Language input is necessary for language development. Importantly, mothers have been shown to speak to infants more than fathers do. My study asks whether this pattern extends to the amount of music that mothers produce or play to infants. Music impacts people neurologically, emotionally, and even physically, and can possibly be used to enhance the linguistic development of infants alongside speech. I am comparing the amount and type of speech and music heard by infants in mother-father families to infants in mother-mother families to isolate the variable of gender and gauge its association with infants’ auditory input. Daylong Language ENvironment Analysis (LENA) recorders are used to record everything in an infant’s naturalistic environment (at home) therefore capturing how many instances of in-person and/or electronic speech or music occur and whether parents’ speaking and/or singing is directed to the infants. Undergraduate students are currently annotating LENA recordings of twenty-one mother-mother families (ages 3-24 months) and twenty-three mother-father families (ages 6-24 months) for the amount and type of speech and music present in infants’ audio environments. Annotators indicate what is heard in 100 randomly sampled 10-second segments from each daylong recording. Using independent samples t-tests, I am analyzing the differences in the average amount of music, the average amount of speech, and the type of music presented to infants of mother-mother families versus infants of mother-father families. I hypothesize that there is significantly more speech and music heard by infants in mother-mother families compared to infants in mother-father families. I also hypothesize there is significantly more singing heard by infants of mother-mother dyads, but a comparable amount of electronic music. If found, these results will point to gender being associated with auditory input variability, expanding the knowledge on environmental factors that influence infant language development.

Geographic Information Systems (GIS) are a powerful framework that enables the analysis of spatial data, or information connected to a location, that can be applied to a variety of fields, such as public health, policy, agriculture, and environmental management. With these frameworks, we can create maps to convey specific data or general information, make comparisons between data sets, and quantify spatial characteristics. In this study, I utilized a GIS framework together with Real-Time Kinematic Global Navigation Satellite Systems (GNSS RTK) methods to collect spatial data regarding hiking trails throughout Saint Edward State Park in Kenmore, Washington. I hypothesize that the positional accuracy of data collected via GNSS RTK is significantly better than positions collected via methods that do not incorporate differential correction. To test this hypothesis, I will collect differentially corrected positional data along trails in the park and compare them to uncorrected data gathered between 2013 and 2015 from the same trails. Comparisons will be made using a t-test to determine whether there is a statistically significant difference between the mean differences of each data set. Furthermore, I will use this spatial data to visualize more accurate trail locations, indicate areas requiring maintenance, and decommission unofficial trails that are harmful to the ecology in the park.

Prostate cancer has a significant heritable component. It is estimated that 10-15% of patients with advanced prostate cancer carry an inherited predisposing genetic mutation, and these patients generally present with a younger age of onset and a strong family history of cancer. The standard in the field of oncology is to conduct short-read DNA sequencing on such patients to find predisposing mutations. While short-read sequencing does well to identify simple mutations that cause disease in many families, our lab concluded that short-read sequencing misses critical mutations in many prostate cancer susceptibility genes. We hypothesized that prostate cancer in many families is due to complex inherited mutations such as genomic deletions, inversions, and mobile element insertions that are not detectable by conventional genomic technologies such as short-read sequencing. To test this hypothesis, our lab specifically recruited prostate cancer patients who, despite having family histories of cancer, did not have any mutations detected via conventional genetic sequencing methods. This project utilizes Nanopore long-read DNA sequencing, which reads DNA in longer fragments and can reliably detect complex mutations. My role is to conduct long-read sequencing on DNA samples from these patients, then analyze the DNA sequence for mutations. I have sequenced 32 patients so far and identified 4 complex mutations through long-read sequencing which were missed by other approaches. These complex mutations include insertions of repeat sequences and duplications which disrupt gene function in BRCA1 and BRCA2. This suggests that, consistent with our hypothesis, some patients who do not have mutations found via conventional sequencing methods do indeed carry causative mutations in well-established prostate cancer risk genes. By finding these mutations, patients can receive more targeted and effective cancer treatment, and undiagnosed family members stand a better chance of catching cancers at earlier stages.

Autism is a neurodevelopmental disorder characterized by differences in social-communication and the presence of restricted and repetitive behaviors and interests. Many autistic individuals engage in “camouflaging” to hide or change their behaviors associated with autism to avoid social stigma. This study aims to explore neurophysiological characteristics underlying camouflaging. Electroencephalography (EEG) is a popular psychophysiological tool that measures brain activity through oscillatory patterns, reflecting various cognitive and emotional processes. Specifically, during “resting state” (when the brain is exposed to minimal external stimuli), theta waves have been shown to have increased activity during periods of increased cognitive load, attentional demands, and task difficulty – mental states that all relate to camouflaging based on qualitative research.  Participants included autistic (n=108) as well as non-autistic adults (n=85), between the ages of 15 and 31 years. Participants completed the Camouflaging Autistic Traits Questionnaire (CAT-Q) which measured three domains of camouflaging in autism: compensation, masking, and assimilation. EEG recordings were taken during resting state and oscillatory activity in the theta frequency band (4-8 Hz) will be analyzed. Our hypothesis is that camouflaging traits will be positively correlated with theta wave activity.  Camouflaging can lead to various challenges for autistic individuals, including depression and anxiety. Thus, identifying the proposed analyses could provide valuable insight into the cognitive and emotional processes of camouflaging, ultimately contributing to a better understanding and potential treatment for mental health challenges faced by the autistic community.

Extensive research has established that certain social determinants of health (SDoH) can significantly influence health outcomes for older adults. Older adults from socioeconomically disadvantaged backgrounds often lack access to a tertiary or quaternary care hospital for complex care for severe chronic illness. Such healthcare disparities have the potential to negatively impact cognitive and mental health for older adults with chronic illnesses, particularly after intensive care unit (ICU) hospitalization. There is limited research examining the associations between SDoH, cognitive function, and mental health in older adult ICU survivors during the early post-ICU inpatient transition period. We aim to gain insight on how to best tailor inpatient interventions to suit older adult ICU survivors from disadvantaged or vulnerable backgrounds. Our project has two overall goals: 1) explore associations between SDoH, self-reported depression and anxiety, and severity of illness; and 2) explore associations between SDoH, selected objective measures of cognitive function, and severity of illness. Data from two ongoing randomized controlled trials of older adult ICU survivors enrolled from the University of Washington Medical Center or Harborview Medical Center are utilized. Participants answer the PROMIS Depression and Anxiety questionnaires to evaluate self-reported depression and anxiety symptoms. Chart review of participants’ inpatient electronic health records provides relevant information regarding SDoH, including home addresses, which provide information on census tracts towards index calculations. Other relevant clinical variables documented by inpatient healthcare providers, particularly severity of critical illness, are used for descriptive analyses. Ultimately, we anticipate that the results from this study will guide the development of future clinical trials. Possible next steps involve creating personalized interventions that integrate insights on SDoH to promote cognitive and mental health in older adult ICU survivors at high risk for cognitive decline.

Rhabdomyosarcoma (RMS) is a rare pediatric cancer that begins growing within soft tissue and becomes deadly with rapid tumor growth, eventually leading to therapy resistance and metastasis. Today, drug therapy options remain scarce in relapsed patients, resulting in significantly lower 5-year survival rates compared to patients with localized disease. Recent research in other cancers has implicated the hyperexpression of an oncogene, YBX1, and an immune checkpoint marker, PD-L1 in driving therapy resistance and immune response evasion of cancer cells. YBX1 is an RNA-binding protein that improves the survival of stem-like cells in RMS. Additionally, PD-L1 is a protein expressed on the surface of tumor cells that binds to PD-1 on T lymphoid cells and aids in the evasion of T-cell-induced cell death. We hypothesize that the combination drug therapy with atezolizumab, a PD-L1 inhibitor, and SU056, a YBX1 inhibitor, will synergistically inhibit tumor growth by increasing immune cell activity. I performed intraperitoneal (IP) transplantation of RMS tumor cells derived from a zebrafish RMS tumor line (‘3.2T’) using a syngeneic line of zebrafish, ‘CG1’. The drug treatment study consists of 4 experimental groups (SUO56 or atezolizumab only, two-drug combination, and vehicle control) receiving two drug injections via IP route over 7 days. I quantified tumor growth over 7 days via ImageJ/Fiji software. Additionally, we use RNAscope (RNA-targeted in situ hybridization) to determine mRNA expression levels of various tumor-associated immune cell markers, including CD4 and CD8 for T lymphocytes and PAX5 for B lymphocytes. Based on our knowledge of YBX1 and PD-L1, we expect to see a greater reduction in tumor growth and an improved immune response with combination therapy compared to single-drug treatments. Completing this research will address the current lack of effective therapies in treating relapsed RMS patients and possible combination therapies for improving RMS patients’ survival.

Understanding the ecology of vegetation systems in Earth’s past in response to past warming events helps contextualize how they might respond to current climate events. Ecological succession is an ecosystem dynamic in which plant species with different life strategies replace each other as plants colonize a disturbed habitat. Reconstructing which successional stage a fossil plant represents is an important step in reconstructing this process in the past. However, fossil plants preserve a limited number of traits. Leaf vein density (LVD) is a trait that relates to maximum photosynthetic rates and can be measured from fossil leaves, but there is limited empirical evidence for how it varies across succession in temperate deciduous forests. To address this knowledge gap, our study measures LVD of modern plant communities across a successional gradient in western North Carolina. We hypothesize that plants in younger forests have greater access to sunlight due to a less established canopy and will therefore have higher LVD to support a higher photosynthetic rate. As succession progresses and the canopy closes, we hypothesize that LVD will decrease with reduced light availability. Samples were taken from five sites in western North Carolina that vary in how long forest re-growth occurred following clear-cut timber harvesting, 4, 21, 44, 94, and roughly 200 years. At each site, leaves were collected and sampled at a community scale and were chemically treated to create images that highlight the veins. We then used ImageJ to measure LVD. The community mean and variance of LVD across succession will be analyzed, using both unweighted and weighted approaches, to test our proposed hypothesis of decreasing LVD through succession. Preliminary results suggest a potential LVD decrease as hypothesized but driven more by understory species rather than dominant tree species. Future work will refine interpretations and consider implications for the fossil record.

Tuberculosis (TB) remains the world’s deadliest infectious disease, claiming over 1.25 million lives annually—surpassing malaria and HIV in mortality. TB’s causative pathogen, Mycobacterium tuberculosis (MTB), continues to spread rapidly due to inadequate access to accurate molecular diagnostic tests. The most commonly used tests include sputum-based and tuberculin skin tests, which require follow-up visits and have suboptimal sensitivity, particularly within certain patient populations. Moreover, these assays cannot identify emerging drug-resistant strains (DR-TB) that have reduced susceptibility to first-line antibiotics. Our aim is to design a diagnostic tool to detect cell-free DNA (cfDNA) in urine and identify the infecting strain to ensure patients receive appropriate antibiotics. To achieve this, we are developing a probe-ligation assay with single-nucleotide specificity. Current implementations are limited by the low specificity of ligase, leading to false positives and an inability to differentiate between mutated MTB strains. We hypothesized that Flap Endonuclease-1 (FEN1) could confer a specificity advantage by integrating a second enzymatic “check” into the process. The protocol involves a ligation reaction with MTB genome-derived targets and two probes, each containing a DNA flap with additional nucleotides. To detect the ligated product, FEN1 must cleave these flaps before the ligase catalyzes the repair of the nick between probes. To experimentally observe this, we carried out several ligation reactions containing FEN1 and ligase with wild-type and mutant targets, followed by PCR or gel electrophoresis to measure ligated product formation. We evaluated the efficiency and precision by analyzing the amplification profiles of WT targets and mutants containing SNPs neighboring the ligation site. Our data about whether FEN1 confers a significant specificity advantage remains inconclusive, but the double enzyme reaction is functional and could be further exploited in future experiments with additional optimization or modifications to enzymes or DNA probes.

Neonatal hypoxic-ischemic encephalopathy (HIE) occurs when the brain receives insufficient oxygen and blood supply before or during childbirth. HIE is a leading cause of neonatal mortality and morbidity that may also affect later brain development, specifically gyrification - folding of the cerebral cortex creating gyri and sulci. The nonhuman primate (NHP) brain is gyrified, similar to humans, making NHPs a highly translatable model to examine brain development after injury, which has not been well-studied in HIE. In our nonhuman primate (NHP) model of neonatal HIE, we induced injury through in utero umbilical cord occlusion (UCO) for 20 minutes, mimicking the cause of HIE in humans. Twenty-two term-equivalent pigtailed macaques (Macaca nemestrina) underwent UCO and were randomized to no treatment (n = 11) or treatment with therapeutic hypothermia and erythropoietin (TH + Epo [5x1000 U/kg]; n = 11), while non-UCO animals served as controls (n = 7). All animals were delivered via cesarian section. Injury severity was determined by physiological parameters (Apgar score), lactate, and pH levels after resuscitation. To evaluate the impact of injury on gyrification, we will utilize magnetic resonance imaging (MRI) taken 6-months post-injury to measure the gyrification index (GI). GI will be calculated by taking brain’s inner-to-outer hemispheric ratio; the inner trace following the contours of the gyri and sulci, and the outer trace following the circumference of the cerebral cortex. We hypothesize that global and regional GI will be altered in animals exposed to UCO, corresponding with decreased brain volume and greater injury. We also hypothesize that treatment will mitigate some of these changes, leading to a GI closer to control. These results will help determine whether hypoxia-ischemia alters the trajectory of cortical development, as well as the association between injury severity, brain volume, and gyrification.

We measured the functional traits of two species of moss (Homalothecium fulgescens and Rhytidiadelphus loreus), native to the Pacific Northwest, to better understand the complex relationships mosses have with other plants in their environment. Mosses play important roles in facilitating other plants around them by increasing moisture and nutrient availability and decreasing the effects of wind. Our research questions aimed to understand how these functional traits of mosses affected seedling growth of 4 native seeds. We measured the water retention and moss thickness of these mosses under control and drought conditions. We watered the control mosses twice a week with 50 mL of deionized water, and drought mosses once a week with 50 mL of deionized water. We measured moss thickness and water retention, then examined the correlation between them. We added seeds (Lupinus succulentus, Pseudotsuga menziesii, Lolium perenne, Castilleja miniata) to these pots to see how mosses and their functional traits impacted seed germination. We found seed germination to be related to less moss mat thickness. We found that more seeds grew in Rhytidiadelphus loreus pots despite having less thickness in comparison to Homalothecium fulgescens pots. We found seed germination to be less related to controlled conditions as our results showed that there were more seeds germinated in Rhytidiadelphus loreus under drought conditions than Homalothecium fulgescens in both conditions. In addition, an unplanned fungus (white rust) infected the moss pots. We examined how quickly this infection impacted seedling growth under drought and non-drought conditions. Homalothecium fulgescens was more prone to the infection compared to Rhytidiadelphus loreus. Less seeds grew under infected condition overall.  

The MYH7 gene links closely to the development of hypertrophic cardiomyopathy (HCM), a condition marked by abnormal thickening of the heart’s left ventricular wall and impaired cardiac function. Pathogenic MYH7 single nucleotide variants (SNVs) account for ~40% of HCM cases. However, ~75% of known MYH7 SNVs are variants of unknown significance (VUS). While clinical and computational data can be used to classify the significance of MYH7 variants as pathogenic or benign, these data are sparse and often inaccurate. The Yang lab has established a β-MHC abundance assay that distinguishes clinically known pathogenic and benign MYH7 variants in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), enabling reclassification of many VUS. β-MHC abundance is not a well-established phenotype, whereas existing well-established phenotypes such as impaired contractility are not high-throughput. Therefore to validate the β-MHC abundance assay findings, I aimed to functionally assess a set of variants with normal or abnormal β-MHC abundances using orthogonal assays. Towards this, I generated transgenic hiPSC lines with heterozygous variants p.Ser866Phe and p.Leu863Pro (abnormal β-MHC abundance) and p.Leu881Met and p.Ser851Phe (normal β-MHC abundance) knocked into the endogenous MYH7 locus using CRISPR/Cas9 gene editing. These variants are evaluated for NPPA and NPPB expression using RT-qPCR, cell size using confocal microscopy, and contractility via traction force microscopy, and are compared to a wildtype MYH7 line as a control. VUS with reduced β-MHC abundance are expected to show increased NPPA and NPPB expression, increased cell size, and increased contractility. Conducting these tests could allow for more confident reclassification of these VUS and other MYH7 variants in the future. Pathogenic variants in MYH7 are clinically actionable, meaning that reclassification of VUS will inform earlier medical interventions that improve health outcomes for patients that develop HCM.

Plants defend themselves against invading pathogens and herbivores using immune receptors that detect molecular signals associated with danger. Two types of plant immune receptors are the leucine-rich repeat (LRR) receptor, receptor-like kinases (RLKs), and receptor-like proteins (RLPs) through which signal transduction may proceed through the cell. Coreceptors like SERK3/BAK1 and SOBIR1 pair with LRRs to facilitate intercellular communication. We studied a specific LRR receptor, the inceptin receptor (INR) that recognizes an eleven amino acid-long peptide chain known as inceptin-11 (in11). Given the close interaction of LRR and its coreceptors, and considering that we still don’t fully understand how INR recognizes in11, we investigated its three-dimensional structure to analyze the mechanism of ligand binding and signal activation. Through predictive modeling in Alphafold of homolog RXEG1, a carboxy-terminal loop out domain was identified as a facilitator in the binding interaction between INR and the BAK1 coreceptor. To explore this mechanism, we introduced targeted mutations in the domain’s K-X5-Y motif to potentially change its ability to recruit BAK1. When a ligand attaches to a receptor, its conformation change allows signals to pass through the cell membrane. We constructed a library of 36 single and double mutants in the K-X5-Y motif and coexpressed them with a luminescence-based reporter construct in Nicotiana benthamiana to screen their activity. We expect that a mutation at K or Y or both will affect BAK1 recruitment, leading to phenotypes that are hypersensitive or inactive. Insight into LRR-RLP coreceptor interactions could open doors to further INR-immunology research alongside better modeling of BAK1 protein binding. Targeting immune-related peptides in this screen could significantly advance cultivation programs for INR expressing organisms.

Brains can somehow maintain functionality despite significant neuron loss. However, we do not yet fully understand what factors contribute to this robustness or under what conditions brains become fragile to neuron loss. Research in our lab has identified two types of neurons: those that, when removed, lead to large changes in the network’s expected activity patterns, and those that do not appear to be so critical. My research aims to address this gap. I study the network properties that confer robustness in an ideal system: the Drosophila fly, the most complex organism with a fully mapped brain at ~140,000 neurons. I am focusing on one particular brain region, the Antennal Motor and Mechanosensory Center (AMMC), because it is a primary sensory region that receives direct connections from the fly’s ear (the antennae) and contributes to auditory-driven behaviors such as courtship, which we can easily measure.  I have found that many anatomically distinct neurons share high level network properties. I hypothesize that the morphological and network properties of these neurons make them special. Here, I investigate the morphological features of several neurons, such as arborization patterns, neurotransmitter profile, and synaptic partners, and also search for genetic driver lines through a large database that will help us test the impact of these neurons in a living fly. Investigating the relationship between neural properties and robustness to removal of a neuron is crucial for developing a deeper understanding of how brain circuitry copes with injuries and the progression of neurodegenerative diseases.

Capillary microfluidics capitalize on surface tension effects encoded in microchannel geometry and chemistry to transfer liquids without external instruments - making them a user-friendly technology for point-of-care tests. For most applications, hydrophilic surfaces (contact angle < 90˚) are necessary to induce surface tension driven flow. Currently, vacuum plasma chambers that alter surface chemistry achieve this. Unfortunately, hydrophilic properties made with plasma processing are temporary, costly, and unstable. An inherently stable hydrophilic 3D-printing resin containing polyethylene glycol diacrylate (PEGDA) and acrylic acid (AA) was developed for capillary microfluidics [1]. Similarly, our group has also optimized printing parameters for resins containing PEGDA and Lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) that are inherently porous, hydrophilic, and have applications for development of engineered living materials (ELMs) [2]. Our objective was to optimize and validate 3D printing parameters and geometries for both resins using a range of liquid crystal display (LCD) printers. Our proof-of-concept prints for the PEGDA-AA resin had average contact angle measurements of 42.8 ± 8.77°. Percent differences between designed and printed channel lengths, widths, and depths were 31.5 ± 0.23%, 28.9 ± 3.41%, and 2.40 ± 13.9% respectively. Additionally, we have demonstrated the feasibility of autonomous flow of fluids in the PEGDA-LAP resin with coefficients of variations (CVs) of <5% for microchannels of widths ≥ 137.6 µm. By exploring innovative resins, we increase accessibility and capability for rapid and inexpensive prototyping of microfluidics to be applied to diagnostic tests. These methods reduce costs and carbon footprints relative to traditional additive manufacturing methods.

Heart attack survivors experience elevated risk of subsequent events and death, as such there is a clinical need for regenerative techniques to rebuild damaged myocardium and reduce risk. Transplantation of human-induced stem cell-derived cardiomyocytes (hiPSC-CMs) into non-human primate hearts has been shown to significantly improve contractile function after heart attack, however, transplanted hearts were also shown to be at elevated risk for developing potentially lethal arrhythmias. Researchers developed a line of hiPSC-CMs to correct this arrhythmia-promoting behavior by inducing a series of four gene edits to prevent the hiPSC-CM spontaneous beating behavior known as "automaticity", these gene edits spawned the cardiomyocyte cell line dubbed MEDUSA (Modification of Electrophysiological DNA to Understand and Suppress Arrhythmia). I have conducted an in-vitro study of the electrophysiological effects of the MEDUSA edits, knockouts of the sodium-calcium exchanger NCX1, the hyperpolarization-activated pacemaker current channel HCN4, the low voltage Calcium ion channel Cav 3.2, as well as overexpression of the potassium channel Kir 2.1. I have employed immunofluorescence microscopy to analyze sarcomere structures, used lentiviral transduction of calcium-sensitive green fluorescent protein to quantify calcium handling, and constructed engineered heart tissue casts to measure contractile force, to compare the electrical and physiological characteristics of MEDUSA CMs and their wild-type counterparts. Characterizing the MEDUSA cell line is essential for identifying issues that could compromise the cells' ability to function in grafts while uncovering its potential for use in regenerative treatments. Here I show that the MEDUSA gene edits create arrhythmia-resistant cardiomyocytes without compromising the integrity of their structure or function, supporting the development of a regenerative therapy future for the field of cardiology.

This research project aims to investigate the perceptions of physical disability among able-bodied college students, with a focus on how the duration of the disability influences these perceptions. Despite increasing awareness and advocacy for disability rights, stigmatization and misconceptions persist within educational environments, potentially affecting the social integration and academic experiences of disabled students. Furthermore, individuals with permanently disabling physical impairments may experience different treatment than individuals who are temporarily disabled by acute injuries. Though anthropological mixed methods research on the perception of disabilities is not uncommon, my research is focused on the difference in the perception of congenital (e.g. cerebral palsy) or acquired permanent physical disabilities (e.g. traumatic limb loss) compared to that of temporary disabling acute injuries (e.g. traumatic fracture). I hypothesize that there is a difference in the treatment of people with permanent versus temporary disabilities; students with permanent disabilities are regarded as “more disabled” than those with temporary disabilities and are considered “less capable” and “less threatening”, and therefore more likely to receive pity and unsolicited help from abled students. They may also be perceived as “less intelligent” despite having no mental impairment. This study's findings fill the gap in the existing body of knowledge on the effect that the duration of physical disability has on its perception. My research employs a mixed-methods approach, combining anonymous surveys and qualitative interviews to explore the attitudes and beliefs of students at UW regarding their peers with physical disabilities. By understanding how the duration of disability affects perceptions, universities can better address the specific needs and challenges faced by disabled students, promoting a more equitable and empathetic academic community.

Plants utilize molecular patterns in order to detect threats to the plant. Through the recognition of molecular patterns by their associated receptor, plants are able to initiate an appropriate immune response, measurable by the increased production of reactive oxygen species (ROS). In the model organism Arabidopsis thaliana, the pathogen associated molecular pattern flg22 is detected by the pattern recognition receptor Flagellin Sensitive 2 (FLS2) to initiate intracellular signaling. The immune signaling kinase Botrytis-Induced Kinase 1 (BIK1) is then phosphorylated by FLS2 to transduce the immune signal, initiating ROS production. However, A. thaliana lacks a group of immune signaling kinases related to BIK1 called Herbivory-Induced Kinase Like Kinases (HLKs), which are present in multiple species including tobacco (Nicotiana benthamiana) and common bean (Phaseolus vulgaris). The goal of this project is to determine the role of HLKs in immune signaling. To investigate the role of HLKs in immune signaling, A. thaliana were transformed with transgenes to express HLKs or overexpress BIK1. Stably transformed A. thaliana lines were then bred to produce progeny that are homozygous for the transgenes. These lines are treated with flg22 in order to initiate an immune response. ROS is used to measure the immune response of each transgenic line, where HLK expressing individuals are compared with BIK1 overexpressing individuals as a control group. I anticipate that HLKs will increase the ROS production when compared with the controls, signifying an increased immune response, since HLKs are related to the native BIK1. An understanding of the role of HLKs in FLS2 immune signaling in the model organism A. thaliana can be applied to crop species that employ HLK mediated immune signaling.

Fuel sloshing in aircraft fuel tanks plays a crucial role in affecting stability and control. This study examines the dynamics of sloshing in wing tanks, integrating theoretical models and practical calculations. The displacement of the fluid’s free surface is analyzed over time, and the resulting shift in the center of gravity (CG) is determined based on liquid distribution. Experimental data were obtained by recording video footage of turbulence simulations and measuring wave heights from the video frames. The measurements were analyzed using manual calculations and Google Spreadsheet functions. Additionally, Computational Fluid Dynamics (CFD) software, LiquiGen, was employed to compare the experimental results. For tanks with baffles, the total liquid mass and CG shift were computed in sections, summing the contributions from all sections to determine the overall shift. The experiments showed notable differences: the total CG shift for a tank without baffles was measured at 1.1 m over 5 seconds, compared to 0.08 m for a baffled tank under identical conditions. Similarly, for CFD simulations, the CG shift was 1.2 m for the tank without baffles and 0.07 m for the baffled tank during the same period. Statistical analysis, including the Shapiro-Wilk test for normality, showed no significant departure from normality for both CFD (p = 0.617) and experimental data (p = 0.116). However, a two-tailed t-test revealed a highly statistically significant difference between the two datasets (p < 0.0001), suggesting that LiquiGen does not accurately replicate experimental results. These results clearly demonstrate the effectiveness of baffles in reducing CG shifts and stabilizing liquid motion. Moreover, they underscore that LiquiGen is unreliable for precise fuel sloshing simulations, which are critical for aircraft stability assessments.

Frontline communities across and beyond the United States are continuously affected by the US nuclear legacy: from mines to test sites to radioactive waste disposal and beyond. Activists advocating for reparations and community safety often have to engage in their own research to provide the evidence the government requires as proof of radioactive harm. Most of this evidence exists only within physical documents that are behind multiple barriers, sometimes only recently declassified, and scattered across US nuclear sites. My research goal focuses on the creation of a living digital archive to help facilitate the digitization and sharing of important documentation among frontline communities. I use GitHub’s Collection Builder platform to create the foundation for an archive that can grow over time: including the first collection, the metadata schema, and information regarding the archive and how to get involved. My work includes selective digitization of documents of the now defunct UW Radiation Ecology Department, which was primarily funded by the US Atomic Energy Commission (AEC) and involved in a multitude of nuclear operations. These materials are housed in the UW Libraries Special Collections. Based on preliminary findings, I expect to find important evidence in this material of neglect to the harm of long-term exposure to low levels of radiation, and the specific testing done and results produced by the UW Radiation Ecology Department. My research also prototypes an archive that community researchers can continue to build and involves community feedback that helps shape the design. It is a step in the process of achieving necessary safety measures that protect people and ecosystems from continued radioactive harm.

Syphilis, caused by the bacterium Treponema pallidum, remains a major global public health concern despite the availability of curative treatment. Cases in the U.S. have increased by nearly 80% since 2018, and congenital cases have skyrocketed by 937% since 2014. Currently, a variety of treponemal and non-treponemal tests exist for syphilis diagnostics. Still, they can be limited by high costs, false positive and negative results, and an inability to distinguish between current and prior infection, depending on the test. Further, the fragility and low protein content of T. pallidum’s outer membrane, coupled with its nature as an obligate pathogen, exacerbates the difficulty of conventional approaches to proteome characterization. Current assays are ultimately incapable of characterizing a high-resolution immune response to T. pallidum in humans. Here, we introduce a phage display and immunoprecipitation sequencing (PhIP-Seq) platform capable of identifying antibody epitopes across the entire T. pallidum proteome. This platform allows for the profiling of antibodies that bind to linear B-cell epitopes. This can further the current understanding of antigenic proteins within T. pallidum, their ability to elicit an immune response in humans, and reveal antigens with the potential as a diagnostic. Utilizing 40 single-draw serum samples from syphilis-infected patients in Peru and Seattle, we characterize how antibody responses differ based on syphilis stage, HIV status, and strain of the infection, and have identified four proteins - TP0136, TP0969, tprK, and arp - as being highly enriched across all patients.

Mutations in glucosidase, beta acid 1 (GBA) are the strongest genetic risk factor for Parkinson’s Disease (PD) and are associated with faster progression of cognitive and motor symptoms. We hypothesize that GBA mutations disrupt the endolysosomal pathway, altering extracellular vesicle (EV) biogenesis and impairing autophagy, leading to faster spread of Lewy pathology from cell to cell in the brain and consequently accelerated disease progression. To study this connection, we utilize a Drosophila model of GBA deficiency that exhibits increased protein aggregation and neurodegeneration. We found that expression of WT GBA in the muscle of GBA mutant flies reduces protein aggregation in the brain, and EVs isolated from these flies have normalized levels of EV-intrinsic proteins that were elevated in GBA mutant flies. These findings suggest that GBA deficiency mediates PD pathogenesis by accelerating the propagation of protein aggregation to distant tissues. To complement this fly model, we differentiate induced pluripotent stem cells (iPSCs) from a PD patient carrying a null GBA IVS2+1 mutation (GBA IVS PD), isogenic wildtype iPSCs generated by CRISPR repair of the IVS2+1 mutation (GBA WT PD), and iPSCs from an unrelated healthy age and sex match control into neurons. To further investigate how GBA influences EVs, I extract EVs from the GBA IVS PD, isogenic GBA WT PD, and sex-age-match control iPSC-neurons to determine if there is a difference in protein cargo in EVs from GBA deficient neurons. I hypothesize that higher levels of aggregated alpha-synuclein will be present in EVs from GBA deficient neurons. I utilize size exclusion chromatography to isolate EVs from neuronal conditioned media. I then conduct western blots to determine protein within EVs. Understanding how GBA mutations influence EV dysregulation and whether EVs act as a vehicle for spread of Lewy pathology could help us uncover new therapeutic targets to slow neurodegeneration.

The objective of the Colgate-Palmolive project is to characterize the organic content and its distribution at the crystalline level in human enamel. This includes determining the distribution of proteins throughout the enamel, distinguishing the differences in composition between young and old adult teeth, and analyzing how protein composition impacts the mechanical properties of enamel within the context of aging. Raman spectroscopy was conducted to analyze the mineral composition of enamels following KOH or NaOCl deproteinization treatments, with control conditions used to isolate the impacts of protein on enamel properties. Additionally, Vickers indentation was conducted to analyze the mechanical properties of enamel following each treatment, with respect to the distance between enamel surface and the dentin enamel junction. These techniques jointly characterized the distribution of proteins within dental enamel and how it impacts mechanical behavior. 

Duchenne Muscular Dystrophy (DMD) is the most common lethal genetic muscular disorder of children and is caused by mutations in the 2.2 MB DMD gene. Absence of the dystrophin protein from the dystrophin-glycoprotein complex, leads to myofibers being highly susceptible to contraction injury, leading to progressive rounds of degeneration and regeneration. There is no cure, and most DMD patients eventually experience cardiorespiratory failure. Utrophin (Utr) is a dystrophin paralog that has long been suggested to be a potential therapeutic for DMD. Here we evaluated 4 novel micro-utrophins (µUtr), with the original micro-utrophin (µUtr1) and micro-dystrophin (µDys5) as controls. Two-week-old mdx^4cv mice were intravenously administered vector genomes of recombinant adeno-associated viral vector (rAAV2/9myo1) with human codon-optimized micro-transgenes, driven by the creatine kinase regulatory cassette (CK8e). At ~8 months of age, we assayed lower limb muscles for contractile performance. After functional testing, I processed heart and gastrocnemius muscle tissues to enable purification of proteins and nucleic acids. I then conducted molecular assays, followed by metabolomics via mass spectroscopy to compare wildtype, mdx^4cv, and treated mdx^4cv mice. These results revealed a leading 6-R µUtr variant (µUtr2) that showed physiological improvements in resistance to contraction-induced injury. We also performed univariate and pathway analysis of ~200 targeted metabolites, revealing fold-changes in tricarboxylic acid (TCA) cycle intermediates along with several genes controlling glucose metabolism. The µUtr2 variant resulted in metabolic and physiological improvements towards alleviating symptoms associated with disease progression in the mdx^4cv mouse model, and may hold promise as a treatment for DMD. Gene delivery of functional utrophin-centric proteins could avoid the adverse immune response events that recently occurred in human clinical trials against dystrophin sequences, potentially providing an alternative therapy for some patients.

Grasses (family Poaceae) are highly diverse (~11,800 species), cover nearly 40% of Earth’s ice-free land surface, and play critical ecological and economic roles. Grasses have evolved a variety of unique traits, including an exceptionally high accumulation of silicon in the form of biological silica bodies (phytoliths) in some lineages. Silicon accumulation confers resistance to both abiotic and biotic stresses, including drought and salinity resistance, herbivore defense, and structural support. Despite the role of silicon in the enormous success of grasses, a clear picture of the exact drivers of silicon accumulation in grasses across species and environments has not yet emerged. I hypothesize that elevated silicon concentrations are primarily driven by environmental stress, most notably high temperatures and low precipitation. To test this hypothesis, I used X-ray fluorescence to analyze the leaf silicon concentration of 482 grass leaf samples, encompassing approximately 200 species across all 12 grass subfamilies. Using occurrence records from online databases, I identified the realized climate niche and its environmental conditions (e.g., temperature, precipitation) for each sampled species. The next step is to collect geolocation data from each individual sample, which will be combined with the climate niche data of its species. By comparing the relationship between a plant’s climate niche, its individual growing conditions, and its silicon concentration, a better understanding of environmental drivers of silicon will begin to emerge. Preliminary results taking into account only climate niche and silicon concentration showed no relevant correlations, illustrating the need for individual growing condition data. Because many of the stresses that silicon helps to alleviate are also those that will worsen under climate change (high temperature, drought, insect herbivory), an improved understanding of the environmental drivers of silicon accumulation will allow us to better prepare for the impacts of climate change on our agricultural and ecological systems.

Cells in the human body contain the same DNA, and yet encompass an incredible range of structures and functions. This is possible due to genetic regulatory circuits that precisely control gene expression in response to external factors. While this is a central aspect of human biology, we lack a mechanistic understanding of how the majority of these regulatory circuits work. In this study, we investigate how the expression of Bcl11b, a gene that plays a large role in T cell development, is controlled by nearby noncoding DNA (referred to as a cis-regulatory element or CRE). We conduct a CRISPR interference (CRISPRi) screen in which a culture of P2C2 cells modified to express Bcl11b coupled to a fluorescent reporter protein is transfected with a guide RNA (gRNA) library that targets locations on the Bcl11b enhancer/promoter, and recruits repressive proteins to these sites. Cells transfected with gRNAs that bind to crucial regulatory locations experience a drop in Bcl11b expression that is measured via flow cytometry. These cells are analyzed using next generation sequencing techniques to determine which gRNA sequences are present, and thus which CRE sequences are the most essential for Bcl11b expression. In preliminary experiments, we have found that cis-regulatory elements at a distal Bcl11b enhancer, containing binding site locations of transcription factors controlling T cell specification, including T cell factor 1 (TCF-1), that play roles in controlling Bcl11b expression. The knowledge obtained from this experiment allows us to conduct future CRISPR gene expression studies in primary immune cells, where by precisely altering Bcl11b expression we can gain a mechanistic understanding of its effect on T cell development. This will drastically improve our ability to program human immune cells, resulting in medical advances such as far easier production of specialized T cells for cutting edge immunotherapies.

This project examines how industries' return-to-work policies influence public transit ridership patterns across various business sectors in the Puget Sound area. By analyzing ORCA card data from local transit agencies and reviewing company policies, the research tracks the impact of policy on industry ridership trends from January 2018 to January 2025, spanning the pre-pandemic, pandemic, and post-pandemic periods. The findings provide insight into industry-specific ridership changes, revealing which industries in the Puget Sound area have experienced the most significant shifts in public transit ridership post-COVID-19, and how return-to-work policies may have influenced these changes. The results of this research can be used by public transit agencies to focus and adapt their strategies to engage low-ridership sectors. This work can also help advance equitable, sustainable, and resilient public transit systems, particularly in the context of evolving remote work policies.

Undocumented youth are a highly politicized group in the United States and Israel, two nations who are themselves composed mostly of recent migrants. Recently, Israel and the United States introduced temporary policies to attempt to address the "issue" of undocumented youth. In 2006, Israel enacted the Temporary Arrangement for Grant of Status for Children of Illegal Residents (TAGS) policy. This one-time program granted legal residence to undocumented youth in Israel. Just six years later, President Obama announced the Deferred Action for Childhood Arrivals (DACA), a policy that granted undocumented youth protections from deportation, a work permit, and sometimes access to certain benefits. In this study, I conduct a comparative analysis of DACA and TAGS to evaluate how both states restrict and expand pathways to citizenship for undocumented youth. I conducted a literature review of the policies and experiences of undocumented youth utilizing large research organizations such as the Migration Policy Institute, academic journals such as the International Journal of Intercultural Relations, and local news sources such as Haaretz and i24News. I argue that through their limited criteria and application, both policies function as a "gatekeeping" mechanism, creating a liminal status of citizenship through their narrow eligibility criteria and temporality. These policies also contribute to state-building through reinforcing a hierarchy of 'deservingness' and belonging in both States (Lee, 2004). As the relationship between the US and Israel continues to evolve and undocumented migration continues to be a politically charge topic, it is pertinent that undocumented youth, one of the most vulnerable groups, continue to be a topic of study.

Within the complex landscape of the human genome, even a single mutation can trigger devastating neurological consequences. The reality is exemplified by a single missense mutation p.G192R in the RAB39B gene causing X-linked dominant Parkinson’s disease (PD) with reduced penetrance in females. Previously, loss of function mutations in the gene were 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 leads to pathogenic protein aggregates and subsequently, parkinsonism and neurodegeneration. To investigate this, we developed a RAB39 G192R Drosophila model which is characterized by neurodegeneration and protein aggregation using western blot, locomotor deficiency, and lifespans. Complementary to the Drosophila model, we developed a human neuronal model by generating induced pluripotent stem cells (iPSCs) from an affected male and similar age unaffected male from a kindred with X-linked PD due to the p.G192R mutation. Neurons differentiated from the iPSCs are analyzed for endolysosomal trafficking alterations by immunocytochemistry, and western blots for evaluating insoluble ubiquitinated protein aggregates and oligomerized forms of alpha-synuclein. Our preliminary results show increased ubiquitinated protein aggregation when a constitutively active RAB39 transgene was expressed in neuronal tissue. The G196R RAB39 adult flies appear morphologically normal, and the G192R mutation does not seem to affect dRAB39 protein expression in Drosophila. RAB39B G196R neurons also do not have altered expression of RAB39B, but have reduced cellular compartment size of p62-stained autophagolysosomes, and Plin2-stained lipid droplets. Understanding mechanisms underlying the pathogenesis of X-linked PD could reveal novel therapies to slow the rate of progression of neurodegeneration and development of PD.

Humpback whales exhibit exceptional maneuverability in water, a trait attributed to the unique scalloped structures (tubercles) on the leading edges of their flippers. This study investigates the influence of such varied tubercles on the aerodynamic performance of wings, using both wind tunnel testing and computational methods. CAD models of the rigid wings were designed for 3D printing. These addressed three variations of the fin morphology, a smoothed base model, one with leading-edge tubercles, and one with tubercles on the trailing edge as well. The fin models feature a swept wing configuration with a concave region before the wing tip, both properties of humpback whale fins. The result of wind tunnel tests at constant, turbulent, wind speeds (Re=10^5) produced plots of the lift and drag coefficients for a varying angle of attack. The experimental results showed that leading-edge tubercles increase the maximum lift and increase the maximum angle of attack before stall occurs at the cost of some additional drag. The addition of trailing-edge scallops reduced drag and raised the overall efficiency to just below the baseline. Computational fluid dynamics (CFD) simulations comparable to the wind tunnel environment and in more turbulent aquatic conditions (Re>10^6) reveal the fluid flow. The tubercles and concave region influence the fluid, reducing span wise flow and the buildup of large tip vortices. The effect of tubercles has already been employed for its influence on stall angle, notably on the rudders of some racing yachts. The studied effect's ability to manage vortices across the wing span may have applications in particle separation, though significant work would need to be done to streamline the necessary manufacturing processes.

The Kodiak Archipelago in southern Alaska has a rich archaeological heritage that has fascinated archaeologists and local communities for decades. Despite the presence of many archaeological research projects, archaeobotanical remains found during excavation have yet to be analyzed. The archaeobotanical remains recovered from the Kodiak Archipelago have often gone overlooked by archaeologists who considered preservation too poor in the wet climate and focused instead on fauna from shell-midden sites or other cultural artifacts. The Tanginak Spring site on Sitkalidak Island in southeast Kodiak was excavated by University of Washington field schools between 1994 and 2003. It is considered one of the oldest identified sites on the archipelago, dating to 7500-6000 cal BP. Sediment samples taken during these excavations were retrieved, floated, sorted, and identified by the archaeobotany class at the University of Washington. This poster presents the initial results of the analysis of wood charcoal and other preserved plant remains from the site, providing evidence to develop new insights into plant use by Kodiak’s earliest settlers.

Since the discovery of DNA in the 19th century, biochemists have been elucidating not only the structure, but unique biochemical environment of each loci. Protein-DNA neighborhoods govern chromatin structure and cellular functions (transcription, replication, etc.). To investigate which proteins and oligonucleotides compose these microenvironments, our lab and collaborators developed DNA oligonucleotide-directed proximity-interactome mapping (DNA O-MAP), a locus purification method using oligo-based ISH probes to recruit horseradish peroxidase (HRP) activity to specific DNA intervals (Liu & McGann et. al. 2024). Once these secondary, HRP-conjugated probes are localized to loci of interest, hydrogen peroxide is added with biotin-tyramide. Hydrogen peroxide activates HRP, forming biotin-tyramide phenoxyl radicals that biotinylate proteins and nucleic acids within 10-75nm. This allows for a scalable, versatile method to investigate these microenvironments. Large scale DNA O-MAP, tiling across several genomic sites, can elucidate insights into biological questions. However, the upstream protocol remains a barrier to its throughput, sensitivity, and reproducibility. In order to ensure this for analysis of tagged proteins, we sought to automate the streptavidin affinity purification protocol onto the Opentrons OT-2 robot. This is where streptavidin-coated magnetic beads capture biotinylated species from lysate. Coupled beads are recaptured with a magnetic rack and pipetting-off of flow-through. Subsequently, several washes cleans up these beads before peptides are digested off via Trypsin/LysC, dried-down, resuspended, and loaded onto a Orbitrap Eclipse LC-MS for proteomic analysis. Purification of streptavidin beads is manually intensive, inherently leading to variation between runs. The Opentrons OT-2 is an open-source liquid handler, allowing our lab to easily transfer methods to others interested in DNA O-MAP. Automating this protocol launches us from technology development to biological application. Here, I present an automated protocol for streptavidin affinity purification and evaluation of its effectiveness via comparison of the automated protocol to our lab's current, manual methods.

Serotonin serves a vital role in the regulation of stress responses, and variance in the release of serotonin was found to contribute to various mental illnesses such as anxiety and depression. Current mental health treatments heavily rely on Selective Serotonin Reuptake Inhibitors (SSRIs). While these medications are generally effective, there are patient populations for whom SSRIs show limited efficacy. Recent studies found that in response to stress, the expression of the FKBP5 gene also increases and modulates many neuronal pathways including serotonin. This experiment determined whether manipulating FKBP5 gene expression in the dorsal raphe (DRN) directly correlates with serotonin release in the basolateral amygdala (BLA), an established neural circuit for fear. To manipulate FKBP5 gene expression, an adeno-associated FKBP5-Cre virus was injected into the DRN of Pet1-Cre transgenic mice, causing up or downregulation of FKBP5 in the DRN. To gauge the effects of FKBP5, fiber photometry was used to measure the release of serotonin following optogenetic activation of the DRN-BLA pathway using a 5HT GRAB sensor injected into the BLA. We hypothesize that increased FKBP5 expression in the DRN will increase serotonin release to the BLA, and as FKBP5 expression is decreased, the release of serotonin to the BLA will decrease. Success in modulating serotonin release using FKBP5 gene expression will expand therapeutic targets in mental illness research, mitigating the gap in treatment efficacy for patients who have undergone ineffective SSRI therapy.

Chlorpyrifos (CPF) is a widely used organophosphate pesticide effective in controlling agricultural pests by inhibiting acetylcholinesterase, leading to the accumulation of acetylcholine and continuous nerve stimulation. CPF exposure has been linked to increasing autism risk and gut microbiome dysbiosis. However, the underlying mechanism linking CPF to autism remains unclear, and the role of the gut microbiome in CPF-induced neurodevelopmental toxicity remains elusive. Using a high-throughput social behavior assay, we found that embryonic exposure to CPF caused lasting social deficits in zebrafish. We then screened seven common gut microbiome metabolites and found that butyrate effectively rescued CPF-induced social deficits. Butyrate is a known inhibitor of histone deacetylases (HDACs). We discovered that valproic acid, an inhibitor of Class I and IIa HDACs, phenocopied butyrate’s rescue effects. Meanwhile, trichostatin A, an inhibitor of Class I, II, and IV HDACs, and nicotinamide, an inhibitor of Class III HDACs, did not. We are currently conducting multi-omics analyses; including metagenomics, metabolomics, RNA sequencing, and CUT & RUN, to further elucidate the mechanisms underlying CPF’s neurodevelopmental toxicity and butyrate's rescue effects. In the long run, our work will help to uncover how CPF exposure contributes to autism risk and to inspire new therapeutic approaches for alleviating autism-related social deficits.

Our sense of touch plays an important role in how we perceive the world. Touch sensation is the result of an intricate interplay between the nervous system and specialized sensory cells in the skin, one such example being the Merkel cell-neurite complex. Within the Merkel cell-neurite complex, Merkel cells (MCs) detect gentle touch signals in the skin and relay them to innervating neurites via synapse-like connections. Many aspects of the MC-neurite complex, including the molecules required for its formation and structure, remain poorly understood. Our lab recently discovered the presence of MCs in the transparent zebrafish skin, making the organism well-suited for study of MC-neurite complexes. Here, we show that Protocadherin-9 (pcdh9), a cell adhesion molecule important in synaptic structure and nervous system organization, is highly expressed in both zebrafish and mammalian MCs. Using a loss-of-function mutation in zebrafish pcdh9, we find a reduction in the number of MC-neurite complexes, but not in the number of MCs, compared to controls. This suggests a role for Pcdh9 in either the formation or maintenance of MC-neurite synapses. Additionally, we observe a higher rate of MCs contacting one another in pcdh9 mutant skin, consistent with a difference in MC spatial organization. In summary, our data indicate that Pcdh9 may regulate one or more aspects of MC-neurite complex formation. We are now in the process of developing tools to further investigate and quantify MC spatial arrangement, and to uncover the ways in which Pcdh9 may affect MC maturation and behavior.

When carbon dioxide emissions are high within a room, short and long-term health effects on humans have been observed. Natural gas stoves are common household appliances that contribute to carbon dioxide emissions. The purpose of this study is to model how the burners of a natural gas stovetop may impact the air quality and potential health risks within households. I measured carbon dioxide emissions from a Samsung gas range with one to four burners on maximum heat in thirty-minute intervals using an Aranet4 Home sensor. The results were observed to have an overall increase in carbon dioxide emissions correlating to the number of burners being used concurrently. Carbon dioxide concentrations reached harmful levels within thirty minutes when two or more burners were used. Knowing how the rates of carbon dioxide concentrations within a room may increase in correlation to use of one, or multiple, burners would provide users of gas ranges with a reference point on how the air quality may change over time. This would allow for a better understanding of the risks associated with natural gas stovetop usage regarding the health impacts of close exposure to high carbon dioxide concentrations. Future analysis can be conducted on the different rates of carbon dioxide emissions for functions of natural gas ovens.

Plasma cell neoplasms are characterized by the secretion of large amounts of monoclonal antibodies into circulation. Monoclonal gammopathy of undetermined significance (MGUS) is a precancerous plasma cell neoplasm that can develop into a cancer called multiple myeloma. Glycosylation of the antigen-binding (Fab) light chain fragment of monoclonal antibodies is a risk factor for MGUS progression to myeloma. However, heavy chain Fab glycosylation may also occur and be a risk factor for progression, but this has not yet been investigated. In this study, I identify whether heavy chain Fab glycosylation occurs on monoclonal antibodies isolated from MGUS and myeloma patient serum, to help determine whether it should also be investigated as a risk factor of disease progression. I isolate intact antibodies to optimize antibody fragmentation and dissect them into light chain, heavy chain, total Fab, heavy chain Fab, and fragment crystallizable region (Fc) fragments. To do this, I purify IgG from serum from patients with plasma cell neoplasms. I either reduce these purified IgG samples using dithiothreitol into heavy chain and light chain fragments, or cleave them using IdeZ enzyme into total Fab and Fc fragments. I separate and purify reduced heavy chain and light chain fragments using high pressure liquid chromatography, and the cleaved total Fab and Fc fragments using an affinity matrix.  I buffer exchange the fragments through dialysis or using filters, and assess fragmentation and purity using SDS Page gel electrophoresis. After optimizing these protocols, I will isolate pure antibody light chain, heavy chain, total Fab, heavy chain Fab, and Fc fragments from MGUS and myeloma patients and healthy controls. This will enable subsequent glycosylation analysis and characterization. We anticipate that our results will lead to an improved understanding of antibody glycosylation in plasma cell neoplasms and provide insight into their potential role as risk factors for disease.

The traditional notion of the nuclear family living in a private home as the cornerstone of the 'American Dream' has become increasingly out of reach. According to the U.S. Census Bureau, Hawaii leads the nation with 8% of its population living in multigenerational homes. The popularity of this living arrangement is driven by the state’s unique demographic composition, high living costs, and deep-rooted cultural practices of caring for Ohana. However, these multigenerational homes are often not readily available, requiring families to retrofit their existing homes, move, or abandon their plans. Fostering multigenerational housing is crucial for preserving cultural values, supporting the diverse needs of Hawaii's communities, and helping to alleviate the housing crisis. This project asks the question: What are the primary barriers preventing the broader adoption of multigenerational housing in Hawai’i and what are effective strategies to address these obstacles through improved design and policy initiatives? It analyzes the historical development and cultural significance of multigenerational living arrangements in the U.S. and Hawai’i highlighting barriers such as insufficient privacy, lack of space, cultural norms, restrictive building codes, and limited developer incentives. These findings culminate into actionable strategies tailored to Hawai’is needs, aiming to make multigenerational living a more viable and sustainable housing solution. The suggestions enhance the accessibility of this living arrangement, recognizing the necessity for diverse policies and designs that cater to Hawaii’s evolving demographic, topographic, and social landscape. A variety of options must be provided, acknowledging that there is no one-size-fits-all solution for families or counties and that flexibility is essential to meet the state's evolving needs. Through case studies, stakeholder interviews, and an analysis of relevant literature, this project synthesizes strategies into a toolkit for policymakers, developers, architects, and families. 

Plastics have transformed the modern materials landscape, as they have a large range of applications at low costs. With this widespread and consistent use it is seen that production will continue to increase, reaching over 1.1 billion tons per year. Of this, only around 16% of plastics are recycled, with complex polymer recycling rates even lower. Remaining plastics are primarily landfilled or incinerated, leading to associated environmental impacts. In response to this issue, chemical recycling methods such as hydrogenolysis have been developed. However, these methods are limited by their ability to separate or otherwise handle mixed plastic waste feedstocks. This work investigates ruthenium-catalyzed hydrogenolysis of mixed polyolefin waste to determine the impact of mixed feedstocks on catalytic activity.  The system of pure polyethylene (PE) and polypropylene (PP) mixtures is studied as PE and PP are two of the most commonly produced plastics, together comprising nearly 50% of all plastics produced. Model compounds tetracosane and squalane are used as analogs of PE and PP, respectively. These compounds are simpler than their polymer counterparts while possessing similar structural elements, therefore making for an easier determination of mechanistic pathways and kinetic parameters. Reactions are performed in 10mL stainless steel reactor systems using a ruthenium on carbon (Ru/C) catalyst. Squalane and tetracosane are mixed at various ratios, maintaining 1g of substrate and 100 mg of Ru/C per reaction. Relatively mild conditions of 225°C, 20 bar of hydrogen are held for 1 hour. Preliminary results find the C-C bonds in tetracosane cleave to a greater extent than in squalane, with much of the squalane preserved. Likewise, reactions with a higher percent of tetracosane result in more bond cleavage compared to squalane-rich reactions. This indicates that branched polymer substrates may decrease overall catalytic activity compared to linear polymer substrates. 

This research aims to enhance the food literacy of volunteers at the University of Washington (UW) Food Pantry by providing foundational knowledge on food security, food policy, nutrition, dietary needs, and allergies. Many pantry volunteers lack a background in nutrition or public health, which limits their understanding of the complex issues surrounding food insecurity and food literacy. Through the Winter 2025 Food Systems, Nutrition, and Health Capstone project, my team and I will create a comprehensive brochure that will be incorporated into the pantry’s volunteer training materials. The brochure will include sections on the pantry’s background and goals, the scope of food insecurity among college students both at UW and across the nation, food policies that affect access to nutritious foods, and basic nutrition education. Our objective is to foster empathy and understanding among volunteers, encouraging a deeper connection to the shoppers they serve and the challenges faced by people who rely on the pantry. At the end of the quarter, my team will present the brochure along with other deliverables to community stakeholders. In Spring quarter, I will administer a pre-survey to pantry volunteers to assess their current food literacy and knowledge of food insecurity. Following the brochure’s distribution and review, volunteers will complete the same survey, allowing for an analysis of any changes in their understanding. The study will evaluate whether the brochure improves volunteers' knowledge of food insecurity, ultimately testing if this intervention leads to better food literacy outcomes. If the brochure proves effective in this regard we also hope it will foster increased empathy among pantry shoppers and volunteers.

Building on the literature of the importance of strategy, this paper explores the relationships between wealth, strategy, and battle related deaths to find out why we may see unexpected conflict outcomes like Ukraine being able to hold back Russian offensives. This paper argues that wealthy actors often use expensive strategies, like strategic air bombing, that are ultimately ineffective at helping win a war and only cause unnecessary casualties, and that less wealthy actors are forced to be more creative with their strategies and this leads to less casualties. To do this, I analyze the relationships between data on rebel contraband (proxy for non-state actor wealth), GDP, and strategies used in war and their effect on battle related deaths. The paper will also cover the ongoing conflicts in Myanmar and Ukraine as a theoretical supplement to this data. Examining these relationships is increasingly important because wars fought in the modern era often see different sides of varying wealth using different strategies. Implementing the discoveries of this paper may give us opportunities to minimize casualties in conflicts by looking at what strategies are the least lethal at what levels of wealth.

There are a vast number of pathogens that impact global public health, necessitating an accessible assay capable of detecting multiple targets simultaneously. Lateral flow assays (LFAs) have the potential to fill this role as a cost-effective, rapid, and simple technology instrumental in the detection of many analytes. However, multiplexed detection using nucleic acid LFAs is difficult due to the increased chance of non-specific binding as more targets are added to the assay. In this work, we aim to increase specificity and multiplexing potential in LFAs. We first showcase the process of developing a nucleic acid LFA by evaluating both fluorophores and gold nanoparticles to generate a visible signal. As fluorophores require a fluorescent light source, we moved forward with gold nanoparticles, which have a readout visible to the naked eye. Additionally, we automated the LFA fabrication process using an Echo Liquid Handler. Finally, we assessed methods to convert double-stranded to single-stranded DNA, required for compatibility with LFAs. In the future, we look to optimize signal visibility while increasing multiplexability. This work highlights the potential of multiplexed LFAs as a robust technology capable of significantly improving public health responses and outcomes.

Asylum policy is constantly evolving in the US and EU in response to shifting public opinion, national interests, and international conflicts. Starting from countries of origin to countries of destination, our task force covers challenges plaguing not only US and EU systems but the world. A side-by-side comparison surveys past and present progress and setbacks influencing asylum processes. We argue that the US and EU share areas for improvement, but can also learn from eachother to enhance their capabilities in tackling unprecedented migration crises. Our research highlights the importance of collaboration multilaterally, bilaterally, and with non-governmental organizations and civil society organizations to implement policies that address the needs of all actors, globally to locally. Using contemporary case studies to demonstrate the real-world impacts of asylum policies, our findings stress the importance of adaptability in asylum policy. The protection of asylum seekers and refugees is not mutually exclusive with promoting domestic interests. Pathways exist that allow the US and EU to ensure their security while adhering to international agreements and standards. Thus, a comprehensive, cooperative, approach proves essential for creating asylum frameworks that both respond to evolving global challenges while supporting national and regional priorities.

It is estimated that ~15% of all cancers are caused by oncogenic virus infections. Two of the top seven cancer-causing human viruses are members of the gammaherpesvirus family: Epstein Barr Virus (EBV) and Kaposi’s Sarcoma Herpesvirus (KSHV). Our lab uses Murine Herpesvirus 68 (MHV-68), a mouse gammaherpesvirus with shares significant genetic homology to KSHV and EBV, as a model system to understand how gammaherpesviruses alter the metabolism of their host during lytic infection to promote their replication. We recently metabolically profiled MHV-68 infected host cells at various time points during the lytic infectious cycle. Our data showed nucleotide metabolism is significantly induced in MHV-68 infected NIH/3T3 cells, revealing a potential antiviral target. This study investigates the antiviral efficacy of Methotrexate (MTX), an FDA-approved nucleotide biosynthesis inhibitor currently used to treat cancer, rheumatoid arthritis, and psoriasis. MTX inhibits dihydrofolate reductase (DHFR), an enzyme crucial for producing thymidylate and purine nucleotides, which are essential for de novo nucleotide synthesis. We hypothesized that MTX can block MHV-68 production and be repurposed as an antiviral drug. To test our hypothesis, we first determined a safe concentration of MTX in NIH/3T3 cells using both qualitative (microscopy) and quantitative (trypan blue exclusion) cell viability assays. Next, we infected NIH/3T3 cells with MHV-68 and treated them with a safe level of MTX or solvent control. After 48 hours, we assessed viral production in control vs MTX treated cellular supernatants via viral plaque assays. Our results revealed that MTX significantly suppressed MHV-68 virion production by ~50-fold. These findings suggest that targeting host metabolic pathways could be an effective antiviral strategy against gammaherpesviruses in humans. Further research is needed to explore the use of MTX as a broad viral therapy against other viruses.

Variations in floral structures influence how plants are pollinated, showier flowers are more attractive to pollinators, while wind-pollinated plants benefit from having smaller, inconspicuous flowers that produce increased amounts of pollen, and have the appropriate morphology to receive pollen from the wind. The genus Thalictrum contains species that range from insect-pollinated to wind-pollinated. Certain transcription factors are known to affect the stigma, the area of pollen reception that consists of papillae, and to increase stigmatic papillae length. By better understanding the genes that influence stigma morphology, this gene could be used in economically important crops to increase their stigmatic surfaces and consequently the likelihood of pollination. In this study, we use in situ hybridization to analyze gene expression of candidate genes for stigma development in the genus Thalictrum, which has had multiple transitions from insect to wind pollination in its evolutionary history. We selected three species representing the range of stigma morphologies found in Thalictrum. I will test the hypothesis that expression of my candidate genes will correlate with stigma morphology, such that the short (capitate) stigmas characteristic of insect-pollinated species will exhibit restricted areas of gene expression just prior to stigma development, while the more elongated stigmas of the wind-pollinated species will show an extended temporal and spatial domain of expression, with mixed-pollinated species lying in between. Thus, this work will provide a connection between developmental genetics and morphology to improve understanding of the wind pollination syndrome.

Westfield Southcenter Mall, located in Tukwila, Washington, is one of the largest malls in the nation, and like many malls, its aging retail spaces and surrounding strip malls are becoming less viable. With rising housing costs and a growing regional population, it is clear that spaces like Southcenter must be transformed into affordable, mixed-use districts that can provide housing, vibrant community spaces, and accessible transportation options. This project explores how Southcenter Mall can be redesigned using New Urbanist principles to foster a thriving community. Using site visits and global case studies of recent successful mall redevelopments, it analyzes the current shortcomings of Southcenter Mall and proposes a more successful mixed-use alternative. The final deliverables include a 3D model mock-up of a revitalized Southcenter that includes affordable housing, ground-level retail, and an overhauled transportation framework prioritizing pedestrians and cyclists.

Most of our chemicals come from petroleum, a nonrenewable resource and a significant source of pollution. Purple non-sulfur bacteria (PNSB) also produce some of these chemicals from one-carbon (C1) feedstocks, however, genetic engineering toolkits are underdeveloped for these organisms. The ability to integrate heterologous genes is a crucial component of genetic engineering toolkits, enabling stable and precise gene expression. Despite their metabolic versatility, PNSB lack well-characterized genomic integration sites, limiting advanced strain engineering efforts. Here, we identify and characterize genomic integration sites in Rhodobacter sphaeroides 2.4.1 that can serve as stable integration loci for heterologous gene expression. Using RNA-Seq transcriptomic data, we identified intergenic regions with minimal transcriptional activity, ensuring that insertions into these regions would not disrupt native gene function. Seven candidate integration sites were selected across the genome, spanning both chromosomes and plasmids. Two-step allelic exchange was used to integrate “landing pads” for Serine Recombinase-Assisted Genome Engineering (SAGE), a site-specific recombination system, into candidate sites. Our next step is to use the SAGE system to integrate fluorescent reporters into these sites to assess positional effects on gene expression. These seven integration sites serve as a testbed, allowing us to validate the workflow for integration into a broader range of genomic locations. Our findings will provide a resource for engineering R. sphaeroides and expand the genetic toolkit for PNSB, facilitating their use in synthetic biology and bioproduction applications.

We are developing a RNA scaffold-based CRISPR activation and inhibition system to controllably tune gene expression in  primary immune cells, which will allow us to manipulate and increase production and function of immune cells, vastly increasing their efficacy in fighting diseases such as cancer. Here we target Bcl11b, a key T cell transcription factor necessary for progenitor cell commitment to the T cell lineage. CRISPR activation and CRISPR interference (CRISPRai) enable activation or repression of targeted genes. Due to the large size of dCas9 activator and reperessor fusions, it is not possible to express the necessary machinery in primary mouse T cells. Thus, we are developing a CRISPRai system where the gRNA (guide RNA) contains an additional RNA hairpin to recruit RNA binding protein-effectors, enabling activation and repression in the same cell. To optimize the efficiency of CRISPRi in T cells, we are 1) cloning and testing a repressor domain for its ability to drive gene silencing and 2) testing alternative RNA base pairs (BP) and hairpin pairs. We are testing these optimizations in a T cell progenitor cell line which has turned on Bcl11b with a downstream YFP (fluorescent) reporter. Here, YFP expression, which we measure using flow cytometry, is directly correlated to Bcl11b expression levels. We hypothesize that an alternative validated RNA hairpin BP in conjunction with a novel compact transcriptional effector will result in decreased levels of YFP expression compared to the existing system.

Nature has long been a source of inspiration when it comes to tackling engineering challenges. Taking this inspiration from nature, our study aims to emulate the behavior of burrowing animals in order to efficiently dig into granular media. The pacific razor clam utilizes its retractable foot muscle to burrow itself while simultaneously expanding and contracting its shell to fluidize the surrounding sand. Our goal is to create a digging device that approaches the digging efficiency of pacific razor clams of 21 J/m. Our hypothesis is that the interaction of vibration and fluidization is key in burrowing performance. We built a cylindrical robotic model composed of both soft and rigid materials, equipped with a vibrational motor and a downward-facing nozzle. We adjusted the energy devoted to vibration and flow to optimize burrowing efficiency as a function of energy per depth.  Preliminary results suggest that a combination of both local fluidization and vibration improves burrowing efficiency over using them independently. We anticipate that different combinations of flow and vibration are optimal for different burrowing stages. By understanding nature's efficient digging techniques, we can create a reliable anchoring system for sensor arrays and underwater vehicles to aid in the study of our oceanic ecosystem.

South Thurston County's car-dependent and low-density built environment can affect the health and economic wellness of its residents. This research investigates the barriers that residents face in getting to work, their daily needs, and things-to-do. What are the best strategies to improve the built environment that provides access to employment, necessities, and entertainment for people in south Thurston County’s SR 507 and Old Highway 99 corridors? A process of observations, community surveys and interviews with government and other agency officials will allow for an understanding of current conditions in the study area, personal viewpoints of community members, and constraints that the government agencies and other organizations might face in making improvements. This project will include investigation into design, policy and economic practices. A free or low-cost guide that would allow rural governments to make economically smart design and planning changes, and help make towns more self-sustainable, will be created after understanding this research. As innovation in planning and re-urbanization occurs increasingly in the United States’ larger metropolitan areas, rural and tribal communities, too, deserve an opportunity at urban core making, simplifying the daily lives of its residents, and maintaining health and wellness for all.

Chronic stress has been associated with maladaptive behaviors in both human and animal research models, but the underlying mechanisms are unclear. In this research study, we sought to define whether stress induces neural inflammation in the ventral tegmental area, the brain region primarily responsible for regulating reward consumption, learning, memory, and addictive behaviors through moderating dopamine release in other brain areas. To do this, male Sprague-Dawley rats were subjected to a chronic intermittent stress paradigm that included stressors such as wet bedding, delayed feedings, social isolation, strobe lights, and forced swims. Following the chronic stress intervention, brain sections were collected from control and experimental groups. Subsequently, immunohistological analysis was performed of microglia and astrocytes, cell types known to mediate inflammatory responses within the brain. By assessing inflammation in the ventral tegmental area through fluorescent microscopy and quantitative morphological analysis of these glial cell types, we will establish whether inflammation in this key brain region regulating motivation may be involved in the harmful behavioral outcomes often associated with chronic stress.

Diabetes has emerged as a leading cause of death in America and can affect the kidney, liver, heart, and lung system. Around 34 million Americans, primarily people of color, are currently diagnosed. Diabetes is a leading comorbidity of SARS-CoV-2 fatalities (~15%), highlighting a pertinent need to establish a human diabetic pulmonary model that may unveil dynamic mechanisms behind this phenomena. This project aims to establish a pulmonary model that reflects how diabetic conditions can affect the cellular phenotypes and morphologies of alveolar lung tissue. We utilized a previously established protocol to differentiate human induced pluripotent stem cells (iPSCs) into lung organoids (LOs). These LOs serve as biologically relevant lung models because they share the same complex, 3D cellular structures as human pulmonary systems. In this project, LOs were differentiated over 25 days and subsequently treated with varying glucose concentrations (5, non-diabetic; 10, diabetic; 21, experimental control; and 80, extreme) mM in growth media for 15 days, when morphological differences appear. The LOs were fixed on day 40 and analyzed using immunofluorescence to quantify lung markers. The primary antibody used was ACE2, the receptor for SARS-CoV-2. Additional markers included surfactant protein C (SFPTC), and NKX2.1, a lung progenitor marker, to compare phenotypic differences across the conditions. Our results demonstrated a pattern of upregulation of ACE2 with increasing glucose concentrations, suggesting diabetic conditions enhance susceptibility to SARS-CoV-2 infection, compared to normal glucose levels. Furthermore, SFPTC (Alveolar Type II cells) and ACE2 co-localize, which may play a key role in increased mortality rates amongst diabetic SARS-CoV-2 patients. Further studies, including qPCR analysis, may provide additional insights into these observations. In conclusion, our model highlights the increased vulnerability of diabetic pulmonary systems to SARS-CoV-2, emphasizing the need for targeted therapeutic strategies and investigation of dynamic disease mechanisms.

Cervical traumatic spinal cord injury (TCSCI) is a devastating condition that leads to tetraplegia, severely impairing essential life functions and independence. Individuals with cervical TCSCI struggle with hand function, reaching, eating, grasping, and writing, significantly reducing their quality of life. In the U.S., cervical SCI is the most common type of spinal injury, affecting over 300,000 individuals, with approximately 17,900 new cases annually. The long-term disability resulting from TCSCI often necessitates continuous medical care, rehabilitation, and assistive technologies to enhance functional recovery. Our preclinical study evaluates upper extremity dysfunction in rats following cervical TCSCI using behavioral assessments, specifically the Forelimb Reaching Task (FRT) and the Irvine, Beatties, and Bresnahan (IBB) test. These tests provide valuable insights into motor impairments and recovery over time. FRT assesses shoulder movement and fine motor control by placing the rat in a transparent box with side slits, allowing it to extend its forelimb to grasp a chocolate pellet. The grasping behavior is scored on a standardized scale. This test primarily evaluates digit precision and reaching ability. IBB provides a broader analysis of forelimb function, including both proximal and distal limb recovery. In this test, the rat is placed in a cylinder with food, and its grasping and eating behavior are recorded. Forelimb function is later evaluated based on elbow position, paw support, forepaw placement, and digit movements. By comparing these tests, we aim to determine their efficacy in assessing functional deficits and recovery post-SCI. This analysis is critical for refining behavioral assessments and guiding the development of new therapies to enhance motor recovery and improve the quality of life for individuals with cervical SCI.

Substance use in Indigenous communities remains a significant public health concern for Indigenous communities worldwide, greatly affecting physical, mental, and social well-being. Substance use in Indigenous communities is nearly double that observed in the non-Indigenous population (Geia, et al., 2018). Such prevalence of substance use among Indigenous populations has been a source of stigma greatly related to colonialism. This study examines the prevalence of substance use among Indigenous populations in Australia, New Zealand, Canada, and the United States identifying patterns within communities and successful intervention strategies to decrease substance use in these communities. A systematic review of literature conducted in the countries mentioned above reveals that Indigenous communities experience high rates of alcohol, tobacco, and illicit drug use. Secondary to trauma and socioeconomic disparities paired with limited access to secular healthcare contributes to the exacerbation of the cycle of addiction (Spillane, et al., 2023). Specific led interventions sculpted around Indigenous culture and various community-driven, utilised participatory approaches show a high chance of decreasing substance use in these communities (Geia, et al., 2018). With such findings, the need for healthcare services that remain respectful to Indigenous culture and overall strengthen community engagement can be put in place to mitigate the prevalence of substance use in these communities and the corresponding risk factors that contribute to such circumstances. My presentation will demonstrate how the high levels of alcohol and substance use can be reduced in Indigenous communities when ethical interventions are implemented that provide long-term care for rehabilitation. This long-term care should include education, harm reduction, and encouragement for partnership between Indigenous individuals and their families and healthcare providers within the programs. The long-term care is to be residential to those struggling with substance use, rather than mainstream outside of reservations, to ensure the patients feel safe.

Remembering ‘when’ things happened is difficult relative to ‘where’ things happened. One reason for this may be because time is an abstract concept while locations are concrete. Many cultures worldwide use space to represent time to get around this problem. A common representation is a mental timeline: a linear mental model of time. For most Western adults, the mental timeline flows from left-to-right: representing the past on the left and the future on the right side of space. Previous research suggests that U.S. adults recall the order of events more accurately when items are presented from left-to-right, rather than nonlinearly or from right-to-left, indicating that adults spontaneously organize temporal order in terms of a mental timeline. However, it is unknown what memory processes influence this benefit. We examine the relation between the mental timeline and temporal memory by assessing both encoding (committing to memory) and recall (remembering) phases. In this study, adults first view triplets of images arranged from left-to-right, right-to-left, or nonlinearly (encoding). Then, images are shown individually, and participants identify whether the image appeared first, second, or third within the triplet (recall). We record participants' responses and their pupil dilation. We predict that item location will affect participants’ errors. For example, they might be more likely to incorrectly remember an image as ‘first’ if it appeared on the left, even if the triplet was presented right-to-left. Additionally, we expect adults to exert greater mental effort when encoding triplets presented nonlinearly or from right-to-left, indicated by increased pupil dilation. These results will provide insight into how the mental timeline interacts with temporal memory beyond memory accuracy measures. The pupil dilation measures will reveal the role of mental timeline in the encoding process, and errors will reveal its role in the recall process.

Since 1947, the Japanese government has been constrained by Article 9 of its Constitution, renouncing war as a means to settle international disputes and preventing the existence of an offensive military force. With the Security Treaty between the United States and Japan signed in 1951, the United States is obligated to defend Japan in exchange for the continued existence of American military bases on Japanese soil. In this research, we explore whether it is in America's interest to encourage Japan to amend or repeal Article 9, considering the security risks stemming from a stronger China and combative North Korea. In our policy recommendation, we used scholarly sources to discover three distinct challenges to regional and global security if a change to the status of Article 9 was to be pushed forward: Japanese public opinion and governmental concerns, the potential antagonization of regional adversaries, and the polarization of American allies with long-standing colonial histories with Japan. To address these concerns, we utilized a qualitative approach, exploring various narratives and opinions on the issue at stake. We found a diverse set of viewpoints, which we used to compile a recommendation for Japan to amend Article 9 to allow for the collective self-defense of its neighbors and allies when attacked. This research has significant implications for Japan's role in the future of East Asia, designating the state as a key military and economic power under America's Free and Open Indo-Pacific. With the culmination of our project, we found a clear path to upkeep and enhance America's time-honored relationship with Japan while maintaining its strategic leadership in the region. As previous efforts to remilitarize Japan have broken down due to several factors, our findings provide insight into the most suitable direction for the United States as a key player in the broader Asian region.

Acetaminophen (APAP) is a widely used over-the-counter drug known for its analgesic and antipyretic properties. Several clinical factors can influence how APAP is absorbed, distributed, metabolized and excreted from the body (i.e., its pharmacokinetics (PK)). APAP is metabolized into several metabolites, including APAP-glucuronide, APAP-sulfate, APAP-cysteine, and APAP-N-acetylcysteine. Therefore, accurately determining plasma concentrations of APAP and its metabolites is crucial for understanding how individuals metabolize APAP and environmental influences on APAP PK. To address this, I am reproducing a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay to determine concentrations of APAP and its metabolites in plasma. I analyzed plasma samples collected from seven healthy volunteers following oral administration of 500 mg of APAP. To prepare the samples, I added 70 μL of diluted internal standard mix into the tubes containing 30 μL of plasma. Samples were centrifuged and the supernatants were transferred clean tubes and dried down with nitrogen gas. Samples were reconstituted in mobile phase for analysis using LC-MS/MS. I analyzed the LC-MS/MS data to calculate the plasma concentration vs. time curves for each analyte. I used Phoenix WinNonlin to estimate key PK parameters, including peak concentration (Cmax), time of peak concentration (Tmax), clearance, half-life, volume of distribution, and area under the curve (AUC). This LC-MS/MS assay provides a platform for precise quantification of APAP and its metabolites, and will be essential for our lab’s future studies on the impact of the gut microbiome on APAP’s PK.

Individuals with Stage II/III severe periodontal disease were recruited from the Graduate Periodontics Clinic at the University of Washington School of Dentistry using an approved IRB (STUDY00016871). Subgingival plaque samples were collected from four tooth sites: an active inflamed site, two adjacent tooth sites, and a distant healthy tooth site. Sterile paper points were inserted into the gingival sulcus for 30 seconds. DNA was isolated and extracted using the Qiagen AllPrep DNA/RNA Mini kit (Cat. #80204), then purified and concentrated using the Zymo Clean & Concentrator (Cat. #D4014) kit. 16S rRNA libraries were generated and sequencing was performed on the MiSeq platform (Illumina, San Deigo, CA, USA) using 300bp paired-end chemistry. Raw reads were processed and analyzed using Qiime2 and the DADA2 algorithm to generate amplicon sequence variants (ASVs), which were then classified using the expanded Human Oral Microbiome Database (eHOMD). Differences between tooth sites were assessed within and across individuals. Correlation between taxonomic levels and clinical data was also assessed. Data analysis is still being performed at this time. Based on previous literature (Pawolski et al, 2005, Kerns et al. 2023), we expect that a subgingival community gradient radiates from tooth sites affected with periodontal disease toward distant healthy sites. We aim to resolve this within individuals using ASVs for the first time. Additionally, we anticipate an increase in disease-associated bacteria within actively inflamed tooth sites, such species within Porphyromonas, Tannerella, and Treponema genera. Furthermore, we anticipate a gradient of perio-pathic disease-associated bacteria will decrease in relative abundance the further away from active diseased sites. We expect that results from our study will highlight the presence of a subgingival microbiome composition and enrichment of specific gram negative perio-pathic disease-associated species within clinically healthy tooth sites in patients with active periodontal disease despite the absence of clinically observed inflammation.

Recent research shows the lux operon utilized with in-vivo bioluminescence imaging to detect infectious diseases in animal models. Modifications to this operon led to the development of enhanced bioluminescence in Escherichia coli cells. However, expression of this operon has not been optimized for expression in other bacteria, such as Staphylococcus aureus. This study aims to optimize the lux reporter gene expression for Staphylococcus aureus, so luminescence is bright enough to register without specialized equipment. To date, the research has explored Gibson Assembly for cloning the gene sequences into a shuttle vector and efforts to modulate gene expression to reduce toxicity in E. coli.

Despite advances in modern medicine in the United States, maternal and child health (MCH) outcomes continue to decline due to the social determinants of health (SDOH), resulting in poor health outcomes and death for mothers and babies. However, community-based models (CBM) of maternal and child health care have been identified as effective interventions that mitigate these negative outcomes by addressing the SDOH. Existing literature identifies CBMs as effective interventions using quantitative methods and analysis. However, my research focuses on qualitative methods and their human-centered real-world applications of CBMs. The goal of my study is to highlight the role that communities play in influencing maternal and child health outcomes and understand the effect of CMBs on participants. To investigate the impact of community-based models on MCH outcomes, I am using two questions to guide semi-structured interviews with expectant families, parents, and community health workers. My research questions are, 1) What are the experiences of providers and recipients of community-based models of maternal and child health care? and 2) Do Black, Hispanic, Indigenous, and White communities in King County experience gaps in maternal and child health care services, and, if so, how do these gaps differ among communities? I am recording demographic data from participants for coding after interviews have concluded. This study will help create a more comprehensive understanding of CBMs of MCH in the field. These narratives will help further legitimize the practice of community care in traditional Western medical spaces as an effective tool to improve maternal and child health outcomes in the United States.

In Adoptive Cell Therapy (ACT), a novel modality of cancer therapy, immune cells can be engineered with T cell receptors (TCRs) to aid in targeting specific antigens presented on the surface of cancer cells. However, TCR-T cells often have limited persistence after transfer into patients, which has hampered the effectiveness of this therapy for solid tumors. Last year, our lab identified LSD1 as a target for drug inhibition, which is an enzyme that alters the epigenome of cells via histone modifications. My project aims to understand the mechanism of LSD1 inhibitor drugs, as well as the effect of these drugs on two types of T cells: cytotoxic CD8+ cells and helper CD4+ cells. In addition to understanding how LSD1 drugs work, I also ask exactly how CD4+ cells enhance the function of CD8+ cells in tumor killing. Which receptors on CD8+ cells are activated by helper T cells, what is the signal phosphorylation pathway transducing the "helping" signal from receptors, and what downstream epigenetic regulators play a role in translating the "helping" signal into better function in CD8+ T cells? To assess these interactions, I will generate a diverse population of CD8+ T cells with targeted receptor knockouts, known as a receptor library. Similar libraries will be generated for epigenetic regulators as well as kinases/phosphatases. The performance of T cells will be assessed via coculture assays, where T cells can kill tumor cells but not fully eliminate the tumor because of periodic addition of new tumor cells. At the end of the coculture period, we will assess gRNAs enriched in dysfunctional populations, which will identify genes critical to CD8+ T cell function. This project aims to provide enhanced function of T cells that are better suited for applications in clinic.

Poor sleep quantity and quality are associated with increased risk of negative health outcomes such as chronic conditions, depression, and anxiety, which ultimately impact overall well-being. The present study aims to investigate the association between perceived stress and sleep quality among college students. A sample of 113 college students in the United States completed an online survey that asked questions about their sleep and perceived stress. Perceived stress was assessed using the Perceived Stress Scale (PSS; Cohen et al., 1983) and sleep quality measured through the Pittsburgh Sleep Quality Index (PSQI; Buysse et al., 1989). Sociodemographic data were also collected. Results indicate that college students with higher perceived stress reported poorer sleep quality. Given that perceived stress and sleep were correlated, we conducted additional analyses to compare levels of stress and sleep quality among different sociodemographic groups. We found that international students exhibited lower perceived stress and better sleep quality compared to domestic students. Furthermore, college students who reported lower family income, and those whose native language was not English experienced higher perceived stress and poorer sleep quality across both groups. Taken together, the data demonstrate an association between perceived stress and sleep quality and highlight differences in both variables based on immigration situation, as well as linguistic and socioeconomic factors. Further longitudinal studies could be helpful to determine the temporal relationships among these variables and the long-term role that they play in health. 

With the recent onslaught of discriminatory executive orders that threaten the safety and rights of queer students, teachers, and their families across the country, cultivating inclusive sexual health education materials in schools that both inform and affirm queer identities has never been more dire. There is evidence that implementing comprehensive, queer-affirming sexual health education can lead to a reduction in homophobic bullying and adverse mental health outcomes for queer students (for review, Goldfarb & Lieberman, 2021). This project applies psychological research methodology to evaluate an intervention summit for inclusive sexual health education. Our three-day summit seeks to provide ten educators from a local Washington school district, including teachers, counselors, and nurses, with the opportunity to come together and develop inclusive, state standards-based sexual health education instructional materials. We hope that this collaboration generates increased educator confidence, leading to improved fidelity of implementation that translates into accurate and queer-affirming sexual health education. This will provide the space for queer-identified children and educators to feel safe in schools. To gauge the efficacy of the intervention, we will assess participant self-efficacy of teaching sexual health and sense of community before, during, and after the summit. To do so, we will collect mixed-methods data, including self-report surveys at baseline and throughout the summit, as well as observational data. The implications of these findings will illuminate key factors that contribute to the understanding of how community engagement can reinforce the importance of community-based learning initiatives, as well as inform the development of instructional materials and networking strategies that will improve the longevity of sexual health education programming. By centering educator development, we combat the fear and misinformation that feeds legislation against queer youth, cultivating safer environments for students and educators to thrive.

Approximately 18% of older adult intensive care unit (ICU) survivors with delirium are diagnosed with Alzheimer’s disease or related dementia (ADRD) within three years of hospitalization. The constant, 24-hour ICU care can cause disruptions to sleep quality and the biological circadian rhythm. These disturbances, which affect up to 80% of ICU patients, can decrease the efficacy and benefits of interventions to manage delirium and improve cognitive function. We aim to explore the roles of individual chronotypes (morning, afternoon, or evening preference) and self-reported cognitive function on acceptability and adherence to a computerized cognitive training (CCT) intervention for older adult ICU survivors.  Data from two ongoing randomized controlled trials (the SLEEP-COG Study and OPTIMIZE Study) are utilized for this project. Older adult ICU survivors hospitalized at University of Washington Medical Center or Harborview Medical Center are enrolled based on the following eligibility criteria: ages 60 and older, functionally independent prior to hospitalization, and within 48 hours after transfer from intensive care to acute care. Participants who are randomized to CCT complete 30-minute daily sessions for up to 7 days or until discharge. Participants answer the Composite Scale of Morningness questionnaire to determine their individual chronotype and complete the PROMIS Cognitive Function & Cognitive Function Abilities instruments to examine self-reported cognitive function. Upon completion of the CCT intervention, participants answer Likert-type surveys and open-ended questions about feasibility, acceptability, and usability.  Analyses will explore trends between chronotype, self-reported cognitive function, and acceptability.  Differences will be described using t-tests, Chi-squared tests, and ANOVA for binary, categorical, and/or continuous variables, respectively. Potential future directions may include the development of personalized interventions that integrate individual chronotype and perceived cognitive ability to minimize delirium and cognitive decline associated with ADRD in older adult ICU survivors. 

Wildfire smoke is a significant environmental health risk for Latino agricultural workers, particularly in Washington State counties with high concentrations of both agricultural activity and smoke exposure, such as Yakima, Chelan, and Douglas. Young workers face unique and poorly understood risks due to potential biological susceptibility and a lack of safety communication tailored to their demographic.

This project explores how social media interventions can reach at-risk populations during smoke emergencies. A short Spanish-language video was developed to deliver protective guidance for agricultural workers in a clear, culturally relevant format. As the first in a planned series of short-form videos, this content promotes protective mask use and introduces key health messages in an engaging, accessible way. Designed for platforms such as TikTok, Instagram Reels, and YouTube Shorts, the video serves as an entry point to a broader educational campaign launching during the 2025 wildfire season.

This pilot will test the reach and engagement of short-form content among younger audiences, who may be less responsive to traditional public health channels. Engagement metrics—including views, likes, shares, and viewer retention—will be collected and analyzed to assess effectiveness and guide refinements in message design and delivery.

Insights from this evaluation will inform a summer campaign focused on improving wildfire smoke safety through practical, culturally grounded messaging. The campaign aims to promote protective behaviors and reinforce employer responsibilities under occupational health regulations. By integrating community-informed design and real-time platform analytics, this work contributes to the development of scalable tools for occupational risk communication and supports ongoing efforts to protect vulnerable workers during wildfire smoke events.

Although being in nature has been associated with benefits including health and well-being, many people do not spend time in nature. Some studies have focused on the reasons why people do not spend time in nature; however, both in this research and in nature and health research, the term "nature" is inconsistently defined. Without a concrete definition of nature, it is unclear what participants consider nature and if this affects their reasons for not spending time in nature. This work is part of a larger experiment looking at students at the University of Washington's reported time in nature, and whether or not having a definition of nature impacted people's responses to nature-related questions. Here, we identify the underlying reasons given for the individual's not spending time in nature. Additionally, we examine whether having a definition of nature which includes urban nature affects reasons why participants report spending no time in nature. Out of 1,573 participants, 273 individuals (17%) responded that they had spent no time in nature in the previous week. They were then asked to explain why they spent 0 hours in nature that week. Common themes included being too busy with school, work, and other responsibilities. Notably, one theme emerged, "feels limited by place" in which participants stated they do not spend time in nature because they "live in Seattle". Preliminary results indicate that when nature is defined, the theme feeling limited by place did not occur, meaning receiving a definition of nature appears to affect the qualitative reasons given for spending no time in nature. This study highlights the importance of defining nature while also providing valuable insights into the reasons why some might prefer to not spend time in nature, as well as barriers that prevent students from spending time in nature.

Electromagnetic launchers currently use a combination of magnetic forces and complex electronic timing to propel objects. One example is the US Navy EMALS system, which uses a linear electromagnetic launcher to launch aircrafts from aircraft carriers. However, using complex electronic timing introduces more failure points within the system, increasing its complexity, making such systems susceptible to being disabled by external electromagnetic interference. Inspired by the design of Tom Stanton, this project explores a new approach that removes the digital-based electronic timing and replaces it with a mechanical timing system that can be used to propel drones or other payloads into the air quickly and efficiently. The goal of reducing the design’s complexity is to create a launcher that is a reliable method for drone and payload deployment. By removing electronic switching and using a mechanically driven circuit closure, this project develops a durable, efficient launch system. The prototype is built using 3D-printed components, powerful magnets, and a coil on a sled with contact arms that touch the conductive rail to complete the circuit. Rather than lining the rail with multiple coils, stationary magnets replace the coils with alternating currents to provide the acceleration when the coil becomes powered. The results allow us to have a competitive design that provides a practical alternative to the typical electromagnetic launchers. Expected results include improved durability, reliable performance due to the simplification of electronics, and reduced energy losses. Our research provides a new way to launch drones or other payloads to be integrated into systems where they would be less susceptible to external electromagnetic interference and jamming.

Tau protein is highly expressed in neurons and other neural cells, including astrocytes. The accumulation of hyperphosphorylated tau aggregates, known as neurofibrillary tangles (NFTs), is a hallmark of neurodegenerative diseases, particularly Alzheimer’s disease (AD). While tau aggregation is thought to advance AD through toxic gain of function, the loss of tau physiological function may also contribute to the adverse progression of the disease. However, how the lack of tau physiological functions in neurons contributes to AD progression remains understudied.  Studies have shown that tau depletion results in minimal phenotypic differences and may even mitigate cognitive decline in AD mouse models. Here, we investigate the molecular consequences of tau loss in both neurons and astrocytes. Using CRISPR/Cas9 gene editing, we generated tau knockout (Tau KO) human induced pluripotent stem cell (hiPSC) lines, which were subsequently differentiated into neurons and astrocytes. We first focused on assessing how Tau depletion affects the hiPSC-derived neurons. Our findings indicate that tau depletion does not impair neuronal differentiation or increase cytotoxicity and cellular stress. However, preliminary data suggest that Tau KO neuronal cultures—composed of 95% neurons and 5% other neural cells—exhibit reduced synaptic firing activity and network burst frequency. These results suggest that tau loss in neurons and glial cells negatively impacts neuronal activity, providing new insights into the functional consequences of tau depletion in AD pathology. To gain deeper insight into how tau depletion negatively impacts neuronal activity, we performed transcriptomic analysis on Tau KO hiPSC-derived neurons using RNA sequencing (RNA-seq). We are currently analyzing and validating the results, which may further elucidate the molecular mechanisms underlying tau loss-of-function in neuronal regulation and AD pathology. In the second phase of this investigation, we will also differentiate the Tau KO hiPSC into astrocytes and assess how tau depletion impacts astrocytic viability and functions.

Non-syndromic orofacial cleft (OFC), comprising cleft lip, cleft palate, or a combination of the two, is among the most common structural birth defects. OFC has both genetic and environmental underpinnings, though the genetic contribution is only partially understood. Members of the same gene regulatory networks associated with OFC are candidates to harbor the “missing heritability” for OFC. Van der Woude Syndrome (VWS) is an autosomal dominant condition characterized by cleft lip and/or palate, and lip pits. Mutations in IRF6, encoding the transcription factor Interferon Regulatory Factor-6, account for approximately 70% of VWS cases, mutations in GRHL3, a transcriptional target of IRF6, account for 5%, and it was recently found that mutations in PRKCi, encoding a kinase, account for 5% more. Published data support that IRF6 is phosphorylated and thereby activated by RIPK4. However, the kinases PRKCi and IKK1 are also required upstream of IRF6 activity. Regulatory interactions within this kinase cascade are unclear. The goal of this project is to shed light upon the kinase cascade encircling IRF6 to illustrate genetic causes of VWS and non-syndromic OFC and ultimately improve patient outcomes. I hypothesize that IRF6 phosphorylation is dependent upon RIPK4, which is activated by IKK1 and downstream of PRKCi activity, and all components are necessary for IRF6 activation. To test this hypothesis, I am conducting experiments injecting mRNA products from our genes of interest into zebrafish embryos and detecting phosphorylation activity within the harvested protein through immuno-precipitation and mass spectrometry analysis to determine protein-protein interactions. I anticipate that in the absence of RIPK4, the pool of Differentially Expressed Genes (DEGs) will align with the pool of DEGs in the absence of IKK1 and IRF6 respectively. Implications of these findings will ultimately give a stronger understanding of the molecular foundations of VWS and non syndromic OFC.

Soldering is a common activity in lab environments that can negatively impact indoor air quality (IAQ) due to the release of airborne particulate matter (PM) and hazardous fumes from solder wire and rosin-based flux. The use of solder wire has been found to increase PM2.5 levels, as well as airborne tin and lead concentrations. Exposure to rosin-based solder flux has been linked to asthma, chronic coughing, and wheezing. For those who spend extended periods of time in laboratories, exposure to air pollutants may lead to an increased risk of respiratory issues and reduced cognitive capabilities. In order to mitigate these risks many laboratories employ air purifiers, however, there exists a lack of recent research on the effectiveness of these air purifiers. This study proposes the use of AeroSpec, an indoor air-quality monitoring system that measures pollutants such as PM1.0, PM2.5, and PM10, to assess the performance of various air purifiers. The AeroSpec system will be used to monitor airborne particulate matter concentrations while a researcher solders under different conditions, testing both with and without various air purifiers in different configurations. Data from the AeroSpec sensors will be used to quantify the effectiveness of different purifiers and examine how the location of the soldering iron relative to a purifier affects its performance. Our project aims to provide both independent verification of the effectiveness of commercially available air purifiers as well as to give guidance on best practices to maximize the effectiveness of air purifiers and to improve IAQ in laboratory settings, therefore reducing associated risks.

Kappa opioid receptor (KOR) ligands have been explored for anti-anxiolytic, anti-depressive, pain, and substance use disorder therapeutics. These therapeutic effects are partly due to biased signaling through the cJun N-terminal Kinase (JNK) pathway, which involves complex molecular interactions and downstream effects that inactivate the receptor by producing reactive oxygen species (ROS). JWT-101, a clinically approved compound, has been shown to produce therapeutic effects for these conditions. We hypothesize that its mechanisms of action are through KOR antagonism. I previously assessed KOR agonist-induced analgesia by measuring the latency of tail withdrawal from 52.5°C water after treatment with U50,488, a KOR agonist. Pretreatment with 15mg/kg JWT-101 24 hours before U50,488 injection effectively blocked KOR-induced analgesia in wild-type male mice. This effect was reversed by the short-acting, KOR-selective antagonist Aticaprant (5 mg/kg), suggesting that JWT-101’s action is mediated through KOR. Further investigation using in-vivo fiber photometry with the novel peroxide sensor AAV oROS-Gr revealed that JWT-101 significantly increases ROS production in KOR-expressing cells. Injection of 15 mg/kg of JWT-101 increases oROS fluorescence compared to control post-injection. Pretreatment with Aticaprant 15 minutes prior to JWT-101, blocks oROS fluorescence, suggesting that JWT-101’s activity is mediated by KOR. Pretreatment with MJ33 (a PRDX6 inhibitor upstream of JNK activity) 50 minutes before treatment with JWT-101 blocked oROS fluorescence, suggesting that this ROS production is through the JNK/PRDX6 pathway of KOR activation. This study provides insights into the mechanism of action of JWT-101 and examines the underlying molecular mechanisms of KOR-associated effects.

Carpels are invaluable floral organs that have undergone myriad innovations in flowering plant evolution, providing fruits and seeds. Carpels consist of a pollination site (stigma), pollen growth area (style), and a region containing ovules (ovary) which become the fruit after fertilization. Understanding carpel development and evolution is fundamental for efforts to increase food production. A decreased abundance of insect pollinators due to anthropogenic climate change has made this pursuit all the more urgent. The plant genus Thalictrum comprises perennial herbs in temperate regions that have repeatedly transitioned from insect to wind-pollination in their evolutionary history. Hence, they are ideal to investigate the adaptations and genetics that decrease reliance on insects for reproduction. A key feature of wind pollination is a longer stigmatic surface that increases pollen capture. I am taking a candidate gene approach, examining homologs of the stigma development gene families STYLISH (STY) and NGATHA (NGA). I am characterizing the role of NGA and two STYLISH gene copies in our model species, Thalictrum thalictroides. Previous work has found that silencing one of the STY gene copies results in the loss of stigma development, while the function of the other copy remains unknown. In other genera, altered NGA expression has been shown to alter stigma development. To characterize the function of both NGA and STY, I am implementing Virus-Induced gene Overexpression (VOX) by infiltrating tubers with Agrobacterium transformed with Tobacco Rattle Virus (TRV) carrying an overexpression construct. STY genes will be overexpressed, and NGA will be both overexpressed and silenced using Virus-Induced Gene Silencing (VIGS). I will characterize the phenotypes resulting from these treatments to determine the functionalization of STY and NGA. By dissecting the genetic basis of floral adaptations to wind-pollination in this system, I hope to contribute solutions to enhance crop production in the face of pollinator decline. 

Previous research has determined that nanoparticle systems require a wide parameter space to effectively conduct synthesis and characterization. As a result, the development of high-throughput techniques is essential for efficiently analyzing the large datasets produced in colloidal particle experiments. These methods enable the rapid assessment of particle properties, such as size, shape, and charge, which are critical for modifying nanoparticles for specific applications. In order to do this, advancements in automated synthesis platforms, such as the Jubilee automated multi-tool system, offer the potential to streamline the fabrication of magic sized clusters. This approach has the potential to accelerate the discovery of novel nanoparticles but also allows for real-time adjustment of synthesis parameters to achieve desired properties with high precision, throughput, and reproducibility. As a result of the optimized synthesis process, characterization using techniques such as small angle X-ray scattering (SAXS) and UV-vis spectroscopy can be done at an accelerated rate. Efforts to enhance the durability and performance of the Jubilee automated multi-tool platform are focused on integrating advanced materials to improve system lifespan. This work will incorporate glass syringes and resin-printed components which offer improved chemical resistance and precision compared to traditional plastic components, extending the utility of the platform to be able to work with solvents and chemicals that are corrosive, volatile, or strong solvating agents for typical plastics. These improvements aim to reduce wear and tear, extend the lifespan of critical components, and ultimately ensure the platform's reliability for long-term use in high-throughput nanoparticle synthesis.

Machu Picchu is an archaeological site in Peru that is an important source of information about the Incas. In the middle of the 15th century, Machu Picchu was established as an ancient city by the Inca Empire. However, their lives were disrupted by Spanish contact and disease. Smallpox spread from 1519 to 1520, which killed the emperor of the Inca Empire, Huayna Capac. Spanish contact in 1532 forced the Inca people to abandon Machu Picchu. My research investigates whether there may have been epidemics before Spanish contact and how they may have affected the Inca people. I applied survival analysis statistics to the raw data on human remains and compared the results to a known plague population from Europe. I also analyzed radiocarbon ages to detect epidemics in earlier time periods. The skeletal data analysis did not find strong evidence of pre-contact epidemics. The radiocarbon analysis suggests potential depopulation due to epidemics. I encourage future scientists to investigate with more data and a bigger range of periods before and after Spanish contact to further explore possible past epidemics in this region.

Surf smelt (Hypomesus pretiosus) are an ecologically and economically significant forage fish species that spawn in the intertidal zone of beaches throughout the Salish Sea. Despite their importance to marine food webs, the environmental factors influencing their spawning site selection and seasonal distribution remain poorly understood. This project aims to investigate the morphological characteristics of beaches used for surf smelt spawning during different times of the year, comparing morphological and spatiotemporal variables that influence spawning. In order to study these characteristics, we will record sediment grain size, slope, wave energy, beach temperature and the water chemistry at verified winter as well as summer spawning sites identified by the Washington Department of Fish and Wildlife (WDFW). We will also sample non-productive sites in order to identify key differences between them and further establish parameters that enhance spawning success. Preliminary research suggests that these key characteristics strongly influence surf smelt spawning distribution​​​. Optimal surf smelt spawning beaches appear to consist of mixed sand and gravel substrates, low levels of wave action, high amounts of shading, moderate slopes and moderate temperatures. Habitat alterations such as shoreline armoring along with sea-level rise in response to global warming could lead to a drastic decrease in the upper inner tidal ranges where surf smelt usually spawn. Consequently, we expect beaches heavily influenced by these factors to be poor spawning sites​​. The results of this study will contribute to a deeper understanding of the environmental variables driving spawning site selection, egg survival, and seasonal spawning peaks. This research will be instrumental in informing conservation projects and supporting policy initiatives aimed at preserving surf smelt populations and their critical spawning habitats in the Salish Sea.

The intricate interplay between different brain cell types is crucial to understanding neural pathophysiological states. This project aims to investigate the effects of the treatment of GHK on glial activity and inflammation using organotypic brain slice cultures. GHK improves tissue regeneration and exhibits anti-inflammatory effects, promoting neural protection. Slices taken from mice mirror the human-brain microenvironment, allowing a better understanding of neuronic health in pathological states. They also preserve the 3-D architecture of our brain, maintaining the intricacies between diverse cell types. First, the brain is collected from an euthanized mouse and rinsed in PBS, then sectioned into 100µm slices to culture, where they are exposed to different levels of GHK. Brain tissue samples are fixed in formalin to preserve cellular structure and stored in PBS. The tissue is embedded in paraffin to support stable sectioning using a microtome, allowing precise slicing into 4µm thick sections for analysis. By employing immunohistochemistry and histological techniques, insights into therapeutic strategies with the comparison of tissue cultures are shown. IHC looks at activated microglia(using IBA1), astrocytes(using GFAP), chronic inflammation(MCP1), and synaptic activity (synaptophysin) and characterizes neurons with cresyl violet staining. MCP1 levels are expected to decrease with GHK treatment. For microglia, there might be a reduction in their activated, proinflammatory state; astrocytes may show a shift towards reduced reactivity, shifting toward a homeostatic role in maintaining brain tissue stability and function. Synaptic activity is expected to improve. Neuronal health is predicted to be preserved, with enhanced structural stability and reduced signs of cellular stress. These results will help demonstrate the potential of GHK in mitigating chronic inflammation and promoting neuronal health. By revealing how GHK influences glial function and neuronal health, this research could pave the way for novel interventions targeting the improvement of neuronal health. 

Xylazine is a veterinary sedative that has become a common adulterant in fentanyl products due to its ability to prolong the euphoric effects of fentanyl. Adulterated drug mixtures containing xylazine have been linked to an increased risk for respiratory depression and fatal overdose. Existing treatments for overdose such as naloxone do not work against xylazine and there is currently no FDA approved reversal agent for xylazine toxicity. Monoclonal antibodies (mAbs) are a promising therapeutic option to reverse drug overdose and can be used to target small molecules by sequestering them in the bloodstream and preventing their passage into the brain. Previously, we made a series of xylazine haptens and formulated them into vaccines where they were found to induce strong antibody responses, reduce brain to serum ratio of xylazine, and mitigate respiratory depression in Sprague-Dawley rats. Based on the protective effects of our xylazine conjugate vaccines, we hypothesized that we can create an effective reversal agent by isolating xylazine specific mAbs. In this study, we immunized mice with xylazine conjugate vaccines to generate antibodies specific against xylazine. We then used the secondary lymphoid organs of the vaccinated mice to establish stable mAb producing cell lines using hybridoma technology. Hybridomas were screened with ELISA and lead candidates were selected and sequenced for further characterization. We will conduct in-vitro characterization assays to quantify binding affinity, functional activity, and thermostability of our lead mAbs. Furthermore, in-vivo studies will assess the efficacy of our mAb lead candidates in rodent models.

Robots must be able to pick objects from densely packed shelves in order to automate industrial warehouses. Dense packing gives rise to challenges in grabbing targeted objects efficiently as the shelves can be clustered, objects can be stacked, and the target object can be obstructed from direct reach. The goal of this research project is to create a new gripper combined with reinforcement learning to manipulate objects within a shelf without multiple attempts or repositioning of the robot arm. The new gripper design includes four fingers that are linear actuators with vacuum units and suction cups attached to the ends of each finger. Additionally, each finger contains a time-of-flight sensor at the tips which provide spatial information for different objects within the shelf. I integrated time-of-flight sensors into the multi-fingered gripper and filtering algorithms for the sensor’s data. I modified the previous vacuum ejector unit which only provided support for one unit to four vacuum ejector units. I also conducted a series of experiments that provided cases where the new gripper design proved to be better than the previous design. We also collected suction cup vacuum data and trained a neural network to predict the success rate of suction cup attachment. The results of this project will inspire new designs for increasing the success rate of robotic grasps within densely packed environments.

The proximal tubule (PT) and glomerulus are vital blood-filtering components of the nephron, the functional unit of the kidney. The components’ micro-scale sizes and intricate three-dimensional structures are critical to kidney function, although accurate in vitro modeling has proven difficult. Limitations in fabrication techniques have forced size scaling and imprecise morphology in models. In this study, we addressed fabrication limitations using multiphoton ablation to etch intricate, three-dimensional proximal tubule and glomerulus vessels in collagen hydrogels. We sought to demonstrate model viability by introducing human proximal tubular epithelial cells (hPTECs) and human umbilical vein endothelial cells (HUVECs), respectively, through cell perfusion. However, we encountered a significant challenge: due to the small diameter and high curvature of the micro-scale channels, the cells tended to aggregate, disrupting cell profusion and cellularization throughout the vessels. Cell aggregation was especially prominent in the glomerulus model due to the more tortuous and complex geometry. While our cellularization trials on native-scale models proved it is feasible to perfuse cells throughout the vessel, we still need to refine cellular profusion and cellularization. To improve cellular profusion and cellularization, we are first studying a 1.5-scale glomerulus model. The scaled model's increased vessel diameter and lower curvature demote cell aggregation and enhance the ease of cell profusion. We anticipate that cellularizing the 1.5-scale model will provide a deeper understanding of the variables facilitating cell profusion that we can use to improve native-scale vessel cellularization. Fabricating native-scale, accurate in vitro PT and glomerulus models is crucial for developing a deeper understanding of hemodynamic influence on kidney function. These findings contribute to the fabrication of more biomimetic in vitro PT and glomerulus models that will pioneer therapeutics and the understanding of kidney physiology and pathology.

The effectiveness of a drug candidate depends on its ability to distribute to its target site of action after administration. Thus, a primary concern for drug delivery labs like the Pun lab is preventing drugs from being cleared from the bloodstream by the body's renal system before they are able to accumulate to therapeutic levels at their site of action. In short, one important goal in drug delivery research is to find ways to extend a drug's blood circulation half-life. Conjugating drugs to the large molecular weight molecule polyethylene glycol (PEG) to slow their clearance kinetics is the current gold-standard method, but a crucial drawback is that PEG's large size leads to its potentially toxic buildup in tissues like the liver. To get around this problem, my project aims to develop a drug delivery platform that will allow small molecule drugs to reversibly bind, or in other words "hitchhike" onto human serum albumin (HSA), an abundant protein in blood plasma with an extraordinarily long half-life. At this point in my project, I have successfully synthesized a novel fatty acid monomer with a methacrylate functionality that can be used to copolymerize the monomer with therapeutic small-molecules or peptides to improve their circulation half-life. The next steps will be to copolymerize the fatty acid monomer with pGmMA, a water-soluble polymer, and use biolayer interferometry to test the fatty acid monomer's ability to coordinate to albumin, which will confirm its efficacy as a drug delivery platform. If successful, this project has the potential to provide a generalizable improvement to the pharmacokinetics of various kinds of small-molecule drugs or peptides, enhancing their potency and overall ease of treatment. 

Cyberbullying is repeated, intentional harm inflicted through digital platforms, including social media, messaging apps, and online forums. Most researchers focus on cyberbullying in adolescence, but it continues into adulthood. Young adults (ages 18–25) undergo major life changes and use social media frequently, which increases their risk of experiencing cyberbullying. LGBTQ+ individuals, especially transgender and non-binary people, face higher rates of online harassment. Cyberbullying can lead to severe mental health issues, including depression, anxiety, and social isolation. While existing research focuses on cyberbullying among LGBTQ+ individuals as a broad group, few studies examine its specific impact on transgender and non-binary people. This study employs a mixed-methods approach to investigate cyberbullying in transgender and/or non-binary emerging adults. A survey of transgender and/or non-binary participants aged 18–25 will be conducted using a modified Cyberbullying Victimization Scale to measure three types of victimization: Verbal/Written, Visual/Sexual, Social Exclusion/In-Person Bullying. Participants will report experiences in public online spaces and private online spaces. The study will collect demographic data such as gender identity, racial or ethnic background, level of outness, and social media usage to examine correlations between these factors and cyberbullying experiences. By integrating statistical trends and personal narratives, this research will provide a deeper understanding of cyberbullying in transgender and non-binary communities. The findings will inform social media policies, platform safety measures, and mental health support efforts for LGBTQ+ individuals navigating online harassment.

Fusion, the process powering the sun, offers a promising solution for deep-space propulsion as it can provide high specific impulse and lightweight fuel. The Flow Z-Pinch lab is exploring the innovative sheared flow stabilized (SFS) Z-pinch technique to mitigate plasma instabilities and enable fusion using axial flows, creating a compact, scalable path to fusion power and space thrusters. Interactions with electrodes often introduce impurities into ZaP-HD plasmas, whose emission can be monitored to measure plasma parameters like electron temperature. However, excessive impurity concentrations can also contribute to radiative losses, degrading fusion performance. Spectroscopy is a key diagnostic tool for analyzing impurities, allowing measurements by examining light emissions from atoms. The ionizations per photon method (S/XB), which correlates emission intensity at a specific carbon line to particle flux using empirical coefficients based on temperature and density, will be used to quantify impurity influx. Our project uses an existing spectrometer and photomultiplier tube (PMT) setup, with initial efforts focusing on absolute calibration to relate pixel intensity to photon flux. This diagnostic is essential for our understanding of impurity dynamics and their migration in ZaP-HD plasmas. The PMT can give us a time-resolved measurement to correlate to other time-resolved diagnostics, especially to characterize the erosion rate of electrode surfaces. Preliminary data analysis will calculate carbon influx using calibrated data and S/XB coefficients, with future work extending the diagnostic system to monitor additional carbon charge states and emissions at varying axial locations. This research provides novel insights into impurity behavior in ZaP-HD, enhancing our understanding of plasma-material interactions and informing us of strategies to minimize impurity influx for improved fusion performance.

The transcription factor LEAFY (LFY) controls the development of flowers in angiosperms, but it is found in all lineages of land plants, including those that do not flower. In the non-vascular plants like moss, LFY promotes the first cell division in the zygote, and in early vascular plant representatives like ferns, LFY controls stem cell activity. Ferns are the sister lineage of seed plants, making them an ideal model to study the evolution of LFY. The model fern Ceratopteris richardii has two copies of the LFY gene, while most angiosperms have one, and the two genes are expressed at different levels across development, suggesting the possibility of sub- or neo-functionalization. In this study, we use transgenic C. richardii plants overexpressing one or both LFY genes to determine their function across development. Since LFY is expressed in sperm cells, I set up assays to observe C. richardii sperm cells during fertilization and determine the role that LFY may be playing in fern sperm development and reproduction. To test whether the two fern LFY proteins interact with each other, I perform yeast two-hybrid assays, which will provide insight into whether the genes play independent roles or share overlapping functions. Determining the function(s) of LFY in ferns will help uncover the evolutionary history of this important plant transcription factor and how it came to control the crucial role of initiating flower development.

During embryonic development, gene expression is temporally and spatially coordinated to control organogenesis and fetal growth. We previously identified a subset of 140 genes that conferred lethal and sub-viable phenotypes in mice and are likely to be haploinsufficient in humans. These genes presumptively play essential roles in fetal development, but their function is unknown. I aim to uncover the role of these genes in mouse embryonic development using Weighted Gene Co-Expression Analysis (WGCNA). Co-expression analysis will be conducted on mouse embryonic stem cell RNA sequencing data obtained at three different stages of in vitro differentiation and across two different genetic backgrounds, creating a subset of nine samples encompassing 12555 genes. Choosing three different time points allows us to see how expression of our genes of interest changes over time, and choosing two different genotypes (wild type and knock-in) allows us to investigate if expression changes due to a single point mutation. We performed dynamic clustering on this RNA sequencing data to identify co-expressed gene clusters. I will map these gene clusters to biological pathways to make inferences about which cellular processes, metabolic functions, or structural components the genes of interest are involved in. This may indicate the role of these genes in fetal development and help reveal why fetal viability is compromised. In future studies, the functional characterization of these genes will generate new ideas and hypotheses about the basis of genetic disease.

Unlawful behavior by both the government and its civilians in Mexico has developed into a crisis of violent crimes and corruption. There is a culture of high disregard of the law that is fueled by distrust in the government, subcultures such as Narco-culture, as well as institutional weakness. The aims of this study are to learn of the experiences Mexican citizens have with unlawful behavior and corruption in their government as well as understand the motivations behind this kind of behavior. For the purpose of this study, unlawful behavior will be defined as any behavior or action that violates Mexican law. Through semi-structured interviews of adult Mexican citizens I will gain insight on their thoughts and experiences. They will be asked questions regarding their participation and experiences with unlawful behavior. Due to travel limitations, the Mexican citizens interviewed for this study will be residents of Washington state. It is Mexican citizens that are facing the consequences of the high crime and corruption which makes it imperative to amplify their stories and understand their experiences and perceptions. The information collected through this research may aid in finding a solution to Mexico's crisis of crime and corruption in hope of providing Mexican civilians with a better quality of life. 

Our goal is to establish water quality baselines and ongoing trends for streams and tributaries of two separate watersheds, both of which drain into Lake Washington through Lake Forest Park, a city in the northwest part of King County, WA. This project is the first intensive multiple-site survey of urban King County watersheds using certified Washington State Department of Ecology methods. In three teams of 4-5 college students, we conducted monthly field tests of 16 sites along tributaries and sub-basins of the McAleer Watershed, and of 6 sites within the Lyon Creek watershed. We used a YSI ProDSS multiprobe meter and a Hach DR 900 colorimeter to collect measurements of air and water temperature, water turbidity, conductivity, % oxygen saturation, dissolved oxygen, pH, total suspended solids, and nitrate and phosphate levels, used Coliform Bacteria R-Cards to measure the quantitative presence of water-borne E. coli. Analysis of these water quality indices for these sites over a 12-month period will allow us to evaluate the overall health of the greater watersheds, and possible causes of poor conditions. Our data will contribute to other conservation research efforts supporting urban watershed health. This work was undertaken as a research project by undergraduates participating in the Urban Stream Ecology Internship and Training (USE-IT) program, funded by a Seattle Waterworks grant to the Stewardship Stream Initiative (SSI), an initiative launched by the Lake Forest Park Stewardship Foundation in 2024.

Malaria, caused by Plasmodium parasites and transmitted via mosquito bite, caused over 600,000 deaths in 2022, making the disease a pertinent public health problem. After injection into mammalian hosts through mosquito bite, Plasmodium parasites travel into the liver and develop in hepatocytes, where they undergo massive replication but cause no symptomatic disease. The parasites then egress into the bloodstream, where they infect red blood cells and cause the clinical symptoms and mortality associated with malaria, along with transmission to mosquitos to continue the cycle of infection. Although the liver-stage of the parasite is clinically silent, parasite infection of the liver results in incompletely understood hepatic immune responses that impact the development of immune memory, which is necessary for protection from future infections. Innate-like αβ and γδ T cells make up a significant proportion of intrahepatic lymphocytes, leading us to become interested in how these immune cells respond to a primary Plasmodium parasite infection of the liver. To address the role of these T cells in combating a primary liver-stage infection, we infected wildtype mice, mice that lack αβ T cells, and mice that lack γδ T cells with Plasmodium parasites and examined parasite density, size, and hepatic localization using immunofluorescence microscopy. Preliminary results demonstrate no significant differences in malaria parasite susceptibility between wildtype mice, mice without αβ T cells, and mice without γδ T cells, indicating that these cell types alone may not mount a significant anti-Plasmodium response upon primary infection. Future work will involve examining T cell localization within infected tissues to determine how T cell localization is impacted by primary infection and characterizing subsets of T cells that are present in infected livers. We hope these results add to a greater understanding of the entire hepatic immune response to primary Plasmodium parasite infection of the liver.

Adoptive cell therapy with CAR-T cells has shown promise in hematological malignancies, but efficacy in solid tumors remains a challenge in part due to CAR-T cell exhaustion and antigen heterogeneity. However, the vast majority of preclinical models do not recapitulate the tumor-immune interactions that produce these barriers. To study CAR-T therapy in a rigorous model that recapitulates tumor-immune barriers, we adapted a Kras^LSL-G12D/+;P53^f/f (KP) genetically engineered mouse model (GEMM) of lung adenocarcinoma. However, adapting the KP-GEMM model for various target antigens, genetic drivers of disease, or interfacing with the vast array of powerful genetic mouse models is resource intensive which prohibits widespread utility. Here, we propose a defined, modular system for generating GEMM for CAR-T preclinical studies using the Sleeping Beauty (SB) transposon system. The proposed system uses polyethylenimine (PEI) to deliver SB transposon encoding oncogenic Kras^G12D and P53^R175H dominant alleles as well as our target antigen hROR1, in vivo to wild-type mice. We demonstrate that in vitro PEI successfully introduces genetic cargo into lung epithelial cell lines, while SB transposons mediate stable integration and expression. Next, we will test this in vivo. This system affords the induction of tumors with specific oncogenic driver mutations and specific tumor antigens on any genetic background. Ultimately, we expect that this approach will streamline preclinical use of GEMM in preclinical research. 

Currently, nearly 10% of Americans have type 2 diabetes (T2D), placing it among the most common chronic diseases in the United States.  Growing evidence points to brain neurocircuits that regulate glucose homeostasis as potential targets for developing novel therapeutics to treat T2D. Recent studies have revealed that the brain can induce sustained remission of hyperglycemia in rodent models following intracerebroventricular (icv) injection of fibroblast growth factor 1 (FGF1). FGF1 stimulates the MAPK/ERK signal transduction system, which translates signals from the activation of cell surface receptors into gene expression changes within the cell nucleus. Following FGF1 injection, tanycytes and astrocytes are sites of transcriptional changes related to the ERK pathway. Inhibition of MAPK/ERK signaling blocks FGF1-induced diabetes remission. Like other growth factors, FGF1 requires integrin signaling to elicit the full spectrum of its cellular responses.  The integrin receptor αvβ3, which is implicated in the chronic cellular response to FGF1 is expressed in hypothalamic neurons and tanycytes and is required for FGF1 to induce sustained activation of MAPK/ERK signaling. To determine if integrin signaling in tanycytes is required for FGF1 to induce diabetes remission, we utilized adult, male, diabetic mice expressing either floxed Integrin β3 (ITβ3) or floxed Integrin αV (ITαV). Mice received a single 500µl injection of TAT-cre or heat-inactivated TAT-cre into the 3rd ventricle. TAT-cre administration inactivates the expression of genes encoding either ITβ3 or ITαV. Next, each cohort received a single icv injection of either FGF1 (2 µg) or saline vehicle. Food intake, body weight, and blood glucose measurements were taken daily throughout the investigation.  Our data demonstrates that the ability of icv FGF1 to induce lowering of blood glucose levels is not blocked by knockout of either ITβ3 or ItαV in tanycytes suggesting that integrin signaling isn't required for FGF1 to induce diabetes remission.

Historical archaeologists understand toys in the material record to explicitly mark past activities of children. This project focuses on the play activities of enslaved children on North American plantations through the toys they left behind. We used data from the Digital Archaeological Archive of Comparative Slavery (DAACS) to explore the material evidence for identity formation and socialization of enslaved children based on existing studies on enslaved childhoods. Specifically, we investigated the following questions: what materials and manufacturing techniques were most frequently used in the creation of the toys of enslaved children on North American plantations? Based on the associated costs of these materials, what is the range of expense exhibited by enslaved children's toys? What role might toys have played in enslaved children’s agency or passivity in socialization? To address these questions, we queried the DAACS repository concerning object specifications like material and manufacturing technology for toys excavated from North American plantations. This data was imported into RStudio, where we used programming packages to clean it and create visualizations to convey trends. Our pilot project (48 toys) found that porcelain and a molding technique were most frequently used, materials with varying expenses were present, and some toys exhibited post-manufacture modifications. We now have data on 599 toys from 41 sites on 13 plantations, greatly improving our representation of enslaved children in the material record. In this poster we present the result of our analysis of this large data set and compare the locations. The findings of this research will help to fill in gaps of a larger conversation about the relationship between childhood play and race, and more broadly contribute to archaeology’s attention to past social dynamics.

Dynamics of activity across the cerebral cortex at the mesoscopic scale – coordinated fluctuations of local populations of neurons — are essential to perception and cognition and relevant to computations like sensorimotor integration and goal-directed task engagement. However, understanding direct causal links between population dynamics and behavior requires the ability to manipulate mesoscale activity and observe the effect of manipulation across multiple brain regions simultaneously. Here, we develop a novel system enabling simultaneous recording and manipulation of activity across the dorsal cortex of awake mice, compatible with large-scale electrophysiology from any region across the brain. Transgenic mice expressing the GCaMP calcium sensor are injected systemically with an adeno-associated virus driving expression of the ChrimsonR excitatory opsin. This strategy drives expression of the blue-excited calcium indicator, GCaMP, in excitatory neurons and red-excited Chrimson opsin in inhibitory neurons. The light channels of the imaging and the opsin do not interfere. We demonstrate widefield single-photon calcium imaging and simultaneous galvo-targeted laser stimulation over the entire dorsal cortical surface and find that the spatial and temporal resolution of the stimulus is suitable for targeting many specific cortical regions in short periods of time. The calcium indicator responded to the laser within 30 ms, and the activity returned to baseline within 100 ms after laser offset. The area of effect was as small as 3 mm² for the lowest laser power or as large as 10 mm² for the largest laser power. Moreover, the preparation is stable over many months and is thus well-suited for long-term behavioral experiments. The ability to stimulate and measure anywhere on the dorsal cortical surface of the brain will allow us to design computational models describing how causal manipulation impacts neural dynamics, especially in the context of designing closed-loop systems to control neural activity and behavior. 

Diatoms are one of the main groups of phytoplankton that form the base of the food web in aquatic systems and make up a large portion of the phytoplankton assemblage in the estuaries and sounds of the Pacific Northwest. Diatoms have a silica-based cell wall and lack flagella, limiting mobility. Due to silica being a significant element of their cell walls, diatoms have a greater silica requirement than dinoflagellates or other phytoplankton groups. As climate change progresses, ocean temperatures rise, and marine heat waves (MHWs) are becoming more frequent. Prior research has found that, in the North-Eastern Pacific Ocean, the relative abundance of diatoms compared to dinoflagellates decreases during these events. Two main hypotheses proposed to explain this trend are that 1) dinoflagellates are more temperature tolerant and 2) silicate availability may decrease with increased temperature, and diatoms are more reliant on silicate than dinoflagellates. For this study, I propose to examine the diatom/dinoflagellate ratio relative to changes in temperature and silicate availability in Tofino Inlet in Clayoquot Sound on the west coast of Vancouver Island BC, Canada from 2013-2023, during which time there were multiple marine heat waves. University of Washington Tacoma researchers have been collecting water property data in Clayoquot Sound in the late summer/early fall annually since 2001 and phytoplankton samples since 2007. These data include CTD profiles of temperature, salinity, density, oxygen, fluorescence and transmissivity and discrete water samples for nutrients and phytoplankton. Phytoplankton 10-meter vertical net tows (20 um mesh) were collected in addition to bottle samples at the surface and 10 m at the same stations every year. These samples were fixed with formalin and counted in the lab using a research grade compound microscope. I will be using these data for my study.

In genomics, large-scale experiments commonly exploit DNA libraries, transforming them into a biological system to assay their effects. In these experiments, smaller DNA constructs provide increased transformation efficiency, such that larger libraries can be screened. Previously, we observed a 3-fold increase in the DNA transformation rate in the fruit fly Drosophila melanogaster when we reduced the size of the DNA construct to approximately 1/6 its initial size. Further improvement of this transformation rate will be dependent on a number of features, including the ease of production of tens of micrograms of minicircles, the ability to create a library of minicircle constructs, and the presence of a PhiC31 attB site to allow integration into the D. melanogaster genome. There are no existing minicircle production methods that address all these needs; thus, we are developing a DNA minicircle production approach that should address them. We use asymmetric PCR to produce single-stranded DNA (ssDNA) that is then circularized. We use these ssDNA circles as templates for rolling circle amplification to generate large quantities of double-stranded DNA (dsDNA). Following debranching and digestion of the DNA, we use a serine recombinase PhiC31 and its recombination directionality factor gp3 to recombine small dsDNA fragments into dsDNA minicircles. To date, we have produced linear single-stranded DNA using asymmetric PCR, ligated the linear ssDNA into circular ssDNA, and performed in vitro recombination using PhiC31 and gp3 to make minicircles from small dsDNA sequences. We are now working to combine all parts of the procedure into one continuous workflow and to measure the method’s efficiency and yield. This procedure will be vital for large-scale genomic manipulations of D. melanogaster, and should thereby improve the scale of these experiments. The approach should ultimately increase our understanding of genetic variation as we can assay it in D. melanogaster.

Children in Low- and Middle-Income Countries (LMICs) are exposed to pathogens and antimicrobial resistance (AMR) genes at high rates due to more exposure to animals and insufficient access to water, sanitation, and hygiene (WASH) resources. This study analyzes the prevalence of AMR gene carriage in young children aged 6, 12, and 18 months living in communities spanning a rural to urban gradient in Northwestern Ecuador. Stool samples were collected from 428 children at ages 6 months (n=327), 12 months (n=373), and 18 months (n=368) in Ecuador. Stool samples were extracted for total nucleic acids and assayed for pathogen and AMR targets via TaqMan Array Cards. This analysis focuses on 15 AMR targets, which include beta lactams, class 1 integrons, fluoroquinolones, and folate pathway inhibitors. There was a very high prevalence of AMR genes, especially SHV (80.7%), sul2 (82.7%), Intl1 (88.9%), qnrB1 (90.0%), sul1 (90.2%), and TEM (98.9%). There was very low prevalence or no detection of KPC (0.0%), NDM (0.1%), and VIM (0.0%). This study demonstrated that there is a high prevalence of diverse AMR genes in children in Ecuador across all community types and ages. This suggests that these children have high exposure rates to AMR genes that could potentially lead to difficulties in determining appropriate antibiotic treatment for future illnesses.

Flowering plants are important sources of agricultural crops and are diverse in flower and fruit structures. To study how this diversity has evolved, I am developing a new plant model system in the order Ranunculales, an underrepresented clade that will help bridge the knowledge gap within dicotyledonous plants, where most of the angiosperm diversity is found. A key tool in model system development is the ability to transform plants to study gene function, therefore, I aim to develop a method for genetic transformation. Specifically, I am implementing the tried and true  “floral dip” transformation method of Arabidopsis using Agrobacterium tumefaciens in Myosurus minimus, tiny mousetails. This is a small, fast-growing plant that produces many seeds, making it feasible to generate and propagate stable transformants. As proof of principle, I am expressing fluorescent proteins (GFP and YFP) and a visual reporter called RUBY, which causes transformed plants to produce red pigment. I found red color on the petals of treated plants, suggesting that transformation is working in Myosurus. I am currently validating this phenotype by planting the seeds on antibiotic selective media. I expect to see that certain individuals are resistant to antibiotics, while others die, which will allow me to identify the genetically transformed plants. By developing this transformation method, I will be able to test the function of candidate genes of interest in this new model plant species, which will further enable the investigation of gene network evolution in flowering plants. Increased understanding of gene function provides opportunities for engineering crop species to have beneficial traits for agricultural purposes. 

Infertility, the inability to conceive after one year of unprotected intercourse, affects about 15% of the global population. Nearly half of couples diagnosed with infertility have a contributing male or sperm factor (male-factor infertility). Preparation of purified sperm is an important component of fertility treatments. Most purification techniques rely on centrifugation, although the effects of centrifugation on sperm motility (movement) and DNA integrity are not well understood. Our objective is to manipulate centrifugation g-force and time settings following sperm preparation to investigate potential impacts of centrifugation on sperm quality for fertile and infertile men. Our hypothesis is that the highest centrifugation g-force and time will result in reduced motility and maximum DNA damage. Participants with normal semen parameters are recruited. A semen analysis is performed to determine volume, concentration, and baseline motility, which is measured using an automated visual sperm analyzer. Motile sperm are isolated via “direct swim out,” a standard purification technique. Nine different centrifugation settings are applied to the sample (time and g-force parameters studied: 7, 30, and 60 minutes, and 250xg, 600xg, and 800xg), and motility is evaluated. For each condition, DNA integrity is assessed with a COMET assay (uses single-cell gel electrophoresis to detect double-stranded DNA breaks). Preliminary results from 11 participants (ages 26-35) demonstrate substantial variability in motility for each participant. We observe that while the highest centrifugation g-force and time condition (60 minutes at 800xg) reduces motility for some participants, it has the opposite effect for others. Data collection for motility analysis and DNA integrity assays are in progress and will continue over the next three months. This project aims to provide high quality evidence to support current laboratory practices in sperm preparation, an essential part of fertility treatments.

In the wake of the Trump and Biden administrations, during which movements such as Christian nationalism and religious deconstruction brought civil religion into the public eye, scholarship regarding religion as it interacts with politics becomes increasingly relevant. My thesis aims to investigate the role of Christianity in the political communication of our two most recent presidents, arguing that public religiosity is not diminishing in America -- rather, it is in a constant state of evolution to fit an ever-changing nation. This content analysis compares Trump's and Biden's usages of Christian language to one another as well as to former presidencies, addressing the question of how religious rhetoric is shifting in American political discourse. Through creation of a codebook of more than one hundred Christian keywords, which are then applied to presidential communication such as State of the Union addresses (accessed through The American Presidency Project), I determine the frequencies and contexts of religious mentions within Trump's and Biden's communication. Using programs such as Atlas.ti, I measure, analyze, and visualize Trump's and Biden's individual invocations of Christian talk. I hypothesize that Donald Trump's use of religious language increased over the course of his presidency as well as being significantly concentrated in self-referential contexts. Meanwhile, Joseph Biden's religious language is expected to be concentrated in contexts of national crisis, mainly taking the form of Biblical quotations. Overall, findings are expected to demonstrate that Christian language in the American presidency is not broadly declining, but rather used in more niche contexts. This project contributes to the vast field of religious and political scholarship by comprehensively analyzing not just the frequency but also the context of religious language in these presidencies, supporting the notion that public religion is a living entity that functions as a sociological and cultural phenomenon.

What explains variations in how ports are acknowledging and responding to the challenges of climate mitigation and climate adaptation?  This research project examines the response of the top 100 global ports (in terms of tonnage) across the world to the growing challenges posed by climate change. Ports face challenges such as reducing emissions from ships and port operations, and adapting to changes like sea level rise, irregular water flows (crucial for ports on river estuaries), shifts in cargo composition, and changing water dynamics. I will explore whether these ports are acknowledging climate challenges and the extent to which they are establishing clear targets and addressing them in their operational strategies with clear timelines. Moreover, how they are preparing for the eventual decline in global fossil fuel trade. In particular, I will pay special attention to coal-exporting and importing ports, analyzing if they continue to make investments in coal infrastructure and whether they have initiated plans for diversifying their revenue streams to prepare for the eventual phase-out of coal. I have already started collecting data by closely examining the websites of these ports. I will create a database of whether these ports have developed climate statements that identify environmental issues, set clear targets with timelines, and outline strategies to mitigate and adapt to these challenges.  I will combine the scores on these dimensions to create a climate score for each port. In addition to my quantitative analysis based on a dataset created by website research, I will conduct interviews with port officials and union leaders at the Port of Seattle, Tacoma, Everette, and Bellingham.

Individuals with Stage II/III severe periodontal disease were recruited from the Graduate Periodontics Clinic at the University of Washington School of Dentistry using an approved IRB (STUDY00016871). Subgingival plaque samples were collected from four tooth sites: an active inflamed site, two adjacent tooth sites, and a distant healthy tooth site. Sterile paper points were inserted into the gingival sulcus for 30 seconds. DNA was isolated and extracted using the Qiagen AllPrep DNA/RNA Mini kit (Cat. #80204), then purified and concentrated using the Zymo Clean & Concentrator (Cat. #D4014) kit. Whole genome libraries were generated and whole genome sequencing was performed on a NovaSeq X (Illumina, San Diego, CA, USA). Relative abundance of species-level assignments represented by clusters were determined, and best matched genomes were then used to generate species pangenomes for comprehensive multi-genome wide read mapping and gene-level analysis. Taxonomic and gene level functional analysis was also performed. Data analysis is still being performed at this time. Based on previous literature (Basic and Dahlén, 2023), we anticipate that the activity of certain microbial metabolic pathways associated with oral disease and their functions will be heightened in the active disease site and lower on the non-affected site within strain-level disease-associated bacteria, such as those of the Porphyromonas, Tannerella, and Treponema genera. Specifically, we expect amino acid fermentation and lipid metabolism activity to be increased in the active disease site. We expect that results from this study will highlight the presence of heightened activity of bacterial metabolic pathways and functions at the strain level associated with specific gram negative perio-pathic disease-associated species within clinically healthy tooth sites in patients with active periodontal disease despite the absence of clinical observed inflammation.

Extreme operating temperatures in rocket engines severely degrades their lifespan, function and reusability. One mitigating approach to help cool rocket engines and extend their lifespans is called Regenerative Cooling, which has been a method actively used in liquid rocket engines (LREs) since 1923. The cooling system utilizes many narrow coolant channels to draw heat away from the liquid propellant near the rocket nozzle. However, experimental research on these channels is rarely done as they are very small and a single channel is difficult to manufacture for basic research testing thereby causing many researchers to look to non-experimentally tested CFD (Computational Fluid Dynamics) simulations to perform their studies. Our experiment aims to fill the gap between simulation and practical testing by testing scaled up models with V-shaped ribs based on a study done by Zhang et al. These scaled up models would allow for more easily obtainable thermal distribution, stress, and pressure data while also being simpler and cheaper to manufacture. We believe our data could offer an alternative to non-tested CFD simulation data and, as access to experimental data increases, result in the expansion of this area of research.

The Filipino American community with its long history of immigration to the United States, has a successful history of adapting to urban environments while also maintaining cultural identity and a sense of belonging. However, the impact of urban spaces on the identity and sense of belonging on second-generation Filipino Americans (SGFAs) is less understood. This study explores the role of urban spaces in shaping the identity and sense of belonging of SGFAs in Seattle, focusing in particular on how SGFAs navigate the challenge of balancing acceptance within the Filipino community while also fitting in as a minority group in the broader U.S. context. Through semi-structured interviews and a community mapping exercise, the study investigates where participants feel a sense of belonging and how they relate to their Filipino identity in spatial terms across the Seattle area. The findings aim to inform a set of policy recommendations for improving urban spaces with high proportions of Filipinos such as ethnic small businesses, community centers, and hospitals. The recommendations focus on changes to zoning codes, transit accessibility, and social programs to support Filipinos' identity reinforcement and foster a stronger sense of belonging within Seattle's Filipino American community.

Sensory signal processes (specifically how visual systems perform) function under the limits imposed by physics. One such physical limit comes in the detection of light. Light is divided into discrete amounts of energy called photons. Because of this division, light is inherently variable and operates under such variance sets on the retina. So far, the only thing studied in low light is if our system can detect the existence of a flash or dim light, but not deduction of movement and time information. Through measurements of the flicker fusion frequency, rod cells have been understood historically as having extremely poor temporal resolution. This suggested that the ability to detect moving objects– which relies on timing information– would be poor at low light levels. Through extracellular electrophysiological experiments conducted using a 512-channel multi-electrode array, I recorded the electrical activity of neural firing from a population of retinal ganglion cells in the primate retina to show that there is high temporal precision in star-light levels due to compensatory mechanisms in the retinal ganglion cells and adjunct circuits. Since we currently do not yet have any definitive understanding of the cellular-level population dynamics that explain the internal mechanisms and physiology in the retina that allow for this behavioral sensitivity and adaptation, developing this understanding of how a population of cells in the retina work together to detect and encode the motion of moving objects during a given time interval will be pertinent to furthering the field’s understanding of the internal mechanisms of the retina.

Background: Premature infants or those with hypoxic-ischemic encephalopathy are at high-risk for cerebral palsy (CP). Early detection of CP can significantly improve outcomes, however, inconsistent attendance at developmental follow-up after UW NICU discharge puts infants at risk of missed diagnoses. Objective: Implementing a process for early detection of CP in the NICU to increase high-risk infant follow-up (HRIF) rates at 3–4 months from 51% to 80% within 10 months. Design/methods: A single-center Quality Improvement initiative. Baseline data included patient characteristics, comorbidities, and CP risk factors. Process mapping and stakeholder meetings began June 2024, informing changes implemented beginning in August 2024. Process measures included % of eligible infants receiving a General Movements Assessment (GMA) and % of infants with a 3-4-month follow-up scheduled before NICU discharge. Outcome measures include % of infants seen at HRIF by 3-4 months corrected gestational age (cGA) and number diagnosed with High Risk for CP or CP before 12 months. Balancing measures addressed % of clinic No-Shows or appointment cancellations. Results: At baseline, 67 infants, discharged from UW NICU between Dec 2024 and July 2024 and met criteria for HRIF; 66 (99%) were very premature, GA ≤ 32 weeks and/or birthweight ≤1500g, 3(5%) were extremely premature, GA 22 to 24 weeks 6 days. 65(97%) infants had a GMA before discharge. No infants had cramped synchronized movements. 97% received referral for HRIF and none were scheduled for a 3–4-month appointment before NICU discharge. Monthly follow-up rate (number of infants seen at 3-4 months cGA / number of eligible infants discharged per month) was 51%. By January 2025, 57/62 (92%) eligible infants were scheduled for HRIF before NICU discharge. Process changes are ongoing. Conclusion: A standardized pathway for high-risk infants in the NICU can improve local follow-up rates, enabling earlier CP detection and intervention.

Neonatal tracheal intubation (TI) is a high-risk procedure requiring careful coordination to minimize complications and improve outcomes. The Personalized Intubation Neonate Safety (PINS) Bundle was developed to enhance team communication and optimize intubation practices for neonates at risk. The bundle incorporates five key domains: patient risk assessment, treatment threshold for intubation, premedication plan, equipment specification, and provider selection with escalation strategies. At the University of Washington Neonatal Intensive Care Unit, eligibility criteria for the PINS bundle include neonates requiring non-invasive respiratory support with >30% FiO₂, those meeting surfactant administration criteria (≥30% FiO₂ and PEEP 6 on NCPAP), already intubated and mechanically ventilated infants, those with difficult airway diagnoses (e.g., craniofacial anomalies, large tongue, micro/retrognathia), and extremely preterm infants (<1000g or <4 weeks old). We will evaluate the impact of PINS implementation by comparing intubation attempts and adverse events before (January – June 2024) and after (July – December 2024) the bundle’s introduction. Data was adjusted to exclude ineligible patients, ensuring comparable cohorts. Primary outcomes include the number of intubation attempts before success and the incidence of tracheal intubation adverse events (TIAEs). We hypothesize that PINS implementation will be associated with a reduction in intubation attempts and TIAEs, reflecting improved preparation and procedural success by the healthcare team. By standardizing an individualized pre-intubation plan, the PINS bundle aims to enhance neonatal safety and streamline team response during intubation. Findings from this study will inform future clinical protocols and may support the broader adoption of personalized intubation strategies in the NICU. Further assessment of long-term outcomes for infants will strengthen the bundle’s clinical utility.

Leigh syndrome (LS) is the most common pediatric mitochondrial disease, manifesting in the first year of life and leading to early death due to a lack of proven therapies. Like other mitochondrial diseases, LS is caused by gene mutations impacting proteins essential for the mitochondrial respiratory chain, including all complexes. Mutations in NDUFS4, a gene that encodes a subunit critical for structure and stability of complex I, have been linked to LS. Mice with the whole-body NDUFS4 KO exhibit major LS symptoms, particularly epilepsy, along with psychomotor deterioration, progressive neurodegeneration and premature lethality (~P60). Our earlier findings showed that mice with Ndufs4 KO specifically in GABAergic interneurons (Gad2-Ndufs4-KO) exhibit the severe epilepsy and sudden death observed in the global KO mice. These mice represent an excellent model of LS epilepsy, isolated from other clinical manifestations of the disease. LS related epilepsy is often very difficult to treat and indicative of poor disease prognosis. Chronic hypoxia therapy (CHT) has previously shown promise in improving survival and reversing neurodegeneration in LS mice. Yet its impact on seizures remains unknown. In this study, we investigated the efficacy of CHT in ameliorating the epileptic phenotype. Mice with LS epilepsy demonstrated a longer lifespan when exposed to  normobaric 11% O2 than normoxia from postnatal day 35 to day 70. Upon return to normoxic conditions, mice kept in chronic hypoxia die within days. In addition, preliminary thermal seizure tests show an increased thermal seizure threshold in hypoxic mice compared to normoxic ones.  Future studies will evaluate CHT impact on spontaneous seizures using video EEG technique. Our study will aid in the development of a novel therapeutic approach for seizures and related death in Leigh syndrome.

Load carriage is a human universal used to transport children and other resources (e.g., water). Load placement (e.g., back or side) can vary, which influences an individual's gait. Previous studies have assessed the energetic cost of center-of-mass placement  (COMP) and head-load placement (HLP), but the kinematic changes during combined loads are less understood. Lumbar bending and arm swing amplitude were collected from participants (N=19) using OpenCap as they walked for four load conditions: unloaded (UL), back-loaded (BL), back-loaded-with-empty-bucket (BEB), and head-loaded (HL). All loaded conditions were 7.3kg, approximately 10% of the participant's mass. Conditions were differentiated as COMP (BL) or HLP (BEB and HL). Participants were not habitual head load carriers and used one arm to support HLP (one arm swing). Normal arm swing was maintained for COM conditions. Using a multifactorial ANOVA, COM induces lumbar flexion while HLP induces lumbar extension (p=0.011). Additionally, a difference in arm swing amplitude was found (p=0.058). Normal arm swing (COM) had low swing amplitude while one arm swing (HLP) had a high amplitude, with the highest arm swing occurring during BEB. These findings imply that even when loaded mass is the same, different load positions are associated with kinematic changes that will have important energetic impacts and the potential for changes in skeletomuscular changes particularly via lumbar extension. These data are specifically relevant to understanding the demands placed on individuals around the world who carry water and/or children for long distances, particularly in places experiencing extreme drying during climate change.

The ability to recognize that others have mental states, separate from us, and that these states are not always accurate portrayals of reality, is central for theory of mind (TOM). This capacity becomes particularly crucial when children explain causal relationships, as they must integrate their understanding of causality with their awareness of other's knowledge states. Skills like this are essential for effective communication and reflect a key developmental milestone in both cognitive and social reasoning. This study examines how children process causal scenarios while considering and tracking the knowledge states of multiple people. We will examine how children (ages 4-7) perform with conjunctive causal relationships, where two separate effects must combine to produce an outcome (e.g., watering a plant and giving it fertilizer causes it to bloom). Children are asked to explain how the outcome happened, and what knowledge each character has. The children will be given four causal events, varying in content, and follow the same causal structure, where each character is only aware of one cause (A or B). After the scenario, children will answer open-ended questions to assess their recall of what each character knows and test their understanding of how the outcome (C) occurred. We predict that younger children will recognize the causal outcome but struggle to differentiate knowledge states, while older children will demonstrate an improved ability to tailor their explanations based on other's perspectives. This study extends beyond previous studies that primarily focused on children's passive evaluation of explanation as our study will investigate children's active role in generating explanations tailored to different character's knowledge states. Our findings will contribute to the understanding of how the development of TOM shapes children's ability to understand and reason about causal relationships.

In the Earth system, land-atmosphere interactions play a crucial role in the development of weather and climate. Rising buoyant air parcels in convective environments create thunderstorms and clouds and can be influenced by energy fluxes at the land surface. However, the role of soil moisture in convective development is still a topic of ongoing debate.  A recent study by Zhang et al. (2023) investigated how soil moisture drydown periods affect the convective available potential energy (CAPE) and precipitation patterns in different regions using satellite data and statistical modeling. Other studies have predicted that CAPE will increase in humid regions and decrease in arid regions due to anthropogenic warming (Diffenbaugh et al., 2013; Taszarek et al., 2021). In our study, we use time series from global climate model (GCM) simulations to compare interstorm CAPE in different regions across current and future warming scenarios. Our research will result in the enhanced understanding of land-atmosphere coupling and how severe weather will respond to a CO2-driven warming climate.

The Aurignacian, a major cultural phase of the Upper Paleolithic, was characterized by remarkable advancement in the social and cognitive capacities of human beings. This period is marked by technical innovations - such as bone tools and weapons, and artistic developments - in the form of cave paintings and portable art. Evidence from this time period also suggests the emergence of social groups spread across Europe. We investigate interactions between these groups by studying the distribution of objects with engraved geometric signs. We use data from SignBase, a large catalog of archeological data from European Paleolithic sites to quantify group areas and site hierarchies, identifying potential ‘checkpoints’ and ‘central places’. Applying seriation, networks analysis, and measures of sign richness and diversity, we examine how the social dynamics change over the four sub-periods of the Aurignacian. We infer patterns of social group boundaries and interactions, improving our understanding of the structure and complexity of Aurignacian social networks.

Tyrosine-kinase inhibitors (TKIs), a class of chemotherapeutic drugs, are a kind of targeted therapeutics which work by inhibiting the human epithelial growth factor, a common site of mutation in many cancers. However, TKIs may eventually fail due to accumulation of mutations leading to resistance and tumor heterogeneity. Drug cocktails, or combination regimens, provide a potential way to combat this problem. Combining different classes of drugs allow for the attacking of the issue from different angles. However, it is imperative to carefully understand these combinations before putting them to medical use. Results from a novel, non-invasive imaging technique--stimulated Raman microscopy (SRS)--quantified chemotherapeutic drug uptake with different transport inhibitors. Results from SRS show that the calcium channel inhibitor Verapamil increases TKI drug uptake until a certain point for Lapatinib but indefinitely for Afatinib when compared to using the drugs alone, indicating that there is likely an optimal range for each TKI-inhibitor combination. For this project, I aim to show that this stands within a cell culture environment, continuing to use the two common TKIs Afatinib and Lapatinib, and determining the difference in efficacy with used in tandem other inhibitors, namely Verapamil and Chloroquine, a drug that inhibits the progress of the cell cycle. To do so, I will be using a bulk cell viability assay, which allows for the observation of the difference in the value for which 50% of cells are inhibited, and observing differences in this point to determine the optimal treatment and concentrations for a TKI-Inhibitor treatment. Combining a single-cell, non-invasive spectroscopic technique and a cell viability assay, we can better understand the mechanisms behind how typical non-cancer therapeutics can be used in tandem with chemotherapeutic treatments to increase drug uptake, while at the same time acknowledging that a balance is needed for the best synergistic effect.

Overdose deaths in the United States have rapidly increased in the past few years accounting for over 107,000 deaths in 2022 with more than half being attributed to the co-usage of opioids and stimulants. Despite the prevalence of polysubstance use, research has predominantly focused on single substance use, leaving a gap in knowledge regarding its neurological effects. Addiction-associated behaviors such as drug-seeking, drug-taking, and relapse vulnerability has been attributed dysregulation of the striatum. We recently found that polysubstance exposure to methamphetamine and fentanyl leads to behavioral differences in methamphetamine-seeking but not fentanyl-seeking relative to single-substance rats, suggesting that polysubstance use causes distinct changes in striatal circuitry. Our overarching objective is to determine to what extent pursuit of methamphetamine and fentanyl involves shared neural pathways. How do distinct striatal neuron subpopulations responsive to either methamphetamine or fentanyl regulate drug-seeking in animals exposed to both substances? To investigate this, rats are placed into self-administration boxes and undergo two phases of daily intermittent access drug self-administration. In the first phase of self-administration, lasting 10 days, a novel targeted recombination in active population (TRAP) technology and recombinant viral vector is used to target inhibitory designer receptors exclusively activated by designer drugs (DREADDs) to striatal cells that are activated during fentanyl- or methamphetamine-seeking. For the second phase of the experiment, lasting 28 days, rats are then divided into groups for either self-administration of methamphetamine, fentanyl or a polysubstance model. Using chemogenetic manipulations in polysubstance rats, we will test the hypothesis that inhibition of the striatal neuron subpopulations active during fentanyl-seeking will decrease methamphetamine-seeking whereas inhibition of methamphetamine activated neurons will have no impact on fentanyl-seeking. These experiments will provide important insights and lay the groundwork for future studies into how striatal circuits regulate behavior during single and polysubstance use of opioids and stimulants.

Privilege and oppression affect many aspects of Asian Americans’ lives. Most notably, Asian Americans are constantly surrounded by discussions that perpetuate harmful messages that lead to discrimination based on their race and gender. Yet based on past research, Asian Americans are caught in a unique position in terms of defining their racial privilege and viewing their societal advantages and disadvantages compared to other racial minorities (Oh & Eguchi, 2022). Additionally, Asian media like K-pop may play a role in giving Asian Americans a chance to embrace their culture and, possibly for Asian American men, give them a chance to push away the many oppressive stereotypes that surround their identities. For our research, we investigated how privilege is felt throughout the Asian American community, how ethnicity and gender play a role in Asian Americans’ sense of privilege, and how Asian media impacts their definitions of privilege. Using a grounded theory approach, we analyzed qualitative data from Asian American college students at UW Bothell. Semi-structured interviews were conducted with five participants so far, and we anticipate collecting data from at least five more participants by the time of the presentation. From our interviews, we have found that the perception and experiences regarding racial privilege vary among different ethnicities among Asian Americans (e.g., East vs. South vs. Southeast Asian groups). Additionally, we have found that struggles in the dating scene shaped Asian American men’s views of lacking privilege. Finally, many interviewees felt that Asian media, particularly K-pop and anime, play a key role in shaping how they view themselves as privileged and increasingly value their Asian American identity. Our study points to the need for future scholarship on the analysis of Asian media’s role within Asian Americans and the different types of privileges felt between different ethnicities and genders.

Behavioral flexibility is the ability of an animal to adapt to environmental changes, crucial decision making and goal directed behaviors. Impaired behavioral flexibility has been linked to psychiatric disorders such as depression, anxiety, and substance use disorders. Emerging research suggests that psilocybin, a psychedelic drug that acts on serotonin receptors, may improve behavioral flexibility by promoting adaptability in responses to new information. Given the growing literature exploring the impact of psilocybin as a therapeutic treatment for psychiatric disorders, this study aims to explore the effects of psilocybin on the use of behavioral strategies on a complex spatial set-shifting task. Long-Evans rats were trained on an elevated plus maze with two start arms, north and south, and two reward arms, west and east. The task requires rats to alternate between two strategies: alternation and place. The alternation phase required rats to switch between reward arms (go West, then go East), and the place strategy required choosing the same arm (e.g. go West) continuously. Rats must determine the correct strategy and implement it; once 12 out of 15 trials are accurately completed, an uncued switch in the correct strategy will occur, requiring adaptation to a new strategy. Once animals were well-trained on the task, a single dose of psilocybin (1mg/kg) or saline was administered 10 minutes prior to a session and their behavior was analyzed. Metrics such as performance, and flexibility scores were assessed to determine the impact of psilocybin on behavior. Preliminary findings suggest psilocybin improves behavioral flexibility compared to the saline group, though overall performance remains relatively stable. These results provide insight into the potential of psilocybin as a therapeutic tool for conditions characterized by behavioral rigidity; even in wild-type animals, psilocybin improves flexibility, indicating the possibility for mitigating more severe impairments in flexibility seen in psychiatric disorders.

The ability to manipulate and ligate proteins has been a driving force in advancing our understanding of the complex regulation of biological processes in space and time. Protein ligation, in which two or more polypeptides are covalently linked, is a powerful strategy in biomacromolecular engineering, enabling precise control over protein modifications, stability, and functionality. This is particularly useful in understanding protein function and interactions, as well as modulating protein activity, including immobilization of protein-based signals within materials triggered by cytocompatible light. One proven system known for its specificity and ease of use is SpyTag/SpyCatcher, a peptide-protein pair capable of irreversible ligation via isopeptide bond formation. Recent work has demonstrated the ability to control SpyTag/Catcher ligation using cytocompatible light due to its non-invasive nature and spatiotemporal (i.e., 4D control) manipulation of protein signals on a biologically relevant timescale. However, the application of this reported photoligation strategy is hindered by the use of genetic code expansion which limits protein yield, entails additional orthogonal protein machinery, and involves translational incorporation of a non-canonical amino acid. To address these challenges, we aim to develop a photocontrolled protein ligation strategy using native protein activity while maintaining spatial and temporal control. We predict this strategy will enable dose-dependent reconstitution of ligation by varying light exposure duration and intensity in native protein systems while sidestepping challenges associated with genetic code expansion. We intend to use this strategy to further assess our capability to control split protein reconstitution and for future applications in directing complex cell fate, which has significant utility in stem cell biology and regenerative medicine.

Fluorophore quencher pairs are commonly used as reporters for DNA reactions due to their low background signal when untriggered and their ability to detect low DNA concentrations. However, these modifications are expensive and require a fluorescent plate reader to detect the signal, limiting their accessibility for point-of-care or low-resource settings. We are developing an alternative reporter using G-quadruplexes, which are guanine-rich DNA sequences with enzymatic activity in vitro. These structures can be utilized in detection assays due to their well-characterized peroxidase activity. Current approaches utilizing G-quadruplex structures have limited sensitivity due to high levels of background activity. Our approach combats this problem using altered G-quadruplex sequences inactivated by sequence-mismatched complexes, later activated by downstream reactions. By making these modifications, we detect DNA concentrations as low as 2 nanomolar. We hope this inexpensive approach can be utilized as an accessible alternative to traditional detection assays due to its colorimetric properties, leading to detection by the human eye and effective point-of-care detection.

Concrete is the most widely used structural material globally due to its strength and durability, with demand steadily increasing. Ordinary Portland cement (OPC) is the primary binder in cementitious materials and participates in hydration reactions that contribute to the compressive strength. Unfortunately, the production of cement is responsible for ~8% of global CO₂ emissions, driving efforts to reduce its use in concrete and other cementitious materials. Consequently, supplementary cementitious materials (SCMs), such as zeolite, are being explored to reduce, but not entirely eliminate, cement in industrial settings. Zeolite is a naturally-occurring microporous pozzolanic mineral that has the propensity for replacing cement through strengthening and densifying mechanisms. It also has capability for carbon capture, which can contribute to further reduction of atmospheric CO₂. However, the effect of zeolite on carbonation reactions in concrete is not well characterized. Pozzolanic and carbonation reactions can synergize to cause concrete and mortar (concrete sans large aggregates) to further densify and strengthen over time, but they compete for the use of calcium hydroxide, a hydration byproduct. This research investigates the effect of replacing up to 30% cement in concrete with natural zeolite on the carbonation of mortar composites over a curing period of 28 days. Carbonation is conducted through elevated pressure and CO₂ concentration, rather than ambient pressure. The effect of zeolite is measured using microCT for porosity analysis, compressive testing for strength, and pH indication of cross-sections for carbonation depth. Results indicate that the porosity decreases while carbonation depth increases with zeolite content. The zeolite addition contributes to the reduction of compressive strength, ranging from 71.6 to 37.7 MPa. Overall, carbonation enhances strength, correlating with carbonation depth, but this trend is more evident under ambient carbonation (2.78% strength increase after four weeks) than under pressurized carbonation, where no distinct trend is observed.

​​​​Bacteria evolve primarily through horizontal gene transfer, the movement of genetic material between organisms that are not directly related. This allows the rapid acquisition of traits like virulence and antibiotic resistance, an increasing public health concern. The mechanisms by which bacteria integrate and control these new traits is incompletely understood. Acquired virulence genes have allowed uropathogenic Escherichia coli (UPEC) to become the predominant cause of urinary tract infections (UTI) throughout the world. A critical step in the UPEC infectious process is the transition from a free-swimming, unicellular, “planktonic” form in the urinary tract to an stationary multi-cellular community, or “biofilm,” when it invades the bladder epithelium. The Fang lab has shown that the transcription factor, MprA, promotes the expression of a horizontally-acquired gene cluster encoding the enzymes required for the biosynthesis of polysaccharide capsule, an important UPEC virulence factor. We hypothesize that this capsule is associated with planktonic growth, and that by regulating capsule, MprA controls the switch between planktonic and biofilm-associated growth. We will test this by growing biofilms of wildtype, mprA, and capsule-deficient mutant strains in the laboratory. Additionally, we will assess their impact on virulence by infecting larvae of Galleria mellonella, the wax moth, with each strain. Understanding how factors like MprA control horizontally-acquired genes can inform the development of future antibacterial therapies.

During neonatal transport, specialized pediatric transport teams closely monitor the status of critically ill newborns. Hyperspectral imaging, a method of manipulating light for medical imaging, can be used for remote monitoring using video of the patient’s physical appearance and to measure vital signs. Appropriate light intensity is critical for clear visibility of the newborn and hyperspectral imaging accuracy, but this must be balanced with safety for sensitive eyes. My previous studies have determined the minimum range of light needed to accurately view the neonate in a transport incubator. My current research is focused on developing a novel method for vital sign analysis by using hyperspectral imaging. A smartphone camera will be used to take RGB photos of a calibration chart and a short video of the wrist of study participants. After taking the RGB photographs, custom MATLAB code will be used to extract physiological data such as hemoglobin and bilirubin content from the skin. Data analysis will compare the vital sign data collected using hyperspectral imaging and using a pulse oximeter to understand the feasibility of hyperspectral imaging for vital sign extraction. The expected result of this study is that the heart rate and blood oxygen levels measured using light and a pulse oximeter will be highly correlated. In conclusion, this research will demonstrate the potential application of hyperspectral imaging to pediatric transport.

The short-term goal of this research project is to integrate and apply machine-learning tools to magnetic resonance imaging (MRI) data housed within a queryable, large-scale MRI database. Long-term goals seek to develop tools to better identify patterns of pathology within multimodal MRI data. Imaging approaches such as morphometric and connectivity analyses provide a means of uncovering the brain’s seizure onset zone in patients with epilepsy, known as the epileptogenic zone. MRI is the standard reference protocol for non-invasive evaluation of epilepsy. This is due to the fact that a majority of epilepsy patients have visible lesions on brain MRI scans. However, a number of epilepsy patients are MRI-negative, defined as having no visible lesion. To improve lesion detection in MRI-negative patients, machine learning has been applied to multimodal MRI as a means to detect subtle patterns of changes. However, these studies are limited by small number of patients or included only a few epilepsy pathologies. Working with a team from the Department of Radiology, I established a structured database integrated with raw MRI data, computational post-processing results, and clinical metadata. In the present study I will examine how available machine learning techniques can be replicated and utilized on the data within this integrated database. As the database accumulates MRI data from a larger number of epilepsy patients with and without visible MRI lesions, we hypothesize that this integration will uncover subtle abnormalities not detected through qualitative MRI. This form of individualized brain mapping we predict will provide new insights into seizure networks and ultimately enhance efficacy of non-pharmacological based treatment approaches such as enhancing accuracy of surgical resection of the epileptogenic zone.

The Campanian stage of the Late Cretaceous (84–72 million years [Ma]) was a dynamic interval for North American ecosystems and included the zenith of dinosaur diversity and the regression and transgression of the Western Interior Seaway (WIS). Most studies that investigate vertebrate biodiversity during this interval focus on dinosaurs, whereas few focus on changes in herpetofauna (lizards, frogs, and salamanders). Herpetofauna are important indicator species of ecosystem dynamics, because they are fragile to ecosystem change. Vertebrate microfossil sites are ideal for studying herpetofauna diversity dynamics through time because they can produce large sample sizes, sample aquatic environments, and are plentiful through stratigraphic intervals. The Judith River Formation (JRF) of north central Montana is rich in vertebrate microfossil sites, preserving ~4 Ma of the Campanian (~79–74 Ma). Here we aim to document patterns of herpetofauna diversity change in the JRF by quantifying herptile taxonomic richness and relative abundances using specimens from three temporally separated microfossil sites: Makela-French 1 (~77.5 Ma), Milkshake (76.5 Ma), and Clamfetti (~75.2 Ma). We collected sediment samples from these sites over four years and processed them via underwater screen-washing techniques. Presently we have studied 470 herptile microfossils (600 planned). Our preliminary results show changes in the taxonomic diversity across the sampled sites. Taxonomic richness of herptiles varies through the formation, first increasing and then decreasing. Salamanders have the highest relative abundance, lizards decrease in relative abundance, and frogs fluctuate. We hypothesize that taxonomic patterns are influenced by the impact of WIS cycles on water supply in ecosystems: amphibians thrive in wetter environments, whereas lizards are more terrestrial. These preliminary results reflect a connection between diversity patterns and extrinsic drivers not observable through the analysis of dinosaur fossils. Our continued analysis will provide more fine-scale resolution of herptile diversity during the Campanian.

Cascadia Subduction Zone, a major fault off the Pacific Northwest coast, has a history of producing powerful earthquakes. These events highlight the need to understand the region's earthquake probability. This study aims to conduct a sensitivity test on the earthquake probability in the Pacific Northwest, evaluating partial and full rupture scenarios. The study analyzes 32 different earthquake chronologies derived from the earthquake catalog and perturbs them to assess how the probabilities vary with changes in data. As a result, we found that the Southern Oregon and Northern California regions face the highest risk, with a 34-49% chance of a partial rupture and a 28-32% chance of a full rupture within 100 years. Within the next 50 years, probabilities are 17-20% and 13-15%, respectively. The relatively small changes in probabilities suggests that the estimates are robust, indicating that changes in data do not significantly skew the overall probability assessment. These findings emphasize the need to implement hazard mitigation strategies to protect vulnerable Southern Oregon and Northern California communities from future earthquake events.

Underground storage organs (USO) allow plants to retreat underground during periods of resource scarcity and/or abiotic stress. These adaptations help plants survive seasonal climates and have evolved repeatedly across the vascular plant tree of life. USOs develop as modifications of various plant tissues, including root, leaf, and stem tissues. The non-model plant Bomarea multiflora (Alstroemeriaceae) offers a unique opportunity to study the development and evolution of USOs as it has two types of USOs (rhizomes and root tubers), allowing for direct comparison. B. multiflora’s dual USOs and its phylogenetic position within the monocots makes it a good candidate for future development as a model species for USO development. Here we analyze a transcriptomic dataset of four distinct tissue types in B. multiflora: aerial shoot, rhizome, fibrous root, and root tuber tissues. We use ZigZag, a recently developed hierarchical Baysian model that determines the probability of active expression for each gene in each tissue type. We found 56 genes differentially expressed between individual tissues and 29 between root vs. shoot tissue. We review these genes and describe avenues for future investigation of USO developmental pathways within monocots.

Human gastrulation is the period of embryonic development during which a single-layered blastula develops the three germ layers, which give rise to all of the specialized cell types in our bodies. Differentiation of these layers is controlled by epigenetic factors, which chemically modify the DNA, therefore activating or repressing a gene’s activity. Very little is known about the role of epigenetic factors in human gastrulation, as it’s difficult to obtain samples at the proper time for study; samples from IVF are too early to study, and fetal samples are too developed. Despite these complications, it’s important to study gastrulation because it’s during this time that developmental disorders and loss of pregnancy can occur. My goal is to delete seven epigenetic factors and determine their role in human embryogenesis using a human pluripotent stem cell derived embryoid model (retinoic acid treated gastruloids). Using CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas9 technology, I plan to perform individual gene knockouts on our genes of interest in human embryonic stem cells. Next, we will generate mutant gastruloids from these knockout cell lines, and phenotype using single-cell RNA sequencing and fluorescent microscopy. From there, we can see how knocking out this gene affects gastruloid cell type composition and morphology. Using this strategy, our lab has successfully knocked out 60 genes. Given the strong phenotypes observed in null mutant mouse embryos for these genes, we expect to observe strong phenotypes from these seven epigenetic factors. With these findings, we can further understand the role of these epigenetic factors in development and disease. These findings can advance the progress of therapies and better understand our knowledge of human embryonic development as a whole.

This project presents an analysis of climate litigation cases on a global level. I have created a unique database of over 969 cases in 74 countries with detailed accounts of the plaintiffs and defendants involved. The primary objective of this study is to assess the spread of environmental litigation cases and the when, where, who, and on what grounds these cases are being brought forth. My preliminary analysis shows that most climate litigation cases have been filed post-2020, with a majority, or 15.04% of cases being initially filed in 2021. Such data emphasizes the substantial growth of climate litigation on a global scale in recent years. Regarding jurisdiction, a majority, or 15.96%, of cases have been filed in Australia. The UK follows closely with 15.84% of climate litigation cases being filed within the country. Additionally, the majority, or 25.28%, of plaintiffs are NGOs with individuals making up the ensuing majority group with around 22.08% involved in bringing forward cases. On the other hand, 49.02%, of defendants are federal government agencies. Regarding the grounds under which these cases are filed, 62.95% of cases have been filed invoking statutory provisions in the relevant national legislation to hold varying actors responsible for climate inaction. I also find that around 36.22% of cases have utilized international law established by custom or treaties recognized by nations. Importantly, my data suggests a new, and growing, trend emerging in climate litigation as plaintiffs invoke human rights laws to seek stronger climate legislation with around 25.39% of cases having relied on this legal framework. Going forward, such a comprehensive climate litigation database will serve as a valuable resource for scholars, practitioners, and climate advocates seeking to navigate and understand the complex landscape of climate litigation. 

I investigate the effects of scopolamine-induced cholinergic disruption on the working memory performance of rhesus macaques. Working memory plays a critical role in cognitive function and relies heavily on cholinergic signaling in the brain. To explore this relationship, I designed and implemented a delayed match-to-sample (DMTS) task to optimize the accuracy and reliability of the data collected. The DMTS trials involve three phases: stimulus, delay, and choice. Monkeys are presented with a sample stimulus they must memorize. After a variable delay period of 5 to 30 seconds, they select the target image from a set of options. Performance is tracked by calculating the percentage of correct, incorrect, and omitted responses. Daily DMTS sessions provide longitudinal data on memory performance, allowing me to analyze patterns before and after scopolamine administration. I am also learning to analyze spike-spike coherence to investigate changes in neuronal synchronization associated with memory performance. Upon inducing amnesia-like conditions under scopolamine, an M1 muscarinic antagonist, the delayed match-to-sample task evaluates memory performance under baseline and drug-induced conditions. The primary objective of my project is to understand how scopolamine-induced disruptions affect working memory performance and to investigate the underlying cortical mechanisms involved. Based on existing literature and preliminary observations, I anticipate observing a measurable decline in memory performance following scopolamine administration, with older macaques likely exhibiting more pronounced deficits compared to younger macques. This project aims to contribute to the understanding of how cholinergic mechanisms influence working memory performance and to provide insights into the cognitive impairments associated with neurodegenerative diseases.

My research is focused on the question of "Why do first-generation Indian Sikh immigrants return back to their origin country?" I am researching about why some migrants return and others do not, specifically the factors that influence the decision-making. My initial research showcased economic motivations being parallel to initial migration as well as return to one's origin country, along with the the influences of the labor market. I hypothesize that return migration in India is driven by the same realities that drive emigration, namely opportunities to develop economically. I conducted qualitative interviews with Indian Sikh immigrants, both those who had returned back to India along with those currently living in a destination country. The subjects were identified through established social personal networks and participants were screened through questions about their migration journey and if they identified as Sikhs. My project is still in progress, however I have completed more than half of my intended goal for qualitative interviews. My preliminary findings suggest that the primary reasons for migration out of an origin country and returning back relates to economic motivations. However social factors, such as conflicts within the origin country and discrimination in the destination country also impact the decision-making processes and intentions of return. The findings of this research are impactful within migration studies, especially as return migration is a relatively new field that is constantly developing. The case study of Sikh Indian Immigrants has not been researched extensively in the past, showcasing why identifying the various factors influencing motivations and intentions is crucial when understanding the patterns of migration. 

Older adults recovering after hospitalization in an intensive care unit (ICU) are at high risk of functional decline. Older adult ICU survivors often report symptoms of cognitive impairment, sleep disturbances, and pain, which can complicate functional recovery. In addition, certain social determinants of health (SDoH) are associated with worse cognitive and physical outcomes that lead to functional decline in older adult ICU survivors. Little is known about the relationships between patient-reported symptoms (e.g., cognitive function, sleep quality, and pain intensity), selected SDoH, and functional recovery for older ICU survivors during hospitalization. This project has two overall aims: 1) explore associations between patient-reported symptoms of cognitive function, sleep quality, pain, and SDoH; and 2) explore associations between these variables and successful participation in physical therapy and/or occupational therapy (PT/OT). We collected and utilized data from two ongoing randomized controlled trials. Older ICU survivors hospitalized at University of Washington Medical Center or Harborview Medical Center were recruited and enrolled. Participants answered the PROMIS Cognitive Function Abilities, Sleep Disturbances, Sleep Related Impairment, and Pain Intensity questionnaires to evaluate self-reported cognitive function, sleep quality, and pain intensity. We did chart review of electronic health records to gather relevant information regarding SDoH, and length of PT/OT participation in minutes. We examined the differences using t-tests, Chi-squared tests, and ANOVA for binary, categorical, and/or continuous variables, respectively. We used descriptive analysis to explore trends between self-reported pain, sleep quality, and selected SDoH on self-reported cognitive function and participation in PT/OT. Results from this project will inform the design of subsequent clinical trials, which could aid in the development of personalized interventions aimed at reducing functional decline in older ICU survivors. 

The Miocene Climatic Optimum (MCO) (17-14 Ma) represents the most recent significant global warming event and provides valuable insights into the future of our planet with higher CO2 levels and warmer temperatures. The Mascall Formation in central Oregon contains a fossil plant assemblage that reflects the vegetation present during the height of the MCO. Despite over 50 years of research in this formation, there is still much to learn about the ancient plant community. For instance, a fossil specimen, consisting of several leaves, that was collected recently exhibits similar trait to bamboo, which represents a new fossil finding in this formation. This project seeks to confidently assign this specimen to the bamboo subfamily Bambusoideae. By analyzing morphological and vein architectural features of the leaves using various microscopic techniques and digital photography. In addition to studying the specimen itself we explore the fossil plant silica bodies (phytoliths) also present in the surrounding substrate to provide independent evidence that bamboo was present in the region. The phytoliths can then be compared to those of current Native American bamboo to find evidence for relatedness or if it was part of some other lineage of bamboo, whether extinct or still present in South America or Eastern Asia. If the specimen turns out to be bamboo, it would have implications for the climate and ecology of eastern Oregon during the MCO as bamboo was not assumed to have previously been present.  

Cancer immunotherapy, specifically Adoptive Cell Therapy (ACT), has revolutionized treatment approaches using genetically modified T cells to recognize and eliminate cancer cells. However, tumors combat this by creating an immunosuppressive tumor microenvironment (TME) blocking effective antitumor immune responses.  Dendritic cells (DCs) are innate immune cells that act as messengers between the innate and adaptive systems.   In the Oda lab we have designed Dual Costimulatory Receptor (DCRs) that combine a FLT3L or CD40L ectodomain with different costimulatory endodomains (e.g. CD40, 4-1BB, OX40), to provide both T cell-extrinsic and -intrinsic costimulatory signals.  These DCRs are expressed on the surface of antigen specific T cells, and the combination of these signals allows for enhanced tumor antigen presentation and dendritic cell activation, leading to an increase of the immune response to target and destroy cancer tumors. I will investigate how incorporating DCRs on T cells will enhance ACT effectiveness. I hypothesize these DCR signals on T cells will enhance dendritic cell function in the TME, allowing for increased T cell activation and antitumor immune responses. To test this, I will conduct in vitro coculture experiments to determine how DCR-T cells, dendritic cells, and pancreatic cancer cells interact together. I will study the interactions of these immune cells using live cell imaging technology such as the Incucyte. Additionally, I will analyze the phenotypes of our distinct cell populations via flow cytometry.  This research aims to enhance the development of immunotherapy for Pancreatic and all solid cancers by improving the recognition of cancer cells from the immune system. These results could help pave the way for improving solid tumor cancer treatment.

Mycoplasma genitalium (MG) is a common sexually transmitted bacterium that causes serious health problems such as pelvic inflammatory disease, urethritis, and pregnancy complications. The efficacy of antibiotics has significantly decreased due to antibiotic resistance; about 50% of US strains are resistant to azithromycin, a common treatment path, and resistance reaches 100% in high-risk populations. Preliminary research done in our lab has shown that MG is susceptible to tinidazole (another antibiotic) in vitro. We hypothesize that tinidazole is effective against MG because it creates superoxide radicals that MG cannot detoxify. To test this, the sodA and katA genes, encoding enzymes that detoxify reactive oxygen species, were introduced into the MG genome. The insertion site was determined by whole genome sequencing, and we selected two mutants with insertions unlikely to affect other genes. These two strains were compared to the parent strain in time-kill experiments to measure susceptibility to tinidazole. Cultures of these strains were incubated for 10 days with two-fold dilutions of tinidazole, plating aliquots onto agar plates each day to quantify surviving MG. The individual colonies present on the plates are counted and graphed, allowing us to compare the efficacy of tinidazole on the separate strains. To confirm the enzymes are being expressed, we used a hydrogen peroxide assay to measure the levels of hydrogen peroxide, which is formed from the radicals released from the cells. In conclusion, we hypothesize that the radicals produced by tinidazole kill MG by inducing DNA damage. We, therefore, measured the susceptibility of 10 DNA repair mutants to tinidazole and found that deletion of MG_360 enhances susceptibility. Results from these experiments can be used to understand the mechanism by which tinidazole and other nitroimidazoles kill MG. This data is critical in the battle against antibiotic resistance and can improve treatment options globally. 

Stigma against individuals with substance use disorders has been shown to negatively affect their health outcomes. Interpersonal stigma has been shown to further perpetuate intrapersonal stigma within this population. The prevalence of this phenomenon has implications in nearly every society, which manifests as structural, societal and interpersonal impacts. This literature review examines how stigma directed at drug users directly affects their health and well-being. The implications are great in that by affecting the health outcomes of drug users, the well-being of an entire population suffers. Data was procured and reviewed using four databases: JSTOR, PubMed, Science Direct and Elsevier. Parameters for the inclusion of data stated that the article must address stigma and how it affects the health and overall well-being of drug users. Additional articles were used to provide background information on the topic. Data suggests that stigma is not only a contributing factor to perpetuated use, but also exacerbates barriers to treatment, fosters a mistrust of healthcare professionals and discourages engagement with evidence-based interventions. Additionally, these factors lead to social isolation and a reduced sense of self-worth. This systematic review highlights some of the gaps in current knowledge pertaining to how stigma negatively affects those who use drugs. By addressing this issue, especially on the systemic level, within areas such as policy reform and health care professional education and training programs, outcomes for those who use drugs can be improved in such a way as to benefit all of society.

The erythrocyte sedimentation rate (ESR) is used as an indicator for inflammation caused by infectious disease, rheumatic disease, and other conditions. When certain proteins become elevated, erythrocytes aggregate at low shear rates, which can be quantified by the ESR. ESR has historically been determined by sedimentation in a vertical tube, but modern technologies aim to make the measurement in different ways. Recently, the industry has turned towards point-of-care testing for patients who need frequent monitoring or are far from hospitals with the necessary equipment. This project seeks to develop a microfluidic ESR measurement device that is suitable for point-of-care usage for long-term inflammation monitoring. The final device will be competitive because of its low cost, speed, and ease of use. Further, this research will improve the collective knowledge and techniques of ESR measurement and blood microfluidics to supplement future investigations in this area.

Parents play a significant role in shaping adolescent alcohol use. The specific role of adolescent decisional freedom—the extent to which adolescents independently make decisions— has yet to be examined. I conducted a brief literature review on adolescent decisional freedom, and this study explores its links with alcohol use within distinct developmental periods. Data from baseline to year 6 follow-up of the National Consortium on Alcohol and NeuroDevelopment in Adolescence study (N=831) were tested for cross-sectional associations between decisional freedom and alcohol use phenomenology (i.e., drinking days per average month, past month alcohol consumption, quantity of drinks during average drinking period, and binge drinking days in the past year and month). Separate models were tested for early adolescence (ages 12-14), middle adolescence (ages 15-17), late adolescence (ages 19-20), and emerging adulthood (ages 21-24). Adolescent decisional freedom was measured using a self-report 5-point Likert scale across distinct domains (e.g., spending money, drinking alcohol, time spent with friends). Gender and age of first drink were covariates, and each model examined associations between decisional freedom and gender. Regression results consistently linked lower decisional freedom to lower alcohol use across all phenomenology in early adolescence (bs ≤ −.41, ps < .01). For middle adolescence, the protective effect of limiting decisional freedom only emerged for drinking days per average month (b = −.29, p < .01) and quantity of drinks during average drinking period (b = −.46, p = .001). In early and late adolescence, the association between lower decisional freedom and fewer past month binge drinking days was stronger for boys (bs ≥ 0.43, ps < .05). These findings suggest that increasing control over early adolescents’ decisional freedom, particularly for boys, and flexibly granting greater decisional freedom over time may protect adolescents from harmful decisions associated with alcohol use.

Understanding the impact of the solvent environment on the electronic properties of modified DNA is essential for their use in nanoelectronics and in medicine. We modeled the impact of counterions in metal modified DNA (mmDNA) using ab-initio density functional theory to model wet and dry conditions. The orbital wavefunctions and charge transport properties were compared for a variety of test conditions, looking at effects for a single basepair as well as a longer DNA chain, using the Thymine-Mercury-Thymine mmDNA basepair as a case study. Preliminary results from single base pair calculations indicate that chlorine counterions in wet DNA do not significantly affect the highest occupied molecular orbital (HOMO) and lowest unoccupied molecular orbital (LUMO) energies, while in dry cases, orbital wavefunctions are more localized and at lower energies, albeit with a similar bandgap. In most calculations, the LUMO localizes on the central metal atom. These findings suggest that longer DNA molecules could potentially form a channel for electron transport along the metal atoms, effectively functioning as a nanowire with a conductance dependent on solvation and counterion presence. With a more accurate model of DNA as a nanomaterial for bioelectronics, it will be possible to develop smaller, more efficient devices operable in biomolecule-friendly environments.

Social norms are informal, shared rules that dictate people’s behavior, influenced by social expectations and potential consequences. Teachers' social norms can influence their willingness to adopt evidence-based practices (EBP) in schools. Although previous research has shown that social norms affect teachers’ behavior with decision-making and prioritization of tasks, less is known about how these norms impact successful implementation of EBP. Understanding how social norms affect teachers’ ability to adopt EBP in the classroom is important in understanding the barriers and facilitators in the school environment, which can in turn guide more effective implementation strategies. These strategies are crucial, as these planned approaches help to promote the adoption and integration of EBP into school settings through training, support, and other resources. This study investigates the relationship between teachers’ perceptions of social norms and their fidelity in implementing EBP. Data were drawn from a larger randomized control trial. Perceptions of social norms were collected via self-report surveys from 324 teachers and observed fidelity was collected via trained school personnel from 39 observers across 25 schools. In future analyses, we will examine the relationship between teachers’ perceived social norms and observed fidelity through Pearson’s correlation coefficient using data from timepoint 7, collected two months post-training. Successful EBP implementation can benefit both the teacher and the overall school environment, which in turn may contribute to positive student outcomes. Findings will contribute to the understanding of how social norms influence EBP implementation in school settings.

Driven by the growing commitment to environmental stewardship and climate action, the aviation sector is adopting more creative technologies to move towards cleaner and more eco-friendly propulsion systems. The substantial CO2 emissions from current jet engines make creative solutions for environmentally friendly air travel a necessity, such as electrifying said engines. However, there are several obstacles that prevent current electric motors from being used in aircraft. Overheating is a serious problem that jeopardizes dependability, efficiency, and safety. Because of these drawbacks, existing electric motor technologies are not a viable substitute for many aircraft propulsion systems. To overcome these obstacles, this study optimizes electric motor designs using cutting-edge temperature control and energy-saving techniques. The suggested method makes use of thermoelectric modules (TEM) and high-temperature superconductors (HTS) to efficiently control thermal energy, lessen overheating, and improve motor performance. By integrating these innovative materials, this work aims to create reliable, efficient, and scalable electric propulsion systems that meet the rigorous demands of the aviation industry.

The first-in-class capsid (CA) binding antiretroviral, Lenacapavir (Len), inhibits viral spread at multiple steps in the viral life cycle. Structural studies show that Len interacts with an FG-binding pocket between the N-terminal and C-terminal domains of adjacent CA monomers resulting in destabilization of the CA core lattice. Three key binding functional groups within Len that interact with CA were identified. Subsequently, six Len analogues (Lenalogs) were designed and synthesized. These Lenalogs vary by the removal or replacement of one of the identified functional groups. My work investigates the impact of Len and Lenalog binding on CA assembly rate, as well as, the structure of the assembled protein. Using an IPTG  E. coli expression system and ion exchange chromatography, I have expressed and purified CA protein. I induced in vitro assembly of the purified CA protein by the addition of inositol hexakisphosphate (IP6) in both the presence and absence of Len or the Lenalogs. Relative to Len, LL-10.4 and LL-15 promoted assembly, LL-14 was similar, while LL-11, LL-19 and LL-20 promoted assembly to a lesser extent. Samples with LL-10.4 and LL-15 were chosen for cryo-EM analysis as these promoted assembly to a greatest extent. CA was assembled on lipid vesicles (templated CA-like particles or CLPs) by the Dick lab (Emory University, Atlanta, GA), and these were subjected to cryo-EM data collection and analysis. Both LL-10.4 and LL-15 bound to the FG-binding pocket like Len. Negative stain transmission electron microscopy and light scattering will be used to further assess the effect of Len and Lenalogs on assembly kinetics. My work will be used to inform the design of next generation CA-targeting antiretrovirals.

Plastic is a prevalent and useful material, experiencing a 230-fold increase in production from 1950 to 2019. However, problems revolving around plastic production and disposal are becoming increasingly clear. In 2015 virgin plastic production was responsible for 4.5% of global greenhouse emissions, with the United states collecting only 9% of plastic waste for recycling. For polystyrene, a common plastic in food storage containers, less than 1% was recycled. The decomposition of polystyrene can also release dangerous chemicals, such as additives Bisphenol A (BPA) and polystyrene oligomers, directly linked to human health hazards such as diabetes, breast cancer, reproductive harm, thyroid regulation issues, heart diseases, and liver problems. To prevent both the need for new plastic and decrease plastic decomposition products from harming both humans and the environment, it is critical to develop a circular plastic economy, where plastic waste is broken down back into the chemical building blocks, or monomers, which can be re-manufactured into new high-quality plastics. The use of basic heterogeneous catalysts, such as zinc oxide (ZnO) can improve the yields of polystyrene deconstruction back into monomers compared to other chemical recycling techniques such as pyrolysis and acid-catalysis. To further improve yields, it is critical to understand the impact of relative basicity of ZnO on styrene yield. To examine base strength effects, the relative basicity of ZnO will be systematically varied through alterations in pretreatment methods. Reactions were tested in a mini-batch reactor at 623K over three hours using a 10:1 substrate to catalyst ratio, with product distribution examined using Gas chromatography-mass spectrometry (GC-MS) and a flame ionization detector (FID). 

The activation of inflammasomes is a crucial component of the early immune response to pathogens and initiates a form of inflammatory programmed cell death called pyroptosis. During infection, a cytosolic inflammasome-forming protein sensor detects a pathogen to assemble the inflammasome complex, which subsequently activates the protease Caspase-1 (CASP1). CASP1 processes Gasdermin D (GSDMD), inducing pyroptosis through pore formation in the plasma membrane, while also facilitating the release of proinflammatory cytokines, such as IL-1β and IL-18. Adenovirus (AdV) is a common pathogen that causes inflammatory symptoms by infecting multiple mucosal epithelial tissues in the respiratory tract and intestinal tract such as the nose, mouth, and eyes. We wanted to test whether AdV infection could activate one of the main inflammasome sensors in human conjunctival epithelial cells (hCjE cells), which is NLRP1. However, we found that upon AdV infection, NLRP1-mediated cytokine release is absent, suggesting that CASP1 signaling is suppressed. Interestingly, despite the loss of IL-1β and IL-18, pyroptosis remains unaffected. Recent studies indicate that in the absence of CASP1, inflammasomes can activate Caspase-8 (CASP8), leading to the cleavage of Caspase-3 (CASP3) and Gasdermin E (GSDME), resulting in an alternative, incomplete form of pyroptosis. Thus, I hypothesize that during AdV infection, host cells are still able to induce pyroptosis by activating the alternative CASP8-GSDME pathway. To test this hypothesis, we generated and validated genetic knockouts of CASP8, GSDMD, GSDME, and CASP3 in hCjE cells to assess their roles in pyroptosis during AdV infection. These findings will provide new insights into viral immune evasion strategies and inflammasome regulation in epithelial cells.

Copepods are an essential link between micro- and macroscopic trophic levels in Salish Sea food webs. The distribution of copepods across spatial and temporal scales is well-known and is partially attributed to physical oceanic processes. However, few studies have characterized copepod densities at different tidal heights in the San Juan Channel. This study investigates copepod densities at depths of 0-25m and 0-50m during high and low slack tides when tidal currents are at their lowest velocity. Higher overall copepod densities were expected during slack high tide due to the influx of oceanic water from the Strait of Juan de Fuca. Samples were collected over four days in the northern San Juan Channel during September 2024. Twelve vertical plankton tows were performed with six replicates for high and low tides respectively. Samples were diluted and randomized using a plankton splitter, creating 5ml aliquots to calculate copepod density. Copepod densities from 0-50m depth during slack high tide were significantly higher (p-value < 0.001) than all other samples. No difference in copepod density was found between 0-25m and 0-50m depths during slack low tide. Increased tidal height resulted in higher copepod densities for low and high slack tides. Our findings suggest the physical processes within the San Juan Channel, such as the influx of colder, saltier waters during slack high tides increase copepod densities in the top 50m of the water column. These shifts in copepod densities may impact the feeding behavior of higher trophic levels in the San Juan Channel food webs.

Alzheimer’s disease (AD) is a neurodegenerative disease that impacts millions of people and costs billions of dollars annually, with both estimates increasing as our aging population grows. Women are diagnosed with AD at a 2:1 higher rate than men, although the biological drivers of this difference remain elusive. Previous studies have demonstrated that changes to the function of microglia – the brain’s immune cells – observed during AD may be driving disease progression. Furthermore, microglia morphology is related to its function. Thus, we seek to characterize differences in microglia morphology between men and women with and without AD. We hypothesize that microglia from women have, on average, a more disease-associated morphology than those of men, and that differences are exacerbated in individuals with AD. We obtained tissue from the dorsolateral prefrontal cortex of 48 individuals who donated their brains to AD research at UW. We conducted immunohistochemistry (IHC) to stain for microglia markers (IBA1) and two markers of AD pathology (AT8 to stain for phosphorylated Tau and a pan-amyloid β stain). I imaged the samples on a Leica SP8 confocal microscope at multiple depths, which allowed us to compose a 3D rendering of the tissue through an image analysis software called IMARIS. Using IMARIS, I quantitatively measured key aspects of each microglia, such as volume and branching details. Using the data from 12-20 microglia per person, we used multiple regression to test for differences between men and women in both healthy and AD cohorts. We anticipate there are differences in the various measurements of microglial morphology between men and women with AD, which may partially explain the discrepancy in AD rates between sexes. This research is important to better understand the role of sex in AD pathology and help contextualize molecular differences observed in the larger project to which it belongs.

The fidelity of evidence-based practices (EBPs) for social-emotional learning in education is influenced by the attitudes of teachers. Teachers who believe an EBP is valuable are more likely to follow its guidelines and implement it as intended. However, the impact of years of teaching experience on these attitudes remains unclear. While experienced teachers may resist EBPs in favor of familiar methods, others may be more flexible and willing to use their experience to implement EBPs effectively. Experience may also foster positive attitudes towards EBPs as teachers feel more confident in their ability to implement them. To address the gap in understanding how years of experience influence teacher attitudes and EBP fidelity, this study will explore the role of experience as a moderator. Data were collected from a larger randomized control trial with a sample of 276 K-8 teachers from 46 elementary schools. This study focuses on three timepoints: 4, 7, and 9 months after training on Positive Greetings at the Door (PGD), an EBP designed for all students. At each timepoint, teachers completed online surveys, and their attitudes were measured using the Evidence-Based Practice Attitudes Scale (EBPAS). Fidelity of PGD delivery was assessed through observations of teachers by trained school personnel. Pearson’s correlation will be conducted to examine the relationship between teacher attitudes and PGD fidelity, while multiple regression analyses will assess how years of teaching experience moderates this relationship. The findings will provide insight into factors that may act as facilitators or barriers to EBP delivery, particularly implications for addressing barriers through selection and tailoring strategies used to improve integration of EBPs, as well as developing teacher training to support high EBP fidelity in schools.

Periodontitis, the most severe form of periodontal disease, affects ~50% of Americans and is expected to continue increasing as a major public health concern globally. Fusobacterium nucleatum (Fn) and Porphyromonas gingivalis (Pg) have been identified as synergistic oral pathogens that play a key role in advancing periodontitis via immune subversion; however, bulk RNA sequencing fails to elucidate the genuine synergistic interactions among these populations due to culture heterogeneity. To capture the true cell-cell interactions within complex polymicrobial communities we are utilizing microbial split-pool ligation transcriptomics (microSPLiT), a cutting-edge high-resolution single-cell RNA sequencing approach to illuminate novel interactions between Fn and Pg. To optimize microSPLiT for oral bacteria, this study explores quantifying RNA and DNA within Fn and Pg mono-cultures to pinpoint ideal sample populations and library preparation conditions needed for accurate single-cell gene expression. Qubit fluorometric quantification was used to quantify RNA and DNA. Pg possess known mechanisms that confer resistance to assorted antimicrobial agents; therefore, an increased concentration of enzymatic reagent may be necessary for permeabilization steps within microSPLiT. These findings are expected to help optimize microSPLiT for Fn and Pg while directly advancing our understanding of in vitro interactions between two pervasive oral pathogens.

This project aims to compare and contrast Little Free Pantries (LFPs) and Community Fridges (CFs) as models of micro pantries for those facing food insecurity. The primary focus is on the ability of each to safely provide nutrient-dense food. There is extensive research on what a nutrient-dense diet is and its importance on a person's health as well as budding research detailing the quality of food provided by food pantry organizations. However, this research is limited in the scope of donation programs being researched. Furthermore, there is a lack of understanding how different donation programs differ in their ability to provide nutritional food to those utilizing their services. This project compares five CFs to five geographically matched LFPs within the City of Seattle. To assess the nutrient-density of food donated to CFs and LFPs two dimensions of food nutrition are analyzed utilizing the FAST score and NOVA categorization methodologies. Each micro pantry is briefly assessed on its food safety standards as well. CFs are expected to provide food with higher nutritional quality than LFPs, and in a safer manner. This is due to their ability to be temperature controlled, allowing for a wider variety of foods to be collected and stored safely. This research will be presented in a final paper and poster detailing the findings of the project as well as any recommendations drawn for the City of Seattle. This project is significant because it addresses the nutrition of food made available to those facing food insecurity. Understanding the difference in quality of food offered between the CFs and LFPs will allow for an assessment regarding how accessibility to nutrient dense food for all residents of Seattle falters and how it can be improved.

Depression is one of the leading psychological disorders worldwide. Neuroscientific research in art therapy has focused on measuring brain activity involved in emotional processing associated with psychological disorders rather than treatment outcomes. One of the reasons art therapy models have not been tested in treatment is traditional EEG measurements are prone to interference when participants move while making art. The recent development of Mobile Brain/Body Imaging (MoBI) is a promising way to measure brain activity during art therapy tasks. This research proposal aims to test the hypothesis, that art therapy treatment models which elicit an emotional response will work more effectively than models that stimulate a cognitive response, on individuals with major depression. This prediction is based on the outcomes of meditation studies, which demonstrate affective experiences can interrupt default rumination, a key barrier to engaging in higher cognitive functions which can improve symptoms of depression. This study proposal utilizes art mediums commonly applied in art therapy practice known to produce both affective and cognitive experiences. The context in which the medium is used will be manipulated to provide either experience. Changes in depression symptom severity between art therapy modalities will be compared within subject’s pretest and posttest. Study results could be used to identify effective treatments for depression, optimize currently used treatment models, and possibly predict treatment outcomes on individuals with major depression.

Although 911 is a universally accessible emergency service for anyone in the United States, civilians utilize the service at different rates. Some people are quick to call over minor issues better suited for the non-emergency line, whereas others are hesitant to call 911 even if a crime has occurred. Existing research suggests that comfort with using 911 and other emergency services varies by social and neighborhood factors, yet the mechanisms shaping this variation remain underexplored. Additionally, social disorganization theory, a theory in the criminology field that hypothesizes that community factors contribute to crime,  suggests that socially disorganized neighborhoods, characterized by low economic opportunity, high residential mobility, and ethnic heterogeneity, contribute to crime and deviance. Drawing on 911 dispatch data from the City of Seattle and neighborhood-level socioeconomic indicators from the American Community Survey (ACS), this study empirically tests the relationship between median neighborhood income and 911 call rates, with homeownership as a potential moderating factor. Using regression analysis, this research evaluates whether social disorganization influences emergency service use in a way that mirrors its impact on crime, with an expected pattern of higher reporting rates in both lower- and middle-income neighborhoods This study is crucial because findings can inform police reform, improve community outreach in Seattle, and provide the basis for future research to further inform the relationship between report rates and neighborhood characteristics in other major metropolitan cities in the United States.

Washington State is known for its progressive policies and support for transgender and nonbinary (trans) populations (i.e., trans-progressive). However, some studies suggest that there are still barriers to healthcare access. This study explored trans Washingtonians’ perspectives of these policies and experiences in access to non-discriminatory healthcare. The Priority Assessment in Trans Health Study conducted semi-structured interviews in August 2023 – March 2024 trans adults (n=24) living in Washington State. Interview topics included access to sexual health services, policy awareness, and community resilience. Themes were identified via inductive thematic analysis using Dedoose. Participants were more willing to visit hospitals that have anti-discrimination policies over those that do not, citing safety as a main contributing factor. Although, many were not aware of the anti-discrimination policies that their clinics had in place.  Some noted the presence of pride flags and trans health posters were signals of safety for trans individuals in these settings. However, some participants believed these endorsements feel performative without actionable follow through and described their personal discriminatory experiences in these hospitals. Additionally, participants noted that the lack of necessary training for trans healthcare makes obtaining care, particularly emergency care or intersectional care  difficult. Lastly, participants emphasized that the national structure of the American healthcare system as a barrier to accessing trans-specific care (e.g. need for multiple referrals). While Washington’s state-level policies supporting trans healthcare create an initial sense of safety, they often fail to address the root structural inequities which perpetuate healthcare inaccessibility. To bridge the gap between performative action and genuine inclusion of trans adults, accountability must be prioritized at all levels: from hospitals holding their employees accountable for discriminatory behavior to reforming medical education.

Traumatic brain injury (TBI) results from a blow to the skull that causes shearing forces in the brain. Elevating intracranial pressure (ICP) at the moment of impact may protect the brain from TBI by stiffening the brain tissue and decreasing shearing. When they expect an impact, humans naturally brace and perform a Valsalva maneuver (exhaling against a closed airway), which momentarily elevates ICP. In a ferret TBI model, we conducted abdominal compression using a blood pressure cuff to induce a Valsalva-like response (VLR) and determine whether VLR resulted in neuroprotection. The ferret model was chosen for its gyrified brain structure and white to grey matter ratio that closely resembles the human. TBI was induced using a CHIMERA (Closed-Head Impact Model of Engineered Rotational Acceleration) device, which is designed to deliver high-energy, controlled skull impacts. Initial work showed that the abdominal compression procedures increased ICP. The TBI study involved a total of 36 adult ferrets of both sexes randomized into three groups: (1) a sham control group exposed to isoflurane with a cuff but no compression, (2) a TBI group with a cuff but no compression, and (3) a TBI group with a cuff and abdominal compression. Baseline behavioral assessments (CatWalk, Novel Object Recognition, Swim Test, and Open Field) were conducted one week prior to injury. Post-injury behavioral testing, using the same assessments, was performed at 24–48 hours and 8 days post-TBI to evaluate functional outcomes. On day 8, ferrets were euthanized, and their brain tissue was collected and assessed for neuropathological outcomes. We hypothesize that abdominal compression will mitigate deleterious TBI outcomes. If these findings are supported, this intervention could improve the lives of those at risk of TBI and contribute to ongoing research in the field.

Pregnant women infected with influenza A virus (IAV) are at higher risk of morbidity, mortality, and poor fetal outcomes. However, the difference in the pathogenesis of IAV between pregnant women and non-pregnant women remains inadequately understood, primarily due to the lack of animal studies that use a translational model of infection. I hypothesized that higher IAV viral load and Type I interferon concentrations would be observed in the lungs and bronchoalveolar lavage of pregnant pig-tail macaques compared to non-pregnant macaques, and that correlating these metrics would yield different results across groups. We inoculated pregnant (n=11) and non-pregnant female (n=18) pig-tail macaques (Macaca nemestrina) with IAV H1N1 (A/California/07/2009) and euthanized them at 5 days post-inoculation, when we expected to observe peak lung pathology. We tested pulmonary function at baseline and study endpoint and conducted clinical assessments daily. I extracted RNA and performed quantitative polymerase chain reactions on the samples to calculate viral load. I also performed enzyme-linked immunosorbent assays to quantify concentrations of Type I interferons (IFN-α, IFN-β). Lastly, I analyzed pulmonary physiology data and clinical assessment scores as a reliable measure of disease severity. A bi-modal distribution of viral load was observed in the lungs of pregnant animals (high>9e5 copies/mg; low<2e4 copies/mg), which was not observed in non-pregnant animals.  When correlating viral load at 5 days post-inoculation with Type I IFN in the lung of the pregnant animals, I found a significant positive correlation between IFN-β and viral load in both the lungs (ρ=0.8, p=0.03) and BAL (ρ=0.9, p=0.02). These results suggest that despite a strong IFN-β response in the lung, a high viral load persisted in the pregnant animals. Next steps could explore whether the kinetics of the pulmonary innate immune response is delayed in pregnancy, which impairs viral clearance. 

This study aims to understand users’ needs in designing a mobile application that assists Cardiopulmonary Resuscitation (CPR). This study is designed through a two-stage online interview, beginning with a screening survey that asks about participants’ occupations and experience with CPR, followed by the main interview that asks about their challenges when performing CPR and their opinions on the most efficient type of feedback. Eight participants who completed the survey included CPR instructors, paramedics, firefighters, and medical students with experience ranging from 3 to 20 years. The participants rated maintaining compression depth as more challenging (7.63 out of 10) compared to maintaining compression rate (5.25 out of 10) when performing CPR, with 10 representing the highest level of difficulty. Participants also reported that visual feedback would be more beneficial for compression depth and auditory feedback for compression rate. Therefore, these findings suggest that an effective assistive CPR device should have real-time visual feedback to aid in-depth consistency while auditory cues support rhythm accuracy similar to a metronome. By leveraging these findings, the proposed application has the potential to enhance CPR effectiveness, improve user confidence, and ultimately increase patient survival rates.

Decision During Drinking (D3 study) is currently developing a mobile app aimed at reducing negative alcohol-related outcomes with a focus on young adults. D3 offers modules to educate users on identifying cues that influence their decisions, understanding their values after drinking, and engaging in practices to avoid such experiences. I have aided in the design of the prototype of the D3 program, both creating new features and adapting previously developed features. The current study assesses core aspects of human centered design on two of the D3 study features I helped to create in Figma. The first is an activity that compares participants’ perception on alcohol expectancies to alcohol’s physiological effects. The second is a “roadmap” planning tool that guides users to create action plans to cope with specific drinking cues, linking each plan to personal values. By using an iterative design approach from research participants’ feedback, I refined the user interface (UI) to improve the features. I am adapting measures on usability (i.e. System Usability Scale) to assess the metrics for these features specifically. Data is being collected from online surveys on the usability of these features. I predict that the final usability score would be higher than the industry-standard threshold (e.g. score of ≥ 70), so the features will provide a seamless and intuitive user interface. Based on the improvements, if deemed usable, these feature designs could be applied to other clinical app developments. 

Hypertrophic Cardiomyopathy is the most common form of hereditary heart disease affecting ~1:500 individuals, characterized by progressive thickening of the left ventricular wall. The first mutation linked to this disease was the heterozygous R403Q mutation in human beta-myosin heavy chain (β-MHC). Conflicting reports of contractile kinetics between human myectomy samples vs transgenic mouse and rabbit models motivated us to study the molecular mechanisms of altered contraction in a CRISPR/Cas9 gene edited human inducible pluripotent stem cell line. Following differentiation to cardiomyocytes (hiPSC-CMs) and maturation in culture, we isolated sub-cellular contractile organelles called myofibrils. Myofibril contractile kinetics from this line had slowed force development and cross-bridge detachment, with reduced maximal force compared to the WT line. hiPSC-CMs were cast into fibrin matrices to form three-dimensional, engineered heart tissue (EHT) for measures of twitch force and contractile kinetics. At 1Hz stimulation, heterozygous mutation EHT’s exhibited a hypercontractile phenotype compared to WT EHTs, with slowed relaxation kinetics. Since the penetrance of our heterozygous R403Q hiPSC-CMs is unknown, we are now studying a homozygous iPSC-CM line where 100% of the β-MHC is mutated. This will allow us to assess the direct contribution of the mutation to the disease contractile phenotype. We will repeat the myofibril and EHT measures of contractile properties and perform stopped flow kinetics analysis on isolated myosin to determine ATP turnover and ATP hydrolysis product release rates. This will provide molecular mechanistic insight of the contractile abnormalities, allowing development of therapeutic interventions that specifically target the mechanisms that alter contractile function. 

Optical coherence tomography (OCT) is a powerful non-invasive imaging technique, widely used for high-resolution 3-D structural imaging in research and clinical settings. Current variations, such as Point-Scanning OCT (PS-OCT), offer detailed structural images but require significant computational power for dynamic signal extraction, crucial for monitoring functional information like blood flow and cellular movement—key factors in disease diagnosis and treatment monitoring. The Biophotonics and Imaging Lab (BAIL) has developed an OCT angiography (OCTA) method for dynamic blood flow imaging. However, the OCTA system faces limitations due to its high computational demands. In this project, I propose an alternative approach using a Line-Scan OCT (LS-OCT) system, which samples lines within the region of interest to acquire simultaneous cross-sectional data. I aim to maintain dynamic signal extraction while reducing computational load and minimizing cell activity noise. If successfully developed, LS-OCT can revolutionize clinical melanoma diagnostics by providing real-time, non-invasive imaging of the epithermal and dermal information, thus identifying disruptions caused by tumor growth and angiogenesis without the need for traditional biopsies. The system will also have significant potential for real-time observations of drug effects on cancer cells, optimizing therapeutic testing by eliminating the reliance on histological processes. Specifically in this project, I will focus on designing an experiment to acquire live tissue data and process signals using the first LS-OCT system developed by BAIL. The goal is to compare dynamic image results with those obtained from PS-OCT systems, potentially enhancing future cell movement analysis research and supporting other PS-OCT-based projects requiring dynamic cellular information within living tissues.

Kaposi’s sarcoma-associated herpesvirus (KSHV) is an oncogenic gammaherpesvirus known to cause Kaposi sarcoma (KS), a cancer of the soft tissues, and several other diseases. KSHV has two distinct replication cycles: a latent and lytic cycle. During latent infection, only a small section of the viral genome, the KSHV latency-associated region (KLAR), is expressed. Spindle cells, the main proliferating cell type in KS tumors, are thought to be of endothelial origin and are primarily latently infected. Due to lowered viral gene expression during latency, these cells have few viral factors to target therapeutically. Cellular factors required for infected cell survival, like altered metabolic pathways, are potential therapeutic targets for latently infected cells. One metabolic pathway altered by latent infection is fatty acid synthesis (FAS). My research focuses on understanding how KSHV infection induces FAS in cells. Since previous research has shown that expression of KLAR is sufficient to increase lipid droplet formation, a measure of FAS induction, I hypothesize that expression of one of the four genes present in KLAR is likely what upregulates this metabolic pathway. To test this hypothesis, I infected telomerase-immortalized microvascular endothelial (TIME) cells with lentivirus containing a viral plasmid overexpressing one of the four KLAR genes. I then measured lipid droplet formation across each transduced cell population using a flow cytometer. This project is still in progress; however, if the increase in lipid droplet production in transduced cells is similar to the increase observed in cells latently infected with wild-type KSHV, then I will conclude that the over-expressed gene was sufficient to up-regulate FAS. Identifying which viral gene induces FAS in infected cells will provide a new direction for future mechanistic studies and aid in identifying potential therapeutic targets for KSHV-associated diseases.

The Campanian stage of the Late Cretaceous (~84–72 million years [Ma]) was a dynamic interval for North American ecosystems including the evolution of angiosperm plants and the regression and transgression of the Western Interior Seaway (WIS). These dynamics likely impacted terrestrial fauna across the continent. Most studies investigating biodiversity in western North America during the Campanian focus on a single group (e.g., dinosaurs or herpetofauna), whereas few investigate diversity patterns of multiple taxa. This approach is imperative because comparing diversity patterns among taxa can provide rare insight into the synecology of animal communities. Vertebrate microfossil sites are ideal for such a study because they preserve large sample sizes, multiple taxonomic groups that likely coexisted, information about environmental conditions, and they sample multiple stratigraphic intervals. The Judith River Formation (JRF) of north central Montana preserves ~4 Ma of the Campanian (~79–74 Ma) and large regressive and transgressive cycles of the WIS. This formation, which is contemporaneous with the Dinosaur Park and Two Medicine formations, is also rich in vertebrate microfossil sites. To examine vertebrate diversity patterns through this critical interval, we compared taxonomic richness and relative abundances of Dinosauria, Squamata, and Lissamphibia from the JRF from three temporally distinct microfossil sites: Makela-French 1 (~77.5 Ma), Milkshake (~76.5 Ma), and Clamfetti (~75.2 Ma). Over four years, we collected and screenwashed fossiliferous, bulk-sediment samples from these sites. Thus far, we have recovered 998 and aim to recover 1,200 specimens total. We use our data and knowledge from the literature to evaluate the extrinsic factors (e.g., seaway regressions) that drove diversity changes in the JRF fauna.  Our preliminary results suggest a connection between diversity patterns and WIS cycles. We observe shifts in relative abundances and richness near the onset of the WIS transgressive cycle. 

TDP43 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 TDP43 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, TDP43 autoregulates its own mRNA splicing, resulting in multiple TDP43 isoforms. Although some of these isoforms go through nonsense mediated decay, other isoforms result in unique proteins with differing C-termini. This leads 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 different isoforms and interacts with aging and ALS-like symptoms, the Darvas Lab created a novel approach to overexpress human TDP43 (hTDP43) via Adeno-Associated Virus (AAV) delivered through retro-orbital injection, leading to ALS-like motor deficits. We tested this AAV in young and old mice cohorts. Then, 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-control 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.

My project intends to discover a DNA aptamer, a single stranded DNA oligonucleotide, that binds selectively to the protein Interleukin-6 (IL-6). IL-6 has an important role in the immune system response and in excess it is known to cause inflammation. Aptamers exhibit binding affinities like that of antibodies but are ~50 times cheaper to produce. The method of aptamer discovery is through SELEX (Systematic Evolution of Ligands by Exponential Enrichment) which involves the selection from an aptamer library that contains 52N random nucleotide region and constant 5’ and 3’ 18 base pair regions for PCR amplification. Positive and negative selection are completed by incubating aptamer libraries with IL-6 or random protein immobilized on magnetic beads respectively. After each round selected aptamer sequences are amplified with a polymerase chain reaction (PCR) with primers that anneal to the constant regions. Reverse primer has biotin on 5’ end that is used later for strand separation with streptavidin agarose. After each round aptamer pool is going to be sequenced using nanopore sequencing platform till the enrichment of IL-6 specific sequences is observed. Binding will be tested through an enzyme-linked immunosorbent assay (ELISA) using the fam on 3’ end. The end goal of this project is to design a cost-effective method of IL-6 depletion from patients blood, allowing for cost-effective method of treatment for overactive immune system inflammation in sepsis patients.

Stress has a profound impact on reward-seeking behaviors, increasing the likelihood of relapse in individuals with substance use disorder. One of the molecular mechanisms underlying this stress-induced shift in behavior involves the activation of kappa opioid receptors (KOR) and the downstream signaling pathways that influence both dopamine (DA) and serotonin (5HT) neurons. Specifically, the activation of KOR by the endogenous neuropeptide dynorphin triggers an arrestin-dependent pathway, resulting in the recruitment of p38ɑ mitogen-activated protein kinase (MAPK), which mediates the aversive effects of receptor activation. This project aims to explore the role of pharmacological and  stress-mediated KOR/p38ɑ  MAPK signaling in DA and 5HT neurons. To determine KOR/p38ɑ  MAPK signaling in both dopamine and serotonin release in the NAc during pharmacological activation of KOR or during stress (rFSS), we utilized CRISPR technology to manipulate p38ɑ MAPK signaling in the ventral tegmental area (VTA) and dorsal raphe nucleus  (DRN) respectively. To monitor both DA and 5HT tone in the NAc we used the novel GPCR-based DA and 5HT sensors (GRAB-DA and GRAB-5HT). Both serotonin and dopamine tone were decreased after KOR agonist administration compared to control in the NAc. Cocaine, our positive control robustly increases serotonin and dopamine compared to control.  During periods of swim stress,  serotonin fluorescence robustly decreases which can be blocked by p38ɑ excision from the DRN or pretreatment with the KOR antagonist norBNI. This study provides insights into the molecular mechanisms underlying stress-induced changes in reward-seeking behavior and identifies potential therapeutic targets for substance use disorder.

Direct Analysis in Real Time Mass Spectroscopy (DART+ MS) is a chemical identification tool that uses a superheated gas stream to ionize chemical samples, producing a distinct chemical signal that can be used to identify the composition of an unknown sample. DART+ MS is used reliably in fields like forensics, food safety, pharmaceuticals, and more recently, environmental protection. At the Wasser Research Lab, at the Center for Environmental Forensic Science, we work to protect endangered species such as African Elephants. Using Direct Analysis in Real Time Mass Spectroscopy, we seek to find if elephant ivory from different regions in Africa has distinct chemical signatures, allowing us to geolocalize ivory samples based on their DART+ MS signatures. Current methods of elephant geolocation include genetic testing, but results can often be ambiguous; By using this completely different, complementary approach, we could improve our estimates of these inconclusive tests. If there is a chemical difference in the ivory of Elephants from the Savannah and Forest regions of Africa, then we can trace the origins of ivory obtained from illegal seizures, aiding in the conservation efforts of African elephants. Chemical distinctions aside, we also hope to answer questions about the effects of certain chemical preservatives on ivory samples and whether the DART+ MS signal varies along the length of the cut of the tusk, establishing best practices for sampling. Ultimately, our goal is to determine if DART+ MS proves to be a reliable and quick method of identifying elephant ivory for conservation efforts. By bridging cutting-edge technology with conservation science, we hope this research will make a significant impact on efforts to combat the illegal ivory trade and wildlife crime. 

Abiotic stressors are an important driver of organism survival and community structure of epifauna in aquatic environments. Stressors present in estuaries are found in a gradient, as freshwater from rivers mixes with marine water. Marine epifauna have been shown to have both bottom up and top-down controls on ecosystems, which can promote the health of seagrass meadows ecosystems. In Washington state, eelgrass (Zostera sp.) grows across a gradient from coastal waters to estuaries; do abiotic stressors within this gradient determine what epifauna communities are supported in eelgrass? To consider for structure area, 30cm x 30cm artificial turf squares were placed in eelgrass beds overnight at two sites along an estuarine gradient in Willapa Bay (Washington, USA). Epifauna were collected by rinsing turf mats with freshwater, then preserved in ethanol and identified to family level. Following identification, this data will be used to explore the difference in community abundance (univariate statistics, ANOVA) and community composition (multivariate statistics, PERMANOVA). I hypothesize that there will be a difference in epifauna community structure in eelgrass beds between sites, specifically a shift in community composition with higher richness and abundance at the site closer to the ocean. Knowledge of how abiotic stressors influence community structure over estuarine gradients may act as a model for climate change and what communities are expected to be found as abiotic stressors change. How these communities may change as environmental stressors do is important, as epifauna are prey for many bird and commercial fish species.

Visual word interpretation involves both phonology, sound-based processing common in alphabetic languages, and orthography, visual pattern recognition common in logographic languages. Previous research suggests that word recognition in alphabetic languages, such as English, activates the left inferior frontal gyrus (LIFG), and logographic languages, such as Mandarin, engage the visual word form area (VWFA). The goal of this study is to investigate the neural and behavioral differences between native English speakers and native speakers of logographic languages when processing English phonemes. I hypothesized that native English speakers would show more activation in the LIFG whereas native logographic language speakers would show more activation in the VWFA. Participants completed a rhyming judgment task based on English phonemes while undergoing functional magnetic resonance imaging (fMRI) to measure neural activation. English proficiency was recorded through both self-report and the Nelson-Denny Comprehension test. Behavioral results suggested that native logographic language speakers had significantly longer response times and lower accuracy compared to native English speakers. Both self-reported and objectively measured English proficiency were negatively correlated with response times and positively correlated with accuracy. Although the neural analysis did not yield statistically significant results, there was a trend of increased VWFA activation in native speakers of logographic languages compared to English.

Chloroplasts are central to plant immunity and act as a hub for immune signalling and defence-related hormone synthesis. The essential chloroplast-localized protein FtsHi1 is a component of the FtsHi import motor and is vital to translocating proteins across the chloroplastic membrane. Viral-induced gene silencing (VIGS) of FtsHi1 in Nicotiana benthamiana results in a bleached phenotype, indicative of decreased translocation of essential chloroplastic proteins and decreased chlorophyll synthesis. Previous work identified herbivore-induced kinase 1 (HIK1) as a potential interactor of FtsHi1. HIK1 is a receptor-like cytoplasmic kinase (RLCK) implicated in the immune response to caterpillars. FtsHi1 contains a predicted RLCK phosphorylation site, indicating possible phosphorylation of FtsHi1 by HIK1, which could promote defence signalling over photosynthesis. This project aims to test the impact of phosphorylation on FtsHi1 function and its role in protein translocation across the chloroplastic membrane. This work utilises engineered FtsHi1 variants, which mutate the serine phosphosite to either mimic FtsHi1 phosphorylation (Ser→Asp), prevent it (Ser→Ala), or recapitulate the wild-type protein sequence. To test the phosphosite mutation effects, FtsHi1 VIGS knockdown of N. benthamiana leaves will be transiently modified using Agrobacterium tumefaciens bacteria to express WT or phosphorylation-modified FtsHi1 constructs. The resulting colour phenotype for each construct will then be compared to the bleached phenotype of TRV2:FtsHi1 plants and the phenotype of wild-type plants. I hypothesise that the FtsHi1 phospho-null mutant will result in a rescue phenotype similar to wild-type FtsHi1, whereas FtsHi1(phospho-mimic) will retain the bleached phenotype.Examining FtsHi1 phosphorylation enhances our understanding of its potential interaction with HIK1 in herbivory-induced plant defences. Future studies will explore FtsHi1's role in defence mechanisms, with implications for engineering herbivory-resistant crops. 

TBC domain containing kinase (TBCK) is an understudied protein with three domains: a pseudokinase; Tre-2, Bub2, and Cdc16; and rhodanese, and is highly expressed in the brain. Homozygous mutations in TBCK cause a rare neurodegenerative disorder in children, which clinically presents as syndrome infantile encephalopathy, brain atrophy, cerebellar hypoplasia, and muscle hypotonia. Two mutations in particular, Arg126Stop and Arg511His in the pseudokinase and TBC domains respectively, are commonly found among TBCK patients. The progression of the disease is characterized by a global regression in brain development, severe intellectual disability, and premature death in acute cases. The pathogenic mechanism underlying TBCK syndrome is unclear, but past studies show that TBCK patient neurons demonstrate aberrant metabolite buildup in the lysosome likely resulting from abnormal lysosomal activity. Immunoprecipitation mass spectrometry was performed for wild type TBCK in both N terminal and C terminal tags, revealing a preliminary list of both known and unknown interactors for TBCK. To further investigate the early developmental implications of mutant TBCK, CRISPR/Cas9 directed mutagenesis is being used to generate two induced pluripotent stem cell (iPSC) lines harboring the Arg126Stop and Arg511His mutants for subsequent differentiation into neural progenitor cells (NPC) and neurons. Immunofluorescent imaging of the mutant NPCs will confirm the recapitulation of growth and lysosomal defects present in patient cells. To analyze the effect of TBCK mutation on lysosomal function/content, we will immuno-isolate lysosomes through lysosome immunoprecipitation (Lyso-IP) and identify proteomic changes through mass spectrometry. While providing a crucial in vitro cell model of two common patient mutations, these experiments will offer critical insight into cellular dysfunctions that contribute to TBCK disease states.

Cave formations, also called speleothems, are natural archives of past climate conditions. Analysis of these speleothems can provide information about environmental changes over thousands of years. Church Mountain Cave is in the Northeast Cascades and is distinctly characterized by its unusually predominant speleothems, which is relatively unusual for western Washington. This project explores the potential of Church Mountain and other caves as paleoclimate archives. I begin with a characterization of Church Mountain Cave, focusing on cave morphology, stratigraphic and structural position of the cave, and forming a hypothesis of cave formation and fault activity. This classification will allow for a detailed view on Church Mountain Cave and the factors impacting speleothem formation. We also characterize a range of speleothems from other caves using trace element and isotope analysis, with the eventual goal of using uranium-thorium dating to establish precise ages and stable isotope analysis with oxygen isotopes to infer past temperature, precipitation patterns, and other environmental factors. By refining regional paleoclimate reconstructions, we can improve predictions of future climate trends and extreme weather events. Additionally, this study underscores the importance of caves as climate archives and highlights the need for conservation efforts to protect these valuable and fragile environments for continued scientific research. 

In 2023, the United States saw the highest number of climbing gyms in operation, with Washington hosting 26 of the 622 gyms nationwide. While the health benefits of bouldering—mental, physical, and cognitive—are well-established, the sociospatial implications of these recreational spaces remain underexplored. This research examines how climbing gyms in Seattle contribute to both gentrification and population displacement, while also fostering a sense of community. To understand the interplay between these dynamics, the study contextualizes gentrification and community through a theoretical framework rooted in sociological research. The project combines demographic analysis using U.S. Census data with an ethnographic approach to capture the lived experiences of individuals building community within these spaces. Ultimately, the research aims to determine whether the impacts of gentrification and community building outweighed one another, and offers insights into how climbing gyms can equitably coexist within their communities. 

Adaptive algorithms used in brain-computer interfaces (BCIs) adjust to user strategies by dynamically adjusting how BCIs decode neural data throughout an experiment. Current adaptive algorithms continuously update the decoder using all available data during training. However, if users are unfocused or inattentive, it is likely that some of the training data is unhelpful towards decoder training and could lead to poor decoder performance. Unfortunately, determining attentiveness in a subject is difficult. Non-human animals cannot self-report attention levels, and even in human trials, self-reporting often leads to subjective data that varies between subjects. A non-invasive estimate of subject attentiveness could improve data selection for decoder training. Pupil size is correlated with a participant’s perception of task difficulty, and participants involved in attention-grabbing tasks display blinking rate-inhibition (Kucewicz et al., 2018; Maffei et al., 2019). I hypothesize that these eye data could be used to estimate a subject’s task engagement. I explored data from a novel task where both human and non-human primate subjects controlled a cursor on a 2D screen with 3D hand motions through some unknown mapping. Due to the unknown 3D-to-2D mapping, this task required constant cognitive attention in order for subjects to succeed. I looked at the subjects’ data to identify trends in pupil size and blink frequency across multiple days of task performance. In the future, I aim to build engagement classification models to better select training data for adaptive algorithms and apply these algorithms to realtime BCI experiments.

Taiwan’s agricultural sector faces systemic challenges rooted in colonial and post-war strategies, prioritizing short-term growth at the expense of local adaptability, leaving producers vulnerable to market dependency, environmental instability, and knowledge erosion. As global trade pressures and climate risks escalate, farmers need to adapt new flexible approaches to be sustainable. Corporate Social Responsibility (CSR) initiatives have emerged as potential solutions to promote sustainable practices and empower primary producers. Government and private-sector efforts provide resources and collaborative platforms, helping producers diversify beyond raw produce sales, aiming to foster innovation, reduce dependency, and promote long-term self-sufficiency. Despite some success in stabilizing agricultural industries during crises, government-led CSR initiatives, such as subsidies and export promotions, often overlook structural issues that drive dependency. This raises critical questions about power dynamics, producer agency, and the sustainability of CSR-driven innovation. This study investigates the impact of CSR on power dynamics, resource distribution, and producer agency within Taiwan’s agricultural sector, focusing on three successful case studies: pineapple export diversification, Milksha’s dairy innovation, and pomelo value-added products. The methodology uses process tracing to identify critical turning points through data from semi-structured interviews, policy documents, and corporate reports. Additionally, interactive timeline activities during interviews allow participants to illustrate key events and decision-making processes, supported by a codebook for iterative data analysis. This research contributes to the investigation of CSR by offering practical recommendations for policymakers to improve existing frameworks and explore new models that prioritize long-term sustainability and producer agency. The findings underscore the importance of equitable resource distribution, shared decision-making, and social network engagement in achieving sustainable agricultural development in Taiwan. 

Periodontitis is a severe inflammatory disease that damages the gingiva and has been linked to systemic diseases, such as diabetes, heart disease, and respiratory disease. These serious health complications express the importance of studying oral microbiota and their interactions. This research investigates the growth dynamics of Fusobacterium nucleatum and Porphyromonas gingivalis, two gram-negative anaerobic bacteria that play a significant role in inducing the progression of periodontitis, under mono- and co-culture conditions. By collecting monoculture and co-culture growth curves along with cell counts, and LIVE/DEAD BacLight staining we are identifying their optimal growth phases and interactions. With this information, we seek to optimize these cultures for downstream experiments, including bulk and single-cell RNA sequencing, to identify unique genes signatures from cell-cell interactions implicated in periodontal disease progression. Understanding these dynamics will contribute to future studies on the persistence of periodontal infections and broader research on gene expression.

Can early adverse experiences possibly enhance cognitive functions? The current literature pertaining to children and youths with adverse experiences suggests that they often present extensive deficits, potentially showing less working memory, later language disorders, and lower impulse control (Ellis et al., 2020; Dannehl et al., 2017; Dixon et al., 2023; 2016; Snow, 2021). However, this deficit-based perspective is incomplete – the hidden talents framework suggests that children possess adaptive strengths of unique skills that emerge under stress and in particular, adverse environments. Children and youths with experiences of adversity show social and cognitive adaptations, revealing enhancements in certain domains (Ellis et al., 2020). Here, we examine a facet of their adapted strengths: cognitive flexibility in adulthood as a response to childhood unpredictability, defined as the rate of alterations or instability in the individual’s childhood environment. We propose that adults with prior experiences of childhood unpredictability develop enhanced cognitive flexibility to adapt to unpredictable environments. In this experiment, we will recruit a normative sample of 180 adults and measure their cognitive flexibility along with experiences of childhood harshness and unpredictability using the Perceptions of Childhood Harshness and Unpredictability scale and the Number-Letter Task. We predict that participants who experienced a high level of unpredictability will demonstrate more cognitive flexibility compared to those who experienced high general childhood harshness. Preliminary analyses from the UW psychology participant pool indicate similar trends, suggesting that higher childhood unpredictability correlates with increased cognitive flexibility, while higher childhood harshness is associated with lower cognitive flexibility. Framing cognitive flexibility as a form of adaptation from unpredictable environments is imperative to pivot the current narrative toward children’s hidden talents – revealing that children are resilient, adaptive individuals with unique abilities to overcome adversity. 

With the increasing prevalence of large wildfire events, especially near urban areas, the identification and quantification of combustion-derived air pollutants is critical. The collection, quantification, and identification of particulate air pollutants, also known as aerosols, provide important insights into air pollution sources, evolution, and health effects. To improve capabilities in this area, with funding and support from the Beckman Foundation, we have designed and developed an aerosol collector that uses high-voltage Dielectric Barrier Discharge plasma and Electrostatic Precipitation (DBD-ESP) to collect and desorb target nanosized aerosols (30 nm – 800 nm) containing components typical of woodsmoke, such as levoglucosan and phenolic compounds, among others. The DBD-ESP is integrated with an Ultra-Portable Time-of-Flight (TOF) Mass Spectrometer (UP-ToFMS) to produce online, near real-time analysis of the chemical composition, mass, and size of target aerosols. Based on preliminary collection and desorption testing, we expect to have a 75% collection and desorption efficiency of submicron aerosol particles within 5 minutes. The DBD-ESP has an internal volume of less than 2 cm3 and an overall external volume of around 1 L. Once coupled with the UP-ToFMS, the overall ultra-portable system will be no larger than 0.095 m3 and weigh around 35 kg, enabling the remote sampling of aerosols from wildfire events while still yielding high-quality mass spectral data.

Organs maintain consistent shape, form, and volume through complex processes, one of which is cell-cell adhesion. E-Cadherin, a key cell-cell junction protein, is critical for cell shape, arrangement, and tissue structure. In this study, I investigate the role of E-Cadherin in the morphogenesis of the Drosophila Malpighian tubules, a model system where I can manipulate E-Cadherin expression and use fluorescence microscopy to observe the effects on organ growth. Previous work involved fixing and staining embryos to track E-Cadherin localization using fluorescent imaging to measure its intensity. I will further analyze E-Cadherin localization spatiotemporally by constructing a fluorescent fly line for live imaging during development. I expect E-Cadherin concentration to increase during elongation and to be enriched in looped regions of the tubules. To assess the requirement of E-Cadherin in organ formation, I will reduce its expression using RNAi and degradFP, expecting significant developmental defects due to the protein's vital role in morphogenesis. These defects will be quantified by comparing changes in cell and organ shape in control and E-Cadherin-reduced tubules. Additionally, I will help develop Python tools for 3D image analysis, including cell segmentation, creating a 3D model of E-Cadherin in tubular cells, and extracting protein intensity. Developing these tools not only enables our work in these tubular organs but also allows for comprehensive image analysis of other tubular 3D organ forms. 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. 

Each year, approximately 84,100 adolescents and young adults (AYAs) in the United States are diagnosed with cancer. Beyond the physical challenges that come with cancer, they also face significant psychosocial barriers, including gaps in health literacy, limited access to professional mental health support, and uncertainty about the future. Addressing these unmet needs requires innovative approaches, and social media, particularly Instagram offers a unique opportunity to bridge the gap between traditional healthcare services and the psychosocial needs of AYAs. With 2 billion users, Instagram is widely used for connection and information sharing, making it a valuable platform for health-related support and education. This study qualitatively analyzes Instagram posts under popular pediatric cancer hashtags to: (1) explore how AYAS and caregivers use social media for health information and support, (2) identify barriers they face in healthcare, and (3) examine how social media can enhance health literacy and education. A direct content analysis of 300 posts was conducted using a newly created Instagram account to minimize algorithmic bias. AI tools, including ChatGPT, Perplexity, and Microsoft Copilot, helped identify commonly used hashtags, which were then cross-referenced on Instagram. The most frequently used hashtags include #childhoodcancer, #childhoodcancerawareness, #pediatriccancer, #stupidcancer, #fightlikeakid, and #morethan4. An iterative coding process, using sets of 5–10 posts, was employed to develop and refine a codebook based on existing literature. Posts were categorized by metrics, user profiles, content types, health-related quality of life, and social support. My coding team consisting of myself, and 3 other investigators will code the data using REDCap, with descriptive statistics analyzed in R Studio. Findings from this study will highlight how Instagram can serve as a powerful tool to improve health literacy, education, and mental health support for AYAs with cancer, ultimately bridging critical gaps in healthcare accessibility and education.

Our brains have evolved to navigate survival and respond to danger, but trauma dysregulates these systems, causing the brain to misinterpret everyday experiences as threats. This dysregulation results in hypervigilance, which can manifest as panic attacks, dissociation, and other debilitating symptoms. Current treatment options for trauma often focus on symptom management, overlooking the physiological impacts of trauma. These treatments can be expensive, inaccessible, and may have side effects. This literature review examines holistic, non-pharmaceutical, neuroplasticity-based (NPNB) approaches, such as breathwork, nutrition, exercise, and sleep, to challenge traditional methods and advocate for integrating holistic interventions into mainstream trauma care, emphasizing accessibility and autonomy for trauma survivors. As we explore the increasing need for mental health care, we look at the interplay between psychological trauma and physical health by exploring the mind-body connection and trauma-induced inflammation. Additionally, this exploration aims to understand how these treatments can reshape neural pathways, improve emotional regulation, and enhance psychological and physiological well-being. It also examines potential paradigm shifts in trauma care and advocates for increased accessibility to alternative treatments, particularly for individuals who cannot access conventional therapies. We expect to find that NPNB treatments are underutilized in the treatment of trauma and, if expanded upon, would have the potential to improve accessibility, reduce or eliminate side effects, and help survivors regain a sense of autonomy.

Silica nanoparticles have diverse applications in catalysis, imaging, and drug delivery. Tailoring these nanoparticles for specific applications requires precise control over their size, surface chemistry, porosity, and polydispersity. These properties are controlled by a wide range of factors such as reactant type and concentration, pH, reaction temperature, and other synthesis parameters. Due to the large parameter space, determining the optimal reaction conditions for synthesizing silica nanoparticles with the desired size and morphology is time-consuming and challenging. An accelerated experimentation platform integrating automation and artificial intelligence can streamline the selection of reaction parameters for synthesizing silica nanoparticles with targeted size and morphology using machine learning-based iterative design of experiments to optimize material properties. This system uses the Science Jubilee flexible laboratory automation platform to carry out sol-gel synthesis. Small-angle X-ray scattering is used to characterize the sample. The data collected is used to optimize the reaction condition for synthesizing the targeted nanoparticle. We have successfully carried out sol-gel processes and synthesized silica nanoparticles with various sizes and polydispersity using the platform. Currently, we are working on optimizing the selection of sample synthesis conditions.

Rotated orientation patch count (OPCr) is a measurement used to quantify the complexity of a 3D surface. OPCr has previously been used to analyze tooth complexity, showing a correlation between complexity and diet in lizards, crocodilians, and mammals. We applied this technique to toothless taxa, with the goal of determining if there is a correlation between the complexity of the occlusal surface of a given species of turtle and its diet category. OPCr is determined by analyzing a 3D mesh of the occlusal surface of turtle specimens, with meshes based on both photogrammetry and CT scans of turtle skulls. Photogrammetry and CT scans are fundamentally different. Photogrammetry is a 3D mesh created from a series of surface images of an object, where the lighting and shadows cast on the object potentially distort its complexity. CT scans are not subject to these errors, and are typically more consistent provided the scan is made properly. However, there is little research analyzing the impact of different scanning techniques on the surface complexity of the resulting mesh. This project is therefore a comparison of photogrammetry and CT scans: do models made from these different methods produce significantly different OPCr scores? Nineteen specimens previously digitized using photogrammetry have been CT scanned. I created 3D models from the CT scan data and analyzed their surfaces using OPCr. I then compared the OPCr values produced by the CT scan models to the photogrammetry counterpart of each specimen. We hypothesize that statistical analyses will show no significant difference between the two methods of digitizing specimens.

Gene arrangements are observed in human diseases such as cancer and developmental disorders. In developmental disease, gene triplication with an inverted central copy is observed, while a hallmark of cancer are palindromes, or inverted repeats of DNA. Therefore, testing one mechanism of gene arrangement in an easily studied organism like the  budding yeast Saccharomyces cerevisiae may shed some light on the human genome and disease generation. Yeast cells grown for >200 generations in sulfate limiting media are enriched for triplication of the high affinity sulfate transporter SUL1 with the center copy inverted, as amplification confers a selective advantage. To explain how such a triplication occurs, the Brewer and Dunham Labs proposed a model called Origin Dependent Inverted-Repeat Amplification (ODIRA). The ODIRA mechanism requires a DNA origin of replication and short, inverted repeats flanking the SUL1 gene. During DNA replication, an error of the replication fork, or fork regression, causes annealing of leading and lagging strands to create a hairpin intermediate. The intermediate then replicates and recombines into the genome, forming interstitial triplications, with the middle copy inverted. While the proposed mechanism explains the observed triplication in yeast, the specific proteins involved are not yet known. The genes I've chosen to test are the DNA helicases MPH1 which prevents cross-over between ectopic sequences, and RRM3 which relieves replication fork pauses. Both are predicted to regulate necessary steps of the ODIRA mechanism, making them good candidates for genes that may be involved in these triplications. By deleting each gene, I can then measure the frequency of ODIRA events in those strains and compare them to wild-type strains. Increased ODIRA events in the knockout strains may implicate their role in the ODIRA mechanism and prompt further study of these genes and how they might affect copy number variation in humans.

This study investigates the mechanical properties of A500-C steel round hollow structural sections through tension testing of coupons cut from tubes with various radii and thicknesses. Testing was conducted following ASTM A370 standards to evaluate relationships between carbon equivalent, strength ratios, and ductility. The carbon equivalent was determined using the International Institute of Welding equation with chemical compositions obtained from mill certifications. Strength ratios that were analyzed include measured tensile-to-yield strength, measured-to-mill certification values, and measured-to-nominal values, with comparisons to the ratios prescribed for design in the American Institute of Steel Construction Seismic Provision. The results indicate that most strength ratios and ductility metrics showed no significant correlation with HSS thickness or carbon equivalent. However, some trends were observed. Negative correlations were found between the ratio of measured to nominal ultimate strength and thickness, the measured tensile-to-yield strength ratio and thickness, the ratio of measured to mill-certification yield strength and carbon equivalent, and the ratio of measured to nominal yield strength and carbon equivalent. A positive correlation was observed between the measured tensile-to-yield strength ratio and carbon equivalent. These findings help provide insight into the variability of A500-C steel properties and their dependence on chemical composition and wall thickness, with potential implications for design assumptions in the structural design code.

Cortical tracking, a method that examines how neural activity encodes the dynamic features of the incoming speech stimuli, allows for the study of naturally produced continuous speech. Successful encoding of acoustic features is fundamental for language processing and comprehension. Studies show that cortical tracking of at least some acoustic speech features is already robust in the first year of life. However, it is unclear whether bilingual infants exhibit enhanced cortical tracking of non-native languages compared to monolingual infants, consistent with the idea of having a "bilingual advantage" as suggested in prior research. To investigate this, we recorded neural responses from 11-month-old English learning monolinguals, English-Mandarin learning bilinguals, and two mature comparison groups of English monolingual and English-Mandarin bilingual adults, while they listen to naturally produced, continuous, infant directed speech using electroencephalography (EEG) in three conditions: English, Mandarin, and Vietnamese. Stimuli were presented at an overall level of 70 dB SPL in a sound-attenuated booth. Using a combination of machine learning and linear modeling (i.e., Multivariate Temporal Response Function approach), we analyze the EEG signals using a multivariate encoding model consisting of acoustic features including envelope, envelope derivative, word onset, and phoneme onset. We hypothesize that both bilingual adults and infants will exhibit enhanced encoding of acoustic features in Vietnamese compared to monolingual adults and infants, indicating bilingual advantage in processing a third language. Additionally, we anticipate the bilingual advantage to be more prominent in infants than adults. These findings will contribute to the understanding of how bilingualism influences neural encoding across different languages and provide neural evidence of bilingual advantage in processing and acquiring a third language. I participated in study design, recruitment, data acquisition and analysis.