Chronic pain affects 30% of children and adolescents, putting them at risk for physical and psychological impairments. Given poor access to pediatric pain care, psychological treatments such as cognitive behavioral therapy are more accessible through digital health interventions. One intervention our lab created is Web-based Management of Adolescent Pain (WebMAP). In this mobile app, youth with chronic pain develop pain management strategies by progressing through interactive modules. Although digital health interventions like WebMAP are transforming accessibility to chronic pain care, limited information is available to understand how best to enhance youth engagement in a mobile app or whether there are differences in engagement among youth with different background characteristics. Our project aimed to 1) identify barriers to engagement with WebMAP among subgroups defined by user demographics and 2) develop recommendations for enhancing engagement with WebMAP and extending its reach to target underserved populations. In a previous cluster randomized controlled trial, participants assigned to the WebMAP intervention were 73 youth aged 10 to 17 (84.3% female) with various chronic pain conditions. We analyzed the app metadata to assess module completion rates across various subgroups, including groups defined by age, race, sex, and annual household income. We also coded interview and questionnaire responses from users to identify app preferences and areas of improvement. Results indicated that although users liked WebMAP’s design and content, they suggested changes to its display and features. By viewing our findings on usage and perception through a health equity lens, we summarized the accessibility of WebMAP and outlined recommendations to enhance its cultural relevance to different groups. Ultimately, our research identified factors that impacted engagement with WebMAP and offered valuable insight into optimizing digital health psychological interventions for chronic pain management. Moving forward, we hope to apply these findings as we develop the next release of WebMAP.

Skin is a densely innervated sensory organ, populated with various somatosensory receptors that help us perceive touch stimuli. Frequent injuries to the skin cause axon damage and lead to axon degeneration. Degenerating axons leave behind debris that must be cleared before reinnervation can occur and sensation is restored. We use adult zebrafish as a model to study injury and innervation because they have homologous cells and structures to humans and have transparent skin, allowing for high-resolution microscopy. Previous experiments in our lab revealed that skin-resident immune cells known as Langerhans cells (LCs) use highly motile protrusions to engulf axonal debris in the zebrafish epidermis. How are these dynamic protrusions regulated at a molecular level? Calcium signaling regulates phagocytosis and cell motility in other cell types, but the role of calcium signaling in LCs is unstudied. Through scale pluck assays and fluorescent microscopy, I have established a model for monitoring calcium signaling in LCs. I observed transient calcium flashes in LCs that varied in frequency, intensity, and subcellular location. I perturbed calcium flux using a calcium chelator and observed decreased flash intensity and protrusion length in LCs, suggesting that calcium signaling is required for protrusion dynamics. To investigate how calcium signaling affects engulfment, I imaged calcium flux in LCs during phagocytosis of apoptotic cells after laser injury. In contrast to the transient flux normally observed, LCs exhibited sustained high concentrations of cellular calcium during corpse engulfment. Because of the effects of perturbed calcium flux on intracellular calcium and protrusion motility, I hypothesize that perturbing calcium concentrations will inhibit LC phagocytosis. Identifying the molecular mechanisms underlying debris removal, such as calcium signaling in LCs, is relevant to understanding skin repair and disease states in which axon homeostasis is altered, including diabetic and chemotherapy-induced peripheral neuropathies.

From previous clinical cases, doctors found that children with Leigh syndrome are sensitive to volatile anesthetics. Leigh syndrome can be caused by defective Ndufs4, a subunit in complex I of the electron transportation chain. Previously, we used the Ndufs4 knockout C57Bl/6 mice to model Leigh syndrome and noticed that individuals with Ndufs4-knockout astrocytes go under anesthesia at a normal concentration, but require a lower concentration to emerge. We also found that isoflurane inhibits the effect of norepinephrine in astrocytes. Since norepinephrine is associated with individual emergence from an anesthetized state, this might help us to understand the mechanism behind our observations of the astrocyte specific Ndufs4 knockout. We furthered our study by assessing the role of gliotransmitters in the mechanism of mitochondrial disease and volatile anesthesia. Gliotransmitters are chemicals released by glial cells and used by astrocytes to modulate neuronal information processing. A pilot study was conducted in vitro in the wildtype mice assessing the release of different gliotransmitters, including cyclic adenosine monophosphate, adenosine, adenosine triphosphate, gamma-aminobutyric acid, serine, glutamine, and glutamic acid. Previously we found that the level of cAMP increased significantly after the norepinephrine treatment. Thus, we expect to see a larger acceleration in cAMP concenration in knockout mice, compared to wildtype. I am continuing this project studying the effect of the gliotransmitters in astrocyte cell cultures from the Ndufs4-knockout mice, to compare to the wildtype counterparts. Each astrocyte culture will be randomly assigned to a treatment: with or without norepinephrine. The amount of each gliotransmitter released per unit of protein is measured. I will analyze the data generated and compare to the wildtype data. Future study includes the assessment of the relationship between the metabolites within astrocytes, and the response to isoflurane and norepinephrine. This study would accelerate our understanding of the mechanism of volatile anesthesia, as well as the mitochondrial disease, which in turn will benefit numerous patients with safer and wiser treatment plans.

High throughput single cell/nuclei RNA sequencing (scRNA-seq/snRNA-seq) has been used to characterize the gene expression at the cellular level in disease control studies. Differentially gene expression analysis aims to emphasize the biological variation between samples without unwanted technical variation. Batch effect correction is occasionally performed where cells from each individual sample are treated as being generated from a batch. We analyzed snRNA-seq data from ApoE neutral and detrimental mouse models of Alzheimer’s disease (AD) to test whether batch correcting the data using all cells from each individual biological sample as representing a batch would result in 1) loss of disease relevant associations, 2) loss of biologically relevant cell types, and 3) reduced association between cell types and phenotypes. We performed scRNA-seq analysis of seven samples from two ApoE genotypes using Seurat workflow applying Harmony batch correction, using each sample as a batch, and without correction. Since we measured two disease-related phenotypes in mice from the two genotypes, we asked whether cell cluster membership associated with genotypes are also associated with unit changes in brain-diseases related phenotypes. After applying individual sample batch correction, we found differences in number of cell types (clusters) before and after batch correction, we showed loss of cell types/clusters specifically from the detrimental genotype, shrinkage of the differences in cell cluster membership across samples and reduced association between cell type membership, genotypes and phenotypes. In conclusion, batch effect correction should be applied consciously based on the experimental design to avoid over correction of biological variability and an appropriate design should help to avoid unwanted technical variation.

Lck is a lymphocyte-specific tyrosine kinase involved in T cell activation, which is essential for the human immune response. Upon antigen engagement with the T Cell Receptor (TCR), Lck phosphorylates the CD3ζ chain of the TCR, transducing intercellular signaling that activates T cells. Recent studies have demonstrated that Lck’s phosphotransferase activity is not only important for T cell activation, but that Lck also plays a critical role in scaffolding the interaction between the phosphorylated CD3ζ chain of the TCR and the kinase ZAP70 using its regulatory domains. Lck’s phosphotransferase activity has been shown to be toxic to yeast, with increased activity correlating with decreased yeast-growth rates. Using a yeast growth-based deep mutational scan (DMS), we calculated the activity scores of ~5,000 single amino acid variants of Lck’s kinase domain. Through this DMS, we identified all positions on the kinase domain that are amenable to substitution without perturbing kinase activity. In particular, we focused on positions where we could install cysteine residues on the kinase domain without perturbing kinase activity. Currently, we are expressing these cysteine variants in primary T cells, and applying parallel chemoselective profiling methods to quantify changes in the electrophilic reactivity of the cysteine side chains upon TCR stimulation. The expected changes in alkylation of the cysteine side chains upon TCR stimulation will provide insight into changes in the conformational flexibility of Lck, accessibility of the substituted residue sites, and intramolecular protein-protein interactions (PPIs) of Lck upon TCR stimulation. Ultimately, this insight into the conformational dynamics of Lck can be applied to deepen our understanding of basic immunology and the T cell activation signaling cascade.

Research suggests that recruitment, the number of fish entering a population in a given year, is influenced by the environment. However, environmental drivers are not currently used to refine the recruitment estimates of most statistical models used in fisheries management (hereafter, population assessment models). This is increasingly relevant as fish populations experience long-term productivity shifts due to climate change. One major goal of the National Oceanic and Atmospheric Administration (NOAA) in recent years has been to incorporate environmental drivers into population assessment models. This is achieved by using time series data of environmental drivers to inform model estimates. One ongoing challenge is determining which environmental drivers have potential to improve model estimates. In this project, we aimed to determine how correlated an environmental driver time series must be to historical recruitment deviations to improve key model estimates – recruitment deviations and population depletion – in recent years for a range of species. We used R to simulate mock environmental driver time series with varying correlation levels to recruitment deviations by randomly sampling data from a normal distribution. We then compared errors in estimates between population assessment models fit with and without the simulated environmental data. Our results suggest that the more correlated an environmental driver is to historical recruitment deviations, the more accurate estimates of both recruitment deviations and population depletion become. However, our results also reveal that the correlation level necessary for environmentally-driven models to consistently produce more accurate estimates than the original model varies across species. These correlation thresholds are also generally higher than observed correlations between recruitment deviations and environmental drivers in actual fish populations. We suggest fisheries scientists run similar simulation experiments to determine which correlation levels have the potential to improve population assessment models for their target species.

Current sociopolitical climates for LGBTQ+ individuals in South Asian countries are hostile, and South Asian communities within America will often reflect similar beliefs. Likewise, there might be an unfavorable atmosphere for LGBTQ+ individuals at faith-based institutions. As such, for South Asian college students enrolled in Christian institutions, their religious context and South Asian cultural practices and values might combine to shape their attitudes towards LGBTQ+ people. To explore this understudied topic, I utilized semi-structured interviews with South Asian students at a Christian institution. Thus far, I have interviewed four participants and intend to complete at least eight interviews to meet the qualitative analysis standard. Preliminary examination reveals our participants’ tendency to avoid conversations about the LGBTQ+ community, and they connected this tendency to South Asian culture. In contrast, they noted that LGBTQ+ topics were actively discussed on their Christian campus. Our study hopes to expand on the research surrounding intersectional identities and their impact on the individual.

Peptide cancer vaccines have had limited clinical success despite their safety, characterization, and production advantages. We hypothesize that the poor immunogenicity of peptides can be surmounted by delivery vehicles that overcome the systemic and cellular drug delivery barriers faced by peptides. We introduce Man-VIPER, a self-assembling, pH-sensitive, mannosylated polymeric peptide delivery platform that targets dendritic cells in the lymph nodes and facilitates endosomal release of antigens through a conjugated membranolytic peptide melittin. We evaluated polymers with both releasable (Man-VIPER-R) or non-releasable (Man-VIPER-NR) D-melittin. Both Man-VIPER polymers exhibited superior endosomolysis and antigen cross-presentation compared to non-membranolytic D-melittin-free analogues (Man-AP) in vitro. In vivo, Man-VIPER polymers demonstrated an adjuvanting effect, induced the proliferation of antigen-specific cytotoxic T cells and helper T cells compared to free peptides and Man-AP. Remarkably, antigen delivery with Man-VIPER-NR generated significantly more antigen-specific cytotoxic T cells than Man-VIPER-R in vivo. As our candidate for a therapeutic vaccine, Man-VIPER-NR exerted superior efficacy in a B16F10-OVA tumor model. These results highlight Man-VIPER-NR as a safe and powerful peptide cancer vaccine platform for cancer immunotherapy.

The Stimulator of Interferon Genes (STING) pathway is a promising target for cancer immunotherapies. However, STING agonists have poor cell membrane permeability and also off-site toxicity that limit their therapeutic application. Our motivation is thus to develop a novel method for the delivery of non-cell membrane-permeable STING agonists. We propose incorporating the STING agonist in a self-assembling polymer, which then transports the drug into the cytosol, thereby avoiding membrane permeability issues. The first step is the incorporation of the STING agonist into Man-VIPER, a cytosolic drug delivery system developed by the Pun Lab. Man-VIPER mediates cytosolic delivery via endosomal escape, releasing the STING agonist cargo into the cytosol. Currently, I am working on synthesizing and purifying the small molecule STING agonist called SR-012. I tested many possible reaction pathways to make SR-012, which is unavailable commercially and has only been previously synthesized once. I developed a novel alternative reaction pathway and purification that works consistently. I am also currently synthesizing the polymer building blocks for the Man-VIPER copolymer that will deliver SR-012. After the optimized drug-polymer complex is completed, I will work with collaborators to quantify the max tolerated dose. We hope to demonstrate that specificity is improved through lower toxicity, allowing for a higher maximum tolerated dose than my control, the membrane permeable STING agonist SR-717. Next, I will assay the immunotherapeutic effectiveness of Man-VIPER delivered SR-012 using in-vivo tumor models.

Alzheimer’s disease (AD) is a progressive neurological disorder associated with protein deposits in the brain, and causes a decline in memory, cognition, and social skills. There is no cure for AD, and as the disease advances, complications lead to incapacitation and death. Insulin-like growth factor 1 receptor (IGF1r) is a transmembrane receptor that responds to the hormone insulin-like growth factor 1, and is associated with healthy aging. An IGF1r variant in exon 6 (Arg-407-His) was found to be enriched in centenarians compared to shorter-lived relatives. Characterizing the role of this IGF1r variant in the development and progression of AD would provide information into a novel therapeutic target. Older-aged C57BL6 mice expressing a CRISPr-generated human Arg-407-His variant of the IGF1r gene were intravenously injected with a neurogenic AAV vector, consisting of the pathogenic components Aβ42 and p301L tau, or SHAM. After 3 months, mice with the IGF1r variant found escape holes more quickly than mice without the variant, using a spatial navigation learning task. Following humane euthanasia, brains were collected and fixed in formalin for immunohistochemistry (IHC). Brain sections were stained with biomarkers for Aêžµ42, ptau, and inflammatory microglia, and digitally imaged for quantitative analysis. Brains from IGF1r variant mice showed a decrease in expression of Aêžµ42 and ptau in line with improved cognition, and an increase in inflammatory glial cells. These preliminary observations help establish IGF1r Arg-407-His variant mice as a model to better understand the role of IGF1r in AD neuropathology, and provide insight into new potential therapeutic approaches for older human patients with AD.

Studies have revealed evidence that some humans have increased midfoot dorsiflexion, or a midtarsal "break", which decreases rigidity of the arch structure during gait. This "break" suggests that more of the sole of the foot is in contact with the substrate in terminal stance, effectively shortening the lever arm during propulsion. This prompts us to wonder: does arch stiffness predict the magnitude of lever arm distance or the forces of peak braking and peak propulsion? The Arch Indices (AI) of 77 people were calculated in dynamic conditions. Locations of the center of pressure (CoP) of the foot at heel strike, peak braking, and peak propulsion were recorded using a pressure-sensitive mat (RSScan International, Olen, Belgium). I calculated the distance from the heel strike CoP to those of peak braking and propulsion. I normalized peak propulsion and peak braking forces by dividing each by weight. All statistics were calculated in STATA (Statacorp, College Station, TX, VBe17). Dynamic AI has a relationship with lever arm distances for heel strike CoP to those of peak braking and propulsion (p=0.042, p=0.005, respectively). Dynamic AI did not demonstrate significance with peak braking and peak propulsion forces (p=0.758, p=0.194, respectively). My results demonstrate an association between AI and the distance of the functional lever arms, suggesting the presence of a midtarsal “break” as flatter feet exhibited shorter lever arms. Midfoot dorsiflexion and loss of rigid arch structure associated with the "break" would result in a decreased lever arm for propulsion, since the anterior midfoot and forefoot are in contact with the substrate at a later point during gait. This analysis has broader implications for foot morphology and midtarsal “break” research. Future work can utilize infrared markers and force plates to create models of the human foot with ground reaction force (GRF) data.

 Subduction zones are regions where two tectonic plates converge, and one is forced underneath the other. They are the primary driver of plate tectonics, and a source of major earthquakes. These earthquakes occur at shallow depths where plates slide past one another quickly, but at greater depths, with higher pressure and temperature (PT) conditions, rocks behave more ductilely. The transition zone between brittle and ductile regions hosts slow slip events (SSEs), which accommodate motion between the plates during events that last months to years, as opposed to the seconds over which earthquakes occur. SSEs are an important mechanism for accommodating plate motion at depth, thereby affecting the occurrence of larger devastating earthquakes, but currently the processes which facilitate SSEs and the rocks that host SSEs are not well understood. By studying rocks formed in and ancient subduction zone from Santa Catalina Island in California, we can learn more about our own modern counterparts, as the rocks record the conditions where slow slip may have occurred. Using optical petrography and data from x-ray spectroscopy, I examine thin sections of epidote-rich blueschist from SSE PT conditions. Optical petrography allows me to characterize the mineralogy of this rock, and the x-ray data provide the chemical compositions of individual minerals. Using image-analysis software, I will pair these two datasets to estimate the bulk-rock chemical composition of my sample. These data will allow me to constrain the starting material (protolith) of this rock before it was metamorphosed in order to determine if it was originally a sedimentary or basaltic component of the subducting oceanic plate. Doing so will improve our understanding of the way in which rocks at those pressure-temperature conditions deform and chemically change to create the context in which modern SSEs occur.

Quantum computing presents a much different challenge than controlling a classical computer. The main difference lies between the methods of control of quantum bits (qubits) and classical bits. Unlike their classical counterparts, which have discrete states defined by voltages, qubits are defined by some quantum state and are not so easily manipulated. Nitrogen-vacancy (NV) centers are defects that occur in diamond that can be utilized as a two-level system, showing promise for a qubit platform. To use NV centers in this manner, microwave excitation must be applied to control their spin states. An external magnetic field is applied to the NV center to lift state degeneracy, allowing us to define a two-level system. Applying resonant microwave excitation causes oscillatory population transfer between the states defining that two-level system. To apply this excitation, we will construct an antenna that is capable of aligning a microwave field to the NV center. These antennas will be designed to maximize the Rabi frequency at lower input powers. To do so, we maximize microwave field strength and optimize power transmission through antenna geometry and impedance matching. Positioning of the antenna on the sample close to the NV center assures the high field strength excites it. We make use of field simulation software to model and simulate various antenna designs. Select designs are chosen for initial fabrication runs on copper, and are used to verify that simulated results align with physical testing. Once that is verified, we finalize designs for fabrication onto diamond via photolithography. We will physically test the antennas by characterizing them using a vector network analyzer. We expect to be able to fabricate and characterize a broadband gold antenna on a diamond sample that matches simulations, and use it to demonstrate spin manipulation through resonant excitation of an NV center.  

Every winter, those residing within the eastern half of the U.S. are slammed by powerful storms that pound cities with snow – costing millions of dollars in damage, halting travel, and impacting businesses and schools. Yet, like a thumbprint, each of these winter storms are unique and they can possess a range of tracks, structures, and intensities. This research project aims to provide a greater understanding of the development of such storms and the causes of precipitation variability within them by focusing on a single storm that hit the Midwest on 17 February 2022 as a part of a research flight conducted during the Investigation of Microphysics and Precipitation for Atlantic Coast-Threatening Snowstorms (IMPACTS). I analyzed data from multiple sources to explore the large scale environmental conditions and the fine scale precipitation structure. The environmental conditions included a strong thermal contrast, or a frontal boundary, that provided the lift needed for precipitation as well as sub-freezing surface temperatures that allowed for precipitation to fall as snow. Analysis of the vertical cloud and precipitation structure from radar data collected during the flight revealed regions of higher reflectivity where snow was heavier than in other portions of the storm. Microphysical properties, such as particle sizes, shapes, and ice water content were different within the region of high reflectivity than outside it. Additionally, the strengthening front at the 700-hPa pressure level coincided with the localized region of enhanced reflectivity observed by the ground-based and airborne radars. The high-resolution radar and microphysics data collected by the IMPACTS airborne instruments are used to help identify precipitation-defining processes within these storms, and ultimately will increase the accuracy of snow prediction and remote sensing of snowfall from space-borne instruments.

Sickle cell disease (SCD), the most common genetic blood disease in North America, is characterized by recurrent episodes of acute severe pain due to blockages of red blood cells. In past studies, digital cognitive-behavioral interventions have been shown to be beneficial in other chronic pain conditions through teaching pain-management skills. Our research investigates the effectiveness of iCanCope SCD (iCC-SCD), a web and mobile-based pain-management program for SCD pain in youth ages 12-18 years. The final enrolled sample for the study was 137 participants, of which 26 participants were excluded because they did not complete pre-treatment assessments. Thus, the final sample consisted of 111 adolescents (107 caregivers), 54 randomized to Education control, and 57 randomized to iCC-SCD. The iCC-SCD program includes modules teaching coping strategies, symptom and goal-tracking, and peer-based social support, while the attentional-control contains static education about SCD. The efficacy of the program is determined through self-report scales at pre-treatment, post-treatment (2 months), and follow-up (6-months) periods, targeting the primary outcomes of adaptive coding, pain reduction, and pain-related disability. A statistically significant effect of treatment group (iCC-SCD vs. Education) on change over time in average pain intensity from baseline to 6-month follow-up was found. While most youth engaged with the program (40/57, 70%), the overall usage was highly variable. Therefore, I will explore the differences between participant website and app engagement from this study, feedback on why participants may or may not have been able to complete the iCanCope program and determine areas to enhance engagement. The information collected in this analysis can help to improve web- and mobile-based interventions for not only youth coping with SCD pain but also those with other pain-related conditions, given the flexibility and universality of cognitive-behavioral frameworks.

The widely accepted theory for how Earth’s moon formed begins with an impact to Earth by a Mars sized protoplanet. This impact creates a disk of debris around the Earth, and the moon is subsequently formed as the debris collides and coalesces. Previous studies have modeled the debris disk in a hydrodynamic environment, but results have found that moon formation is uncommon. This research project models the debris disk after it has cooled and condensed into a collection of solid particles using an N-Body simulation. These simulations are run using the ChaNGa code developed by the University of Washington’s N-Body shop and processed on the Hyak supercomputer. Simulations begin with an initial conditions file that we generate with different parameters, including particle resolution, angular momentum, and coefficient of restitution. From each initial conditions file the simulation runs time-progressions that model each particles position and velocity at every time-step, calculating the collective gravitational forces between all of the particles and recording any collisions that occurred. I analyze the data using plots detailing the eccentricity, mass, and semi-major axis of objects that form. Previous results from work I have completed on this project appear to show robust moon formation, with roughly lunar sized objects forming around the Roche limit. Future work on this project will include running more simulations with similar initial conditions to determine how common moon formation is, as well as analyzing the data using plots of semi-major axis vs. number of collisions/bodies accreted to determine if there are specific regions of the debris disk where the moons’ mass is originating from. While previous studies have found that a moon is only formed at very specific impact angles and sizes, this study looks to see if the moon formation mechanism may be more robust when modeled using N-Body simulations.

In the United States, Black adolescent (10-19 years old) girls have the highest prevalence of pediatric obesity. Despite public attention and public health interventions on obesity, current research has yet to evaluate what weight management recommendations are prevalent in mediums most frequently used by Black adolescent girls. Previous scholars show that Black adolescents primarily consume content on Instagram and Tiktok compared to all other social media platforms. Our research seeks to compare the weight management recommendations present in Instagram and Tiktok with those of public health organization websites. Using the rhetorical themes identified, we move to assess the feasibility of health messaging from public policy organizations compared to social media platforms. Ultimately, we seek to evaluate the efficacy of the recommendations based on the availability of the infrastructure accessible to Black adolescents. We analyze weight management messaging on Instagram, Tik Tok, and public policy organization websites. These primary sources are excerpted through simple random sampling and coded for common weight management themes. We seek to determine the feasibility of such messaging as well as points of convergence and divergence between the platforms. We hypothesize that the weight management themes found in social media will not demonstrate consistency with the recommendations made by public health organizations. In addition, we predict that neither the social media nor public policy weight management recommendations will be attainable for Black adolescent girls when assessing their feasibility in relation to the environmental and socioeconomic resources available to them in the United States. In conclusion, there are insufficient resources for Black adolescent girls to make informed weight management decisions and available resources are not effectively communicated.

Plasmodium vivax (P. vivax) is the most widespread malaria species. P. Vivax can lay dormant in hepatocytes and cause recurring malarial infections. Tafenoquine (TQ) is an antimalarial drug approved by the FDA in 2018 for the radical cure of P. vivax. Unfortunately, 8-aminoquinolines like TQ cause hemolytic anemia in G6PD deficient patients. Due to this contraindication, genetic G6PD screening is required before TQ administration. These additional tests pose a significant challenge for broad administration in resource-constrained countries. Our polymer prodrug conjugates (drugamers) are designed to improve TQ delivery to the liver and reduce red blood cell exposure. Using RAFT polymerization, drugamers can be optimized for delivery efficiency and avoidance of hemolytic anemia. Polymer architectures can be further enhanced by utilizing a variety of enzyme cleavable linkers, monomers, and receptor binding cofactors. We previously demonstrated increased liver-to-blood area under the curve ratios in mice using a variety of different polymer architectures. Mice received an IV or subcutaneous dose of drugamer and relevant organs were harvested at previously determined time points up to 48 hours. Pharmacokinetic profiles were created using a plate reader fluorescence assay and tandem LC/MS/MS with tissue samples. These drugamers represent an ongoing effort to iteratively improve our drug-polymer design. The improvement in TQ delivery through our drugamer vehicles could allow for more liberal administration guidelines, completely removing the need for G6PD testing for the treatment of P. vivax.

Climate change has resulted in environmental changes that pose direct and indirect challenges to marine organisms. One such organism that has been widely studied is the common murre (Uria aalge), which has experienced decreased reproductive success and adult survival in response to climate change-induced ocean warming. Because common murres molt flight feathers relatively synchronously, they may be especially vulnerable to environmental stresses, such as the challenge this may pose on obtaining food. In this study, we explore the degree to which shifting environmental conditions impact adult survival of common murres at a physiologically sensitive point: flight feather molt. The data we used for this study include: monthly expert-verified observations of bird carcasses from the Coastal Observation and Seabird Survey Team (COASST) citizen science program collected from the outer coast of Northern Washington south down to Humboldt, California; significant wave height data from the National Data Buoy Center (as a proxy for storminess); spring transition date (from the Columbia Basin Research website) as an indication of annual production potential; and Bakun upwelling index (from the Pacific Fisheries Environmental Laboratory website) as a measure of upwelling strength and production potential. We annualized both the bird and environmental data over the years 2003-2021. For this project, my role was to conduct statistical analyses using the statistical program R. I employed generalized additive mixed models to determine the relationship between the proportion of adult carcasses in molt and the environmental variables. To select models, I used Akaike Information Criterion corrected for small sample size (AICc). Our research will give insights into how the combined effects of physiological and environmental stressors may impact upper trophic seabirds as climate change continues to intensify.

From playing sports to driving a car, our brain constantly receives a barrage of complex visual information. Previous work shows that recognizing two things (e.g., words, animals) presented simultaneously is harder than recognizing one, suggesting that the brain has a limited ability to attend to multiple visual objects at once. In this project, I examine whether American Sign Language (ASL) experience affects sign recognition. Specifically, I hypothesize that signers and non-signers may differ in their ability to attend to multiple signs at once. Participants look at the center of a monitor where a reference letter sign appears, followed by test letter signs on both the left and right. On each trial, either the left or right sign is relevant (single-task condition), or both signs are relevant (dual-task condition). Participants respond, with a button, whether the relevant test sign(s) match the reference. The dual-task deficit (the difference in performance between single- and dual-task conditions) measures how well people can attend to multiple simultaneously presented signs. I compare the dual-task deficit across hearing non-signers, hearing fluent signers, and deaf signers. Previous work has shown a large dual-task deficit for written English letters and a small deficit for objects. I hypothesize that signers’ will have similar dual-task deficit to those of written English letters, while non-signers’ dual-task deficit will be small, similar to objects. So far, preliminary results from two non-signers support this second prediction. This project will reveal how the different sensory experiences and language demands of deaf and hard-of-hearing populations alter how their brains process visual information. Understanding these differences can help improve accessibility for these marginalized groups in public spaces. For example, it can help improve methods for presenting sign information to Deaf students to improve learning outcomes.

Chronic kidney disease (CKD) is an incurable, progressive condition that affects up to 700 million people globally. Progression of CKD currently leads to a deteriorating quality of life on dialysis, often resulting in terminal end-stage renal disease (ESRD) and increased risk of cardiovascular disease. A hallmark feature of CKD progression is epithelial-to-mesenchymal transition (EMT), a process by which kidney cells obtain malignant properties like increased mobility and resistance to apoptosis. Despite the pressing burden of CKD, current therapeutics like angiotensin-converting enzyme (ACE) inhibitors and angiotensin-receptor blockers are unable to halt the fibrotic progression of CKD. In recent years, epigallocatechin-3-gallate (EGCG) has been shown to be a promising candidate to inhibit EMT in kidney tubular epithelial cells (TECs). In this project, we conjugated EGCG to a low molecular weight polymer previously engineered at the Pun Lab for enhanced localization to the kidney TECs. In a TGF-β1 induced fibrosis model in human kidney cells, we showed that this polymer-EGCG conjugate (poly-EGCG) could diminish the RNA and protein expression of mesenchymal markers compared to untreated controls. We also confirmed that poly-EGCG was well-tolerated across a broad range of concentrations through in vitro cell viability assays. Finally, immunohistochemistry staining of mouse kidney samples injured with anti-glomerular antibodies displayed partial fibrotic recovery when treated with unconjugated EGCG. Future in vivo studies will aim to optimize the efficacy of poly-EGCG treatments compared to unconjugated EGCG treatments by assessing histology and urine samples for markers of kidney dysfunction. Through improved delivery of EGCG to the kidney TECs, this novel polymer-EGCG conjugate has the potential to halt the progression of EMT for future patients with CKD.

Cognitive decline with increasing age is an aspect of growing old. Age-related cognitive impairment (ARCI) entails the early stages of decline and is extremely common, affecting millions of older people. However, little is known about why some people have ARCI and some are cognitively normal at older ages. A recently characterized mouse model of naturally occurring ARCI, showing a distribution of affected and non-affected animals similar to older humans, was used to interrogate brain samples for transcriptomic profiles generated by RNA sequencing (RNA-seq). Hippocampal brain samples were collected from 22-month-old male and female C57BL/6 mice with and without ARCI (as determined by a spatial navigation learning task). RNA-seq was done by the NovoGene UC Davis sequencing Center. Preliminary data show a stronger presence of pathways of neurodegeneration and oxidative phosphorylation in the hippocampus of mice with ARCI compared to mice without ARCI. Detailed computational analysis will be done to investigate gene-expression quantifications using sequencing pipelines aligning Differential Gene Expression, KEGG orthology pathways, and Star RNA-seq read mapper in order to more accurately identify unique transcriptomic profiles in the brains of mice with and without ARCI. These findings will help identify genetic pathways that could be therapeutically targeted to ameliorate and possibly reverse the effects of ARCI, and provide insight into internal brain factors responsible for an increased risk in developing more severe conditions of neurodegeneration and dementia such as Alzheimer’s disease.

The negatively-charged nitrogen-vacancy (NV) center in diamond, consisting of a substitutional nitrogen atom and an adjacent vacancy defect, can provide an ideal platform for quantum computing. The NV center has many useful properties such as its optical stability and long spin-coherence times. However, due to the high refractive index of diamond, the light-collection efficiency of NV centers is very low. To solve this problem, I registered the positions of the promising NV center candidates in diamond and fabricated solid-immersion-lenses around the pre-selected NV centers. As a next step towards building a diamond quantum processor, here I present the construction of a cryogenic confocal microscope, which consists of a confocal imaging system with 515 nm laser and a 637 nm laser. Through optical pumping by the 515 nm laser and on-resonance driving by the 637 nm laser, this setup will allow for single-shot readout of the NV electronic spin state. This paves the way for our futhure implementation of quantum protocols and small-scale quantum algorithms for both pedagogical and research purposes.

An effective peptide-based cancer vaccine requires efficient intracellular delivery of antigen peptides to activate tumor-killing immune responses. However, the localization of peptide antigens during endosomal release to optimize the immune response remains under-investigated. The Pun Lab has developed the self-assembling Virus-Inspired Polymers for Endosomal Release (VIPER) that can induce endosomal escape of peptide antigens. The current VIPER formulation employs reducible disulfide bonds for antigen conjugation. We hypothesize that the controlled antigen peptides release mediated by endosomal proteases can result in more effective antigen presentation that leads to more potent tumor-killing cell responses. In this project, I replace VIPER’s antigen conjugation strategy with the pentafluorobenzyl (PFB) moiety (VIPER-NR) and introduce an enzyme-labile linker in the antigen sequence to determine the optimal peptide release kinetics and localization for optimal tumor-killing cell responses. I synthesized a library of enzyme-labile linkers for the ovalbumin antigen peptide of VIPER-NR. I utilized a dendritic cell cross-presentation assay to screen for linkers associated with that efficient antigen presentation in vitro. Subsequently, I conducted enzyme-mediated drug release assay and red blood cell lysis assay in vitro to evaluate the cleaved antigen profile and endosomal escape induction capacity of VIPER-NR with cleavable linkers compared to VIPER-NR. We expect early release of antigens during endocytosis with these modified antigen sequences in VIPER-NR in vitro and significantly elevated cytotoxic T-cell response against the model antigen in vivo. This project could improve the effectiveness of the VIPER system as a peptide vaccine delivery platform. It would also provide further understanding of optimal antigen release profiles for a better tumor-killing cell response in cancer vaccine applications.

Hydrogen sulfide (H₂S) impacts biological systems in multiple ways, including the arrest of aerobic respiration, and thus is mechanistically similar to cyanide. Unlike cyanide, however, H₂S can accelerate as well as end cell growth, including in plants, where it drives germination rates when administered in micromolar concentrations. However, the limited research to date leads to a need to better quantify and contextualize chemical composition changes in plant tissue following H₂S-induced plant growth. This study resulted in the ability to quantify the biophysical impacts of H₂S-induced growth in plants. The novel use of δ³⁴S and δ¹⁵N isotope ratios produced at the IsoLab in University of Washington represent the results of this sampling and its subsequent analysis. They may represent a new means to understanding the effects of H₂S on plant growth, including during the crucial phase of plant germination. The effects were observed on hypocotyl tissues from seedlings of Pisum sativum (pea), Phaseolus vulgaris (bean), and Zea mays L. (corn), all grown in hydroponic H₂S solutions, ranging from 0-100μM. These specific isotopic methods may allow comparison between modern and fossil material, because these isotopic species are known to have been preserved across a wide diversity of plant fossils. This novel application of these classic staples in the biochemical toolbox may have further implications for better understanding past events, because many major mass extinctions have now been linked to excess oceanic and atmospheric H₂S (compared to today), and may also, paradoxically, present new paths toward increased crop yields.

The association of eukaryotic DNA with histone proteins serves not only to package entire genomes into the nucleus of a cell, but these histone-DNA functional units, called nucleosomes, are hubs for biochemical signaling that regulates gene expression. In the Chatterjee lab, we are fascinated by the transcriptional biology of the small ubiquitin-like modifier protein 3 (SUMO-3), a posttranslational modification that occurs on histones and has been correlated with reduced gene expression. Previous members have demonstrated that SUMO-3 stimulates the activity of transcriptionally repressive enzymes by binding with a scaffolding protein called CoREST1. Hence, my focus has been to understand the functional details of the SUMO-CoREST interaction, particularly how cancer-associated mutations in the SUMO-interacting motif (SIM) of CoREST1 affect its ability to bind SUMO-3. To answer this question, I started by using solid-phase peptide synthesis to prepare truncated CoREST SIM peptides bearing the mutations of interest. I then utilized these peptides, along with SUMO-3 enriched in nitrogen-15, for two-dimensional nuclear magnetic resonance spectroscopy. By comparing the chemical shifts of [¹⁵N]-SUMO-3 with and without the presence of each peptide, I could assess the effects of mutations on the proportion of bound and unbound species in solution. Of special interest was an acidic residue in the hydrophobic core of the CoREST SIM, which distinguishes it from canonical SIMs found in other proteins. Excitingly, my results indicate that substitution of this amino acid with lysine, a mutation found in gallbladder cancer, ablates binding. I observed a similar effect for other mutations in the hydrophobic core of the CoREST SIM. Using these results to guide studies with full-length CoREST in biochemical assays, my research will identify the effects of these mutations on downstream biochemical pathways that may be misregulated in human cancers.

Deep learning methods for protein sequence and structure generation have shown remarkable success in many design scenarios when combined with structure prediction networks such as AlphaFold2. Despite this advance, many design challenges such as de novo binder design still haven’t been fully solved. Diffusion-based models have demonstrated considerable success in image and language generation yet their application in protein design has not yet been fully explored. Recently, the development of a protein diffusion model called RoseTTAFold Diffusion (RFdiffusion) has shown significant success in protein design and enabled us to explore the challenging problem of designing protein binders. Here I demonstrate utilization of RFdiffusion towards generation of de novo binders to disordered major histocompatibility complex (MHC) peptides. Specifically, we took an MHC peptide from KrasG12D and used RFdiffusion to generate a diverse range of structures that can bind this peptide. To optimize the sequence of these structures we used ProteinMPNN. We used AlphaFold2 to predict the structures of these optimized binders in complex with the peptide and saw promising interaction metrics. Further, structure prediction of the designs in complex with Kras wild type (WT) peptide resulted in lower AlphaFold2 confidence metrics of the interaction occurring. This is a promising preliminary result that RFdiffusion can generate fully de novo MHC-mimics, which can differentiate between neoantigens and WT peptide. Many cancers are caused by a single point mutation such as KrasG12D, thus, designing protein binders with point mutant specificity is exciting as it allows for targeting of disease causing proteins over healthy WT proteins. 

Melusin, a chaperone protein expressed in cardiac tissue, induces a protective hypertrophic response in response to chronic mechanical stress. This protective hypertrophic response prevents the progression of cardiomyopathy into heart failure. In previous work done in wild-type (WT) and melusin knockout (melKO) mice, the absence of melusin was correlated with a hypertrophic response indicative of heart failure. I plan to investigate the biomechanical role of melusin in humans using human-engineered heart tissues (EHTs) created from human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) that lack melusin and their isogenic controls. EHTs are more representative of the human heart, making them an ideal model for studying the role of melusin in humans. I hypothesize that WT EHTs subjected to mechanical stress, i.e., high afterload, will outperform the melKO EHTs. To measure this, I will increase the stiffness of the EHT posts and measure contractile force. I have successfully differentiated high-purity WT and melKO cardiomyocytes from iPSCs, essential for creating healthy EHTs. I will cast both WT and melKO tissues on a bed of silicone posts that can be stiffened to varying extents to induce different amounts of mechanical stress on the cells. I will conduct a western blot on EHTs from all treatment groups to determine the level of melusin expression and examine the expression of Heat Shock Proteins (Hsp) 70 and 90, due to their coregulation with melusin. The EHTs that undergo mechanical stress are expected to express melusin and these results will work to establish whether melusin expression in humans is activated by mechanical stress. I will measure and compare the contractile force between the WT and melKO tissues. Improving our understanding of the role of melusin in humans can lead to further research into therapies and treatments for heart failure.

The Religious Land Use and Institutionalized Persons Act (RLUIPA) of 2000 was passed to protect the rights of incarcerated individuals in the US to freely practice their religion in federal prisons. However, even 22 years after RLUIPA placed a strict scrutiny standard on reviewing inmates; free exercise claims, community organizations have had to repeatedly advocate for civil rights violations in prisons. This project aims to figure out why the free exercise rights of religious minorities are still threatened in the face of strong legislative protection. I will do so first by determining whether there is a bias in RLUIPA decisions for claims made by majoritarian religions, leaving religious minorities with a lower likelihood of success. Then, I will analyze whether the difference in success rates can be explained by a plaintiff's access to resources. Most incarcerated individuals represent themselves in court as they often lack the financial resources necessary for professional legal representation. Pro se plaintiff's often lack the preparation and legal knowledge that goes into a successful court claim. I hypothesize that prisoners with access to more resources, whether it's financial or community-based support, will be more likely to have a successful free exercise claim. My data will be collected from RLUIPA judgements from 2000 to 2022 and compiled into a dataset that includes variables such as the plaintiff's religion, type of representation, and the outcome of the claim. I expect to find that religious minorities will have lower rates of success, and that they will be less likely to have private representation. I also expect that plaintiffs with private representation will have higher rates of success with their RLUIPA claims as compared to pro se plaintiff's. The findings of this research will reveal the difficulties that individuals face in prison because of their religious identity and speak to broader themes of the trust and authority that our political systems give to prisons at the expense of incarcerated individuals' rights. 

Political scientists have rarely considered the intersection of religious organizations and foreign policy. Currently, there is no literature that investigates why religious organizations decide to either support or oppose American wars overseas and why this support can vary within each organization across conflicts. I attempt to answer these questions by testing whether religions are more likely to favor wars with other countries that have smaller groups of same-faith adherents. The social psychology theory of in-group favoritism and an “open-door” incentive to potential proselytization provide a plausible argument for such a relationship. To test this, I analyzed three different denominations of Christianity with a significant presence in the United States and their public position toward specific military conflicts throughout U.S. history. I created an original dataset containing these public positions of support or opposition to a given war along with the existence or size of like-faith communities (i.e., how many of their own church members were present) in the exterior target state of the conflict. I then utilized a multivariate regression analysis to test my theory and determine the relationship between a church’s support for war and the size of same-creed presence in each target state. I expect to find a positive relationship between these two variables, supporting the theories of in-group favoritism and “open-door” proselytization. These findings will contribute to current literature on American public and religious interest group foreign policy attitudes and offer methods for anticipating faith group positions on future military operations.

The American prison system is notorious for the mistreatment and abuse of the incarcerated people left in their care. In the past ten years there have been large pushes for prison reform; yet, these programs have had varying results across states. Why is it that some states have a much higher frequency of sexual assaults in prisons then others? I expect to find that prisons in Democratic-controlled states will have lower rates of sexual assault then their Republican-controlled counterparts. Literature on racial and social justice initiatives point to several differentiating factors between the two parties that aids in explaining this issue. Democrats are more likely to support social welfare and the Black Lives Matter movement, because racial justice activism is important to the Democratic coalition and subsequently their voter base. Additionally, Republican leadership supports tax and social welfare cuts while prison reform depends on subsidization from the government to work. In order to test this theory, I gathered data on sexual assaults in prisons from each state and compared them against each other, controlling for certain factors such as age, population and prison demographic. I ran a multivariate regression to see if Republican-controlled states experience greater frequencies of assault. This study focuses on an underserved population in the United States which greatly deserves thought and attention. Additionally, learning what factors contribute to violence in prisons helps provide a framework for improvements in our judicial system. Through research my goal is to contribute new information to a critical issue that has so far been largely understudied.

International criminal justice is often criticized for being unfairly selective in prosecuting cases. For instance, all suspects brought before the International Criminal Court in The Hague have come from Africa. This is despite the fact that human rights violations have been reported in many other parts of the world during this same period of time. Although restrictions in time and resources limit international courts’ and tribunals' capacity to prosecute all meritorious cases, the criteria used by the Prosecutors of these courts to determine which cases to pursue and which to disregard remain largely unknown, compromising the legitimacy of international criminal law in its application. This raises the question of why international criminal cases were only initiated in certain cases and not in others, and against particular suspects? To address these questions, this research examines four of the most prominent theories on selectivity in the application of international criminal law, including state capacity, popularity (that the courts will single out people from countries that have a bad reputation in their global affairs while granting leniency to others), crime severity, and victor's justice (countries with powerful positions within the international arena are most likely to escape international scrutiny). Most empirical research examines a single theory in isolation and employs a case study approach. However, I compiled an original dataset that encompasses all cases of international criminal law, from the Nuremberg Trials to the most recent cases of the International Criminal Court in order to test all four major theories systematically. I expect multivariate analysis to provide substantial support for the theory of victor's justice by showing that the power of a country correlates with the likelihood of international interference in the prosecution of international crimes committed in the country.

As of 2022 there are 103 million displaced people, of which 32.5 million are refugees, and 4.9 million are asylum seekers recognized by the United Nations High Commissioner for Refugees. As the number of displaced people grows, it becomes even more imperative that the United States offers asylum seekers a fair trial in court to pursue refuge in order to comply with the Universal Declaration of Human Rights, as well as to uphold strong ethical obligations based on foundational liberal values. However, today the right to seek asylum in U.S. immigration courts is undermined by the stark variation of adjudication rates between immigration judges. In this paper, I use original data from the Transactional Records Access Clearinghouse at Syracuse University to examine asylum adjudication variation systemically. Building from social identity theory, I hypothesize that asylum adjudication variation occurs due to the implicit bias exercised by judges based on their personal identities. I define implicit biases as unconscious learned biases that usually take the form of stereotypes and narratives that one automatically associates with particular people. Using a multivariate regression, I examine how the race, gender, political leanings, and religious affiliations of 314 judges correlate with the judges’ adjudication rate as well as the racial makeup of their asylum applicant pool. I expect to find that judges that have the characteristics of white, male, conservative and Christian will be more likely to deny asylum applications and I posit that this will have a disparate impact on asylum seekers who are black, brown, or are coming from majority Muslim countries.

Donald Trump’s presidency and its divisive legacy on the American government and its citizens continues to plague our society years after he has left office. It is crucial that policymakers and social scientists understand what values or grievances resonated with Trump’s voters and drove the rise of the Make America Great Again movement, in order to ensure that his voters do not continue to feel alienated from mainstream US politics. Through this paper I explore the electoral motivations of the US white working class, building on existing research that studies voting patterns and trends in previous presidential elections. I test two popular competing theories, pocketbook voting and social identity voting, and apply these theories to the context of the 2020 US presidential election. Pocketbook voting posits that voters respond to individual economic incentives and vote for who would benefit them economically, while social identity voting theorizes that individuals vote based on their subjective sense of belonging to an in-group and a desire to distinguish their group from others. I expect to find that white working class Trump voters were motivated by social identity voting, voting for Trump because of racial grievances and status anxieties. To conduct this study, I use a racial threat index and an economic well-being index, drawing from survey data to compare the social and economic grievances of white working class Trump voters. I expect to find that social identity voting, driven by factors such as status anxiety and racial threat, is a stronger electoral motivator than economic incentives, such as changes in wages or employment. Understanding such motivations lets us fight polarization and address the concerns of disenfranchised voters in future elections and in politics more broadly.

Year after year, catastrophic hurricanes rip through the mainland United States and its island territories. And yet, year after year, we hear stories of communities going without electricity or running water for months, disastrous temporary housing units and evacuations, and months-long wait times for immediate needs like food and medical care. Why has the Federal Emergency Management Agency (FEMA) failed to improve its distribution of disaster relief, even after three major reform bills? This paper seeks to provide an answer by investigating how corporate lobbying may be influencing FEMA’s allocation of relief funds between public and private actors. Considering the lucrative nature of post-disaster contracts awarded to corporations, they may use their lobbying powers to pressure the government to allocate more funds for private contracts and less to its Public Assistance (PA) program, which provides grants directly to state and local governments. FEMA’s preference for private contractors, despite their extensive history of performing inadequate work and defrauding the government, may be a significant part of the agency’s overall dysfunction. I conduct a multivariate regression analysis to determine the relationship between lobbying expenditures from top contractors and FEMA’s allocation of relief funds among public and private sectors between 2004 and 2021. I expect to find a positive relationship between these variables, suggesting lobbying does interfere with the delivery of post-disaster aid. Identifying the private sector’s influence in this area is critical to ensuring communities are properly supported after hurricanes and other natural disasters, which will only increase in frequency due to climate change.

 The spread of misinformation has increased rapidly in the last few years on many social media platforms. Our understanding of its effects, strategies, and influence is growing along with the information in real time. During political elections, we have seen that misinformation can become contagious and pose harmful threats to many aspects of our society and our political environment. How exactly does misinformation disseminate online, and are these social media posts used as a political strategy? To delve deeper into this study, I examine the relationship between false information online and legal cases that challenge election results. Using a mix of qualitative and quantitative methods, I analyze articles, data, and social media posts concerning the 2020 and 2022 elections in Maricopa County. With this data, I identify recurring narratives and influential political figures, using Python visualizations and codebooks for the empirical evidence found online. I anticipate that my findings reveal a pattern of legal cases being used to spread false political narratives that mislead the public about the voting process in Maricopa County. Since Maricopa County is the fourth most populous county in the United States, this study provides insight into how online users receive information about political elections and voting processes. I also anticipate that utilizing courts and election lawsuits can be an effective strategy to uphold and spread misinformation. Further research into other counties may demonstrate similar patterns and narratives with misinformation and U.S. elections.

Negative life events play an etiological role in posttraumatic stress disorder (PTSD) and depression. Social support and disclosure are associated with resilience following negative events. This secondary analysis examined the relationship between disclosure to specific individuals following negative events and current depression and PTSD symptom severity. Female-identifying undergraduate students (N = 101) were asked to describe "the most negative event" they experienced in the past year and identify individuals to whom they disclosed (e.g. friend, partner, parent, therapist, law enforcement, teacher, and university employee). Using path analyses, I examined whether: 1) for female-identifying undergraduates (n = 89), the identity of the individual to whom participants disclosed their negative memory was correlated with current severity of depression symptoms (Quick Inventory of Depressive Symptoms) and 2) for those who experienced a DSM Criterion A trauma (n = 12), identity of the individual to whom participants disclosed the event was correlated with current PTSD symptom severity (Posttraumatic Diagnostic Scale). Disclosure of negative events to professionals (law enforcement, teachers, university employees, and therapists) were associated, at a trend level, with higher depression severity (B = .56, p = .07); driven by an effect of disclosing to a therapist (B = .96, p = .04), while no strong association was found for disclosure to personal confidantes (friends, parents, and partners). Disclosure of Criterion A traumatic events to partners was associated with higher depression (B = 1.33, p = .07). These findings complicate literature on the mental health correlates of disclosure, which may be associated with increased depression and PTSD symptom severity following negative events, and emphasize the need for trauma-informed care for mandated reporters and others working with individuals who may disclose negative events. 

 Historical discourse has dangerously misrepresented and normalized the positioning of Black women in various states of vulnerability. Such discourses have then been embodied by society and further perpetuated within institutions- shaping the realities of Black women. This study explored how social services at a Seattle-based women’s shelter affect the experiences of Black women utilizing social services, while also understanding how Black women thrive utilizing such services. In this community-based qualitative research project, I used ethnographic methods to conduct semi-structured interviews with Black women utilizing social services at a Seattle-based womens shelter. I also conducted participant-observation methods to collect data on the shelter’s environment and the dynamic between Black women seeking services and employees providing services. Finally, I reviewed relevant theoretical frameworks such as intersectionality, the criminalization of poverty, and racial capitalism to assist in my analysis of how Black women seeking services at a Seattle women’s shelter have been positioned to experience vulnerability. Key findings include 1) social services have been socially constructed to sustain the experience of vulnerability within the Black community, specifically by Black women utilizing services; and 2) through various levels of gratitude, Black women find spiritual peace and grow new resiliences to systemic challenges. These findings draw awareness to the ways social services have deepened Black women’s dependency to them. Such awareness could serve as a critical framework for reshaping the way social services within the greater Seattle area assist Black women to promote prosperity, dismantle racist rhetoric, and aim to eradicate, not perpetuate, homelessness.

One way to obtain plasma is by using a Direct Current (DC) discharge. Plasma is an ionized gas, meaning the separation of positive ions and electrons in a gas. There are three main variables when it comes to a DC discharge configuration. A gas forms into a plasma in an isolated space of low pressure between 2 electrodes, a cathode and an anode. Voltage must constantly be applied across the cathode and the anode to maintain the plasma. The initial voltage needed to initiate the separation of electrons and protons in a gas to produce a plasma is called the breakdown voltage. Our study investigates the configuration of a DC discharge plasma and the correlation between electrode separation, breakdown voltage, and pressures in a DC discharge environment. We constructed an environment consisting of long oval glass tube housing an anode and cathode on each side. A vacuum pump is attached to the glass container to extract air to reduce pressure in our glass tube. To maintain an ideal pressure, we established a concealed air tube connected to our glass tube with a fine adjust valve to let air into our glass tube at the same rate as our vacuum pump extraction resulting in a stable low pressure in our experimental configuration. We designed and conducted a series of tests to investigate the properties of a DC plasma formation. Moreover, we wanted to establish evidence of the Paschen Curve, which relates the breakdown voltage and the product of electrode distance and pressure in DC discharge. We experimentally determined the optimum pressure and electrode separation distance product for plasma breakdown in air and Argon gas. DC plasmas can be utilized as sputter sources to deposit thin films for solar panels; characterizing the breakdown voltage is significant at low pressures and short spacing to control the sputtering rate.

Racial discrimination can heighten stress for people of color. The self-medication model suggests that people who experience racial discrimination consume alcohol to cope with negative emotions elicited from these experiences. Most existing studies have tended to correlate general alcohol use behaviors with recalled experiences of stressors such as racial discrimination. These methods are limited in understanding the causal effects of stressors on people’s intention to drink. There are very few experimental studies that considered the impact of racial discrimination and other daily stressors on drinking behavior among people of color. This study was aimed to examine the effects of racial discrimination and daily hassles (work, academics, and finances) on people of color’s intentions to drink. Using data from a between-groups experimental study, we used virtual reality technology to simulate typical interpersonal exchanges at a house party where alcoholic and nonalcoholic beverages were available. Participants were recruited from a private four-year university and the general community (N=184; M_age=23.9; 48.9% community adults; 47.8% women; 40.8% Latinx/Hispanic, 28.3% Asian American, 19.6% African American/Black, 11.4% other). They were randomly assigned to experience racial discrimination (n=141) or daily stressors unrelated to racism (n=43) in the simulation. Participants’ responses during the simulation were audio recorded; we coded whether they requested to drink an alcoholic or nonalcoholic beverage immediately before and after exposure to the stressor. Logistic regression will be used to assess group differences in intent to drink. We expected more participants in the racial discrimination condition to express intent to drink alcohol than those in the daily stressors condition. Results can illustrate the unique effects of racial discrimination on drinking intentions among young adults of color, and enhance knowledge on predictors of alcohol use and ways to mitigate negative drinking-related consequences.

Pacific salmon are both an ecological and cultural keystone species, essential to Pacific Northwest ecosystems, Indigenous Peoples, and the economy. Road salts (primarily NaCl), an increasingly common toxic constituent of stormwater runoff, adversely affect salmonids in vulnerable early life stages and mobilize heavy metals found in roadside soils. Dissolved copper (Cu), a ubiquitous nonpoint source pollutant, has been correlated with the impairment of olfactory systems in salmon; the negative effects of Cu decrease pre-smolt survival rates when compounded with NaCl. Our study investigates the concentrations of Cu and Cl^- in urban and rural watersheds home to Pacific salmon. We collected water samples from the Cedar River watershed and the Thornton Creek watershed in the greater Seattle area following storm events. Ion chromatography (IC) and inductively coupled plasma-mass spectrometry (ICP-MS) analysis were performed on our samples to determine the concentration of Cl^- and dissolved Cu, respectively. Our results show levels of Cl^- that have been shown to increase the mortality rate of Atlantic salmon alevin, whereas concentrations of Cu are undetectable in Cedar River and at very low levels in Thornton Creek. Annual returns of salmon runs across the Pacific Northwest are in rapid decline and although the cause is neither singular nor isolated, water quality and habitat degradation are leading concerns. Our results provide insight into how road salts and metal-containing road dust contaminate streams and rivers, altering water quality conditions for salmonids from fertilization to swim-up. We relate our findings to current ecological management practices and research examining the effect of salinization and copper toxicity on salmonids in critical early life stages.

Alzheimer’s disease (AD) is characterized by aberrant cleavage of Amyloid Precursor Protein (APP) leading to toxic β-Amyloid (Aβ) peptide aggregates. Presenilin 1 (PSEN1), and presenilin 2 (PSEN2) are intimately involved in this process. We found that alternative splicing of PSEN2 leads to the increased inclusion of a cryptic exon (part of the RNA that codes for a protein), in the typically non-coding intron 9 of PSEN2, titled exon 9B (PS2x9B), which has elevated abundance in individuals with sporadic AD. My aim is to uncover the link between PS2x9B and sporadic AD. I designed a PCR approach to measure the ratio of PS2x9B inclusion compared to non-mutated Wild Type (WT) in cDNA generated from parietal and temporal lobe tissue samples. I distinguished PS2x9B inclusion by amplicon size on a gel and measured the ratio to WT via band intensity using ImageJ. I found that the ratio of PS2x9B inclusion was significantly elevated in sporadic AD cases in the parietal lobe (p = 0.02) and the temporal gyrus (p = <0.001). Next I transfected SH-Sy5y cells with plasmids containing either WT or PS2x9B PSEN2 joined to a Flag-tag. I extracted proteins from these cells and control empty plasmids, and used Western blot to examine size differences between each plasmid’s protein products to research the impact of PS2x9B inclusion on PSEN2 protein levels. I hypothesize that inclusion of PS2x9B will lead to truncated proteins due to an early stop codon. Additionally, I am working to establish the consequences of PS2x9B on APP processing and Aβ cleavage. I expect that PS2x9B could cause either an increase in Aβ production or modify its length. These findings will allow us to expand our understanding of PSEN2 and alternative splicing in sporadic AD which could guide development of novel gene therapy treatments for AD.

 

Across 200 psychology undergraduate programs in North America, only 6% makes multicultural courses a requirement for majors. Research remains unclear about the effectiveness of a lower-level and content-general course on undergraduate students; critical understanding of cultural diversity in human Psychology. Our research goals are to examine the effects of multicultural instruction on students' appreciation for diversity, and whether race moderates this effect. Data came from undergraduate students enrolled in Research Methods in Psychology course at a mid-sized, predominantly White private university (N=133, Mage=19.83, 79.8% women; 75.9% White/Euro American). As part of the course, they completed surveys assessing variables such as enrollment motivation, ethnic identity, and self-reported importance of cultural diversity in Psychology in the first and last two weeks of the semester. A separate class is employed to be the control group covering contents in open science rather than cultural diversity as part of the quasi-experimental design. Regression analyses were conducted to test whether students in the cultural diversity condition reported greater appreciation for cultural diversity in their post-term assessment than students in the control condition, with pre-term assessment entered as a covariate. In addition, whether changes in appreciation for cultural diversity were greater among White students than students of color was also tested. Students in the multicultural condition scored higher on critical awareness of current limitations in diversity representation in psychological science at the end of the term than those in the control condition. Students across conditions did not differ in appreciation for the importance of cultural diversity considerations in psychology. Implications for how to integrate multicultural training in undergraduate psychology curriculum will be discussed.

Irritable Bowel Syndrome (IBS) is a common, chronic gastrointestinal disorder consisting of three common subtypes; IBS-Constipation (IBS-C), IBS-Diarrhea (IBS-D), and IBS-Mixed (IBS-M) — a combination of the two prior categories. IBS is associated with abdominal pain and changes in bowel habits. Patients may also experience other symptoms such as nausea, anxiety, and depression. It is unknown, however, how the gut microbiome impacts signs and symptoms. This study aims to compare the gut microbiota between adults with IBS and healthy controls and describe the relationship between symptoms and gut microbiota taxa. Fecal microbial communities are examined through stool samples collected from women aged 18 to 45 with 16 S rRNA gene sequencing analyses. We hypothesize that compared to healthy controls, those with IBS will have higher quantities of pathogenic gut bacteria such as Clostridium difficile (C. diff), Escherichia coli (E. coli), and other Firmicutes. Comparing the gut microbiome between those with and without IBS can improve screening of the disease, leading to improvements in symptoms, treatments, and quality of life.

The severity and prevalence of alcohol consumption and alcohol use disorder among the Hispanic/Latinx population in the United States have steadily increased for the past 10 years. With the rise of high alcohol use and the development of alcohol disorders, there is a focus on research for the white population, leaving a gap for group-specific factors and how there is minimal research on efficacy differences in population and risk factors. This literature review aims to examine previous research on Hispanic/Latinx populations with alcohol use and alcohol-related issues and to bring to light the gaps in the literature. The question this literature review is answering is “how do group-specific risk factors hold the Hispanic/Latinx population from seeking alcohol use disorder assistance, and how to increase treatment efficacy for this population?” A critical literature review was conducted using PsychINFO and Google Scholar for articles published in the last 25 years with the following keywords: Hispanic/Latinx, alcohol use disorder, interventions, and treatment barriers. Preliminary results indicate that many people in the Hispanic population face more attitudinal barriers than logistical ones. Additionally, results indicate that there is a lack of Hispanic representation in psychological research, thus, making it difficult for evidence-based intervention to be useful for these populations. Finally, results indicated potential adaptations to existing evidence-based therapies to make them more effective for Hispanic populations. Through the findings from the literature review, we found it necessary to address the risk factors and disparities the Hispanic/Latinx population faces and highlight the urgent need for more culturally adapted interventions and treatment plans. We will discuss future research directions concerning the need for more data from the Hispanic/Latinx populations on group-specific treatment efficacy.

The Developmental Origins of Health and Disease (DOHaD) hypothesis evaluates how the prenatal environment affects health after birth. The placenta is a multi-faceted organ that sustains life during human development and is key to evaluating the DOHaD hypothesis. Glial Cells Missing Transcription Factor 1 (GCM1) is a transcription factor that plays a critical role in placental development. Our goal is to understand the downstream effects of GCM1 on various genes necessary for placental development by evaluating gene expression after GCM1 knockdown. The BeWo choriocarcinoma cell line is a model of placental syncytiotrophoblasts cells which undergo a cell fusion process called syncytialization in the placenta to form multinucleated cells that help exchange nutrients. Previously, we knocked down GCM1 in full-term primary placental cells that spontaneously syncytialize and assessed gene expression using RNA sequencing. We identified 10 differentially expressed genes. Based on those findings, we hypothesized that GCM1 plays a greater role during early pregnancy leading us to repeat the GCM1 knockdown in BeWo cells. BeWo cells were treated with 20µM, 50µM and 100µM forskolin (FSK) for 48hr to induce syncytialization which was confirmed via qPCR of syncytialization markers GCM1 and Syncytin-2 and through fluorescence microscopy. GCM1 expression increased 3.15, 1.3, and 1.2 fold respectively after treatment with 20µM, 50µM, and 100µM FSK, whereas Syncytin-2 increased 78.1 fold after 50µM FSK treatment. We then performed an siRNA knockdown of GCM1 in unsyncytialized BeWo cells with two concentrations of siRNA (25nM and 50nM) for 24hrs and observed a 70% and 80% reduction in GCM1 expression, respectively. Next steps include optimizing the siRNA procedure for syncytialized BeWo cells and comparing these results to our previously conducted experiment. Overall, this will improve understanding of how GCM1 coordinates gene expression in the placenta during pregnancy.

Road salts are commonly applied as deicer during the winter months in Washington and can enter freshwater systems through run-off. The salinized waters can harm aquatic ecosystems. Zooplankton, specifically Daphnia, play a crucial role in providing food to other trophic levels in many ecosystems and could threaten the stability of these systems if they are unable to tolerate salinized waters. We hypothesized that Daphnia exposed to low levels of salt through several generations would better adapt to salinized water than an untreated population over time. Two populations of Daphnia were cultivated in the lab, one control with standard media and another with low levels of additional salt. We then placed these two groups in varying salt concentrations for one week. Under each condition, the survival rate of Daphnia was recorded. Heart rate was also observed as an indicator of physiological stress. We expect the pre-treated Daphnia to adapt to the road salt while the non-treated will have higher mortality rates. Understanding the impact of road salts on Daphnia can help us predict the possible effects on the overall health of aquatic ecosystems.

As part of cellular metabolism, all organisms produce reactive oxygen species (ROS), such as (O₂^– ) and hydrogen peroxide (H₂O₂). ROS can damage multiple cellular organelles and processes, disrupting normal physiology. Proteins, lipids, and nucleic acids can be damaged by ROS molecules, resulting in cell death. Also, several human diseases have been linked to an imbalance between ROS production and antioxidant defenses. Plants' chloroplasts and mitochondria contribute a substantial amount of ROS, as they are responsible for photosynthesis and aerobic respiration. In maize, Whirly ssDNA-binding proteins help to maintain the stability of the plastid genome. We recently demonstrated that maize seedling development is accompanied by increasing oxidative stress to the demise of chloroplasts, mitochondria, and their DNA. We also showed that maize seedlings grown in the light had higher levels of oxidants and lower levels of antioxidants than dark-grown seedlings. Here, our objective is to elucidate the role of Whirly proteins in oxidative damage. We isolated organelles from maize seedlings and performed oxidant and antioxidant assays for both wild-type and whirly-mutant plants. We found a significant difference in oxidation levels between wild-type and mutant plants. Our study should provide a better understanding of the role of ssDNA-binding proteins in oxidative damage to organelles.

Climate change coupled with inadequate infrastructure has disproportionately impacted people from underserved communities, as is the case for one village located in rural Alaska. Due to the increasing frequency of microbial water contamination exacerbated by temperature rise, this community experiences water insecurity. Previous research also illustrates that water samples collected from the Kuskokwim River, which is used for subsistence, had excessively high mercury levels compared to the regional baseline levels. Furthermore, our team's preliminary findings from informal interviews with community members indicate that their use of a rainwater cistern is well received by many households. However, community leaders have questioned the cistern's operation from September to March. When the temperature falls below zero degrees Fahrenheit in the winter, the cistern must be shut down to avoid damage to the system, and because it is not possible to collect water. Our work will develop and administer an online household-level survey to understand perceptions about local drinking water conditions. We implement principles derived from Indigenous research methodologies and Community-Based Participatory Research approaches. * Preliminary results gathered from conversations with the tribal council suggested a positive perception of the water quality from the installed water cistern, but we hope to quantify the water quality from survey data. * Our research focused on two areas: we will evaluate the utility of an online survey in collecting data from one remote community to understand their water needs and use the data to help guide future decision-making to improve the cistern operation under extreme temperatures. This work is part of a larger initiative to underscore the self-determination of under-resourced communities in Alaska, offering insights into Indigenous research methods and inspiring future design in the field. Note: * means revison per feedback, but will still need to get tribal approval as they approved for previous version as is

Illegal, unreported and unregulated (IUU) fishing violates the rights of Indigenous peoples to traditional fishing grounds; compromises food security of legitimate fishers and coastal populations; and facilitates human labor trafficking. Once caught, tails, fins, and heads are cut off for storage and transportation, which makes identification by phenotype impossible. Polymerase Chain Reaction (PCR) and electrophoresis can detect different tuna species but are time-consuming and require access to a fully equipped laboratory and trained personnel. Thus, I developed a novel biological assay and a 3D printable portable device to detect Ahi tuna, one of the species of interest among three tuna species. The biological assay consists of three steps: DNA extraction by crude lysis method; DNA amplification by Recombinase Polymerase Amplification (RPA); and detection by fluorescence using Clustered Regularly Interspaced Short Palindromic Repeats associated (CRISPR Cas) 12a. qPCR verified Ahi tuna DNA extraction by crude lysis method, generating 10⁶ copies of DNA per reaction comparable to the standard silica-based capture method. Nested qPCR and Tapestation verified RPA’s successful amplification of Ahi tuna DNA extracted by crude lysis method. Quantification of fluorescence by qPCR verified that one-pot RPA and CRISPR Cas12a reactions could generate up to 6000 Relative Fluorescence Units above the negative control within 15 minutes. The isothermal device keeps the samples at a constant temperature (37-42℃) for the RPA and CRISPR Cas12a reactions. I successfully demonstrated a <50 min sample-to-result assay to detect Ahi tuna DNA, and this protocol will be further adapted for testing other types of tuna including Bluefin tuna. By developing such a rapid and affordable isothermal biological assay and a point-of-need 3D printable portable device for tuna identification, I aim to contribute in helping individuals and ecosystems impacted by IUU fishing.

The protozoan parasite Giardia lamblia infects hosts via the ingestion of cyst-contaminated water. We recently identified EncystR, a 7-trans membrane protein at the cell surface that acts as a negative regulator of encystation and responds to encystation cues by internalizing. By following the localization of EncystR through an encystation timecourse we discovered a novel compartment of unknown function. The Giardia endocytic pathway was only believed to include hybrid endosome/lysosome compartments statically positioned beneath the plasma membrane. We suspect this newly identified compartment may be a lysosome-like compartment. Using EncystR as a marker for this compartment we found that the compartment is highly acidified based on the florescent reporter pHluoren2. Canonically endomembrane compartments are marked by specific phosphatidylinositol phosphates (PIP). Relevant to endomembrane compartments, PI(3)P marks endosomes and PI(3,5)P2 marks multi-vesicular bodies and lysosomes. To test if this newly identified compartment is evolutionarily related to lysosomes, we will generate reporters for these phosphoinositides. Namely, we utilized the FYVE protein’s PIP binding domain as a sensor for PI(3)P which is located on endocytic membranes, the pH domain of PLC delta as a sensor for PI(4,5)P2 or PIP2 which is necessary for endocytosis and membrane-based cytoskeletal protein regulation, and ML1N for PI(3,5)P2 which is localized to lysosomes. Additionally, a mutant variant of the ML1N protein which is incapable of binding to PIPs is utilized as a negative control. These protein sensors were fused to the fluorescent protein mNeonGreen and imaged via fluorescent microscopy. While the experiment is currently in progress, we hypothesize that the localization of PIPs to the novel compartment will likely feature PI(3,5)P2 due to the previously found acidic nature of the compartment, implying a lysosome-like functionality. This possibly novel or conserved compartment could give insight into the evolution of eukaryotic cellular organisms and could potentially offer treatment routes for Giardia.

Alcohol use disorder affects 5.8% of adults in the U.S., incurring a yearly cost of $249 billion annually. The repeated and prolonged use of alcohol creates a variety of physiological and behavioral issues. Acute alcohol withdrawal syndrome encompasses seizures, delirium tremens, and in some cases, death. The dependence and withdrawal cycles lead to neuroinflammation, worsened withdrawal symptom severity, and impaired neuromodulation. The striatum plays an important role in the chronic aspect of addiction. This project focuses on how alcohol alters microglial function in this key brain region. At the Neumaier lab, we found that alcohol withdrawal increased the expression of genes involved in the unfolded protein response (UPR) in striatal microglia, the brain’s immune cells. The UPR is activated when there is an increase of misfolded proteins in the endoplasmic reticulum; which contribute to impaired cell function. The UPR is a protective mechanism in moderation, but when left unchecked, has been shown to increase cell death. We predict that removing CHOP will lead to a decrease in withdrawal symptoms and regulate the need to consume alcohol. Knocking out CHOP involves breeding CHOP fl/fl |Cx3cr1 CreER /eYFPI mice. We observe changes in the offspring via emotional behavioral tests after a 5 week period of CIE (chronic intermittent alcohol exposure). A second cohort’s brains are analyzed using cryosectioning and immunohistochemistry (IHC). My role is preparing the brains for analysis. The first step is removing and perfusing, which prepares the samples for cryosectioning. This is my other specialty, which is freezing and slicing the brains. Lastly, I conduct analysis with IHC, and look for physiological changes in the striatum. We anticipate our findings will have a positive and large impact on treating alcoholism. Our hope is to reduce the stigma surrounding addiction, and instead offer compassionate and scientifically based care.
 

Giardia Lamblia is an intestinal parasite known for causing the diarrheal disease giardiasis in host organisms. It commonly infects humans and companion animals such as cats and dogs. Giardia’s life cycle is defined by its infectious cyst stage and proliferative trophozoite stage. GlRac is a small Rho-family GTPase which we recently determined to play a role in regulating encystation in Giardia, the process of transition from a trophozoite to a cyst. Guanine nucleotide exchange factors (GEFs) and GTPase activating proteins (GAPs) are key regulatory proteins of GTPase activity. The GEFs and GAPs that regulate GlRac activity are currently unknown, but four candidates for GAPs and three candidates for GEFs have been identified. We are testing these GAP and GEF candidates in Giardia with a two-phase approach. First, we are using the protein-protein interaction reporter NanoBit to determine if the candidate GAPs and GEFs interact with GlRac. We are verifying results for NanoBit assays, a split NanoLuciferase reporter that indicates protein-protein interaction. Nanobit can reveal interaction but not the localization of the interaction, so we are performing co-localization with deconvolution microscopy. Candidate GAP and GEF proteins are being visualized using a mNeonGreen fluorescent tag while GlRac is being followed using the halogenase (HALO) tag labeled with Janeliafluor 646. The microscope assays allow us to determine if and where GlRac co-localizes with candidate proteins. Subsequently we will test the role of the identified GAP and GEF proteins in encystation through transcriptional repression using CRISPRi and translational repression using morpholino-modified antisense oligonucleotides (MOs.) If the candidate proteins are truly GAPs and GEFs, the knockdown cells should not progress through encystation as normal. GAPs and GEFs are potential drug targets for treatment of Giardiasis, as well as important in understanding the process of encystation in Giardia.

Non-Human Primates (NHPs) have gained importance in neural engineering preclinical studies as their brains are relevant models to investigate and better understand neural function. The Yazdan lab uses optogenetics to control neuronal activity in order to develop stimulation-based therapies for neurological disorders such as stroke. These experiments require the implantation of various devices such as headposts, cranial chambers, electrode arrays, and optical windows. The use of head posts and cranial chambers requires customization to the curvature of the skull prior to implantation in order to prevent gaps that could introduce complications, including infection or decreased stability. Using an in-house method of NHP neurosurgery preparation that processes MRI data, we can develop 3D brain and skull models. This technique has allowed for chambers to be customized and implanted chronically in two NHPs. My project builds off of this implementation by creating custom chambers for future implantation surgeries and designing custom-fit headposts, which had never been done before. In order to design these components, I extracted the skull and brain using custom Matlab code which allowed for the craniotomy location to be determined and provided a footprint for the chamber and headpost implants. I then imported the skull extraction into a design software where the chambers and headposts could be built off of to ensure a tight fit to the skull. With the components designed, I will 3D print the brain, skull, chamber, and headpost to be assembled together. This platform will simulate the surgical and experimental setup, which provides a template for various experimental components to be modified and tested. It will provide a simple and affordable solution for neurosurgical planning, reducing in-surgery and in-experiment complications. This model's versatility, ease of use and low cost allow for further expansion to other labs and to a wider scope of surgeries.

Organic photovoltaic (OPV) solar cells present promising solutions in photovoltaic technology due to their lower cost and the abundance of materials compared to earlier solar technologies. As energy costs rise, OPV’s are increasingly of interest as a source of energy. The development of new curricula using a socio-scientific issues (SSI) framework can encourage students to consider careers in organic chemistry to fill these critical needs in global energy. The SSI framework also allows students in the developing stages of their STEM pathway to engage more deeply in traditionally ‘weed-out’ coursework and develop skills which will allow them to persist through STEM. We have designed a laboratory experiment using a SSI framework to allow undergraduate organic chemistry students to explore OPV’s current energy. Students synthesize poly(3-hexylthiophene) (P3HT), the active layer of an OPV cell and a promising polymer in OPV technology due to its stability and scalability. Undergraduates also build and strengthen skills of fundamental processes of organic chemistry using Grignard monomer formation and gain insight into the benefits and current challenges of organic solar cells, increasing their scientific literacy. Synthesis is conducted without the use of an inert atmosphere, lowering the barrier to implementation in under-resourced learning environments. This laboratory protocol exposes students early in their STEM careers to SSI-based learning in OPV technology and allows them to see connections in coursework to broader global issues.

Crowded farmworker housing conditions are associated with stress and adverse mental health effects. The H-2A program, administered by the Department of Labor (DOL), grants employers the ability to apply for foreign workers to perform temporary agricultural labor. H-2A employers are required to provide housing for workers at no cost. We sought to describe the distribution of H-2A worker housing and crowding within Region 10 (Washington, Oregon, Idaho, and Alaska). We defined crowding as the occasion when the reported total workers certified exceeds the reported total occupancy (the total occupancy capacity for all housing units per the employer H-2A application). Using data gathered by the DOL over 2020 and 2021, we examined 2,315 applications, representing 72,151 workers (54,873 in WA). Across Region 10, 22% (n=15,892) of all workers lived in mobile homes, 12% (n=8,476) in houses, 11% (n=7,832) in temporary worker housing, 9% (n=6,397) in stick-built homes, 8% (n=5,970) in labor camps, and 6% (n=4,054) in apartments. The definition of crowded housing was met by 24% of applications (n=567/2,315). A total of 36,101 workers (50% of total) lived in crowded housing conditions, with 10,569 workers in crowded mobile homes, 6,256 in temporary worker housing, 3,289 in houses, 2,931 in labor camps, 2,534 in stick-built homes, and 1,471 in apartments. Our results suggest crowding is prevalent in Region 10 H-2A worker housing and is especially of concern in mobile homes and temporary worker housing. Our work helps to guide efforts to prevent crowding and its adverse health effects among vulnerable workers.

People embody the joy and pain of nature through their own experiences, and have been sharing and cherishing this interconnectedness from time immemorial through paintings, stories, songs, dance, and more. I am a migrant living in a foreign country. i was eating sushi that day…. is a solo dance and an ethnographic research project, exploring ways in which I connect to, resonate with, and embody nature and personal experiences through performance. The awe-inspiring habit of salmon migration sparks my curiosity about the complex and multifaceted nature of human migration. As anadromous fish, salmon spend their juvenile life in rivers, and migrate to the ocean where they spend their adult life. They return to the upstream rivers to reproduce when they reach sexual maturity. Salmon are able to precisely return to their natal river, and even to the very spawning ground of their birth. This creative research explores my questions around identity and the idea of belonging through examining migrating salmon, and my own migrating experiences. What is the definition of home? Is it where you were born, reproduce, and die? Or where you mature? Salmon migrate to optimize their chance of reproduction as it defines their success. What about humans? What are we migrating for? Are we ultimately going to return to where we were from? My creative process starts with producing a soundscore combining text, breathing, and waves. The choreography is inspired by and generated through filmed improvisations as I experiment with different ways to interact with the soundscore and the props, always keeping my research questions in mind. The piece is a product of my desire to understand and reify the fear, confusion, exhaustion, excitement, and hope in the process of migration.

Coastal fog levels along the Pacific Coast have been declining over the past century, likely due to climate change. Fog plays an important role in coastal ecosystems and supports many species native to the Pacific Coast. Plants native to the California coastline have been shown to benefit from the higher humidity and lower temperature conditions associated with fog. Little research has been done of the implications of fog decline on native vegetation in Washington State. This study measures the effect of varying fog levels on photosynthesis rates in Washington State native plants. Plants were divided into a fog and control group in two isolated chambers. The fog chamber was exposed to humidity typical of Washington coastal conditions. Photosynthesis rates were measured by gas exchange (carbon dioxide uptake) and chrorophyll a fluorescence using the Li-Cor LI-6800 Portable Photosynthesis System. The results suggest decreasing fog could cause a decline in the productivity of coastal vegetation, which in turn affects other members of the ecosystem. A better understanding of the impacts of coastal fog decline can inform habitat restoration to include techniques such as fog collection to help mitigate the impacts.

The drug rapamycin can increase lifespan in a variety of model organisms by repressing the activity of the mTOR complex, a cellular component required for growth and development. Other than one small study that looked at genetic variation in the effect of rapamycin on lifespan in fruit flies (Rohde et al., 2021), little is known about how natural genetic variation affects the response to rapamycin. Previous work by the Promislow lab, utilizing developmental time to indicate rapamycin’s affect, has shown that some strains of fruit flies are completely resistant to rapamycin while others are sensitive. These genotypic differences are also reflected in the metabolome, the complete set of small molecules and metabolites present within cells. Metabolome analysis of these strains revealed significant differences in metabolite concentrations between resistant and sensitive lines. Interestingly, when we treated sensitive strains with rapamycin, their metabolome profiles were like those of starved larvae. I hypothesize that rapamycin is affecting larval ability to take up nutrients and that the starved metabolome is a result of actual starvation. To test my hypothesis, I am designing a starvation assay to compare the death rate of sensitive and resistant larvae. Measurement of resistance to starvation is taken two days after rapamycin treatment by transferring larvae to nutrient deficient food. The duration of time for individual larvae to die is recorded, and the death rates between the two populations are compared. If my hypothesis is true, the sensitive larvae will have a higher death rate than the resistant larva. If death rates are similar however, that could mean that rapamycin does not cause a nutrient deficit and there is another explanation for its effects on the metabolome. This study will provide insights to the underlying mechanisms of sensitivity to rapamycin, and why it might differ between individuals.

Giardia lamblia is a gastrointestinal parasite which causes diarrheal disease and hinders nutrient absorption. G. lamblia colonizes the small intestine by a two-phase life cycle: the reproducing trophozoite stage and the transmissive, infective cyst stage via a fecal-oral route. How G. lamblia detects encystation signals in the encystation process is unknown. Our laboratory discovered EncystR, a seven transmembrane protein which compartmentalizes after perceiving cholesterol depletion and increased pH. Knockdown of EncystR promoted encystation, indicating negative regulation. We hypothesize that EncystR is responsible for perceiving encystation stimuli and de-repressing cAMP signaling to promote encystation. However, the mechanism responsible for triggering cAMP signaling is uncharacterized. This project hopes to identify EncystR transient interactions using proximity labeling and mass spectroscopy (LC-MS/MS). Proximity labeling of EncystR over an early encystation time course will identify proteins involved in downstream signaling. To determine time points of interest, EncystR was endogenously labeled with mNeonGreen (mNG), and imaging of non-encysting cells displayed EncystR-mNG at the plasma membrane. To delineate the EncystR trafficking pattern, we induced encystation and followed EnystR localization for several hours using fluorescent microscopy. Individual cells were categorized by localization to peripheral vesicles, a novel acidic compartment, or non-responsive cells where localization remained on the plasma membrane. Percent peripheral vesicle localization peaked at 1h and percent compartmentalization stabilized after 3 hours. Proximity labeling proteomics at these time points can identify connections to cell trafficking to the novel acidic compartment. While EncystR does not tolerate TurboID proximity labeling, miniTurbo produced significant biotin labeling after 30 minutes in 50 mM biotin. Biotin-tagged proteins are sequestered using Streptavidin-coated columns through liquid chromatography, then cataloged using mass spectroscopy (LC/MS-MS). Proximity-tagged proteins using miniTurbo can inspire drug inhibition candidates of Giardiasis. Due to Giardia’s model nature to other parasites, such as reliance on cholesterol, parallel drugs may be found as well.

The migratory songbird Gambel's White-Crowned Sparrow (Zonotrichia leucophrys gambelli) experiences drastic seasonal shifts in its song production and stereotypy, driven by changes in distinct song circuit nuclei, namely increased neurogenesis in the forebrain nucleus HVC and increased electrical excitability in nucleus RA. Manipulating the photoperiod and hormone levels of these sparrows in a laboratory setting results in changes very similar to the seasonally-induced behavioral and neurophysiological changes they experience in the wild. We are interested in reducing the number of birds necessary for our studies by developing a protocol to produce seasonal effects in vitro via organotypic slice culture. Organotypic slice cultures preserve the neural connections and functions involved in the seasonal changes we are interested in, so they can be more directly observed and manipulated. We maintained slices of neural tissue from white crowned sparrows, including HVC and RA, on a membrane that allowed for exchange with the media, a technique initially developed by Stoppini et al., in 1991. After the tissue was cultured, it was fixed and resectioned into thinner slices so it could be Nissl stained and imaged to test for the presence of healthy cells. Organotypic culture is an established technique for neural tissue from juvenile animals; we are attempting to use it on tissue from adult, wild caught, white crowned sparrows. There is currently no protocol for organotypic cultures of this tissue, and the nature of the tissue itself poses a challenge. Adult tissue is less plastic than the juvenile or neonatal tissue that is usually used with this technique, so it has more difficulty surviving the shift to culture. Once the protocol is developed, we plan to manipulate the hormonal environment to try to mimic changes that occur during their breeding season.

Engineered genetic circuits provide an environmentally friendly path to chemical industries, including fine chemicals and therapeutics. To effectively modulate genetic circuits, a programmable tool to control multiple genes is necessary. CRISPR-mediated gene activation (CRISPRa) is an emerging tool suitable for this purpose. In CRISPRa, a nuclease-deficient dCas9 protein is used to deliver a transcriptional activator domain (MCP-SoxS) upstream of genes of interest. A complementary guide RNA (gRNA) enables dCas9 recruitment to any DNA target. Despite the programmability of CRISPRa, the number of genes that can be simultaneously regulated remain unexplored. In this work, we aim to experimentally investigate the number of gRNAs limitation in the chemical bioproduction context. First, we designed CRISPRa circuits with an increasing number of guide RNAs encoded on plasmids constructed with a scalable and high-throughput technique via Golden Gate Assembly. CRISPRa circuit performance was then evaluated by simultaneously regulating multiple fluorescent proteins as a proxy for multi-enzyme cascade in biosynthetic pathways. Increasing the number of gRNAs was found to decrease CRISPRa activity, suggesting competition of CRISPRa components. Furthermore, we applied the constructed circuits for metabolically engineered pathways in P. putida regulating production of p-aminocinnamic acid (pACA), a precursor for polymer synthesis vital in photovoltaic and biomedical applications. Bioproduction of pACA in P. putida was enabled by simultaneously regulating 9 heterologous genes. The outcome of CRISPRa circuits will be analyzed via High-Performance Liquid Chromatography (HPLC).The implication of this work will allow us to construct large scale CRISPR genetic circuits and optimize multi-gRNA CRISPR circuit integrations into other systems such as non-model organisms and cell-free systems, which will expand metabolic engineering capabilities and chemical productions beneficial in a wide range of biosynthetic applications.

Current smart home devices understandably prioritize the needs of a primary user/owner, who is also typically the purchaser of the device and a corresponding subscription plan. Yet smart home cameras and other smart devices with microphones, location tracking, and other spatial sensing capabilities invariably impact the privacy of people nearby, such as family, friends, guests, neighbors, and domestic workers. We refer to these affected nearby people as adjacent users (or adjacent subjects) because they may interact with smart devices but do so with relatively little or no direct awareness, consent, access, control, or benefit. Although work in privacy and security has begun to address the privacy needs of adjacent users, there is little design research that has responded with either concrete interventions like proposals or prototypes. We present a novel speculative design of a smart home camera called Arca with a physical camera prototype and a mobile application. A significant insight of our empirical and design research is that the most common issues with smart camera privacy is the interpersonal tensions and conflicts stemming from inadequate disclosure, consent, autonomy, and transparency from primary owners. Whereas traditional privacy/security research often focuses on harms from improper disclosure of personally sensitive information, our research suggests that many adjacent users do not necessarily mind being recorded, they do mind the lack of “communication,” “respect,” and “professionalism” from primary users. Furthermore, our studies reveal that even if our specific privacy modes and access sharing features are not regularly used, they may nonetheless function as mechanisms to facilitate better, more open conversation between primary and adjacent users. We continue our work with the goal to enhance adjacent user privacy and experience with privacy-sensitive camera features and reduce tension between adjacent users and primary users.

Smartphone detection of anemia using patient photos has the potential to provide a non-invasive method of measuring hemoglobin levels, introducing the possibility of increasing the accessibility and cost-effectiveness of current practices. While traditional methods of anemia detection require a complete blood count by a trained healthcare professional, smartphone detection instead relies on the user to take a high quality picture of their fingernails. However, it currently lacks the ability to provide feedback to the user on the quality of their image. For example, an overexposed image or one with low fingernail visibility can lead to inaccurate predictions of hemoglobin levels. We propose that machine learning classification methods can analyze these patient images to estimate the image quality and predict the effectiveness of smartphone detection of anemia for a given image. With various classical machine learning models, we demonstrate and compare the capabilities of each in classifying images of patients’ hands as being of “good” or “bad” quality (or on a more granular numerical scale) when given features of the images. Preliminary results show that a logistic regression model reaches 91.4% accuracy labeling images when compared to empirically assigned labels, and we expect iterative models to achieve improved performance. When completed, we would propose that this classifier could be used in the field to identify if patient image is of high enough quality to produce an accurate measurement of hemoglobin levels in real-time, providing feedback on the phone to adjust or correct the image-taking process.

Ophthalmic drug administration has been increasingly prevalent in recent years, with eyedroppers being utilized to administer costly medication like that for glaucoma. There haven’t been many solutions addressing eyedropper instillation for those with preexisting conditions like arthritis, who often deal with a host of problems when administering them: producing the necessary force to distill a drop, aiming the drop into the eye, and contamination of the eyedropper tip. We are testing the question of whether accessible eye drop aids can significantly improve eyedrop compliance and distillation for the elderly. Solutions to eye drop administration can save money and make the overall process easier for many patients. Existing solutions on the market seem to address the issue of contamination using apparatuses that press onto the lower eyelid, but there is still much to be desired with the force and aim required. Many require the use of gripping or squeezing, motions that many elderly patients can’t apply as much force with. I propose a couple of solutions to these problems in the form of eyedropper aids that each make use of a few different methods, including translating the motion, applying the force with different limbs, and even mechanizing the force required. Through a quantitative study, I hope to eventually test these prototypes through an ophthalmology clinic among a wide variety of elderly. Assessing these prototypes through both questionnaires and observation, I hope to notice an increase in effectiveness from previously existing apparatuses. We will use a survey to ask a variety of questions to around 100 elderly patients with varying expertise in eye drop instillation. The survey will ask whether the tool was more useful, easier, how hard it was to assemble, and we will also monitor quantitatively whether the accuracy of drops actually instilled was better. This work hopefully saves patients money from medication cost from a reduction in wastage, allows for better administration of medicine, and eases the process of distillation.

In this research project, our multidisciplinary team is developing environmental forensic technology to identify illegally caught seafood, with a focus on high-value species such as bluefin tuna. To tackle the difficult task of immediately detecting illegal, unregulated, and unreported (IUU) fishing, we are developing a rapid, affordable, and portable detection device that uses isothermal amplification and blue LEDs to detect resultant green fluorescent products indicating the presence of tuna DNA. As the lead Electrical Engineer, I designed an electrical circuit to precisely and stably control the temperature of a resistive heater using a microcontroller, thermistor, and PID algorithm along with an LED circuit. The requirement for the process involves the utilization of isothermal amplification technique at a fixed temperature of 37°C (33-42°C range) and green fluorescence detection (at 550 nm) in DNA using blue LEDs (at 470 nm) and an orange acrylic filter to filter the blue wavelength. The result is a circuit that meets the requirements for the biochemical process and enables real-time feedback without the need for shipping samples to a laboratory. With the development of the device finished, the next step is to streamline the user experience. I am leading the software development effort to create a phone application that facilitates the assay setup process and automates image capturing and analysis. With a capable research team, including colleagues shaping the user interface, a junior researcher connecting the device to the phone application, and a Ph.D. candidate in Computer Science and Engineering developing a proprietary camera software, our team's effort culminates in a user-friendly phone app that streamlines the assay workflow and provides real-time sample analysis. With the completion of this project, we will be able to make a difference by helping communities and marine ecosystems.

Starting in European folklore, antisemitism has gone largely unnoticed in fairy tales. Antisemitism present within our modern fantasy standards stems in large part from European folklore, further manipulated by rising antisemitic propaganda in the late nineteenth century, coalescing in Nazi propaganda, and internalized as stereotypes by storytellers, thus leading to a double standard of otherizing and assimilation in Western culture. As folklore evolved into the current fantasy genre, these stories transferred antisemitic and stereotypical of villainous Jewish characters into our modern-day media. Antisemitic caricatures populate our media, and gentiles, or non-Jews, often don’t realize it because of how pervasive these stereotypes have become in the main body of Western culture developed over the millennia. Drawing connections to the villainized images of a Jewish-coded character throughout fantasy stories and films, this paper uncovers the systemic and institutional problems Western media pushes onto an indifferent public, where thinking critically about our media doesn't extend to antisemitism. From case studies including Tolkien's work, Barbie movies, and the Harry Potter series, this paper identifies antisemitic stereotypes that have become impossible to avoid in media. Through widespread acceptance of these stereotypes, we perpetuate the misconceptions of our predecessors and otherize the Jewish body. Through perpetuation of hateful stereotypes in modern media and a lack of acknowledgement of ongoing antisemitism, we normalize the person of Jewish descent as the villain. Spotting these stereotypes is the first step in overcoming this imagery. Subversion becomes the next step to undo the damage to Western culture and modern media; reinterpreting and rewriting antisemitic stories as aspirational stories is a way to subvert the ideas of what a hero and a villain can look like.

The peculiar behavior of vowel hiatus resolution in Biloxi provides a useful test case with which to consider the relative merits of two prominent theories of morphologically conditioned phonology (MCP): Cophonology (Inkelas and Zoll, 2007) and Constraint Indexation (Pater, 2009). Biloxi, an extinct or critically endangered (sources vary) language of the Siouan language family, has received little to no phonological study since the 1974 grammar this project is based on (Einaudi, 1974). Vowel-vowel sequences are generally avoided in Biloxi. When vowel-vowel sequences are formed by affixing, this sequence is repaired by deleting the first vowel, as in (1) /ohi dani-axehe/ → [ohi danaxehe] ‘three sitting on ten’ (=13). When such a sequence is formed by the dative prefix /ki-/ and a vowel-initial verb root, glide epenthesis occurs instead, as in (2) /Ø- ki- ε -tu/ → [kijetu] ‘they said to him’. Yet another exception is observed when /ki-/ specifically precedes the verb root /õ/ ‘to make, do’, resulting in glide fortition and an additional /k/ in the surface form, as in (3) /Ø- ki- õ -tu/ → [kikõtu] ‘they made for him’. This analysis lends support to whichever model of MCP better accounts for the complex vowel hiatus pattern of Biloxi. As an understudied language, a framework’s ability to fit Biloxi, in addition to previously considered languages, gives that model greater explanatory power. Avenues for future research include the consideration of a learnability model for Cophonology as compared to that proposed by Pater (2009) for Constraint Indexation. An updated grammar of Biloxi with more detailed phonological analysis would also extend the scope of modern studies of the language. This work brings new attention to the Biloxi language, and contributes to the ongoing debates over morphologically conditioned phonological processes.

Creative coding refers to a method of computer programming that prioritizes artistic expression. Remixing is the iterative behavior of altering an existing artifact. Frameworks such as p5.js support sketching with creative code. Given the focus on expressivity over functionality, code reuse in creative coding practice is distinct from other programming contexts. Additionally, remixing facilitates iteration on existing code, but we have yet understand how creative coders use remixing in practice. To understand creative coder remixing strategies, we studied the community of OpenProcessing, a site dedicated to sharing code-generated artworks. To begin, we conducted a network analysis to determine which datasets of original sketches (also referred to as the antecedent sketch) and their remixes to use in our study. Our visualization consisted of a social network graph in which the nodes represent individuals, and edges showing their relationships. We found that 30% of the 1.2 million sketches in our dataset were involved in remixing. For data analysis, we utilized a code-diff tool to showcase ways the antecedent sketch's code differs from the remix and categorize various types of remixing strategies. Over time, these categories became increasingly focused on changes made visible on our code-diff tool. We present on the diversity of ways that authors remix to curate projects, annotate process, explore variations, and transform existing sketches. Through remixing, artists have already begun to tailor, customize, and explore different ways to use their creative tools, in ways system developers may not have foreseen. We find that remixing also encourages exploratory programming and experiential learning. As creative code is increasingly used to support computational education, we can consider the implications of remixing for understanding and facilitating informal learning. At last, we reflect on the prevalence of these remix types and how future systems could support a multiplicity of remixing strategies for creative work.
 

T cells are a crucial part of the human immune system that play a role in the response to diseases from type 1 diabetes to COVID-19. All T cells undergo genomic recombination to create distinct T cell receptor complex (TCR) cell surface molecules that recognize antigens presented by major histocompatibility complex (MHC) molecules, thereby triggering a series of signaling pathways that culminate in T cell activation and cell division. Afterwards, daughter progeny share the same unique TCR molecules. In my project, I will be examining the role TCRs shared within a single individual (private) and between individuals (public) during COVID-19 infection. While the existence of these classes of TCRs is well documented, their functional roles remain unclear. My hypothesis is that the presence of public TCRs is linked to the strength of an immune response and, hence, to different disease severity in humans. We reason that common evolution may have led to the rise of these public TCRs because of their major role in the human body’s antiviral immune response. This hypothesis predicts reduced levels of public TCRs results in an inability to fight disease, whereas high levels of public TCRs will be associated with less severe disease. To test this, I will use a mixture of single cell RNA analysis and statistical tests on a dataset of TCRs acquired from blood samples of 254 SARS-CoV-2 patients and 16 healthy patients. Using a multiomic approach will allow me to also find genetic links between cells of interest. The data includes baseline and acute readings for each patient, and can be subsetted to cells with expanded TRA and TRB junctions individually. This experiment will help advance our knowledge about the function of public TCR chains and their role in fighting disease. Furthermore, our data could have translational applications for disease biomarkers.

Cannabis use has dramatically increased in response to legalization in the U.S., with total sales in the U.S. jumping 46% from 2019 to 2020. ᐃ9-tetrahydrocannabinol (THC) is the primary psychotomimetic compound in Cannabis and has been shown to modify memory and motivation, processes mediated by the prefrontal cortex (PFC) brain region. I sought to test the effects of THC on PFC activity during appetitive Pavlovian conditioning in mice- a behavior in which a subject learns to associate a non-rewarding stimuli to a reward. THC acts on the endocannabinoid (eCB) CB1 receptor (CB1R), a presynaptic signaling protein responsible for modulating neural activity throughout the brain, with robust expression in the PFC. To monitor neural activity during behavioral trials, we implanted optic fibers into the PFC and virally expressed biological sensors: GCaMP6f to track Calcium activity, and the novel GRABeCB2.0 to measure eCB activity. VGAT-Cre and VGLUT1-Cre animals were presented with a house light prior to a sucrose reward to observe the neuronal GABAergic and glutamatergic activity during the conditioning, respectively. After 5 days of conditioning, I administered vehicle or THC (i.p., 5 mg/kg) to observe behavioral and neural effects of THC. We observed neural activity that transferred from the sucrose reward to the house light cue suggesting these neurons encode for this learning. Endocannabinoid activity also transitioned from sucrose reward to the house light cue suggesting cannabinoid involvement in regulating this association. THC pre-treatment reduced licking and motivation for sucrose while modifying neural activity without eliminating it. This provided much needed insight into the formation of memory during learning and reward motivation under the effect of THC.

Massive stars place powerful constraints on stellar evolution and are observed in a menagerie of exotic evolutionary phases. These objects play a crucial role in regulating their environments. They drive the chemical evolution of their host galaxies, and set the energy balance of their surroundings via feedback processes. Due to the importance of massive stars, placing constraints on their evolution serves as a key to understanding galactic ecosystems. Notably, stellar variability is a powerful probe of the poorly-constrained physics of massive star evolution. In particular, variability studies on ensembles of evolved massive stars can significantly constrain stellar evolution. We aim to understand the variability of hot massive stars through a census of these objects. We accomplish this using data from the Gaia mission, cross-matched with light curves from the Zwicky Transient Facility (ZTF). We expect to characterize the evolution of massive star variability timescales and amplitudes along the main sequence and beyond. Our results will place key constraints on the evolution of massive stars. 

Active galactic nuclei (AGN) found at the center of galaxies are a compact region of space that produce significantly higher than normal luminosity. AGN are powered by the accretion of matter into a supermassive black hole and sometimes present winds called outflows. Whether outflows affect the evolution of their host galaxies is still a topic of research. Outflows with speeds of more than 10% the speed of light, called Extremely high-velocity outflows (EHVOs), have not been thoroughly explored, and due to their large energies, they might play a large role in star formation. In the 16th data release of the Sloan Digital Sky Survey, 98 new quasars with EHVOs were identified. With this new sample, I will present the results of my findings on the following questions: (1) What does the average EHVO quasar look like? (2) Do these extreme outflows vary more or less than other previously explored outflows? (3) Can we detect these EHVOs at higher speeds? To answer the first question, I will present a composite of the 16th data release EHVO cases found. To analyze if they vary more often or less than other EHVOs, I will present the results of a longitudinal variability study on the 51 cases that were observed on multiple occasions in the EHVO sample. For the last question, I will show the results of applying code our team has created to remove the Lya lines from our spectra, so the intrinsic ions (such as NV, Lya, OVI) will be easier to analyze at higher speeds.

My project studies Extremely High Velocity Outflows (EHVO) in Quasars. Quasars are particularly interesting due to the fact that they are one the most luminous cosmological objects that we have observed in the universe. They are systems at the centres of galaxies that host supermassive black holes--which have masses that range from millions to billions of solar masses--surrounded by an accretion disk of superheated plasma, gas, and dust. The phenomenon that we are studying are the EHVOs in quasars, which are gas outflows traveling at above 10% of the speed of light from the active central region. My project attempts to answer the question of whether quasars with EHVOs show distinct physical characteristics when compared to Broad Absorption Line Quasi-Stellar Objects (BALQSOs), which are quasars with outflows that show a broad absorption width but at lower outflow speeds, as well as compared to the general parent sample of quasars. We hypothesized that quasars with EHVOs show distinctive physical characteristics when compared to the other quasar categories. To study quasar physical characteristics to test our hypothesis, we used data from the Sloan Digital Sky Survey (SDSS) in the latest data release. Previously, our group has found the measurement values for all quasar samples to be overestimated. Thanks to data provided in Rankine+2020, we developed software and used the data to cross-correlate the values of physical parameters, such as bolometric luminosity (Lbol), Eddington ratio and black hole mass (Mbh). I will present the preliminary results of the analysis on EHVO quasar’s physical properties when compared to BALQSOs and the parent sample. These results suggest that quasars with EHVOs exhibit larger values of their Lbol and Eddington ratio when compared to the other two classes of quasars, while the Mbh parameter does not show significant differences.


 

Lead perovskites have attracted the interest of the industry for optoelectronic devices applications due to their strong and tunable absorptions. However, their stability and environmental toxicity impose a challenge in its use for commercial application. Bismuth perovskites are a promising alternative due to their similar ionic radius to lead with long-term stability and lower toxicity. To replace lead-based perovskites with bismuth-based ones, it is necessary to increase the photoluminescent quantum yields and extend the emission wavelength range. The crystalline structure of bismuth perovskites can be altered with a dopant to redshift the usually blue light emission. We doped bismuth-based perovskites with rare-earth metals via sonication. The produced materials were analyzed by ultraviolet-visible spectroscopy and photoluminescence spectroscopy. The collected measurements determine if a redshift was produced on the emission spectra. It is expected to see a redshift from the current 400 nm wavelength on the doped perovskites. However, the current results do not show a redshift, instead they show a change in intensity. The comprehension of how to efficiently produce redshifted bismuth perovskites can propel the industrial level use of this less toxic alternative.

Stereotypes are generally harmful, regardless of the content being positive or negative. However, people may perceive certain stereotypes are more acceptable or less harmful than others. We are interested in seeing the varying degree of how people perceive the social acceptability of the stereotypes about women’s interest or ability in computer science. We hypothesize that stereotypes about women being less interested in STEM fields are more socially acceptable than stereotypes about their ability in computer science. For this purpose, we intend to conduct a survey study including 4 scales measuring how much participants, students from University of Washington, believe it is socially acceptable to assume gender stereotypes about women’s interest and ability in computer science. Data will be analyzed using a dependent t-test. We hypothesize that people believe it is less socially acceptable to talk about the stereotypes about women having less ability than being less interested in computer science. The finding may stop the adverse intuition that expressing some stereotypes are tolerated and further motivate women to express their interest in computer science as the stereotypes about their interest should be just as unacceptable as any stereotypes regarding their ability. Future work could develop interventions to spread the awareness that having either types of stereotypes about women on computer science topics, no matter woman’s interest or ability, should be considered harmful.

The trapping of individual ions has allowed physicists to control and observe otherwise inaccessible phenomena. Ion traps have enabled the most precise measurements of fundamental physical constants, mass spectrometry for chemical characterization, atomic clocks that would only lose a fraction of a second over the entire age of the universe, and the direct observation of many core concepts in quantum mechanics. Many crucial developments in ion traps occurred here at the University of Washington in the group of Hans Dehmelt, who shared the 1989 Nobel Prize in physics for that work. Today, techniques in ion trapping continue to be developed because trapped ions are one platform for creating qubits in quantum computers. With the growth of quantum information science in academia and industry, there is a need for inexpensive, scalable educational labs to introduce students to concepts in quantum computing. To fill this need, we developed a reproducible lab, which demonstrates key concepts in ion trapping. Our process utilized, first, a comparative approach with reference to literature and, second, iterative improvement on built components. The lab consists of two, independent quadrupole traps: a four-rod trap and a planar five-rail trap. To reduce cost and complexity, we trap charged particles with 25 µm and 50 µm diameter, rather than atomic ions. The particles are trapped in air, at atmospheric pressure. Due to the damping forces provided by this background gas, the trapped particles are easy to control. The result of our project is a lab capable of several experiments, including controlling the number of particles trapped through voltage modulation at a constant frequency, studying the phase transition between one- and two-dimensional Coulomb crystals, exploring micromotion compensation, observing two- and three-particle secular modes, and demonstrating particle shuttling along the trapping axis of the planar trap.

In regions with high HIV prevalence, periods of pregnancy and postpartum are associated with higher risk for HIV acquisition. The WHO recommends oral tenofovir (TFV)-based pre-exposure prophylaxis (PrEP) as an effective method of reducing HIV acquisition risk for pregnant people. Although most pregnant patients in Kenya with identified HIV risk factors accept PrEP when offered, >50% discontinue PrEP within the first 30 days.​ Few studies to date examine factors that contribute to PrEP discontinuation in this population and there are no existing intervention studies aimed at improving PrEP adherence during pregnancy/postpartum. We utilized data from the ongoing Mobile Women and Children (mWACh)-PrEP study, a randomized control trial testing a two-way short messaging service (SMS)-based platform to facilitate communication between peripartum patients taking PrEP and remote nurses. In the parent study of 379 HIV-negative, cisgendered Kenyan women taking PrEP, 188 were randomly assigned to the intervention (mWACh-PrEP) group. The purpose of our sub-analysis was to evaluate message contents and identify frequently discussed topics. System messages were tagged into categories based on subject. Of weekly automated messages, 42.1% were responded to by participants, with an average response time of 87 minutes; topics of responses included PrEP concerns (55.1%), antenatal concerns (22.4%), maternal health concerns (17.8%), and infant concerns (17.2%). A total of 366 spontaneous messages were received, with >50% of participants sending at least one spontaneous message; topics of spontaneous messages most frequently included antenatal concerns (22.9%) and PrEP concerns (19.7%). These results indicate that a two-way SMS based intervention provides important support to pregnant and postpartum patients taking PrEP; the mWACh-PrEP system could be an effective solution not only to promote PrEP adherence within this population, but also to increase access to maternal healthcare.

The focus of our research is on digitizing linguistic material from two understudied and endangered Brazilian Amazonian languages, Panãra and Kawaiwete. Panãra (ISO code: kre) is a Northern Jê language with about 600 speakers, and Kawaiwete (ISO code: kyz) is a Tupí-Guaraní language with about 1,000 speakers. Our project focuses on the digital archival of linguistic materials from these communities, the creation of a dictionary, and the study of the morphophonological grammar of these languages. This talk describes the process of data digitization thereby making it usable to a variety of research questions to be answered in future work. Data elicited by Professor Lapierre from field trips in 2018 and 2019 to Kawaiwete speaking communities, and from 2015 to 2019 to Panãra speaking communities forms the foundation for this project. Materials utilized for this research project include elicitation recordings and transcriptions, and field notes obtained through in-situ fieldwork. The initial stage of our research was digitizing handwritten field notes over the course of ten months so the notes could be fully searchable in an online database. We transcribed a total of 16,128 words for the Panãra notebooks and 12,860 words for the Kawaiwete notebooks. The second stage of the project, which is our current stage, is entering lexical items into FieldWorks Language Explorer (FLEx), a software that stores and analyzes lexical and grammatical information. We currently have 351 words entered into the Panãra database, and plan on entering sentences in order to analyze parts of their grammar. The work of this project responds to the lack of an effective writing system in Panãra and the trending language shift towards Portuguese in Kawaiwete. A digital dictionary will help both the creation of written materials by Panãra speakers and the preservation of the current Kawaiwete language.

Early-onset Alzheimer’s Disease (EOAD) patients have a greater risk of developing seizures. Consequently, EOAD patients who develop seizures have worsened comorbidities, including mortality and behavioral outcomes. The causal relationship between chronic seizures and AD is still unknown. Neuroinflammation has been postulated as one of the links between these two disorders. Our lab has previously demonstrated that young mice with a presenilin 1 (PS1) genetic variant and amyloid precursor protein (APP) overexpression, mimicking an EOAD phenotype and genotype, subjected to chronic seizure are at high risk of mortality. This behavior was not observed in young mice with an EOAD-associated presenilin 2 (PS2) genetic variation. We hypothesize that chronic kindled seizures evoked in 2 months-old APP/PS1 mice worsen neuroinflammation and other neuropathological hallmarks of EOAD, including β-amyloid (Aβ) accumulation in the brain. To address this, 2-month-old male and female APP/PS1 and PS2 mice were subjected to seizures through a corneal kindling model of chronic seizures. The right brain hemisphere was collected before and after mice achieve the kindled criterion (five consecutive 5 Racine scale scores). Brains were harvested and cryopreserved for immunohistochemistry. My role in the project was to process tissues for immunohistochemistry. 20-µm thick sections of hippocampus from each mouse were sectioned on a cryostat and slide-mounted before processing with commercially available antibodies for molecular markers of neuroinflammation and neuropathology. Photomicrographs were collected and images were analyzed as the number of immunoreactive cells for each molecular and protein marker. We anticipate increased neuroinflammation and Aβ accumulation in kindled APP/PS1 mice versus their respective wild types and non-kindled littermates. We do not expect differences in PS2 variant animals. These results add to a larger research study in the laboratory to suggest that targeting seizures, and its inflammatory response, may be a potential therapeutic strategy to mitigate the behavioral and neuropathological burden of AD.

My project involves making new tools for the genetic manipulation of Staphylococcus aureus. S. aureus is an important and pervasive human pathogen, however, much is still not known about its pathogenesis pathway and virulence genes. To better understand the genes and mutations that make S. aureus successful at causing disease, it is necessary to induce specific genetic changes and assess their impact on the organism’s virulence. However, currently available tools for bacterial genome engineering have yet to be optimized for S. aureus. Building on a successful recombineering system, previously developed by the Salipante lab, that is able to edit the S. aureus genome, we aim to address this need by using a dominant negative mutant protein of the mismatch repair (MMR) system to achieve suppression of DNA repair. Recombineering is the process of incorporating mutagenic DNA molecules into a host genome through the recombinase enzymes. Dominant negative mutations in the highly conserved MutL protein have been shown to disrupt the DNA base MMR pathway in several other microorganisms but have not yet been evaluated in S. aureus. My project aims to construct a temperature sensitive vector that highly expresses a ssDNA recombinase that is active in S. aureus along with a dominant negative mutL mutant able to bypass the MMR pathway. We anticipate that WT mutL should be able to repair a single base mutation since it has a functional MMR, reducing the number of successful recombinants relative to experiments performed with the multiple base pair mutations. The dominant negative mutL strains should not be able to correct a single base pair mutation, resulting in a similar number of drug-resistant transformats when recombineering with either a single base pair or many base pair mutations. This will allow us to better generate custom mutant strains and subsequently test the functions of various S. aureus genes towards a variety of clinically relevant phenotypes.

Urolithin A (UA) is a metabolite produced by gut bacteria from ellagitannin and ellagic acid, which is present in pomegranates, berries, and walnuts. Previous studies have shown that UA supplementation improves skeletal muscle health in both mice and humans. UA supplementation also appears to be beneficial to the heart, resulting in both systolic and diastolic functional improvement in old mice. This study aims to determine a mechanism for the improvements in the aging heart. We hypothesize that the mechanisms behind improved heart function from UA supplementation are related to the pathways that directly improves mitochondria, contractile properties and mitophagy. To investigate the mechanism behind improved heart function with UA supplementation, we split the treatments into two staggered cohorts with 20, 24-month-old mice per cohort (10 control/10 UA treatment). Both the control and the treatment group were kept on the control diet during week 1 for acclimation. In the following 8 weeks, the UA treatment group were moved to a UA 50 mpk supplemented diet. Echocardiography measurements were done pre and post supplementation. At the end of the study, all mice were euthanized under anesthesia after an overnight fast and sample collection was performed. Heart samples were collected from each mouse. Western blots of heart samples on mitophagy, mitochondrial and cardiac contractile protein markers indicate that those pathways remain unaffected by UA supplementation. In future work, we will explore other informatory pathways in order to determine the mechanism behind UA supplementation improvement of heart function.

A primary stressor of college students is financial insecurity. Research estimates that 36% of college students are food insecure, 36% lack reliable housing, and 9% experienced homelessness in 2018. Sources of financial stress for college students typically include housing, tuition and other academic expenses, credit card debt, familial responsibilities, and employment status. Furthermore, research has also shown that psychological stress correlates with decreased sleep quality. During periods of intense stress, an individual may have more frequent disturbances during sleep, sleep for fewer hours, and adopt later waking times (Galambos et al., 2013). This study investigates the relationship between financial insecurity and sleep quality of students attending a 2-year college in Washington state. More specifically, this study aims to determine the impact of a sleep wellness workshop on the sleep quality of college students. Data collection will occur through an online Qualtrics survey before and after the sleep wellness workshop. The Qualtrics survey includes modified Sleep Quality Scale questions, the Beck Anxiety Inventory (BAI), and the Perceived Stress Scale (PSS). It is hypothesized that experiencing financial insecurity will positively correlate to poorer sleep quality. It is predicted that this relationship is mediated by stress and that the sleep wellness workshop will improve sleep quality in college students. Understanding factors that contribute to poor sleep quality is vital as sleep plays a key role in maintaining proper cognitive functioning. The results of this study are intended to improve current resources, as well as implement new resources, concerning sleep quality and stress of college students.

Aging is an important problem in biomedical research. Given the increased risk of death with age, techniques to delay aging hold substantial promise for human well-being. The premise of my research is that rapamycin, a drug commonly used in transplant patients which is hypothesized to have development-slowing effects, can slow development of fruit flies. One finding from a previous study, however, is that there is enormous genetic variation within the Drosophila population (akin to the variation between dog breeds) which results in varying sensitivity to rapamycin. Earlier measurements indicated that strains of fruit flies that were sensitive to rapamycin had higher levels of histamine, a molecule that participates in metabolism, when on the drug. We set out to determine whether histamine could effectively extend sensitivity to the drug to a wider range of genotypes within Drosophila. Thus, I hypothesize that introducing histamine to the food that fruit fly strains that are genetically resistant to rapamycin are consuming will increase sensitivity. To test this hypothesis, I added solutions with different concentrations of histamine to food with or without rapamycin. I then placed eggs from a resistant strain onto the food to observe the time it takes from egg to pupa. Because I hypothesize that increasing histamine levels will make a resistant strain of Drosophila sensitive to rapamycin, the strain should become sensitive and consequently show an increase in development time compared to the conditions without histamine. Each test condition of a histamine solution and rapamycin has a corresponding control with no rapamycin. Hence, if the histamine-treated conditions show slower pupation times than the histamine-free controls, the hypothesis is validated. If validated, this work could help researchers understand ways to provide the benefits of rapamycin to individuals who might otherwise be genetically resistant to its impact in both aging and medical contexts.

This presentation discusses a unique grammatical phenomenon surrounding person marking on transitive verbs in Kawaiwete, an endangered Tupi-Guarani language spoken in Mato Grosso, Brazil. Like many related languages, Kawaiwete exhibits complex variation in how it conjugates verbs (i.e. changes verbs to fit grammatical context). Transitive verbs, i.e. verbs that need both a subject and an object, require a prefix on the verb. This prefix denotes the role of the sentence’s component parts, called arguments. Whether the subject prefix or object prefix is selected depends on the grammatical role of the arguments and word order of the sentence, which is flexible. I analyzed data collected by Professor Lapierre by comparing sentences with first and third person arguments (i.e. “I/me” and “he/him”). I surveyed (i) the verbal prefixes used, and (ii) the order of the sentence’s arguments. Consider sentences (1a) /miara jemiwara je a-esat/ ‘I see the jaguar eating’, (1b) /miara jemiwara je w-esat/ ‘I see the jaguar eating’, (2a) /a-esat je miara jemiwara/ ‘I see the jaguar eating’, (2b) */w-esat je miara jemiwara/ (ungrammatical). When /je/ ‘I’ appears before /-esat/ ‘see’, its role as the subject is unambiguous, meaning the verb may take either the prefix /a-/, matching with the subject (1a), or the prefix /w-/, matching with the object (1b). However, when /je/ does not precede /-esat/ (2a), the verb must take /a-/, marking /je/ as the subject (2a). This implies that in situations where the role of the sentence’s arguments is unambiguous due to word order, the verbal prefix may select to match either of the verb’s arguments. Crucially, when word order results in ambiguity, the verbal prefix must match with the subject to disambiguate the sentence. This finding suggests that Kawaiwete verbs mark more than simply the verb’s subject or object, a unique phenomenon rarely observed.

Most of the current literature on relationships and marriage in the US overlooks the ethnic, religious, and generational variations within the black population. This study explores the nuance in views and attitudes among African American and African immigrant young women toward marriage, especially hypergamous and interracial marriages. This study follows a deductive approach through testing hypotheses developed through four existing theories. First, symbolic interactionism theory suggests African Americans and African immigrants hold different attitudes toward marriage because they developed different meanings of marriage. Second, social exchange theory posits that because people tend to marry those of similar social and economic backgrounds, interracial and hypergamous marriages represent a social exchange of status. Third, classic assimilation theory suggests as the generational status of African immigrants increases, they increasingly develop similar marriage views and attitudes to African Americans. Finally, segmented assimilation theory holds that Muslim African immigrants are selective in their assimilation, thus, would not have similar views to African Americans. This study uses qualitative data collected through interviews, and quantitative data from the public use microdata series, IPUMS USA. The mixed methods approach compares interview participants — black female college students — with the trends in the nationally representative microdata. The expected findings would confirm all of the aforementioned hypotheses. This study aims to enrich the current body of the literature and provide greater depth and breadth into the ethnically, generationally, and religiously diverse black population. A better understanding of marriage views and attitudes in the black community can help shape public policy that fosters more transitions into marriage. Moreover, marriage is among many predictors of the integration of immigrants. Thus, comparing immigrants to Americans can highlight how immigration may play a role in disruptions and delays in the integration process.

Inequities still exist despite efforts to address racism and reduce racial disparities in mental health care. One reason for this may be clinician’s limited knowledge and guidelines on addressing and exploring the consequences of racism in clients' lives. The current study is a mixed-method study using secondary data from the Applying Theatre-Based Training Methods to Address Racism in Community-Based Mental Health Services, which surveyed 119 community mental health clinicians across Washington State in 2021. The first aim of this analysis was to examine qualitative data on clinician's perceived barriers and facilitators in broaching racial issues with clients. The second aim was to explore how these barriers and facilitators are correlated with quantitative measures of multicultural counseling knowledge and awareness. Three significant barriers emerged: 1) Clinicians not engaging in racial topics unless clients initiated; 2) Clinicians avoiding discussions of race and racism because of feeling uncomfortable or unconfident; 3) Clinician's assumptions that their racial identities would be a barrier to discussions. Three significant facilitators included: 1) Client's initiation, emotional openness, and availability to have racial conversations; 2) Relevant social events that provided an avenue to prompt discussion; 3) Clinician's willingness and intentions to address racism through asking or actively listening. There was no statistically significant correlation between either the count of perceived barriers or facilitator themes and clinician's multicultural counseling knowledge and awareness. The present research can supplement and support efforts to train clinicians in broaching racial discussions and inform clinical practice with clients from diverse backgrounds.

Benzalkonium Chlorides (BACs) are widely used antimicrobial disinfectants in a variety of settings, including large scale food processing and consumer environments. Persistent usage of BACs raises concerns about the potential disruption of the gastrointestinal microbiota, an increasingly recognized regulator of an individual’s health. Furthermore, the gut microbiota has been shown to regulate drug metabolizing enzymes (DMEs) and the Gut-Liver Axis is a known prominent crosstalk pathway. Previous work in our lab has found BACs are capable of altering gut microbiome composition in BAC exposed C57BL/6 male and female mice, with notable differences between the male and female sexes. Therefore, we hypothesize that exposure to the BACs can alter the composition of gut microbiota, leading to sex specific changes in bile acid homeostasis as well as the metabolic phenotype and DME expression of the liver. In this study, we exposed male and female mice to C12- and C16-BACs at 120 ug/g/day for one week via oral dosing. Additionally, through a targeted bile acid quantitation analysis, we found sex specific decreases in secondary bile acids in BAC-treated mice. This finding is supported by decreases in bacteria known to metabolize primary bile acids into secondary bile acids, such as the families of Ruminococcaceae and Lachnospiraceae. We also aim to elucidate both transcriptomic (RNA sequencing) and functional (enzyme activity assays) analyses of the harvested livers from both male and female cohorts. Upstream pathway analysis from the results of these analyses is expected to yield sex specific differences in the downregulation of genes responsible for a variety of pathways such as protein digestion and absorption and transcriptional regulation in cancer. This study is expected to provide novel insights into the sex specific alterations in the relationship between the gut microbiome and liver caused by BAC exposure and the mechanisms underlying BAC toxicity.

Uridine-5'-triphosphate (UTP) is a precursor for RNA synthesis. Ura3 catalyzes the conversion of orotidine-5'-phosphate (OMP) into uridine monophosphate (UMP) and is commonly used as a selection marker to characterize mutation rates of S. cerevisiae. URA3 can be positively selected for by growing cells in the absence of uracil and can be selected against by growing cells in the presence of the toxic fluorinated UTP precursor, 5-Fluoroorotic acid (5-FOA). While mutations in URA3 make up the majority of 5-FOA-resistant mutants, mutations in a small number of other loci can also cause this phenotype. We whole genome sequenced the 5-FOA-resistant mutants with a wild type URA3 and identified mutations to URA6 in each of these individuals. URA6 is an essential gene that encodes an enzyme that catalyzes the conversion of uridine monophosphate (UMP) into uridine-5'-diphosphate (UDP). Here, we describe 41 non-synonymous mutations to URA6 that permit growth in both the absence of uracil and in the presence of 5-FOA. It remains unclear how the URA6 mutants can maintain a functioning UTP synthesis pathway while remaining resistant to the toxic fluorinated precursors. We hypothesize that these mutations alter the protein structure in a manner that decreases the affinity for fluorinated substrates while maintaining the affinity for UDP. To test this, we will evaluate the structural changes to URA6 resulting from these non-synonymous mutations. Our goal is that our findings can benefit our understanding of the UTP biosynthesis pathway and encourage further investigation of the mechanisms involving fluorinated substrate analogues.

Past research found racial diversity in the workplace can increase equity (e.g. Edelman et al., 2001), but could proximity to diversity make White leaders less likely to be held accountable for bias? I conducted two studies examining how having racial diversity near, but not within, the work team may influence people’s perceptions of workplace equity. In Study 1, participants recruited through Prolific (N = 252) rated how equitable they think a White leader would be in two conditions: one where all other leaders are White and one where the other leaders are racially diverse. Participants were informed that the White leader works entirely independently from the other leaders. Results show that participants rated the leader as more equitable in the diverse condition as compared to the all-White condition. Study 2 examines whether proximity to diversity could shield White leaders from accountability in an ambiguous situation where racial bias may have played a role. Results show that participants (N = 234) rated the leader's decision as less likely to be driven by racial bias in the diverse leadership (vs. all-White) condition. Mediation analysis shows that perceiving the leader as more equitable in the diverse leadership condition helps explain the present effect. Implication suggests that people may be less sensitive to detecting bias in diverse environments. Future work should disaggregate the experiences of people of color, examining whether there may be differences among racial groups in how the effect operates. Individual differences may also be moderating factors for the investigated effect. 

Atrial fibrillation (AFib) is the most common sustained cardiac arrhythmia, contributing to significant morbidity and mortality worldwide. Patient-specific computational models of the left atrium are currently studied to predict characteristics of reentrant activity that promotes fibrillation. However, current models’ patient-specificity is limited to anatomical structure and the distribution of disease-related remodeling (fibrosis), whereas electrical properties of cells and tissue are based on literature values. In cases where patients are clinically known to present with either AFib or atrial flutter (AFl), this lack of personalization can lead to inaccuracies in simulation outcomes (e.g., AFib-like behavior in simulations for a patient who actually had AFl, or vice-versa). My goal was to derive parameter sets that favor the initiation of one type of arrhythmia or the other (AFib or AFl). Ten fibrotic left atria were reconstructed from late-gadolinium enhanced (LGE)-MRI scans and the bioelectric parameter space (comprising ion channel expression levels and impulse propagation rates) was explored using a Taguchi L27 Design of Experiments (DoE) approach. Arrhythmias were induced by initializing four atrial regions to different phases of the action potential under each parameter permutation. I ran 300 simulations and manually classified each arrhythmia episode as either AFib- or AFl-like based on prior definitions. I pinpointed a pro-AFl parameter set – bioelectrical conditions under which 89% of all induced arrhythmias were AFl and only 11% were AFib. The pro-AFib parameter set in these preliminary simulations was comparatively less robust (61% vs. 39% for AFib vs. AFl inductions, respectively). My future work on this project will establish stronger relationships between model configurations and simulation outcomes by probing a wider array of possible parameters in a larger population of patient-specific models. Data from the present study will guide future simulations to accurately tailor models to represent the arrhythmic state in patients predisposed to AFl.

The extra cellular matrix (ECM) is a complex, heterogenous environment that plays an important role in cellular functions such as proliferation, signaling, movement, and differentiation. The mechanical properties of the ECM vary spatially and temporally, across and within tissues, i.e., during development and disease progression. 3D biomaterial platforms, such as hydrogels – water-swollen polymeric networks—provide a greater understanding of matrix-cell interactions and can be used to study drug delivery and basic disease mechanisms. My research works to create a double network (DN) hydrogel system that allows for spatial control of ECM mechanics in 3D. Our system contains two different polymer networks, one of which uses light polymerization. I have optimized concentrations of multiple gel components and gel light exposure conditions to allow for accurately patterned stiffnesses within the gels. Currently, I am encapsulating live cells to study the amount of cell spreading and movement in the stiff and soft regions of the gels over the course of a week. I then fix, stain, and image each gel to quantify relative cellular spreading. Additionally, I have synthesized multiple components necessary for gel formation, cultured enzyme producing bacteria to degrade formed gels, and performed western blotting to analyze cellular protein concentrations. Imaging results have shown the DNs and the patterning process are cytocompatible. Current experiments have shown differences in fibroblast spreading between stiff and soft regions; future results are expected to show differences in protein expression within mechanosensitive pathways between patterning conditions. Using multiple, intertwined hydrogel networks, I have engineered a dynamic, heterogenous model of the ECM, enabling me to study cellular responses to mechanical stimuli. Accurate modeling of the ECM will allow for a better understanding of how diseases such as breast cancer progress based on differences in environmental stiffness and provide an in vitro platform for future cellular response research.

Quasars are compact regions at the center of a galaxy that have much higher than normal luminosity. Gas outflowing in winds is common in quasars, and it might play a role in regulating the black hole growth and star formation in the host galaxies. In particular, the ones with extremely high speeds (>~0.1c) have not been thoroughly studied and might pose the biggest challenges to the simulations of how these outflows are driven. A 2020 study by Rodríguez Hidalgo et al. identified 40 quasars with Extremely high-velocity outflows (EHVOs) in the 9th data release of the Sloan Digital Sky Survey by detecting absorption in quasar spectra. The C iv emission properties of this sample were examined in a 2022 study by Rodríguez Hidalgo & Rankine. Compared to the general population as a whole, these extreme outflows seem to be more predominant in quasars with large blueshifts of the C iv emission line, an indication of outflows in emission, but the sample was small. My project aims to answer the question: do EHVO quasars show distinct emission features compared with the general population of quasars? To answer this question, I carried out an extension of a 2022 study, but using a larger sample of 98 new EHVOs that we identified from the latest data release. I built on the python code that our team has previously created, which cross-correlates the sample of 6743 quassars from the 2020 study with measurements calculated by a 2020 study by Rankine et al and allows us to place the EHVO quasars in the C IV emission line parameters space (blueshifts vs emission intensity). Potential connection between the maximum EHVO velocity and increasing C IV blueshifts is also explored.

Increasing mental illness among college students continues to be a critical issue. College life is often a time of great transition, contributing to and potentially exacerbating students' rising stress, anxiety, and other mental struggles. Poor sleep habits may develop during college, and poor sleep quality can amplify stress. To explore how institutions might help students struggling with stress, we conducted a small pilot study examining whether a sleep-focused workshop would reduce stress levels in students at a Pacific Northwest community college. For this pilot study, I helped generate an online pre- and post-workshop survey of demographic questions, the UCLA Loneliness Scale, and the Perceived Stress Scale (PSS). Our results indicated that participants' workshop attendance reduced their stress levels. Before the sleep workshop, participants scored an average of 20.27 (SD=7.41, range=2 to 30) on the PSS, while two weeks after the workshop, the average PSS score had reduced to 16.58 (SD=7.65, range=4 to 27). These results suggest benefits of a sleep-focused wellness workshop for college students. In our present study, we replicated the pilot study. I examined student stress in correlation to sleep quality and assessed these factors through an online survey of demographic questions, sleep quality questions, the Beck Anxiety Inventory, and the PSS. We distributed this survey to participants before and after a sleep workshop run by the Bellevue College psychology department. I hypothesized that poor sleep quality correlates to higher student stress levels. I also hypothesized that the intervention of a sleep workshop would reduce stress levels. I hope that our results provide insight into the utility of wellness workshops for students and whether they are an avenue to help students manage stress and improve their sleep quality, which could improve student mental health by lowering stress and anxiety and improving quality of life and education overall.

The innate immune system must have a rigorous response to many pathogens in order to successfully defend host cells against infections. During infection, inflammasome forming sensors detect pathogen-specific features which release proinflammatory cytokines such as IL-1B for immune activation. The inflammasome forming sensor NLRP1 directly detects multiple signals indicative of infection, including viral protease activity. Recently, it was shown NLRP1 is indirectly activated by bacterial toxins and UV irradiation that disrupt host protein synthesis, demonstrating it can detect environmental stimuli to cause inflammation. However, much of this activation pathway is poorly understood with only one nonpathogenic cause having been investigated. Our goal is to identify the host proteins required for NLRP1 activation and determine whether NLRP1 can broadly detect disrupted protein synthesis. We hypothesize NLRP1 detection of disruption in protein synthesis is a broad strategy to combat infection and may have an important role in causing inflammation in other diseases associated with disrupted protein synthesis, including cancer and neurodegeneration. Knockout cell lines of proteins suspected of activating NLRP1 are produced through lentiviral transduction of Cas-9, a gene editing tool which cleaves off specific nucleotides corresponding to the target gene of each protease sensor in the inflammasome activation pathway. Each cell line is confirmed to be absent of the target sensor via genotyping, and is followed by a functional assay of each knockout line which induces cellular stress targeting the activation of each cleaved protease. We predict that inflammasome activation as defined by IL-1B concentration will be significantly decreased in knockout cell lines targeting key proteases in the inflammasome signal cascade, showing that the overactivation of these proteases is sufficient for inflammasome activation. These results could provide key targets for drug discovery in the treatment of multiple diseases which cause disruption of protein synthesis, including cancer and neurodegeneration.

The Middle Miocene (23-5 Ma) represents a period of rapidly changing climate and active volcanism, particularly in the Pacific Northwest. Our understanding of the structure and composition of plant communities during this timeframe is complicated by a limited or degraded leaf fossil record. Plant fossil assemblages are also often time averaged, representing accumulation of plant material over an extended period. A plant community that was preserved because of a single, short-lived event can provide insight into the composition and structure of that community in life. The Watersnake locality of the Sucker Creek Formation in southwestern Idaho contains two thick (8-14 m) ignimbrite tuffs (volcanic ash layers) that preserve charcoal fragments, reflecting plants that were burned when the tuffs were deposited. In order to identify the woody taxa that made up this community, thin section microscopy is used to examine the preserved cellular detail of the fossil charcoal fragments from the lower tuff. Since ignimbrites are deposited as a part of a single event, the impacts of time-averaging are minimized. Although these deposits reflect a short time, they are likely to integrate across space, providing a view of the broader landscape. The results of this study will reveal the structure of this plant community immediately prior to the eruption that caused the ash flow. I hypothesize that the taxa identified within the ash will be very similar to those of the leaf fossil record found in the shale beds below. A lowland community consisting primarily of wetland Glyptostrobus oregonensis, and Quercus simulata found near flowing water will likely be represented. Upland vegetation consisting of conifers (Pinus, Tsuga) were also likely incorporated into the ash flow as it moved downhill. This study will provide insight into the dynamics of plant community change during the Miocene in relation to volcanic disturbance.

In humans, the KRAS protein normally acts as a switch to regulate cell growth. Acquisition of certain mutations result in constant activation of the protein, leading to uncontrollable cell growth. The most frequent such mutations are at the glycine-12 residue of the protein, including changes to valine (G12V). Cells with this mutation can be recognized and eliminated by T cells engineered to express an antigen-specific receptor (TCR), but posttranslational modifications, such as methylation, can interfere with the ability of such engineered T cells to recognize G12V mutant KRAS. The objective of my project is to identify and eventually disrupt mechanisms that lead to methylation of KRAS mutant antigens. I hypothesize specific enzymes are responsible for methylation of mutant KRAS. To accomplish this, I will generate CRISPR-Cas9 gene knockout libraries to screen for potentially responsible enzymes. The CRISPR library will target human genes encoding methyltransferase and demethylase enzymes. I will co-culture these CRISPR-Cas9 knockout cancer cells with T cells engineered with a TCR targeting the unmethylated KRAS mutant antigen. From the tumor cells that have survived this coculture, I will sequence genomic DNA to determine which knockouts are enriched/depleted after the co-culture. Preliminary results show certain gene knockouts are significantly enriched/depleted in co-culture compared to baseline. The sgRNAs that appear from the high-throughput knockout library most likely to be involved in the KRAS methylation pathway will be individually evaluated in additional co-culture experiments. This should allow me to assess the proliferation/susceptibility of these cancer cells in more detail. For confirmation of mechanism, I will evaluate the methylation status of the KRAS mutant antigen in cancer cells rendered susceptible. My goal is to determine the process leading to methylation of KRAS antigen and then target it to allow for more effective targeting of KRAS-driven cancers using TCR-T cell immunotherapy.

Microchimerism (Mc) occurs when a small amount of genetically different cells (or DNA) is acquired from another individual. Mc is acquired naturally during pregnancy due to exchange between the mother and child and can be detected decades later. Maternal microchimerism (MMc) is when a person harbors Mc from their mother. MMc is frequently detected in healthy adults but is increased in individuals with some autoimmune diseases, including scleroderma. Few studies have investigated MMc, especially whether it changes in a woman after her own pregnancies. One study tested MMc in peripheral blood of women during the time they were pregnant and occasionally detected MMc, but not if the woman had preeclampsia. No study has addressed whether MMc prevalence and quantities change in healthy women according to the time since the woman’s own childbirths or number of childbirths. This study addresses this knowledge gap. MMc was assayed using a panel of polymorphism-specific real-time polymerase chain reaction (qPCR) assays on DNA from peripheral blood. Most assays employed human leukocyte antigens (HLA)-specific primers and fluorogenic probes targeting non-inherited, non-shared HLA sequences. Each woman and her mother were HLA typed to identify an appropriate target. In total, 142 women were tested, and 266 qPCR experiments were run. Preliminary analysis found evidence of MMc in 59 of the 266 samples and a trend of MMc prevalence being highest within the first year postpartum and ten years after childbirth. Prevalence and quantities of MMc are being analyzed in collaboration with a biostatistician. Pregnancy and childbirth are known to affect some autoimmune diseases and cancer risk. Addressing the knowledge gap about MMc according to the time since birth and the number of births in women could provide further insights about some autoimmune diseases and cancers.

Adoptive T cell therapy is a promising therapeutic strategy for the treatment of many hematologic malignancies, however, its efficacy in solid tumors poses several challenges. Some of these challenges include the limited infiltration and activation of cytotoxic T cells due to the effects of a diverse immunosuppressive environment within the solid tumor. One of the main suppressive immune cells present in several solid tumors are regulatory T cells (Tregs) and high numbers of Tregs within the solid tumor have been correlated with poor prognosis. Therefore, there is a clinical need to develop strategies targeting the suppressive immune cells that limit the efficacy of adoptive T cell therapy. Tumors have highly dysregulated metabolism, which results in the secretion of multiple metabolites into the extracellular space. This allows a buildup within the tumor microenvironment, which may have an effect on the infiltrating immune cells. Our group has identified one metabolite, succinate, that enhances Treg numbers within the tumor microenvironment. Here we further explored the effect of succinate on the function of Tregs. To examine this, we identified tumor cell lines that produce succinate (lung and melanoma) and further altered these to modify the levels of succinate secreted by these cells. We then co-cultured high succinate secreting tumor lines with healthy donor CD4+ T cells that were isolated from PBMCs. We screened these cells and found that the higher levels of succinate resulted in higher numbers of Tregs and increased anti-inflammatory function, as evidenced by TGFb levels in Tregs. Future experiments will validate these findings in in vivo mouse models with the aim of developing synergistic approaches to enhance adoptive T cell therapy.

Fire is a fundamental disturbance that drives changes in biome structure. Knowledge of ancient fire regimes may help predict future fire regimes resulting from anthropogenic climate change. Charcoal morphometry (quantified shape of charcoal), particularly charcoal aspect ratio (length:width), is an emerging proxy of ancient fuel type wherein higher mean aspect ratios are associated with grassy fuels. This study aims to experimentally validate this proxy method. Thirty-four modern plant species were sampled from UW Herbarium collections, separating leaf, stem, and reproductive body tissues for each species. Each sample was burned at 500°C for 20 minutes, crushed in a water slurry, and imaged under a binocular microscope. Charcoal particles were enumerated and morphometrics were measured using ImageJ with charcoal particles 125-250 μm and >250 μm analyzed separately to account for differences due to differential particle breakage. Strong evidence was found in the 125-250 μm size fraction through an analysis of variance test (F = 2.66, p = 0.03), that aspect ratio varies as a function of taxonomic group. The strongest evidence for a difference in aspect ratio is found, through Tukey's Honestly Significant Differences to be between graminoid and conifer charcoal (p = 0.03). Evidence is even stronger for a taxonomic effect on aspect ratio in the >250 μm size fraction (F = 3.64, p= 0.007). This variation seems to also be driven by a difference between graminoid and conifer charcoal (p = 0.002), corroborating earlier findings. Future validation of this methodology will be focused on the potential effects of burn temperature and charcoal transport in charcoal morphometric records. Rigorous verification of charcoal morphometry as a proxy of fuel type will help increase confidence in paleo reconstructions of fuel type. 

Hemagglutinin (HA) is a glycoprotein found on the surface of the influenza virus. HA binds to sialic acid receptors on host cells and mediates membrane fusion, allowing the virus to enter the cell. This project investigates how mutations associated with species crossover affect viral fusion mechanisms in influenza HA. HA from the past outbreaks of H5N1 influenza strains in Vietnam in 2004 (VN04) and Indonesia in 2005 (IN05), are being compared to an HA from an on-going avian influenza outbreak using a strain isolated in Colorado in 2022 (CO22). Through site-directed mutagenesis, mutations that affect acid stability and affinity for the human receptor were added to HA based on the VN04 and IN05 strains; we are studying these effects in recombinant protein rather than on infectious virus. These mutations are believed to enable the virus to increase transmissibility among mammals including humans. To compare how HA from these H5 isolates with and without the adaptive mutations behave, hydrogen-deuterium exchange mass spectrometry (HDX-MS) is being used to measure changes in deuterium incorporation on the protein backbone for specific peptide segments, giving a profile of local dynamics and structure throughout the HAs. By comparing the dynamic profiles for each as pH is lowered, mimicking acid-activation in host cell endosomes, we can probe how their structure in important fusion and human receptor-binding regions change as the fusion protein becomes activated. By comparing the structural dynamic changes of the WT and mammal-adapted, mutated IN05 and VN04 HA to the new CO22, we will be able to increase our understanding of the effect of the mutations that are associated with species crossover and hopefully be able to gain insight into the potential of this new avian influenza strain’s ability to become transmissible among humans.

The “Taiwan issue,” a potential breaking point for U.S.-China relations, is essentially about Taiwan’s highly controversial political status and national identification. Historically, the Taiwanese have identified themselves in different ways, as some self-identify as Chinese and some as Taiwanese. However, there has been a significant change in Taiwan’s national identification in recent decades, as people are increasingly self-identifying as Taiwanese instead of Chinese. This paper tries to answer what has led to changes in Taiwan’s national identity. I explore theories of democratization, authoritarian control in mainland China, and elite manipulation through discourse as potential causes for the shift in Taiwan’s national identification. I use democratic indices from the V-Dem dataset to measure degrees of democratization and the political rights and civil liberties indices from Freedom House to measure levels of authoritarian control in mainland China. To measure elite manipulation through discourse in Taiwan, I conduct a content analysis of key concepts related to both identities on Taiwan’s newspapers. I employ these data to test each of the theories in relation to survey results in Taiwan from 1990-2021. To test each of the theories, I conduct a multivariate regression analysis to determine if there is a relationship between each theory and change in Taiwanese national identity. I expect to find a positive correlation between the three potential causes and change in Taiwan’s national identification, which would indicate that democratization, elite manipulation through discourse, and authoritarian control in mainland China has led to the increasing trend of identifying as Taiwanese. Understanding the cause of change in Taiwan’s national identification and how support for the identities change over time provides insights into why and to what extent Taiwan has separated itself from mainland China and helps inform policy implications for both China and the U.S.’ Taiwan policy.

The onset of the 21st century brought with it an increasingly authoritarian world. Since 2005, countries making democratic declines outnumbered those making democratic gains. Why is it that, despite seemingly disparate conditions in individual countries, democratic backsliding is globally on the rise? Although this sudden and alarming decline has drawn the attention of many political researchers, there currently lacks a strong scientific consensus on the major catalysts for democratic backsliding. This research project explores whether political and affective polarization influence the quality of democracy experienced by a country. I hypothesize that the amount of polarization within a state significantly impacts the extent to which democratic backsliding occurs. This hypothesis comes from my observations of the United States’ current cultural climate, where high levels of polarization have contributed to political dysfunction on a national level. Polarization appears to undermine democratic norms and principles by discouraging bipartisan cooperation, encouraging politicians with autocratic policies to run for elections, and incentivizing voters to choose partisan interests over democratic ones. To test this theory, I run a multivariate regression analysis of the relationship between political and affective polarization on over 100 countries, all of which were considered to be some form of democracy in the year 2013. I test this analysis against multiple potential confounding variables, such as the income inequality within a state, public support for democracy, and rate of immigration. I expect my evaluation to show a significant positive relationship between a nation’s level of polarization and the rate of democratic decline. Erosion of democracy has real-world significance for citizens. Since this period of democratic backsliding began, election freedom and checks on human rights abuses have deteriorated. It’s important to identify a catalyst for 21st century backsliding because it provides us with a method for preventing further liberalism to occur.

Non-evangelical denominations of Protestantism have experienced unsettling declines in funding and membership since the 1960’s. Seattle churches are no exception. A collective of “progressive” identifying Seattle churches are tackling the organizational issues they face. Together, they are asking questions such as: “How can we disrupt the entrenched Christian elitism that separates our church from the community landscape?” and “How can we reteach our congregations what it means to be in community with other people?” Political scientists and religious scholars have offered many theoretical explanations for why some churches, denominations, or religions thrive when others do not. This paper seeks to quantitatively assess mainstream explanations for progressive Protestant decline by using survey data collected from this local collective of churches. The survey utilizes likert scale questions, which will be analyzed using a logistic regression to understand how individuals relate their political beliefs to their religious beliefs in the context of their congregations.This paper advances three hypotheses: progressive Protestantism decreases theological and political cohesion among their congregations; progressive church-goers develop political beliefs that are less influenced by the church than their conservative counterparts; and lastly that progressive churches have weak self-conceptions of their theological teachings. Progressive theologies inherently encourage open-mindedness and individuality: beliefs that are often corrosive to the ideological strength of a church because they undermine the church's moral authority. I believe that progressive Protestants generally-- and these Seattle churches specifically-- have so diluted the coercive influence of the church, that their congregations are struggling to maintain a cohesive theological outlook. The Christian church undergirds many Americans’ political beliefs. Understanding how a church’s adoption of a progressive theology impacts the organizational strength of a congregation, and the ideological beliefs of the individual are crucial considerations for a country increasingly divided by political polarization.

In face-to-face surveys containing sensitive questions, Social Desirability Bias (SDB), respondent’s tendency to provide socially acceptable responses rather than truthful ones, can compromise data accuracy. Randomized response techniques (RRT) are survey models that allow respondents to provide scrambled responses, thereby circumventing SDB. In this study, we introduce a mixture optional quantitative RRT model that combines the elements of both the Pollock and Bek (1976) additive RRT model and the Greenberg et al. (1971) unrelated question quantitative RRT model. We examine the utility of the proposed mixture model using a unified measure of efficiency and privacy introduced by Gupta et al. (2018) that provides a metric of both predictive accuracy and respondent privacy. We also account for the lack of trust in RRT models. Both empirical and theoretical results show that the mixture model outperforms the two component models. The proposed optional quantitative mixture RRT model provides a survey technique that can account for not only SDB but also respondent lack of trust, leading to more accurate and interpretable data used to inform decision making that does not compromise the privacy of respondents.

Radon in high concentrations has been proven to be one of the world’s leading causes of lung cancer. Climate change is an ongoing problem that affects the environment and human beings; however, there is not yet a widely known relationship between radon exposure and climate change. Our research aims to find a correlation between climate change and risk of radon exposure. We are reviewing how radon exposure potentially increases through the lens of greenhouse gas emissions, melting ice caps, and human habits based on increased global warming effects. In our literary review, we are comparing how radon is measured, finding the limitations of each technique and which technique is best suited to measuring radon in air, soil, and water. We are also working with our collaborators overseas to understand the radon concentrations in soil in Hebron, Palestine. We will investigate our hypothesis that radon exposure will increase with climate change.

In this project, I conducted a literature review comparing the strategies the governments of Vietnam and Thailand are implementing to mitigate and adapt to climate change. Southeast Asia is projected to be severely impacted by climate change. In particular, rising sea levels are likely to encroach on lowland coastal regions that account for much of the population and serve as centers of economic activity. Additionally, nations in this region continue to undergo rapid economic growth, and therefore face a dilemma in that the carbon emissions which have largely fueled their quick growth are also contributing to climate change imperiling their populations. The purpose of this study is to compare the climate response of two major growing economies in this region in terms of mitigation and adaptation, and in this manner discover how differences in political structure, national wealth, and the level of specific risk faced by each nation has shaped their climate policies. I conducted research by reviewing policy reports published by these governments, non-governmental organizations, and scholarly articles. My review suggests that both governments have tended to downplay the scale of climate adaptation needed to prepare for the massive population resettlement and economic adjustment from the inundation of several major cities. Additionally, the findings show that Vietnam has undertaken more extensive mitigation efforts than Thailand, in part stemming from a varying response to the 2008 financial crisis. In contrast to Thailand, which responded by further developing fossil fuel infrastructure to pursue greater energy independence, Vietnam worked toward the same aims by expanding green energy sources like wind and hydropower. By understanding the causes of these varying climate responses, we can better identify areas in which further action is needed to bring policy in line with scientific projections and understand how institutional structures influence the success of climate policy.

Climate change is a growing threat to communities worldwide, with extreme weather events like droughts and wildfires causing food insecurity and affecting the lives of millions of people. Despite the availability of research that describes the consequences of climate change, there is a lack of urgency in the response to this crisis. To understand the barriers that inhibit action on climate change, a study was conducted on 206 STEM students at the University of Washington. The study surveyed the students to identify factual and conceptual barriers to addressing climate change. We distributed questionnaires through social media and undergraduate classes. I analyzed survey responses to compare and contrast the concern levels for ecocentric and anthropocentric consequences of climate change. The findings of the study indicate that environmental education was not associated with more climate change knowledge, and students were more concerned with ecocentric impacts and anthropocentric consequences that directly impact basic human needs like water, food, and shelter. We aim to develop a teaching tool that addresses the conceptual barriers identified in their research. The results of the study emphasize the need to shift the focus towards addressing the immediate impacts of climate change that affect human well-being. The lack of urgency in the response to the climate crisis highlights the need for more education and action to mitigate the effects of climate change.

Climate change has two dimensions: mitigation and adaptation. In spite of progress towards net-zero emissions, climate change is already underway, and its effects are visible through extreme weather events including hurricanes. Many argue policymakers should focus on adaptation along with mitigation. Given that some climate effects are location-specific, adaptation requires that households carefully think about where they buy property, given that an investment in a home is the most important source of household wealth. To minimize property damage, households could take steps to enhance the resilience of their property, which should be reflected in property value. In the absence of federal regulation, only 29 states of the 50 U.S. have enacted laws mandating property owners disclose flood history and risk to a property. Louisiana was among the first states to enact these regulations, while in 2022, Texas strengthened its legislation. What explains this variation in flood disclosure legislation over time and space? Collecting data from federal and non-profit sources, I will examine a slate of factors: state partisanship (control of the legislature and the gubernatorial office) from 2017-2019; percentage of Trump votes in 2016; the number of water-related disasters from 2013-2018; percentage of flood hazard area; in-migration; length of coastline; percentage of forested area; and the number of flood related deaths from 2013-2018. In R, I will conduct linear regression models to estimate the effect and significance of these factors on the response variable: the grade/strength of flood disclosure legislation. I will also conduct a Cox Proportional hazard model to examine the effect of these factors on the passage of flood disclosure legislation. The results of the analysis will provide evidence as to why certain states are more likely to have strong flood disclosure legislation, and what events could result in a state passing flood disclosure legislation.

In the past year climate activists have engaged in increasingly disruptive tactics to try and draw attention to the looming climate crisis. To raise awareness for their cause, activist groups have utilized a variety of confrontational and controversial protest tactics, such as sit-ins, hunger strikes, road blockades, and occupations of crucial industries, for example, airports. However 2022 featured a unique new form of climate protest: art attacks and museum vandalization. This radical new protest tactic is designed to force public attention on the climate crisis by visually destroying and threatening precious artworks. A slew of climate art protests swept news headlines in 2022, drawing both condemnation and admiration. For this project I created a dataset of art attacks and museum vandalism focusing on when, where, and what group executes such attacks with the goal of understanding the context of this new protest phenomenon. Preliminary data was collected by tracking news reports and activist social media posts to determine the relevant data points. The preliminary findings from this project find that all instances of museum vandalism occurred in a 7-month period in 2022, with attacks concentrated in the days leading up to COP27 in November. In total, 39 protests occurred in 11 (primarily European) countries, with the majority occurring in England (7), Italy (8), and Germany (8). Established climate activist groups conducted the majority of these protests, with only a handful of these incidents being perpetrated by individual actors. The activist groups most implicated in the dataset are: Just Stop Oil (England), Letzte Generation (Germany), and Ultima Generazione (Italy), which aligns well with the countries in which the majority of the protests took place. The results of this project could indicate the future of climate protest tactics- and public reception- as the climate crisis worsens and activist groups become more desperate.

The marine intertidal is subject to challenging hydrodynamic forces. Organisms that reside within these zones; such as purple (Strongylocentrotus purpuratus) and green (Strongylocentrotus droebachiensis) sea urchins, must have strong attachment to prevent dislodgement, ambulate, and feed in intense environmental conditions. Sea urchins attach to the substrate via their podia (tube feet). The disc found in the distal part of tube feet has a duo-gland that releases specialized adhesive compounds and de-adhesive enzymes to achieve adhesion. Adhesive performance was evaluated to determine if purple sea urchins, that are predominantly found in high wave exposure areas, have better adhesive performance than green sea urchins, which are abundantly found in areas protected from wave action. We evaluated the following adhesive performance variables: stem mechanical properties, tube foot disc tenacity, attachment area, speed of detachment and whole animal adhesive force. We found that for most adhesive performance metrics, purple urchins performed better than greens, except for detachment speed. These results suggest that, in the field, behavioral changes may play an important role in determining risk of detachment for sea urchins. Understanding how morphology, behavior and mechanical properties of tube feet work in conjunction to provide strong attachment, is critical to predict sea urchin fitness in challenging environmental conditions.

Avian scavengers perform a critical ecosystem service by breaking down dead and decomposing organic matter and recycling nutrients. However, the scavenging dynamics of birds have received relatively little attention compared to the dynamics of scavenging mammals. Carcasses provide an ephemeral food source for avian scavengers, which presents them with the unique challenge of locating the carrion. This makes detection and consumption efficiency a topic of interest as not all avian scavengers follow the same pattern of arrival at a carcass. Here, we examine avian scavenger succession – the order in which species arrive and consume carcasses – through monitoring ungulate carcasses in Washington State. We found that turkey vultures outcompeted all other avian scavengers when they were present in summer months, while black-billed magpies and common ravens outcompeted golden and bald eagles in winter months. This confirms previous studies that have found that turkey vultures have morphological and genetic advantages that enable them to readily find carcasses and that body size does not determine succession in this scavenging guild. Our study highlights how social scavengers outcompete solitary scavengers and reach carcasses sooner, regardless of the physical characteristics of a species.

An estimated 5-8% of the American population have cerebral aneurysms, showing higher rates of development in patients with common risk factors like hypertension, smoking, and family history of cerebral aneurysms (CA). This study focuses on understanding the causes of aneurysmal subarachnoid hemorrhage (aSAH), where a CA ruptures, resulting in bleeding in the brain. Endovascular coiling is a minimally invasive surgical treatment method for aSAH. Unfortunately, up to 30% of endovascular coiling treatments are unsuccessful, leading to aneurysm recurrence, growth, or rupture. The risk of these outcomes can be predicted using Computational Fluid Dynamics (CFD), a tool that quantifies the hemodynamic environment by solving the equations of motion for a fluid. The CFD simulations calculate factors significant in predicting the effectiveness of coiling treatment including flow rate, wall shear stress, and pulsatility. In this project we have studied the effect of using patient-specific blood viscosity values (the resistance of the blood to fluid flow), that have typically been standardized for all patients in CFD simulations. We have analyzed the effect of using patient-specific blood viscosity on pre-treatment patient-specific computational fluid dynamics simulations of endovascularly-coiled cerebral aneurysms. Preliminary results show that there is an expected improvement in CFD simulation predictive power of treatment effectiveness when patient-specific blood viscosity values are used. We hope to improve the predictive power of CFD simulations regarding the treatment outcome of aneurysm coiling, allowing us to better predict aneurysm recurrence, and eventually guide treatment outcomes.

Cyanobacteria are tiny photosynthetic microbial organisms responsible for producing roughly an eighth of the oxygen we breathe. Synechococcus is a model cyanobacteria, meaning the species has characteristics making it easy to study and modify. As a scientific community, we don’t know the function or purpose of many genes expressed by Synechococcus. The goal of this project is to determine the function of particular genes hypothesized to be important to the adaptive survival of Synechococcus in different environments. We are approaching this by building a start-to-finish gene characterization method, starting with computational analysis to identify genes of interest, followed by knocking out, or disabling these genes and observing the effect on the growth of the culture. On the computational side, I’m now analyzing residual gene expression, using that information to characterize gene clusters, and analyzing external data to infer genetic context. On the laboratory side, I’ve characterized the growth of the un-modified base strains and developed procedures for genetic modification. Identifying the function of Synechococcus genes allows scientists to better study the response of Synechococcus to varying environments, which is especially important in a changing climate. Increasing understanding of the molecular mechanisms of Synechococcus also opens the door to genome engineering for the production of biofuels, plastics, and other commodities, or for using Synechococcus as a tool for bioremedial carbon sequestration. Additionally, the genes of Synechococcus are similar to those in other related oceanic microbes such as Prochlorococcus, the most ubiquitous photosynthetic organism in the world. For all these reasons, Synechococcus is an important model organism, and a deeper understanding of its biology will bolster our sparse understanding of marine genomics.

(Bobola et al. 2020) has shown that ultrasound pulsed at a certain frequency and applied to the brain of an AD mouse model reduced the burden of plaque in the brain. This was key as plaque are tied to long-term poor clinical outcomes. Further research has demonstrated the ability of transcranial ultrasound to enhance cerebrovascular flow through upregulation of endothelial nitric oxide synthase (eNOS) - Eguchi et al. 2018. eNOS is an enzyme that produces the vasoprotective molecule nitric oxide, which Eguchi et al demonstrated caused a reduction in plaque buildup. In addition, up-regulation of eNOS might be applied to treat not only AD but also vascular dementia. We seek to demonstrate that transcranial applied ultrasound to wild-type and AD brains after TBI can delay the onset of AD and reduce the extent of associated symptoms. I contributed to this research project by performing (1) the surgeries to obtain a model of TBI in mice, (2) the transcranial ultrasound treatments on these animals, (3) computational analysis of plaque burden and brain activity, (4) and behavioral tests on the treated animals. Preliminary results indicate that differing protocols of US treatment can decrease amyloid beta plaque burden. Intraneuronal eNOS has increased with ultrasound treatment, of potential therapeutic utility for advanced AD. Finally, our results depend upon the age and total plaque burden, unexpected findings that will motivate more research.

In recent years the field of global health has been plagued by accusations of “neocolonialism.” Rather than genuine involvement in health research, physicians and public health researchers in Global South countries are relegated to menial positions in the research pipeline, and local priorities are not reflected in the research that receives the bulk of international funding. The multilateral donors which fund and manage this research are said to devalue the contributions of Southern researchers, who, many studies of global health journals have shown, are not allowed to take on leadership roles in public health research “collaborations” with Western institutions. Moreover, a large critical literature has characterized the field of global health as taking a narrowly conceived, “vertical” view of health in developing countries. This biomedical/technological bias leads donor-funded research to neglect the social determinants of health and illness. The current project contributes to our understanding of the origins and practical manifestations of these institutional biases of health research in Uganda. A historical analysis of colonial medicine in Uganda was conducted and it was found that medical research in Uganda indeed has historically neglected to address the social determinants of health such as poverty, labor conditions, and the presence or absence of social infrastructure (e.g., roads and sanitation systems). In addition, a review of recent COVID-19, Ebola, and HIV/AIDS research confirms the hypothesis of other scholars that donor-driven public health research tends to treat health in isolation from the larger political economy of development, and is biased toward a highly biomedicalized view of health. I conclude by offering an interpretation of these biases and how they vary by individual donor, as well as some suggestions as to what can be done to make Ugandan health science genuinely equitable and responsive to local, rather than Western, needs.

Merkel cell carcinoma (MCC) is an aggressive skin cancer with high risk of metastasis. Recent developments of PD-1/PD-L1 immunotherapy significantly improved treatment outcomes of metastatic MCC. While approximately half of patients respond to immunotherapy, the other half of the patients do not benefit. Thus, it has become increasingly important to identify factors that potentially impact immunotherapy response. Multiple studies in melanoma have demonstrated that patients who are older or have a higher body mass index (BMI) show better immunotherapy response. The purpose of this study is to explore the effects of age and/or BMI in MCC patients to immunotherapy response. I helped create a cohort of 183 patients who had undergone immunotherapy initially identified in a longitudinal single center registry. I then helped collect information about age and BMI at the start of immunotherapy along with other clinical features. Treatment response, disease-specific and overall survival were analyzed in 183 patients using cox regression and natural cubic spline models. During this process, I assisted in distinguishing the number of splines we would like to use for our analysis, as well as choosing the best model to accurately represent data for different variables. After adjusting for age, sex, and stage, BMI did not have significant impact on overall survival (p=1.0), objective response (p=0.5), or disease progression (p=0.8) while on immunotherapy. A nonlinear relationship between age and immunotherapy response was observed and showed potentially worse response to treatment in older patients. However, this was not statistically significant (p<0.1). Unlike prior studies in melanoma, we found that BMI does not have a significant impact on immunotherapy in MCC. Older age may have a negative impact on the response. This warrants additional research into the difference between melanoma and MCC to further elucidate mechanism of actions.

Birth asphyxia is the inability of a newborn to begin and maintain breathing. Twenty-three percent of neonatal deaths globally are caused by birth asphyxia [1]. Birth asphyxia results in a neurological injury called hypoxic ischemic encephalopathy (HIE). Rapid HIE screening within six hours after birth is crucial to identify neonates at risk. Unfortunately, the diagnostic equipment is impractical for low resource settings because it is costly ($20/test and $5,000 for equipment) and requires technical staff, that are in short supply, to operate. We hypothesize that a cost-effective device can be developed for HIE analysis. pHast Cam quickly screens for birth asphyxia and HIE in infants via a paper-based blood pH sensor. The device combines an inexpensive pH sensitive dye, a smartphone camera, and a fixture that controls the imaging environment to quickly identify acidosis that results from HIE. A low-cost paper-based strip is made with a water-soluble resin doped with a pH-sensitive dye, bromothymol blue (BTB), and a membrane to filter out red blood cells. The fixture removes lighting variation. The smartphone camera records the pH indicator image, and an algorithm captures, reduces noise, and accesses color change. pHast Cam incorporates four features: 1) accurate assessment of acidity within 0.05 pH units, 2) require only a few microliters of blood, 3) use electrical hardware and software only from the smartphone, and 4) affordability. At this stage, we have achieved a regressive linear model that predicts buffered solution acidity. In the future, we will transition from measuring buffered solutions to blood-plasma. Ultimately, we expect pHast Cam to screen for HIE by quantifying plasma pH in neonates so that timely therapeutic interventions and plans to address long-term complications may occur. [1] Diaz-Rosello JGP, Niermeyer S, et al. WHO Basic guidelines on new born resuscitation. 2012.

Half a million people develop drug resistant tuberculosis (TB) each year. Cases of drug resistant TB often result in poorer outcomes both healthwise and economically for patients, and many populations lack access to the resources needed to treat resistant TB. Increased antibiotic resistance has resulted in an urgent need to develop new, cost-effective drugs that are effective against Mycobacterium tuberculosis, the bacteria responsible for TB. In my research, I am using deep learning methods to design peptides that bind to the enzyme ClpP, a vital protease and known antibiotic target in M. tuberculosis. A class of drugs called Acyldepsipeptides (ADEP) have been shown to bind to ClpP and cause cell death in M. tuberculosis by preventing the formation of the ClpP complex with necessary ATPases, resulting in significantly lower proteolytic activity. We used the structure of ADEP as a basis for the peptide design and employed Rosetta, a macromolecular prediction and design software, to generate cyclic peptides bound to ClpP. I then used a sequence based deep learning tool to generate multiple sequences for each backbone design and computationally validated the resulting structures with AlphaFold, a highly accurate, machine learning based structure prediction tool. The structure of the ClpP binding interface resulted in it being a difficult target to design for with current deep learning methods. One peptide binder was predicted to bind to ClpP in our preliminary design rounds. We will chemically synthesize this binder and test it against ClpP in an enzyme inhibition assay. If the binder inhibits ClpP, it can serve as a basis for an effective and low cost drug that targets the ClpP enzyme in drug resistant TB. We will also expand and refine our design pipeline to produce more binder designs that can serve as viable drug candidates.

The epidermis is a multi-layered tissue at the body surface made of cells called keratinocytes that protect humans from infection and dehydration. Keratinocytes undergo a unique program of differentiation while moving upwards in the epidermal tissue. During the final stage of maturation, each keratinocyte must eliminate its organelles and nuclei to allow flattening of the cells to form the uppermost layers that provide a water-tight seal for the body. To understand this crucial biological process, I aimed to train an online artificial intelligence (AI)-based image analysis algorithm called Biodock AI to detect key morphological features of differentiating keratinocytes and of engineered epidermal tissue. I annotated specific cellular and tissue structures in a set of microscopy images to train a new supervised AI model to recognize these features. In brightfield histological images of engineered human skin, I used an area selection tool to identify the boundaries of the epidermis to assess tissue thickness. To train our pipeline to recognize a common pathological feature, I next labeled nuclei that were improperly retained in the cornified layers of various drug-treated tissues. After the algorithm was trained by Biodock AI, our pipeline successfully replicated our tissue outlines and identified retained nuclei. Zooming to the subcellular level, I next labeled organelle features within fluorescence microscopy images of keratinocytes. I selected endoplasmic reticulum (ER) fragments that had broken off the tubular network during differentiation. Our AI-trained image analysis pipeline successfully identified ER fragments with high concordance with those annotated by a lab member. In summary, I successfully trained and implemented AI-based image analysis pipelines to detect tissue and sub-cellular features that characterize the process of epidermal differentiation. Our results demonstrate the potential of AI algorithms to accelerate imaging-based research to understand cellular differentiation and tissue pathology while mitigating bias and error from human investigators.

For Saccharomyces cerevisiae (S. cerevisiae) to survive, they must either synthesize nitrogenous bases or reside in a nitrogenous base-rich environment. If there is a mutated enzyme along the pathway that makes the nitrogenous base uracil, then the organism would be unable to synthesize uracil and die without uracil. When S. cerevisiae is grown in 5-Fluoroortic Acid (5-FOA) media, the URA3 enzyme will catalyze 5-FOA into a toxic intermediate that causes cell death when incorporated into RNA. In URA3 mutations, the intermediate is not produced, and S. cerevisiae can survive. In an experimental setup where we explored mutation patterns in aging S. cerevisiae cells, we used URA3 to select mutants that grow on 5-FOA media when uracil is provided. However, when analyzing the mutant sequences, there were fewer URA3 mutations than expected. We performed Sanger sequencing on the URA3 gene in individual mutants and were surprised to find that some did not have any mutations. We, therefore, sequenced the whole genome of those mutants and found that they had missense mutations in the URA6 gene. We then analyzed the URA6 locus from the original pool. Across the URA6 gene, the mutations appeared randomly spread with the possibility of some mutation hotspots, indicating that they could be loss of function mutations. In contrast with the URA3 mutants, we observed URA6 mutants were able to grow 5-FOA media when uracil was not provided. There may exist an alternative pathway for URA6 S. cerevisiae mutants to synthesize uracil and survive. To further study this observation, we plan on analyzing the function of the URA6 gene in 5-FOA media compared to the URA3 gene and the components of the URA6 pathway. Ultimately, the result of this study could clarify nitrogenous base production pathways in S. cerevisiae and the impacts of 5-FOA on S. cerevisiae.

Sexual assault is pervasive (1 in 4 women experience assault in their lifetime; RAINN, 2019) and holds the highest risk for posttraumatic stress disorder (PTSD) of any type of traumatic event. However, Asian women are underrepresented in media representations and research regarding the prevalence of assault and the etiology of PTSD. The purpose of this study was to examine the frequencies of lifetime occurrences of sexual harassment, unwanted sexual touching, and penetrative sexual assault (i.e., rape) in Asian women and compare these rates to their white peers. College-aged women (N = 201) completed an online version of the Sexual Experiences Survey - Victimization (SES-SFV). 60.86% of Asian and 74.65% of white women reported experiencing sexual harassment, 28.70% of Asian and 39.43% of white women reported experiencing unwanted sexual touching, and 20.00% of Asian and 21.13% of white women reported experiencing rape. Three independent samples t-tests compared rates of sexual harassment, unwanted sexual touching and rape in white women (n = 69) compared to Asian women (n = 101). Results showed no significant differences between occurrences of sexual harassment and rape. However, white women (M = .75, SD = .43) experienced significantly more unwanted sexual touching compared to Asian women (M = .57, SD = .50), t(168) = -2.4, p = .016. These findings indicate that Asian women encounter similar rates of sexual harassment and rape compared to white women. The parity found between Asian women and white women is underrepresented in media and research and may have negative consequences on the believability of and support for Asian women who have experienced harassment or rape. Further research is necessary to understand the unique mental and physical health impact of sexual assault on Asian women. 

Evaluating the risk of developing breast cancer is an important aspect of cancer care as it can allow for more tailored screening strategies and preventative therapies. Clinicians use multiple measures to determine a patient’s risk of developing breast cancer, including breast density on mammography and genetic mutations. Background parenchymal enhancement (BPE) on magnetic resonance imaging (MRI) has shown promise to improve stratification of breast cancer risk in women at high-risk of cancer development. BPE is the increase in signal intensity of normal breast tissue on dynamic contrast-enhanced (DCE) MRI after the administration of contrast agent. Despite BPE having an association with an increased risk of breast cancer development, the biological basis of this increased enhancement is unknown. The aim of this study is to investigate what biologically drives BPE by connecting quantitative MRI measurements with pathological markers from normal breast tissue. Our study cohort includes women that received prophylactic mastectomies and DCE-MRI scans acquired ≤1 year before surgery. From mastectomy specimens, pathological measures of COX-2, VEGF, and Ki-67 are used to measure inflammation, vascular recruitment, and proliferation, respectively. To quantify BPE, I used in-house software to correct pre-contrast images using N4 bias field correction and segment the whole breast. I then applied the breast mask to the pre-contrast MRI and used fuzzy c-means clustering to automatically segment fibroglandular tissue (FGT) from surrounding fat, generating an FGT mask. This mask was then applied to the DCE-MRI series, which includes pre- and post-contrast images, to calculate BPE, which is the mean percent enhancement across FGT. As part of ongoing work, I will obtain more specific measurements in quadrants of the breast from which the pathology specimen was derived. I will then correlate BPE measurements to the pathology measures to determine if any associations exist between BPE and inflammation, vascular recruitment, and proliferation.

HIV infection impairs B cell function, in turn, altering immunoglobulin production and function. Immunoglobulins (Igs) exist as isotypes including IgA, IgG, and IgM. Within the IgA and IgG isotypes there are subclasses IgA1-2 and IgG1-4 respectively; all with distinct functions. Previous studies have shown that HIV infection influences Ig isotype and subclass concentrations in serum, but few have explored their concentrations in the breast milk of mothers living with HIV (MLHIV). Widespread use of antiretroviral treatment during pregnancy has led to an increase in the incidence of HIV-exposed and uninfected infants (iHEU). iHEUs have heightened immune activation and inflammation and display high infectious morbidity compared to compared to HIV-unexposed infants. It is plausible that immune factors transferred in breast milk contribute to altered immunity in iHEU. Therefore, knowing whether HIV infection impacts total immunoglobulin concentrations or inflammatory biomarkers in breast milk is important. I used enzyme-linked immunosorbent assays (ELISA) to measure the concentrations of immunoglobulin isotypes and their subclasses and Luminex to profile cytokines and chemokines in breast milk 4 weeks after delivery. Assays were performed according to manufacturers’ instructions and sample values were extrapolated from a standard curve. I compared these factors between MLHIV and uninfected mothers using Mann-Whitney U test. MLHIV had significantly higher mean concentrations of total IgG1 (36.9 ug/mL versus 26.6 ug/mL, p=0.018) and IgG3 (2.9 ug/mL versus 1.3 ug/mL, p=0.0013). There was no difference in concentrations of IgA and IgM between the groups. Furthermore, MCP-1, MIP-1-β, and SDF-F-α were the most abundant chemokines in breast milk in both groups. However, we found no significant difference in concentrations of cytokines and chemokines in breast milk of MLHIV versus controls. Overall, we observed increased breast milk concentrations of IgG1 and IgG3 in MLHIV. Future work will explore implications of these IgG subclasses on iHEU immunity.

Leishmaniasis, caused by various species of Leishmania, is a global public health burden, with the World Health Organization reporting over a million cases in the last 5 years, leading to over 20,000 deaths annually. Leishmania and closely related organisms have the unique ability to glycosylate thymidine (Ts) residues to form a base termed Base J. Previous work by the lab has shown that de novo insertion of J is initiated by a protein called JBP2 that is targeted to specific regions of the genome by J2-TDP, a Tudor domain protein. Tudor domains bind to methylated arginine or lysine residues that are most commonly found in histones. To determine if targeting JBP2 to a genome location was sufficient to initiate formation of J, we wanted to target JBP2 to a region of the genome that does not normally contain J. To achieve this, we took advantage of the ability of the tetracycline repressor (TetR) to bind to the operator (TetO) by fusing JBP2 and J2TDP separately to TetR and expressing the proteins in Leishmania harboring a TetO cassette in a locus that does not contain J. I constructed a cassette with the necessary components which contain in order- a drug selectable marker, TetO sites, and a GFP, as a reporter gene to monitor any possible effects J has on the cells when inserted. Using CRISPR/Cas9, I inserted this construct into Leishmania that expressed the TetR fusion proteins. I then grew these strains in the presence or absence of Tet and then determined if J was present at this location. The results of these findings will give us a deeper understanding of the molecular mechanism(s) responsible for this specificity as well as this will offer potential opportunities for development of novel therapeutic agents against Leishmania.

Treatment of individuals with HIV using antiretroviral therapy (ART) is highly effective, but effective clinical management depends on maintaining therapeutic drug concentrations. Antiretroviral (ARV) drug concentrations in patients with HIV can vary due to differences in drug metabolism, medication adherence, or interactions between multiple drugs. These individuals may have subtherapeutic or supratherapeutic drug concentrations, putting them at risk of treatment failure, acquisition of drug resistance, and risk of hospitalization or death. Current measurement of ARV concentration is done through liquid chromatography tandem mass spectrometry, which requires expensive equipment and requires a labor-intensive protocol. This restricts accessibility to specialized laboratories, making it difficult for persons with HIV to have routine measurements of ARV drug concentrations. The goal of the project is to develop an assay that is simple to perform and uses standard equipment to increase access to routine clinic-based drug level monitoring to improve HIV care. We designed an assay using a 2-step process of DNA strand transfer and quantitative polymerase chain reaction (qPCR) to quantify integrase strand transfer inhibitors (INSTIs). We tested for dolutegravir (DTG) and cabotegravir (CAB) in both buffer and plasma -- the latter to simulate patient blood samples. We were able to demonstrate that the assay could quantify clinically relevant drug concentrations of DTG and CAB. By developing an assay that can be readily integrated into most clinical laboratories, we will contribute to increasing access to routine HIV drug level monitoring to improve clinical HIV care and maintaining viral suppression in persons with HIV.

Intracranial Epidermoid Cysts (IECs) are benign tumors located in the parasellar region and cerebellopontine angle of the brain. The incidence of IECs is ~1% of all intracranial tumors. These slow-growing, asymptomatic tumors are significantly enlarged when discovered which leads to trigeminal neuralgia, visual and hearing loss, tinnitus, severe face pain, and seizures. IECs are suspected to be a result of disruptions during neural tube closure during embryogenesis. Given their rarity, IECs are poorly investigated, and the mechanism of IEC formation remains unknown. Consequently, there is no drug therapy available up to date and surgery is still the only option. However, because the cyst lining adheres to important brain structures, a gross resection can result in deafness, facial weakness and chemical meningitis. These observations highlight the need to develop additional therapeutic options for these patients. We performed paired-end Whole Exome Sequencing (WES) on DNA isolated from sporadic IECs resected from 6 patients. From these, 4 were investigated with matching peripheral blood and 2 were sequenced without. We processed raw unmapped reads in tumor-normal and tumor-only mode as appropriate, following the GATK best practices for variant calling. Of the 5230 somatic variants found, 233 variants were found in protein coding regions (183 missense, 46 frameshift, 4 stop-gained/lost) suggesting damaging effects on the phenotype. Two mutations were identified in oncogenes (FCGR2B, PABPC1) and 2 mutations were identified in tumor suppressor genes (RSPO2, KMT2C), indicating possible tumor driver candidates. Gene enrichment analysis using the EnrichR interface revealed patterns of downstream microRNAs (miRNAs) regulation and signaling pathways implicated in the development of IECs. The validation of these findings and the study of their role in the IEC pathogenesis are currently under investigation in our laboratory. The identified driver mutations may serve as drug targets for the development of therapies to treat IECs.

Oral tolerance to antigens is a mechanism by which immune responses are inhibited to prevent chronic inflammation and tissue damage in response to common exposures, such as dietary antigens or commensal bacteria. Regulatory T cells (Tregs) are an important cell type in such regulation of immune responses, especially in the intestines. There is, however, heterogeneity within Tregs, including a subset that expresses the transcription factor RORgt. However, the mechanisms by which RORgt+ Tregs carry out their suppressive function are currently unknown. Intestinal Tregs are induced in the mesenteric lymph nodes (MLNs) by antigen-presenting dendritic cells (DCs) that migrate from the gut. Antigen transfer from DCs to Tregs is crucial for the development of oral tolerance and DCs are largely regarded as being upstream of Tregs. However, it has also been shown that Tregs play a role in DC migration via a CTLA-4-mediated mechanism. Because this DC-Treg relationship is not fully understood with regard to Treg heterogeneity, I have examined the changes in DC populations in a mouse model where the RORgt+ Treg population alone has been ablated. To do this, I developed a new panel of antibodies to use in flow cytometry in order to characterize the subpopulations of DCs in the intestines and related organs. I harvested and processed murine spleens, MLNs, Peyer's Patches, and small intestine lamina propria in order to compare the populations systemically and locally. I anticipate seeing fewer DCs in the mice lacking RORgt+ Tregs and more DCs in the small intestine and Peyer's patches. This work furthers our understanding of the intricacies of the intestinal immune system. This knowledge can be applied to vaccine research, as RORgt+ Tregs have been implicated as suppressors of immune response to oral vaccines. Intestinal immunity is also of interest in allergy and gut inflammation research.

Heart disease is the leading cause of death in the United States. Echocardiography is used clinically to highlight cardiac structures, wall motion, and contraction abnormalities to diagnose heart failure. Heart failure is classified into diastolic (relaxation) or systolic (contraction) heart failure using left ventricular ejection fraction (LVEF), a measurement of the amount of blood pumped during each contraction. Global Longitudinal Strain (GLS) analysis, which measures the stiffness and deformation of the heart wall during contraction, has recently emerged as a more sensitive metric of systolic function that predicts cardiovascular mortality in patients. Elamipretide (ELAM;SS-31) is a mitochondrial-targeted intervention that improves aging heart mitochondrial and diastolic function. In this study, we used an aging mouse model to compare sex-specific outcomes in systolic function by LVEF and GLS. We hypothesized that GLS would be a better predictor of systolic dysfunction in mice than LVEF, and that ELAM would repair aging systolic dysfunction. We compared young (4-6 mo) and old (25-26 mo) male and female mice using 2D echocardiography to obtain left ventricular parasternal short and long axis images. A cohort of aged male mice was imaged before and after 8-week ELAM treatment. These images were analyzed using Vevo LAB software using conventional echocardiography to measure LVEF and speckle-tracking echocardiography (STE) for GLS and LVEFStrain. Statistical analysis was performed using GraphPad Prism Software. Limited change in LVEF was observed by conventional echocardiography. Using STE, GLS and LVEFStrain declined with age. Treatment with ELAM restored GLS in aging mice to young levels. Here we show that the more sensitive STE analysis reveals that aging mice exhibit both systolic and diastolic dysfunction. The research supports our hypothesis that ELAM treatment improves systolic function in aging. Future treatments to target systolic dysfunction can be assessed in an aging mouse model using STE.

Diffusion-weighted imaging (DWI) shows great potential for improving breast cancer detection and diagnosis. Primary findings from the ECOG-ACRIN A6702 multi-site, multi-vendor clinical trial indicate that DWI apparent diffusion coefficient (ADC) values may help reduce false positives and unnecessary biopsies. Gradient nonlinearity (GNL) correction was previously found to improve the accuracy of ADC mapping within and across MRI vendor systems. In this study, we evaluated the impact of GNL correction on breast lesion ADC measures in the A6702 dataset. The dataset comprised 81 suspicious breast lesions (28/81 malignant) in 67 women. Standardized DWI scans were acquired across 9 different MRI scanners. ADC maps were created from DWI scans, and ADC values were measured for each lesion. Direction-averaged GNL correction maps were constructed based on scanner-specific gradient specifications. ADC map correction was then performed through pixel-wise scaling by the GNL correction maps using custom software developed in MATLAB. Lesion ADCs before and after GNL correction were compared using a two-tailed z-test. ADC diagnostic performance (benign vs. malignant) was evaluated using area under the receiver-operating-characteristic-curve (AUC), and optimal ADC cutoffs were chosen to maximize specificity while maintaining 100% sensitivity. GNL-corrected lesion ADCs were significantly lower than uncorrected ADCs (1.12±0.29 vs 1.17±0.30x10-3mm2/s, p<0.001). GNL error in lesion ADCs varied across gradient systems (mean ∆ADCvendorA=0.14±0.08, ∆ADCvendorB=0.03±0.02, ∆ADCvendorC =0.004±0.01, p<0.001). GNL correction produced a slightly lower optimal ADC cutoff (1.33 vs. 1.35x10-3mm2/sec). However, no overall difference in diagnostic performance was detected: AUCuncorrected=0.78 (95% CI 0.68-0.88), AUCcorrected=0.79 (95% CI:0.69-0.89), p=0.22, and 18% potential biopsy reduction for both. This study showed GNL substantially affects lesion ADC measures, with significant variability across different vendor platforms. These findings suggest that GNL correction should be implemented to ensure uniformity and consistency in diagnostic breast lesion ADC measures across MRI platforms, especially for multi-center clinical studies.

As of 2021, there were approximately 38.4 million people living with HIV who require routine viral load testing. Viral load testing returns a quantitative measure of viral concentrations and is indicative of antiretroviral therapy efficacy and adherence compliance, with lower viral loads correlated to better health outcomes. Quantitative polymerase chain reaction (qPCR) is the gold standard for measuring viral load, but is not accessible to many clinics and patients around the world due to its long assay times and requirements of specialized equipment and highly trained personnel. As a result, qPCR is limited to centralized testing facilities far from the point-of-care (POC), leading to delayed results or loss of follow-up. Our group has addressed this by developing an HIV viral load test using recombinase polymerase amplification (RPA), which has a 20 minute sample-to-answer time and is more appropriate for POC settings. Our test involves the formation of discrete fluorescent nucleation sites which can be counted to estimate the viral load. However, our test fails to accurately quantify higher HIV viral loads (>3,000cps/rxn). We have difficulties distinguishing between individual sites at these higher copy numbers due to sites merging. In this project, I address the limited dynamic range of this test by performing RPA between two glass slides and investigating the effects of different slide thicknesses and concentrations of polyethylene glycol (PEG), a crowding agent used in RPA reactions. I perform nucleation site analysis using computer vision techniques to measure nucleation site radius and intensity and study how these factors affect site diffusion and amplification. By analyzing nucleation site behavior, we demonstrate potential for an HIV viral load test with a higher dynamic range and gain a better understanding for RPA nucleation site formation, ultimately helping to improve access to testing and treatment for people living with HIV.

In storytelling, mental state terms (MSTs) are words that attribute thoughts, feelings, emotions, and perspectives when describing the internal state of the characters. Although MSTs have been shown to be indicators of literacy and Theory of Mind development among monolingual children, less is known about how bilingual children who were exposed to two languages simultaneously develop MSTs. In this study, we aim to examine the usage and development of MST among Mandarin-English bilingual children by coding the narrative elicited using two wordless-picture books, Frog Goes to Dinner and Frog Where are You? by Mercer Mayer. The data was collected from sixty students enrolled in 1st grade (N = 20), 3rd grade (N = 21), and 5th & 6th grade (N = 19) in a Mandarin-English Dual Immersion program. The research team coded MSTs into three categories (thoughts, desires, and feelings) and examined lexical diversity by analyzing the different meanings of the word "think" in both languages across all three groups. We expect to see more MST usage in older children and also an association between Mandarin and English MST categories. We may also see more lexical diversity in Mandarin compared to English due to cultural and linguistic differences in narrative production. Understanding the trajectory of MST development can further our understanding of how bilingual children acquire languages and the potential cross-linguistic differences between Mandarin and English. This study may also help practitioners in the field of speech-language pathology understand the importance of comprehensive bilingual assessments which measure skills in both languages.

Many curricula today fail to connect STEM with the issues students and their communities face. This shortcoming can lead to increased dropout rates and equity gaps especially in early STEM courses. With the COVID-19 pandemic and the George Floyd uprising, fostering students’ ability to tie STEM into the issues of social justice have never been more important. In this study, we investigate whether a Course-Based Undergraduate Research Experience (CURE) centered around the public health implications of indoor CO₂ can give students a greater understanding of the relevance of STEM to social issues and help them see STEM as a tool to solve problems in their communities. Students were administered a brief pre-survey before beginning the CURE. They were then introduced to the low-cost CO₂ sensors and the public health implications of high indoor CO₂. Students formed groups, formulated their research questions, and collected data. Students then analyzed their data and presented their research to their peers. A post-survey was administered following the CURE. We are currently analyzing the results of the surveys and post-curriculum interviews. The preliminary results suggest that connecting CUREs to relevant social issues in early STEM courses is a powerful tool that not only teaches students to interpret the world around them but also to change it.

The drug rapamycin has been shown to extend lifespan in model organisms ranging from mice to fruit flies, but little is known about how genetic variation affects the response to rapamycin. It is possible that some individuals might not respond to the drug, or may even respond negatively. To study the impact of genetic variation on rapamycin sensitivity, the Promislow lab has been using the fruit fly, Drosophila melanogaster, to test the ability of rapamycin to slow early development in different strains. They found a range of responses, from strains that were completely resistant to those that were highly sensitive. To better understand why resistant and sensitive strains differ, the Promislow lab analyzed their metabolome profiles. The metabolome consists of diverse small molecules, metabolites, that are fundamental to sustaining life in cells, and the relative abundance of metabolites is referred to as a metabolome profile. By comparing metabolome profiles, the Promislow lab found that the metabolome of sensitive larvae treated with rapamycin was similar to those of larvae starved of nutrients. One possible explanation for this observation is that sensitive larvae on rapamycin-treated food eat less food than resistant larvae. To test my hypothesis, I am measuring how much food they ingest, using dyed food, and scoring how much dye each larva ingests over time. If sensitive larvae eat less rapamycin-treated food than resistant larvae, then we can investigate why rapamycin affects feeding. If no significant difference is found between scores for sensitive and resistant larvae, then we can investigate why rapamycin affects early development in sensitive strains when their food consumption is not limited. By determining if sensitivity to rapamycin is accompanied by differences in feeding, then we could potentially manipulate feeding to sensitize flies, and possibly humans, to this beneficial drug.

Cyanobacteria are ancient single-celled photosynthetic organisms, prevalent throughout Earth's oceans. Over billions of years, cyanobacteria have evolved genes that enable them to survive across a diversity of adverse, ever-changing environmental conditions. However, researchers are faced with the problem of not understanding the role of many of these genes. This research project entails tracking down the function of some high variance genes in marine Synechococcus, an important model organism and a genus of cyanobacteria. We will test gene function by generating knockout strains in which a gene of interest is inactivated, and testing the growth of these mutant strains in various conditions. In particular, our project focuses on the importance of the flavodoxin gene, which codes for an electron transport protein that is involved in photosynthesis and is expressed in response to iron scarcity. This gene inactivation is done with a plasmid, which is a genetic structure in bacteria that can replicate itself independent of bacterial chromosomal replication, that’s enabled to knock out the flavodoxin I gene when inserted into Synechococcus cells. We insert the plasmid into our Synechococcus cells and once the DNA is taken up by the cell, we then use CRISPR technology to remove the gene. Successfully creating the flavodoxin knockout of Synechococcus establishes the procedures necessary for generating knockouts of other genes that could be expressed in similar patterns as flavodoxin. Ultimately, this research furthers our understanding of how Synechococcus’ genes allow it to adapt to various environments and contributes to ongoing research on how organisms might withstand the pressures of Earth’s ever-changing climate.

Pathogenic bacteria often promote their growth by introducing proteins into host cells they are infecting. Some of these introduced proteins hijack host cell ubiquitin signaling pathways. Ubiquitin (Ub) is a small protein that is involved in many signaling pathways within eukaryotic cells. In ubiquitin-mediated degradation, the degradation of a target protein is promoted by the attachment of chains of ubiquitin onto the target protein. The attachment of poly-ubiquitin chains is facilitated by a class of proteins called E3 ubiquitin ligases. Salmonella Typhimurium secretes into host cells an IpaH/SspH class E3 ubiquitin ligase called SspH1. My research investigates the mechanism for how SspH1 modifies its target protein, Protein Kinase N1 (PKN1), with poly-ubiquitin chains. I am investigating whether ubiquitin is transferred sequentially onto PKN1 or if a long poly-ubiquitin chain is first formed on SspH1 and then transferred en-bloc onto PKN1. Using biochemical methods, I will compare whether the rate of poly-ubiquitin chain synthesis is faster starting with unmodified PKN1 or mono-Ubiquitinated PKN1, an intermediate in the sequential addition pathway. I hypothesize that the formation of poly-ubiquitin chains onto mono-Ub-PKN1 is slower than the formation of poly-ubiquitin chains onto free PKN1. This would suggest that the mechanism does not occur through the sequential addition of ubiquitin. Learning more about the mechanism of SspH1 will allow us to both better understand the IpaH/SspH class of proteins and better understand how ubiquitin can be transferred onto protein substrates.

The placenta is a crucial fetal organ providing oxygen and nutrients to the developing infant. Researchers typically use placental cell models to study the placenta, which are derived from immortalized cells. The use of in-vitro placental cell models is important because human samples are difficult to obtain, and placental biology is highly species-specific. Despite this, our understanding of the characteristics of these cell lines and how they compare to placental tissue samples is limited. This project aims to determine which placental cell model most directly reflects the gene expression of the human placenta. RNA sequencing data from the placental cell models HTR-8/SVneo, JEG-3, BeWo as well as data from primary trophoblast cells was obtained using the Gene Expression Omnibus (GEO) database or through lab-generated datasets. Data for each cell line was combined into a single dataset of shared genes (n=6835) and individual datasets of genes unique to each cell model. For genes that were unique to each cell model, I performed KEGG pathway analysis and characterized placenta specific genes using the Human Protein Atlas (HPA). BeWo cells expressed the highest number of unique genes (n=355) and shared the highest number of genes with the primary trophoblast cells (n= 1,167). Pathway analysis showed that genes unique to primary trophoblast cells (n=2661) were overrepresented in 24 pathways, unique BeWo genes were overrepresented in 15 pathways, while unique HTR-8/SVneo genes (n=355) were overrepresented for a single pathway- the neuroactive ligand-receptor interaction. Placenta specific genes were expressed within the uniquely expressed genes for JEG-3 (n=1), BeWo (n=1), and primary trophoblast cells (n=17), but not HTR-8/SVneo. Ultimately, the results from this project will provide a tool to evaluate differences in placental cell models and aid the placental biology research community in understanding which cell line is most representative of human placental tissue samples.

During non-REM slow-wave sleep, the thalamus and cortex generate widespread synchronized epochs of action potential firing that repeat at 1 to 7 Hz. This synchronization is thought to be an essential component of healthy sleep. Cortical excitatory synaptic feedback to the thalamus is required to maintain synchronized firing epochs across the thalamus. A major source of cortical to thalamic feedback comes from a subset of layer 5 (L5) pyramidal neurons (PNs) which occur in the largest numbers in the primary motor cortex. There is also evidence that the basal dendrites of those neurons in the motor cortex receive monosynaptic executory inputs from excitatory thalamic neurons but the number and strength of those connections are not known. For our project, we decided to quantify the number of thalamic inputs onto the basal dendrites of those L5 PNs that project back to the thalamus. I used tissue from the motor cortex of thy1 mice which express a yellow florescent protein in the L5 PNs that send axon branches to the thalamus. The tissue was treated with an antibody to the VGLUT2 protein which is selectively expressed in excitatory synaptic terminals from thalamic neurons. The VGLUT2-containing terminals were visualized using florescent immunocytochemical techniques combined with confocal microscopy to count the number of putative thalamic synaptic terminals that were closely opposed to spines (postsynaptic protrusions) on the L5 PN basal dendrites. Preliminary results suggest that between 5-20% of the dendritic spines are closely opposed to VGLUT2-containing presynaptic terminals. The study of this thalamocortical loop will allow for a better understanding of the processes that are necessary for proper sleep and the implications of disrupted neuron synchronization.

The periodontal ligament is a connective tissue that anchors teeth to the bony socket and is crucial in providing nutrition for the survival and functioning of the human body through the mastication of food by teeth. Because the periodontal ligament is anatomically sealed off from the oral cavity, no non-invasive techniques currently exist to investigate it in-vivo, making the development of sound in vitro models critical for periodontal research. With periodontal disease affecting over 743 million people worldwide, in-vitro research to develop regenerative therapies to replace diseased periodontal tissue is urgently needed. To achieve this, we have developed a novel 3D in-vitro model, which closely mimics the in-vivo periodontal ligament. The 3D tissues are fabricated by inverting an array of silicone posts into silicone molds containing a cell-collagen gel mixture in a 24-well plate. Once the tissues have been incubated and the collagen polymerizes, magnetic mechanical force stretches tissues on posts. The post deflection is used to calculate tissue stiffness and contractility. Preliminary data shows a reduction of contractile force in the tissue constructs after 24 hours of mechanical stretching. I expect to find similar outcomes through additional experimentation. Understanding the periodontal ligament’s response to mechanical force is crucial for its effective restoration and ensuring that it is mechanically sound. The novel in-vitro 3D model that we have developed provides a valuable opportunity to better comprehend the ligament's response to these forces. By performing further experiments with this model, we can gain a deeper understanding of the periodontal ligament, allowing for informed decisions when it comes to replacement and repair in patients. This model offers controlled and repeatable experiments, providing more accurate insights into the biology of the periodontal ligament and contributing to the advancement of periodontal disease treatment.

Kānaka Maoli (Native Hawaiian) health practices have been significantly impacted by colonialism, beginning with the illegalization of practices like hula, lā’au lapa’au, ‘ÅŒlelo Hawai’i (Hawaiian language), and the growing inaccessibility of cultural foods. Increasing attention has been paid to creating culturally-grounded interventions that address these disparities, which have proven to be effective in increasing health outcomes in Indigenous communities. This study aims to understand the potential benefits of Papa and Pohaku, a culturally-grounded family intervention created and led by esteemed elder Uncle Earl Kawa’a through Keiki O Ka ‘Ä€ina (KKA). KKA is a non-profit organization built to perpetuate Kānaka Maoli culture for ‘ohana (families) and keiki (children). Uncle Earl Kawa'a, a respected kupuna (elder), leads courses on creating a papa (pounding board) and pohaku ku’i ‘ai (stone pounder) to promote healing and wellbeing through grounding participants in cultural practices. To understand the impacts of the intervention, two focus groups were conducted with participants and KKA staff. Questions explored the benefits of participating in Papa and Pohaku, specifically its impact on relationships and traditional Hawaiian knowledge. Our team used Collaborative Qualitative Analysis, which is a structured and rigorous method of conducting inductive thematic analysis. All research team members identified as Indigenous, consisting of one faculty advisor, one doctoral student, and two undergraduate researchers. Respondents reported the intervention positively impacted the following: 1) participants’ pilina (relationships) with their ‘ohana, partners, and with other participants; 2) appreciation for the huaka’i (journey), or process, and commitment to future growth; and 3) understanding of and connection to mo’oemeheu (Kānaka culture). The findings of this study indicate the various benefits of culturally-grounded family-based interventions and a greater need for the availability of culturally-grounded interventions for Indigenous communities.

Many age-related diseases in humans such as Parkinson's and Alzheimer's involve intracellular protein aggregation, but much is still unknown about the molecular mechanisms behind how this occurs. Characterizing these mechanisms is therefore important for developing effective treatments for age-related illnesses. Our work investigates the relationship between cell life span and aggregation of processing bodies (P-bodies), which are cytoplasmic ribonucleoprotein (RNP) granules that form inside cells experiencing stress and perform several molecular functions that appear to benefit cells experiencing stress. Using GFP-tagged Dcp2 as a P-body marker in S. cerevisiae and microfluidics to study single-cell lifespans, I demonstrated that P-bodies aggregated in aging cells that were not experiencing other forms of stress. P-body aggregation also correlated to the remaining lifespan of any given cell. To investigate this link further, I adjusted cytosol pH and observed a relationship between cytosolic pH and P-body aggregation rate. Slowing of P-body aggregation correlated to extension of cell lifespan. This suggests the need for additional research to determine whether there is a causal link between P-body aggregation and fatal single-cell pathogenesis and if so, whether these pathogenesis mechanisms are conserved in human cells and therefore a possible target for treatment for age-related illnesses.

Nature uses only four nucleobases to store genetic information in DNA. However, additional synthetic bases which use Watson-Crick pairing have been developed and are known as non-standard bases (NSBs). NSBs P, Z, B and S incorporated alongside standard bases A, G, C and T compose DNA strands using a new genetic alphabet. Nanopores offer the potential capability for direct single-molecule sequencing of DNA containing non-standard bases (NSBs). Using a voltage gradient, DNA strands were directed through a nanopore, the biological membrane protein MspA, while we measured the ion current through the pore over time. In studying the effect of NSBs on the ion current through the pore, we observe current measurements corresponding to the Z base have a different noise profile compared to other bases. We hypothesize this noise may be associated with pH-dependent protonation of the base. To test this hypothesis, we conducted experiments with identical sequences in buffers of pH 8 and pH 7, as Z is known to have a pKa of 7.8. I analyzed the noise from the ion current signals to look for signs of protonation. I found increased current noise values associated with the Z NSB in pH 7 compared to pH 8, while the canonical A base had no change in noise values from pH 7 and pH 8, supporting the hypothesis that the increased current noise is due to protonation of the Z base. In addition to indicating potential sensing abilities of nanopores for probing protonation kinetics of DNA, this research contributes to a better understanding of the fundamental mechanisms that control the currents in nanopore sequencing of DNA.

Cystic Fibrosis (CF) is a recessive genetic disease that affects roughly 70,000-100,000 people worldwide. CF is caused by mutations in the CFTR gene, which encodes for a channel that regulates the flow of water and ions into and out of the cell. CFTR deficiency results in mucus buildup and other defects that lead to increased bacterial burden and inflammation. However, whether or not CFTR deficiency itself is sufficient to initiate or potentiate epithelial inflammatory responses is unknown. Inflammasomes are multiprotein complexes that upon activation initiates a lytic form of cell death called pyroptosis and the release of pro-inflammatory cytokines including IL-1B, a hallmark of CF. Previous studies have shown that inflammasomes are expressed in the epithelia of mice and humans. Although CF is largely a disease that affects airway epithelium, other tissues such as the skin and gut are also affected by CFTR dysfunction. Moreover, we and others recently found that inflammasomes play an important role in host defense of epithelial barriers, including the intestinal epithelium. Thus, we hypothesize that inflammation in the gut of CF patients may be caused or enhanced by inflammasome activation. To test this possibility I used both human and mouse intestinal organoids as models and ran a Forskolin-induced Swelling (FIS) Assay to assess CFTR function in presence and absence of a CFTR inhibitor (CFTRinh-172). Having established this model system, I am now evaluating the impact of CFTR function on inflammasome activation. We anticipate that our work may reveal a link between CFTR dysfunction and inflammasome activation which may provide further insight into how inflammasomes affect inflammation in the gut epithelia of CF patients.
 

The mammalian target of Rapamycin (mTOR) signaling cascade plays an important role in a variety of cellular processes, such as autophagy, cell proliferation, and protein synthesis. Previous depictions of signaling through the mTOR pathway have suggested linear signal transduction; however, this does not accurately represent the network of interactions between proteins in complexes of this pathway nor their dynamics in response to stimuli. To better characterize mTOR protein interaction network (PIN) dynamics, SEPS lab has developed a panel of antibodies targeting key proteins in mTOR signaling for use in quantitative multiplex co-immunoprecipitation (QMI), a method of detecting changes in protein interactions using flow cytometry. Following QMI of mTOR signaling proteins in serum-starved and serum-refed mouse 3T3 fibroblasts, I validated changes in select interactions from this pool separately via co-immunoprecipitation and western blot analysis. The lab then applied inhibitors of mTOR pathway constituents, including PI3K, AKT, MEK, ERK, and mTOR, to define modules of interactions that comprise the PIN and observe changes in these interactions with stimulation after application of each inhibitor, which I again validated via co-immunoprecipitation and western blot analysis. Finally, to validate antibody specificity in human cells, I prepared human embryonic kidney 293 (HEK293) cells for short interfering RNA (siRNA) transfection and knockdown of mTOR pathway proteins targeted by antibodies from the initial panel. Assuming these HEK293 cells lack any additional proteins with high affinity for these antibodies, I expect flow cytometry data to reflect specificity seen in the 3T3 fibroblasts. Conducting this validation is critical for ensuring the reliability of the PIN changes observed in QMI analysis. These experiments allow us to evaluate coordinated interactions between mTOR pathway proteins and their dynamics during signaling events, which is highly useful in developing treatment strategies for mTOR pathway-associated disorders.