found that there was not a significantly higher risk of maternal death and premature rupture of membranes in mothers with SCD, although their risk was higher overall. Proper knowledge of the risks of pregnancy with SCD as well as proper management and treatment of the complications associated with it can improve maternal and fetal outcomes.

Lizards in the family Tropiduridae have ventral epidermal gland organs that are involved in chemical signaling and whose secretory mechanism is entirely unknown. This is because, like other epidermal generation glands, 'alpha-glands' lack a pore through which their secretion can be exerted. Chemical signaling is a valuable aspect of tropidurid lizards' social and ecological interactions, and some have been observed territorially scraping their alpha-glands against the substrate. This process has been hypothesized to facilitate the release of chemical signals via abrasion. To investigate this abrasion hypothesis, we analyzed 74 skin samples from 27 tropidurid species, using light microscopy and scanning electron microscopy (SEM). The SEM revealed incredible surface variability in epidermal glands, providing morphological insight. We found that the exposed glandular mass of each gland scale rests atop the oberhautchen layer of the skin's subjacent generation, which indicates that the secretion of chemicals involves exposing a mostly solid glandular material on the outside of the scales. Histological sectioning of gland scales revealed morphological consistency, indicating that the same secretory mechanism is shared across the tropidurid phylogeny. Imaging of histology samples also revealed that the shedding process which exposes the glandular material may be facilitated by the clear layer, found directly above the glandular mass during development. Characterization of morphological patterns in the formatted SEM images and comparison with histological data should provide evidence for or against taxon-specific or ecology-specific alpha-gland structures, and further support the idea of chemical secretion requiring epidermal exposal of glandular material. Investigations of the morphology and functional mechanism of this unique organ provide insight into the behavior and evolution of tropidurid lizards and shed light on factors influencing the evolution of chemical signaling in terrestrial organisms. 

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.

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.

Seattle's Black community faces significant challenges in banking and finance, including a lack of trust, limited access, and systemic barriers. While FinTech presents potential solutions, concerns persist regarding data privacy and cultural sensitivity. In response to these issues, I took on a central role in facilitating a research project that aimed to co-design an inclusive FinTech solution directly with Black communities in Seattle. To create a participatory design approach and host collaborative sessions with a Black community advisory board, my research group worked closely with community leaders and stakeholders. This involved organizing focus group sessions, where we explored three key questions: 1) What challenges do Black Americans face in banking and FinTech? 2) How can FinTech revolutionize their economic outlook? 3) How can community members co-design a culturally sensitive virtual AI assistant for mobile FinTech? Throughout these sessions, I played an active role in observing and leading discussions to ensure that community members had a meaningful voice in shaping the solution. I also analyzed the data collected from these sessions using inductive and deductive coding techniques to identify key insights and inform the process and steps for the next focus group. We anticipate developing a prototype solution designed by and for Black communities, prioritizing trust, accessibility, and empowerment. Preliminary findings suggest that community engagement and co-design processes are crucial for creating effective and culturally sensitive FinTech solutions. Additionally, fostering understanding and collaboration between Black communities, financial institutions, and FinTech developers is essential for paving the way towards a more equitable financial future for all stakeholders involved.

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.

Quadrupedal locomotion in leporids (rabbits and hares) relies on a unique appendicular skeleton that facilitates a propulsive gait. Long hindlimbs compared to forelimbs supports the pushing followed by a tucking motion that is characteristic of a rabbit hop. Leporids inhabit a range of habitats leading to a variety of specified anatomical needs to support propulsive gaits on different terrains. In this study, we investigated how habitat type influences the morphological diversity of the appendicular skeleton in rabbits and hares. We quantified the appendicular skeleton of different species using caliper measurements from osteological specimens held at the Burke Museum and other natural history museums. We then used phylogenetic comparative methods to test three predictions on how different habitats influenced the evolution of these skeletal traits . First, leporids inhabiting deserts or wetlands will exhibit adaptations in the limb bones that facilitate greater muscle attachment because jumping on softer ground requires more propulsive force than jumping on solid ground. Second, leporids inhabiting open grassile environments will have longer legs to support quickly escaping predators in search of limited vegetation for hiding. Third, more closely related leporids will exhibit more similar hind limb morphologies due to having a more recent common ancestor. Overall, this study elucidates the evolution of appendicular skeleton of leporids and informs us of a possible method to predict how the morphology of limbs evolve based on habitat type and close ancestry in mammals.

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.

Gravitational waves are ripples in the fabric of space-time caused by the rotation and merging of black holes deep in our universe. On earth, these waves cause a minute strain that can be measured with the Laser Interferometer Gravitational-wave Observatory (LIGO). We are developing a calibrator that exerts an oscillating gravitational force on the test mass of LIGO to precisely calibrate the strain sensitivity of the interferometer. The calibrator consists of four motor-driven rotors which are placed around the test mass. The four motors have to run at a constant speed and have to maintain an exact phase relationship. In this research, I designed motor-controller software in Python, which rotates the motors with constant speed with a phase uncertainty of less than 2 degrees. The system uses one of the motors as the reference and converts its encoder position changes to frequency. A Proportional-Integral-Derivative (PID) loop locks the encoder’s frequency to a reference frequency. Then, the three witness encoders are locked to the lead encoder. The gravitational force from the calibrator is calculated using a Python program from Prof. Gundlach's team. The code decomposes the calibrator into 3D points and applies multipole expansions to accurately compute the force at the center of LIGO’s test mass. The gravitational calibrator will help to reduce uncertainties in LIGO’s strain readouts.

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.

Apolipoprotein E (APOE) is a polymorphic gene with 3 main alleles: APOE2, APOE3, APOE4.The APOE4 allele is the greatest genetic risk factor of late-onset Alzheimer’s disease (AD). Carriage of the APOE4 allele results in increased amyloid-β (Aβ) accumulation, aggregation, and deposition in the brain compared to other genotypes, thus contributing to AD pathogenesis. Preeclampsia (PE) is a pregnancy-specific disorder associated with maternal and perinatal mortality. PE has also been linked to protein misfolding pathology, characterized by the accumulation of misfolded proteins (including Aβ) in the placenta, urine, and blood. Due to the similarities with AD pathology, we investigated the relationship between APOE4 and the development of PE-symptomatology in a transgenic mouse model carrying human APOE. To assess this relationship, we induced a PE-like syndrome in APOE3 +/+ and APOE4 +/+ pregnant females on gestational day 13.5 using the Reduced Uteroplacental Perfusion Pressure (RUPP) procedure (n=5-6/group). Sham procedures were performed on the same number of females in each group as controls. On gestational day 17.5, differences in blood pressure, proteinuria, kidney integrity, placental efficiency, placental morphology, and fetal weights in response to RUPP were assessed and compared between APOE3 and APOE4 mice. Our results show that RUPP induced significant kidney damage in APOE4 mice (p=0.014), while APOE3 mice maintained normal kidney morphology. Moreover, RUPP was associated with a significant reduction in placental efficiency in both groups, but only APOE4 fetuses developed fetal growth restriction (FGR, p=0.045). APOE3 was also associated with mild changes in placental morphology, but APOE4 was not. Blood pressure or proteinuria did not vary in response to RUPP in either genotype. Our findings suggest that APOE4 confers an increased susceptibility to kidney damage and FGR in response to RUPP compared to APOE3. Further investigation is needed to determine the molecular basis of these genotype-dependent effects.

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.

Primates are known to have relatively larger brains for their body sizes (i.e., encephalization) when compared to other mammals. Previous studies have examined the relationship between food scarcity and endocast volumes in Old World Monkeys, specifically looking at the Cognitive Buffer Hypothesis –which states that greater rates of encephalization are correlated to living in more variable environments. In this study, we examine whether and how the cognitive requirements of social interactions (Social Brain Hypothesis, SBH) and investment from parents (Parental Investment Hypothesis, PIH) influence the evolution of encephalization in primates at the macroevolutionary scale. Social interactions stimulate the brain, causing it to produce more neural connections and as a result, greater encephalization. A higher degree of parental investment renders a safer space for infants to mature and their brain to develop, providing the capability of reaching larger sizes. To test these hypotheses, we first quantified encephalization as the ratio of endocranial volume to estimated body mass. Our data was collected using CT scans of primate crania from the Natural History Museum, London, United Kingdom. We quantified endocranial volume in 3D Slicer. We then used phylogenetic comparative methods to test how encephalization relates to social factors such as group size, social complexity, gestation length, and litter size across various primate species. Group size and social complexity serves to quantify the social interaction variable stipulated in the SBH, while gestation length and litter size correspond to determining the scale of parental investment per offspring. If the SBH and PIH are supported, then each of the four factors will exhibit a positive relationship with encephalization. Our findings will elucidate how social interactions and parental investment influence the evolution of brain sizes in primates.

The Tacoma-Pierce County Health Department assessed six communities in Tacoma and on the peninsula that are facing limited opportunities for optimal well-being; the objective is to investigate health care quality and efficiency. These communities are called Communities of Focus (CoF). Our work explores state data to evaluate the relationship between hospital length of stay (LOS) and cause-specific hospital readmission rates (CSR) in CoF. The current literature has not reached consensus on understanding the designated relationship. Applying a multivariate logistic regression model, we explored the relationship between length of stay and CSR using data from the Comprehensive Hospital Abstract Reporting System (CHARS). Several covariates including race and gender were included in the regression model. One extra day of hospital stay resulted in a 0.5% higher risk of 30-day CSR [Odds Ratio (OR): 1.005, CI: 1.00–1.01]. The control factor substantially increasing risk of readmission, was age 55-65 years (OR: 3.477, CI: 3.19–3.79). The covariate significantly reducing CSR was being female (OR: 0.816, CI: 0.80–0.83). Individuals of Indian race (OR: 1.120, CI: 1.04–1.20, p <0.05) and Hawaiian race (OR: 1.157, CI: 1.06–1.26, p <0.0001) were more likely to experience CSR compared to White patients. Finally, CoF faced higher risk of CSR (OR: 3.973, CI: 3.77–4.19) relative to other regional patients. CSR is re-admission to the healthcare setting due to a complication or an exacerbation of disease. A single day hospital stay yielding higher risk for CSR suggests that further exploration is needed to understand patient experiences while admitted. Additionally, an investigation into the social, economic, environmental and personal factors placing CoF at risk for CSR is warranted in order to reduce risk. Our research identifies the problem; future work is needed to elucidate the source so that approaches can be identified to lower CSR for CoF.

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.

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.

Colorectal Cancer (CRC) incidence is rapidly rising in patients younger than fifty with no prior family history. Recent research has revealed that very low frequency somatic mutations accumulate in normal tissue with aging. Many of these mutations occur in common cancer genes and are positively selected, producing clonal expansions, that might be linked to cancer risk. I hypothesize that the normal colon of patients with early onset CRC might carry clonal expansions driven by mutations in CRC genes, which could be detected with ultra deep duplex sequencing. Duplex sequencing is an error correction method that improves sequencing accuracy through double stranded molecular tagging. I used duplex sequencing to perform ultra deep sequencing (~3000x) of the main driver genes of colorectal cancer (CRC) which includes: BRAF, APC, FBXW7, KRAS, PIK3CA, SMAD4, and TP53. First, I performed adjustments of gene probes by increasing or decreasing the proportion of each probe in each gene pool to achieve comparable depth of sequencing across genes. Then, I sequenced normal colon tissue of two patients with and without CRC. Preliminary data showed deleterious mutations in tumor suppressor genes: APC, FBXW7, and TP53, which occurred at higher frequency in the patients with cancer. The next steps of this research involve analyzing the normal colon from a larger cohort of patients with and without CRC. Our results have demonstrated that clonal expansions are visible in early onset CRC with these sequencing methods, and I anticipate that I will identify more clonal expansions in patients with early onset CRC than in those without CRC. In conclusion, duplex sequencing allows for the detection of cancer driver mutations in normal tissue, allowing the discovery of early somatic events in CRC and offering potential for early CRC detection, prevention, and prediction.

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.
 

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.

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.

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.

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.

During times of increased hospital capacity, unconventional areas (what we have termed “non-standard clinical spaces” such as converted conference rooms, hallways etc.) may be activated for inpatient activities. Studies regarding issues with Emergency Department boarding while awaiting an inpatient bed have been well documented, but the same practices in inpatient spaces have not been well studied. Qualitative semi-structured interviews were conducted between February-April 2023 at a large urban, public hospital to understand how patients experience these spaces. Any adult English-speaking patient admitted to the hospital for at least two days who was physically located in a non-standard clinical space (hallway beds, PACU beds, former Emergency Department bays converted to inpatient spaces) upon approach for interview was eligible to participate. Eight participants ranging in age from 20-70, across diverse demographics were interviewed for the study. Three themes were identified from the interviews: overall experience, unspoken messaging, and impact to care. Poor patient experience was affected by noise related to crowding in their space and a lack of barriers from noise, lack of privacy from close quarters with others, lack of reliable bathroom access, loss of control over their environment, and inability to gain the attention of nursing staff (though, interestingly, this was not universal as some mentioned that they were in closer proximity to their nurses and able to receive more direct interaction). They described perceiving unspoken messaging about being selected for a nonstandard clinical space. Most commented on the kindness and understanding of the nursing staff and did not necessarily perceive a significant impact to their overall care. When these types of spaces are considered for use for inpatient boarding, patient privacy, control, and access should be prioritized. Transparency and communication about the situational context may help mitigate negative effects of these spaces.

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.

Stimulator of Interferon Genes (STING) signaling contributes to tumor immunity. However, treatments targeting the STING pathway are limited by route of administration, insufficient STING activation, and off-target toxicity. We introduce poly-STING, a copolymerized, mannosylated variant of the diABZI STING agonist-3 known to activate the cGAS-STING signaling pathway, promoting the release of type-1 interferons and pro-inflammatory cytokines leading to tumor immunogenicity. The STING agonist-3 is a non-nucleotide molecule that activates the STING pathway, but it has poor solubility, which limits its usage in-vivo. The developed poly-STING platform improves the drug's solubility, is designed to target immune cells, and provides enzyme-triggered drug release upon delivery, which has been shown to induce improved therapeutic efficacy compared to the free drug. The Pun and Stayton labs seek to investigate modalities for optimization of the cGAS-STING pathway activation and characterize the mechanism of action. Specifically, my project will evaluate STING activation by observing macrophage repolarization from type M2, as the mannose from the poly-STING binds to the CD206 receptors on M2 macrophages. This activates the STING pathway, repolarizing the macrophage to pro-inflammatory type M1. To test effects in vitro, I will culture bone marrow-derived M2 macrophages with various formulations of poly-STING, and repolarization will be measured through flow cytometry and RT-qPCR to quantify expression of macrophage markers. We expect to find higher M1 activity in macrophages treated with poly-STING as opposed to the free drug. Next, I evaluate the therapeutic efficacy of the STING formulations through an in-vivo tumor reduction study using murine models of breast cancer and melanoma, expecting to find longer survival of mice treated with poly-STING. The culmination of this project will result in a polymer-based STING agonist delivery platform that solves the solubility and bioavailability issues associated with the STING-3 agonist, with enhanced efficacy and decreased toxicity after systemic administration.

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.

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.

VLA-4 is a surface protein of immune cells that plays an important role in their extravasation into tissues during an immune response. In multiple sclerosis (MS), pathogenic T cells enter the brain and attack nerve cells by using VLA-4 to bind VCAM-1, a cell adhesion molecule on endothelial cells that line blood vessels. Current MS treatments rely on antibodies that bind VLA-4 and block interaction with VCAM-1, preventing a pathogenic immune response. However, antibodies are expensive to manufacture, and their binding cannot be easily regulated to control drug-induced side effects. Aptamers are single-stranded DNA or RNA molecules that fold into sequence-defined structures capable of binding targets with affinities and specificities comparable to antibodies. Being chemically synthesized, they are much cheaper to manufacture and offer no batch-to-batch differences. Unlike antibodies, their binding in vivo is rapidly reversible, which could alleviate some side effects of disease treatments. However, aptamers have limitations in vivo – degradation by nucleases in serum, and rapid clearance into urine. This project designs and assesses modifications to a novel VLA-4 binding aptamer to improve in vivo function, with the goal of developing an alternative for MS treatment. We designed various modifications to the aptamer backbone to prevent nuclease degradation and conjugated the aptamer to a polymer to increase size and reduce clearance. To assess aptamer functionality, an in vitro model of T cell adhesion is used. VCAM-1 coated plates are used to simulate endothelial cells, and VLA-4+ T cells are incubated in the plates to allow adhesion in the presence of modified versions of the aptamer and serum. VLA-4 inhibition by our aptamer designs is assessed by characterizing the extent of cell adhesion inhibition. Successfully designing a modification that significantly improves the in vivo function of aptamers will have broad implications for their clinical translation to in vivo use.

 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.

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.