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

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

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

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

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

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

Two common craniofacial anomalies are cleft lip and palate, where the lip and roof of the mouth do not form completely, and craniosynostosis, where the soft spots of the skull fuse prematurely. Despite the prevalence of these birth defects, the genetic mechanisms by which they occur are still widely unknown. Hedgehog (Hh) signaling is a core developmental pathway that plays many roles in skull development including functioning as a guidance cue for cranial neural crest cells (cells that provide the foundation for bone and cartilage within the head) and regulating bone ossification (the hardening of the bone by osteoblasts developing into osteocytes). I am exploring the mechanism by which elevated Hh signaling changes cell fates, either through neural crest cells or osteoblasts, to influence craniofacial development. I investigated a mutant embryonic mouse model to identify regions where overexpression of Hh correlates to abnormal craniofacial phenotypes. I explored and measured these phenotypes via imaging, utilizing a genetic Hh reporter mouse line and skeletal stained embryos of various ages. My early qualitative and quantitative observations show patterns of widening midline structure that are seen by the increase in eye spacing and incomplete formation of the nasal and palate regions, as well as regions of premature ossification in the parietal regions of the skull. These findings suggest that elevated Hh signals result in abnormal development of the craniofacial region similar to cleft lip and palate and craniosynostosis. Deriving from these findings, I’m continuing to explore how Hh signaling plays a role in craniofacial development as well as gaining insight into the mechanism in which craniofacial anomalies arise.

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

Escherichia coli (E. coli) abundance is commonly used to indicate water quality and environmental health. The pH of water has been shown to affect the survival of E. coli. Possession Sound is an estuary that faces a wide range of pH (around 7.5-9.0) throughout the year due to alkaline salt water from Puget Sound mixing with acidic fresh water from the Snohomish River. Primary production, organism respiration, nutrient runoff, carbon emissions, and currents also affect pH levels. This study aims to analyze the relationship between pH and E. coli abundance in an estuarine environment. PH and E. coli data was collected from 2018 to 2023 by myself and other Ocean Research College Academy students. PH data was collected with a YSI EXO2 Sonde. E. coli data was collected using a Niskin bottle to obtain water samples which were then transferred to Petri dishes for growing and counting E. coli. My preliminary analysis shows that Possession Sound’s average pH range is around 7.5-8.5, with pH being higher in spring and summer than in fall and winter. Early analysis using Spearman’s Rank Correlation suggests that pH and E. coli have a weak, inverse relationship. There is minimal research on the relationship between pH and E. coli in a marine setting, so my study helps to provide insight into the relationship between E. coli and pH in a unique estuary.

The paradigm for manganese (Mn) cycling in the marine environment has shifted over the past two decades to include not only the +IV and +II oxidation states. It is now recognized that dissolved Mn(III) can also exist when stabilized by organic ligands. Mn is critical for sustaining life and influences the cycling of many other bioactive elements. Because of this, further research is needed for understanding how Mn cycles in the environment, both between physical and chemical phases. My project aimed to look at how Mn cycles between dissolved and particulate phases and its three environmentally relevant oxidation states along the salinity gradient of the Mississippi River delta and what role organic ligands play in mediating Mn transformations. I hypothesized that the salinity gradient would influence the availability of organic ligands, which would promote the oxidation of dissolved Mn(II) and particulate Mn oxides (MnOx), making the cycling of Mn during estuarine mixing more complex than previously understood. To test this, I collected water from the Mississippi River and the Gulf of Mexico to conduct a mixing experiment to model the salinity gradient. UV-Vis spectrophotometry was used to analyze particulate and dissolved Mn speciation as well as the characteristics of the organic matter present. Inductively coupled plasma-mass spectrometry was used in analyzing dissolved Mn and Mn flocculants. Preliminary results show increases of dissolved Mn during mixing, and potential loss of particulate MnOx. Combined, these suggest redox cycling of Mn during estuarine mixing impacts its solubility and ultimately transport to the Gulf of Mexico. This region experiences heavy nutrient loading that leads to seasonal hypoxia. Understanding the cycling and solubility of Mn is imperative because it has broader implications for redox processes and element cycling in the Northern Gulf of Mexico, especially during hypoxic events.

Corepressors are proteins found in all eukaryotes that work with DNA-binding proteins to repress many genes. Keeping some genes off, yet ready to quickly turn of if needed, is essential for development and physiology. Our project aims to identify interacting proteins that work with TPL, a conserved plant corepressor, to form a transcriptional repressor complex. To uncover these proteins, we created a visually screenable plant line containing RUBY, a reporter that expresses throughout the plant, turning it dark pink to purple. We next created a synthetic repressor dCas9-TPL and guide RNA (gRNA) construct that binds to and represses the RUBY reporter. Roots of plants with both constructs appeared whitish-pink, indicating dCas9-TPL is transcriptionally repressing RUBY. We then mutagenized 40,000 individuals from this line using the chemical Ethyl methanesulfonante (EMS), which creates new point mutations in random locations throughout the genome. We identified 257 individuals from 129 mutagenized families with dark pink roots, which show that repression by dCas9-TPL has been impaired. Many of these adult plants had phenotypes in addition to appearing pink, including miniaturization, infertility, and irregular growth patterns, suggesting that the mutations we found are affecting other pathways that require TPL. Using Mendelian genetics, we are currently characterizing the mutation types (i.e. homozygosity, recessive, or dominant) as well as establishing complementation groups. We will then backcross the lines with the parent line to eliminate extraneous mutations and perform whole genome sequencing to determine the precise mutation causing loss of repression. This will also tell us if repression was due to mutating a TPL interactor, or mutating one of our reporter or repressor constructs. By finding genes required for TPL to act as a corepressor, we hope to understand conserved mechanisms of corepressor activity across diverse eukaryotes.

Some genes are on all of the time in most cells, and carry out functions that are essential for life. Unsuprisingly, essential genes are difficult to study, as interfering with their function leads to death. One such critical component is the multi-protein Mediator complex, which is found at every eukaryotic promoter where it coordinates activation of gene expression. My project focuses on one of the core components of the Mediator complex, MEDIATOR21 (MED21). While MED21 is required for gene activation, the Nemhauser Lab recently found that it also plays a role in repression of gene expression through interaction with the corepressor protein TPL. I would like to be able to differentiate the role MED21 plays in activation versus repression using the plant model Arabadopsis. This work is made more complicated by the fact that most mutations in MED21 lead to lethal phenotypes. As an alternative I recently developed a new technology called a molecular switch that turns off MED21 in certain tissues or in reponse to addition of a chemical. The molecular switch relies on the expression of serine integrases that recognize, and recombine the DNA between, two specific DNA sequences. By expressing an integrase portein from a promoter that is only expressed in secondary roots, I can study MED21 loss of function in a small pool of stem cells while the rest of the plant is wild type and healthy. Plants that have undergone this cell-type-specfic switch exhibit several abnormal root phenotypes including agravitropism, increased root formation, and more root hairs. My next experiments include uisng a switch from wild-type MED21 to a mutant form incapable of binding to the corepressor TPL. This study will help us better understand the role MED21 plays in repression versus activation, and how state switching contributes to organogenesis.

In response to changing conditions, organisms express genes to optimize the match between their phenotype and the environment. Understanding the mechanisms for how genes are turned on or off is therefore an important research area. One challenge in conducting this research is that many of the proteins involved in regulating gene expression are essential to life, and disrupting their function can lead to death. My research focuses on the essential gene SPT6, which encodes a protein that works with RNA polymerase during the elongation phase of transcription. Recently, the Nemhauser Lab has found that SPT6 also plays a role in transcriptional repression. My project aims to differentiate the role that SPT6 plays in transcriptional activation and repression by disrupting its expression in Arabidopsis. Given that SPT6 mutants do not survive, here I test the use of a new tool that allows me to remove my gene of interest in a particular tissue at a particular time. The tool is based on a molecular switch that relies on serine integrases which can recombine DNA between two specific sequences. So far, I have worked with my mentor to rescue SPT6 mutants with a target that expresses the wild-type version of SPT6. Once the integrase is expressed, the recombination turns off the SPT6 gene and turns on a fluorescent reporter. I express the integrase from a promoter that is active only in the first stages of making a new root, so I can observe the impact of loss of SPT6 function in a cell type unnecessary for plants to survive in lab conditions. This project promotes an understanding of the multiple roles of SPT6 during the transtition from repression to activation, and as SPT6 is highly conserved across eukaryotes, my work in plants may also contribute to understanding human diseases. 

During mitosis, the kinetochore plays a central role in ensuring the proper segregation of chromosomes by connecting them to spindle microtubules, which facilitate the equal distribution of chromosomes to daughter cells. In budding yeast, the Dam1 complex is an essential protein complex that binds the kinetochore to spindle microtubules. The Dam1 complex strengthens the kinetochore-microtubule attachment by self-assembling into a sliding ring around microtubules. This self-assembly occurs at nanomolar concentrations of the complex in the presence of microtubules, but in their absence, appreciable oligomerization occurs at concentrations in the micromolar range. Dimers of the complex predominate in high salt concentrations (500 mM NaCl). This is thought to be due to hydrophobic interactions between the monomers. Yeast strains relying on human histones in place of their yeast histones grow slowly. This slow-growth phenotype is rescued by several different mutations in the Dam1 complex. Preliminary characterization of the mutant Dam1 complexes led to the hypothesis that the mutations that allow the yeast cells to adapt to the humanized histones changed the monomer-dimer equilibrium for the Dam1 complex. To measure the affinity of Dam1 complex monomers for each other, I purified the wild-type and mutant protein complexes and used size-exclusion chromatography and mass photometry to determine the different quaternary structures that arise at different concentrations of the complex. I found that each mutation that enhances growth of yeast strains with humanized histones decreased the affinity of the Dam1 complex monomers for each other. The results of this investigation yielded a greater understanding of the requirements for accurate chromosome segregation.

Almost every form of cardiac disease is characterized by fibrosis, or the accumulation of collagen, an extracellular matrix (ECM) protein, secreted by the cardiac fibroblast. The buildup of fibrosis is a major clinical burden, as it contributes to diastolic dysfunction, or the heart’s inability to relax, and arrythmias, or an irregular heartbeat. In previous studies, the Davis lab has found that in chronic injury, the heart likely undergoes minor offenses along with periods of rest which accrue over a lifetime. Even when exposed to repeat injury stimuli, the heart is able to recover, and the cardiac fibroblasts can transcriptionally regress. Yet, what remains unclear is when the heart experiences repetitive stress, which is common with hypertension, will these once-activated cardiac fibroblasts have a more aggressive response? And if so, are the activation cues stored in the primed external environment, or are they intrinsic to the cell? To address this, we developed a fibroblast isolation and injection protocol that will ultimately allow us to isolate discrete populations of fibroblasts and study them in hearts void of injury. Our results found that fibroblasts from donor hearts that were subjected to a myocardial infarction injury were detectable at 4 and 14 days post cardiac injection but had little proliferation. However, there was an increase in host fibroblasts recruited to the graft site, many of which were proliferating, and fibrosis was found within these same regions. These results demonstrate that cardiac fibroblasts from the same strain can be isolated and adoptively transferred to other hearts, without exogenous ECM. We can apply this baseline protocol to further examine fibroblast memory in vivo in a model of intermittent hypertension.

The hallmark neuropathological finding of Parkinson’s Disease (PD) is the presence of intraneuronal protein aggregates, consisting of aggregated proteins and misfolded forms of alpha-synuclein. These intraneuronal protein aggregates, known as Lewy bodies, are implicated in many neurodegenerative diseases. Lewy pathology spread in a PD brain correlates with clinical disease progression. Glucosidase, beta, acid (GBA) gene mutations, the strongest genetic risk factor for PD, is also associated with accelerated disease progression and altered extracellular vesicles (EVs). EVs play a crucial role in intercellular communication and delivery of bioactive cargos throughout the central nervous system (CNS). I use a human neuronal cell culture model derived from induced pluripotent stem cells (iPSCs) to examine how GBA mutations alter EV composition, and investigate whether EVs truly act as a vehicle for the seeding of Lewy pathology in other cells, potentially accelerating the propagation of Lewy pathology throughout the CNS. To isolate and purify EVs from the conditioned media of neurons, I use centrifugation and size exclusion chromatography. I visualize and quantify the EV’s size and concentration using a ZetaView nanoparticle analyzer. I perform Western Blot Analysis for candidate cargo proteins within EVs, including alpha-synuclein, ubiquitinated proteins, and EV intrinsic proteins (CD-63 & CD-81). I isolate EVs from the media of GBA PD or WT control neurons expressing alpha-synuclein-GFP fusion protein and apply these EVs to GBA PD or WT neurons. I anticipate that EVs secreted by GBA versus control neurons will contain increased alpha-synuclein protein levels and that increased cell death, endolysosomal trafficking defects, and aggregation of endogenous alpha-synuclein will be associated with the uptake of GBA EVs by recipient neurons. This work will provide evidence supporting the role of GBA in influencing Lewy pathology propagation via EVs, which could elucidate a novel therapeutic mechanism that could be targeted to slow the progression of neurodegeneration.

Currently, over 6.6 million Americans use walking canes, rollators, and forearm crutches. However, little work has been done to improve the practicality of mobility aids for users. Prior work on modifying these mobility devices has centered around sensing and monitoring user interactions with their mobility device, without changes to the core structure of the devices. Our project aims to explore a set of mobility aid modifications including aesthetics, comfort, and ergonomics. We conducted over 15 qualitative interviews with mobility aid users using phenomenological interviewing strategies to understand user preferences and experiences better and gain feedback on possible adjustments to mobility devices. After qualitative analysis and creating codes based on patterns observed in the interviews analyzed, we identified and compiled unique experiences amongst mobility aid users into a codebook. We then sought to address these observations using fabrication methods such as 3D printing, laser-cutting, and soldering to modify existing mobility devices and develop prototyping materials. Subsequently, we conducted a follow-up design workshop to have users develop modifications and accessory ideas using the tools and templates we provided. Some modifications considered included interactivity stickers, physical feedback mechanisms, and improved mobility aid tip designs. Ultimately, we gained feedback for modifications in future mobility aids research and produced guidelines from our experiences working with mobility devices that can improve community input in accessibility aid research. This work also contributed valuable insights into approaching mobility aid improvements from a Human-Computer Interaction perspective.

While research on burnout has focused specifically on service-oriented roles (e.g., healthcare), recent trends indicate a growing prevalence of burnout in academic settings. Burnout syndrome (BS) is a set of psychological symptoms arising from interplay of chronic occupational stress and individual factors. These symptoms often manifest as emotional exhaustion, depersonalization, and diminished professional satisfaction. Academic burnout has become a concern due to its association with poor academic performance, dropout rates and mental health symptoms (e.g., depression & anxiety). This study employed a pre-and post-design, assessing burnout before and after a burnout management workshop. An online survey via Qualtrics gathered data from community college students in the Pacific Northwest (N = 28, primarily women (78%), aged 16-20 (63%), including dual enrolled high school students. The survey employed a revised Purdue Burnout Questionnaire and collected demographic information. The burnout management workshop, facilitated by two psychology faculty members, addressed burnout facets and various management techniques. We hypothesized that participating students would show improvement after attending the workshop. Initial analysis revealed a reduction in total burnout scores before and after the workshop. Despite a noteworthy decrease, the change did not reach statistical significance. This nonsignificant trend suggests a positive direction, and further qualitative data, scheduled for collection in Spring 2024. Understanding and addressing academic burnout is crucial as it directly correlates with students' academic performance and mental health. By examining burnout in the college setting, we hope to contribute valuable insights that can inform policies and interventions to create a more supportive academic environment. 

Age-related macular degeneration (AMD) is an acquired degeneration of the retina characterized by the presence of lipid-rich deposits, or drusen, underneath the retinal pigment epithelium (RPE). Development of drusen has been linked to degradation of extracellular matrices and aberrant RPE lipid metabolism. Mutations in tissue inhibitor of metalloproteinase 3 (TIMP3), involved in extracellular matrix (ECM) degradation, have been associated with Sorsby Fundus Dystrophy (SFD), an autosomal dominant inherited disease phenotypically similar to AMD. SFD and AMD share clinical features, such as the presence of drusen, geographic atrophy, and choroidal neovascularization. The aim of this project is to evaluate the hypothesis that increased ECM degradation results in reprograming of SFD RPE metabolism towards increased branched chain amino acid (BCAA) oxidation, resulting in lipid synthesis and deposition. We found that SFD induced pluripotent stem cells (iPSC)-RPE have increased apolipoprotein E deposits, and may have increased lipid metabolism. SFD RPE were found to have decreased levels of FABP7, a lipid binding protein that regulates lipid metabolism by increasing fatty acid oxidation, by proteomics and confirmed by Western blot. SFD RPE cells show increased BCAA consumption and upregulated expression of branched chain amino acid transaminase 1 (BCAT1), an enzyme that catalyzes the animation of BCAAs. Control RPE were treated with BCAT1 inhibitors, BCATc Inhibitor 2 and gabapentin. Treatment of RPE with 50 µM of BCATc Inhibitor 2 resulted in a greater than two-fold decrease in BCAA consumption at 48 hours, indicating effective inhibition of BCAT1. Results from this study will help determine whether enhanced BCAA oxidation results in activation of increased lipid synthesis and lipid deposits in SFD iPSC RPE.

Are we happier when we experience more comfortable weather, or do we prefer seasons that provide increased social opportunities? Current studies have shown improvement in mood due to warmer, more tolerable climates or spending more quality time with loved ones. There is a gap in the relation of these two factors combined towards an individual's mood. We hypothesized change in relationship between mood and social activity throughout fall and winter. After our second survey period, we will review and organize data points based on temperatures and climates, time in and out of school sessions for students, levels of socialness, and those who changed their answers after reflecting on the social aspects of the questions. In this study, we surveyed 260 Washington State residents aged 18-64 (M = 24.46, SD = 10.03), 56% from Eastern Washington and 44% from Western in the fall prior to the holiday season (September-November 2023) and again in winter (January-February 2024) to measure changes in mood changes. Both surveys, administered through Qualtrics, asked participants about weather types experienced during each season, followed by a Brief Mood Introspection Scale (BMIS) regarding their last two weeks and the Self-Assessment Manikin (SAM) scale of weather tolerance (0°F-104°F). Participants then were asked about socialness during each season and potential seasonal affective diagnoses. We had two target populations: participants under 23 and those who are students and those who are not students or are 23 or older. We expect students to favor periods not in school regardless of socialness, as opposed to those who are not students, who we expect to favor the times they spend with those they care about over periods of isolation as they tend to have more defined schedules.

Age-related macular degeneration (AMD), a multifactorial eye disease, is distinguished by drusen formation and thickening of Bruch’s membrane. Early onset macular drusen (EOMD) is a rare inherited retinal degeneration with clinical similarity to AMD. EOMD results from genetic variants that cause decreased protein expression levels of complement factor H (CFH) and factor H-like protein 1 (FHL-1). The precise mechanism of drusen formation is unknown, although there are multiple lines of evidence that complement dysregulation and inflammation play a major role. RPE cells derived from EOMD patient induced pluripotent stem cells (iPSCs) can serve as in vitro models for understanding the effects of altered local complement. The complement system is a cascade of proteins and reactions that modulate inflammatory responses for the removal of pathogens. Specific components, such as C3a, could be involved in inflammatory responses that contribute to drusen formation and AMD. The purpose of this project is to observe how increased C3a impacts inflammatory cytokine secretion in EOMD RPE cells. iPSCs were generated from peripheral blood mononuclear cells collected from two patients in an EOMD family and differentiated into RPE cells. Western blot analysis was performed on EOMD and control RPE cells to quantify C3a, CFH, and FHL-1 levels to determine baseline expression levels. Control and EOMD RPE were treated with C3a, and inflammatory cytokine (IL-6 and IL-8) secretion was measured by ELISA. Decreased CFH/FHL-1 secretion and increased local C3a were observed in EOMD RPE. Preliminary data indicate that increased C3a levels may alter cytokine secretion by RPE, indicating that increased complement may play a role in local inflammatory responses that contribute to the pathophysiology of EOMD and AMD.

Traumatic brain injury (TBI) is recognized as a risk factor for neurodegenerative diseases, but the underlying mechanisms remain unclear. This study aimed to investigate how localized brain injury alters pathologic protein aggregation associated with neurodegenerative disease, focusing on hyperphosphorylated tau (p-tau) and beta-amyloid deposition in brain parenchyma adjacent to chronic contusion. Using brain donors from the University of Washington Brain Repository, cases with a diagnosis of chronic contusion were identified. Beta-amyloid and p-tau deposition were assessed in sections adjacent to the contusion and in contralateral sections without contusion (internal control). Manual counting and HALO imaging software were utilized to quantify the highest density of neuritic plaques/neurofibrillary tangles and overall deposition of abnormal protein in grey matter, respectively. Statistical analyses were performed to compare deposition in contusion versus control sections. Preliminary data was collected in 9 cases, predominantly male with a median age of 79 years. Neurofibrillary tangles were significantly higher in sections with contusion compared to internal controls (p=0.0188), with a similar trend observed for neuritic plaques (p=0.0723). HALO software analysis confirmed increased deposition of both proteins in the contusion compared to control sections (p=0.0391 for both p-tau and beta-amyloid). These findings support that TBI may modulate neurodegeneration by increasing p-tau and beta-amyloid deposition and underscores the importance of further research into the relationship between TBI and neurodegenerative diseases. Next, I will expand our cohort with more cases from the last few years and plan to compare brain contusion in mid-life to contusion occurring after the age of 65. In addition to assessing abnormal protein deposition, I will also examine the neuroinflammatory response. I expect to find greater levels of neuroinflammation as well as increased tau and beta-amyloid aggregation in the older age group as the neuroinflammatory response becomes prolonged during aging.