Deformities of the foot often have serious implications for people's health. In particular, hallux valgus (HV) is a painful condition in both children and adults, in which the medial column of the foot is angled outward. In severe cases, it causes gait problems. Although flat feet are considered to be one of the main causes of HV in the medical literature, the correlation between arch index (AI) and HV severity remains to be explored. The purpose of this study is to explore the correlation between AI and the degree of HV deformity and to investigate how foot arch contributes to foot deformity, using footprints from 79 subjects recorded by a plantar pressure measuring device. I quantitatively analyzed each arch index and measured the angle, using Fiji ImageJ, which is an image analysis software. I drew a line tangent to each side of the footprint and calculated the angle that each creates with the midline. AI is derived by dividing the foot into three equal parts, excluding the toes, and dividing the central area by the total area. I hypothesize that the higher the AI, the more likely the foot is to have a high HV angle. The results of this study may support the development of a system to predict the likelihood of developing HV from the AI derived from the footprint by confirming the correlation between AI and HV severity. Furthermore, finding prospective patients with HV in a community of people with flat feet based on this study could lead to more specific medical prevention ideas to prevent their morbidity and improve the overall health of the community.

In this study I investigate potential changes in plant community ecology in response to Earth’s most recent major global warming event, the Miocene Climatic Optimum (MCO). During the MCO (from 17-14 million years ago) global temperatures increased by approximately 8° C and CO2 levels increased by 300-400ppm. In assessing these ecological changes, I use minor leaf vein density (mLVD), a leaf functional trait correlated with photosynthetic rate, as a proxy for understanding plant community strategies. This trait corresponds with the spectrum of “fast” versus “slow” growing strategies described in plant physiology, with high mLVD in fast-growing plants facilitating higher photosynthetic rates, and low mLVD reflecting slow-growing persistence strategies with lower rates of photosynthesis. I hypothesize that global warming led to long growing seasons that enabled the dominance of ecological strategies that prioritize persistence over productivity (i.e., slow growing strategies), and more favorable climates increased the diversity of ecological strategies present within the community. Currently, I am measuring fossil leaf mLVD from specimens collected in the Pacific Northwest from sites representing before, during and after MCO. I examine the community-level distribution of this trait (mean, variance, kurtosis) and compare these values between sites, and thus across the MCO. I predict that plant community ecological diversity would increase during this global warming event; I also expect to see higher variance in distribution of mLVD values as warming temperatures opened new ecological niches, while mean mLVD would decrease due to an increase in persistence strategies correspondent with low mLVD. This work will help us not only to understand how plant communities responded to rising temperatures in the past but also how plant communities could potentially respond to changing climates in the future.

Massive binary star systems influence the formation and evolution of galaxies through production of energy and heavy elements. These systems can evolve into X-ray producing high mass binaries: a neutron star or black hole accreting material from a high-mass companion. While high-energy mass accretion makes these sources especially important, it makes them exceedingly difficult to model. One extremely useful characteristic for testing models of their formation and evolution is their temporal variability. By observing the properties of these sources over time, we can directly compare observed variability against predictions from models. M33, a large spiral galaxy just under three million light-years away from the Earth, is home to a collection of these X-ray binaries, last catalogued in 2015. With the goal of constraining the variability of the binaries, I have analyzed five separate epochs of Chandra imaging data taken after the production of this catalogue. I compiled a preliminary list of observed X-ray sources in the images and used positions of the previously cataloged sources to correct the astrometry, ensuring consistent alignment among observations. I then selected the most accurate position estimate for each source by-eye. Combining data from all aligned observations, I have now extracted a catalogue of reliable source detections, as well as source properties such as position and flux. With these values, I have assessed source validity and created lightcurves to study their variability characteristics. Through this work, I have found a total of 56 bright sources that meet my criteria for validity, of which 49 sources were previously identified in literature and 7 sources are new, appearing only in our observations. Looking at the characteristics of both new sources and previously observed sources, I will constrain source variability to determine how much the X-ray binaries of M33 changed in brightness over the course of the observations.

Genome Wide Association Studies enable researchers to identify which genes are associated with human diseases. Once genes are identified, gene editing of zebrafish models allows researchers to further examine the link between genotype and phenotype, with the long-term goal of treatment development. One such gene is MEOX1, which encodes for transcription factor Mesenchyme Homeobox 1. MEOX1 mutations in humans have been associated with Klippel-Feil syndrome, a congenital disorder with the most frequent symptoms being a shortened neck caused by the fusion of cervical vertebrae, leading to a decreased range of motion. Previous studies have established that zebrafish with loss-of-function meox1 mutations present with similar phenotypes in the axial skeleton. However, fusion of the vertebral column could also impact other aspects of fish morphology. The goal of this study is to test if a semi-automated screening tool can be used to quantify craniofacial variations in meox1 crispant zebrafish. Here, I utilized micro-CT scans of CRISPR-modified zebrafish with meox1 mutations (n=12) and compared them to wild type zebrafish (N=12) from the same clutch. I used 3D Slicer to manually landmark 23 major anatomical points on each individual wildtype and crispant zebrafish. I then used a semi-automated process to distribute pseudolandmarks on the surface of each zebrafish. Anatomical differences between the groups were quantified using a geometric morphometrics approach. Preliminary results show that meox1 mutations are associated with a wider posterior section of the skull and a shorter skull length. There were also differences found in the degree of asymmetry between groups. This last result in particular aligns with previous human studies of Klippel-Feil syndrome. The findings from this investigation are important for the understanding of how diseases from meox1 mutations present clinically as well as the testing of a semiautomated pipeline that will be used as a screening tool for crispant zebrafish.

Universities hold expectations of students to possess forms of cultural and social capital in order to be successful in their educational careers. This may include knowledge of the financial aid process, an ability to build relationships with faculty, and an understanding of cultural norms of the middle-class. While research shows that underrepresented students may be less likely to possess this capital, institutional expectations regarding social/cultural capital, specifically among first-generation (FG) students, seems largely unexplored in educational research. The goal of our research is to identify the hidden things FG students are expected to learn in higher education, and how they are able to learn those things. We conducted focus groups of students enrolled in LSAMP (a program that supports underrepresented students in STEM) across 5 universities in the Pacific Northwest over Zoom. Our research focuses on how the hidden curriculum, through the lense of cultural norms and the Federal Application for Student Aid (FAFSA), serves as a barrier to self-identified first-generation students in STEM. Focus groups were conducted by LSAMP students across three universities affiliated with LSAMP and researchers at the University of Washington. Focus groups were recorded and transcribed through Zoom, and later coded and analyzed. Our analysis focuses on identifying common barriers among FG STEM students. In our focus groups, FG students spoke about how relationships with faculty and peers were hard to establish because of their FG background, or because of different/potentially intersecting identities, and also how the FAFSA served as a barrier to them. Our findings support the idea that knowledge of the hidden norms and processes may be expected of students by universities, and that FG students may not possess this knowledge. By developing a better understanding of the experiences of FG students with the hidden curriculum, we can better learn how to support these students.

Sexual minority women, and bisexual women in particular, are at greater risk of experiencing sexual assault and other types of traumas and stress, as well as heightened symptom severity than heterosexual women. Among sexual minority women, bisexual women in comparison to lesbian women have shown a higher risk of reporting worse mental health. To our knowledge, only three studies have looked at differences in mental health in bisexual and lesbian women who have been sexually assaulted, but their results have been contradictory. As such, the purpose of this study is to investigate the symptom differences between bisexual and lesbian women following sexual assault to better understand their differences. To do this, I conducted a secondary analysis of an existing national data set that surveyed 1057 sexual minority women. I used bivariate statistics tests to compare lesbian and bisexual women regarding their mental health following sexual assault with a focus on anxiety, depression, and PTSD symptoms. No statistically significant differences were found between the two sexual identity groups as a function of sexual assault, but bisexual women reported greater symptoms of depression regardless of sexual assault status in comparison to lesbian women. A potential explanation for this difference could be that bisexual women may have higher levels of minority stress (i.e., stress resulting from societal prejudice and other stigmatizing experiences) than lesbian women, which has been connected to heightened mental health issues. Future studies should consider minority stress as a factor when studying the two sexual identities to better tailor research and treatment to bisexual and lesbian women’s unique identities.

An ostomy is a surgery performed to create an opening in the abdomen called a stoma. The purpose of this procedure is to divert the body waste in cases of injury or disease in the intestine. Living with a stoma requires that the patient carries a pouch that is attached to the stoma to collect the body waste. A plastic disk called a wafer goes around the stoma to secure the pouch and protects the skin surrounding the stoma from the corrosive output by acting as a barrier. To prevent any leakage, the patient must accurately cut the wafer to fit around the stoma and needs to know the exact size and shape of the stoma. An inaccurate stoma measurement and wafer cut can result in leaking bags, bleeding, infection, pain, diet changes and anxiety in public environment. As a potential solution, the innovation of a telehealth mobile application utilizing 3D imaging in modern smartphones will allow the patients to scan and measure their stoma shape by just using their phone cameras. In my research, I conducted interviews asking questions regarding the daily struggles of an ostomate and where the stoma application fits as a novel and effective solution. As a result, I gained insights showing that while patients with ileostomies (stoma in the small intestine) face more challenges because of increased chances of stoma leakage, patients struggling with obesity, dexterity issues, and disabilities have a hard time carrying out stoma care on their own. Moreover, patients with irregularly shaped stomas as opposed to perfectly round stomas go through the trial-and-error process of accurately cutting a wafer more often. Therefore, a convenient and cost-effective stoma scanning app will make it easier and faster for the patients to have an accurate wafer cut that will fit perfectly around their stoma.

Organisms are required to maintain genome stability for the correct propagation of genetic information. Glycation damage is one of the most important types of DNA damage that cause genome instability. Glycation damage in organisms is caused by the covalent attachment of parts of sugar molecules to proteins and DNA. Accumulation of the Advanced Glycation End (AGE) products may cause Parkinson's disease, cancer, and other oxidative stress-induced diseases. The protein deglycase, known as DJ-1 in plants and Parkinson Disease Protein 7 (PARK7) in animals, can prevent glycation damages in many organisms. In maize, plastid genome stability is maintained by Whirly ssDNA-binding proteins. Our lab recently showed that the demise of plastids and their DNA is associated with increased DNA damage due to oxidative and glycation damages during maize seedling development. Here, our objective is to understand the role of Whirly proteins in glycation damage. Our approach involves quantifying the glycation and deglycation levels in maize plastids. We isolated plastids and their proteins from the wild-type (wt) and whirly (why) mutant maize seedlings and performed glycation and deglycation assays. We find a significant difference in deglycation levels between wt and why mutant plants. Our study should provide a better understanding of the role of ssDNA binding proteins in glycation damage. 

In neonates, hypoxic-ischemic encephalopathy (HIE) increases the risk of impaired neurodevelopmental outcomes and death. In high resource settings, HIE occurs in 1-4 per 1,000 live births, and in low resource settings, 12 per 1,000 live births. Where available, therapeutic hypothermia (TH) is the standard of care for neuroprotection in HIE. However, the protection provided by TH is incomplete, motivating the search for adjunctive therapies. To assist in testing new therapies, researchers have attempted to predict injury severity in animals before ex vivo quantification, but the association between early injury and long-term outcomes has not been studied in the Vannucci model of hypoxic-ischemic brain injury - the most widely used HIE model in rats. This study sought to determine the association of early injury identified by in vivo magnetic resonance imaging (MRI) with weight gain and behavioral testing data. Using the Vannucci model, postnatal day (P) 10 rats underwent ligation of the left carotid artery followed by 3 hours of hypoxia and then 5 hours of normothermia in room air. Animals were then randomized, with half receiving an additional 3 hours of TH while the other half were returned to their cage. On P12, MRIs were performed, and injury quantification was determined using imaging software FSLeyes. Animals were weighed daily for one week following the HIE injury and then every 2-3 days until P43. Behavioral testing was conducted on P42 and P43 and included open field, novel object recognition, and CatWalk. We hypothesize that greater injury on the early MRI will be associated with reduced weight gain after injury and worse performance on behavioral testing.

Metal–organic frameworks (MOFs) are crystalline, porous extended solids that are formed through coordination between metal cations and bridging organic ligands. Since their discovery in the late 1990s, MOFs have been a topic of acute interest in the scientific community due to their intrinsic porosity, high surface area, and precise tunability. However, MOFs are typically insulating, which limits the scope of their applications. The recent development of electrically conductive MOFs has opened the door to exciting multifunctional applications in electrocatalysis, advanced electrochemical energy storage, chemical sensing, and much more. However, a molecular-level understanding of charge transport in MOFs remains lacking. My research aims to address this knowledge gap through the investigation of one-dimensional (1D) metal organic chains. These 1D chains can be thought of as the primary subunit of higher-dimensional MOFs; they allow for high synthetic and electronic tunability, making them ideal model materials for studying the genesis and tuning of electronic properties in conductive MOFs. Here, I will present the synthesis of a series of highly-tunable 1D metal–organic chains that exhibit delocalized π systems and high electrical conductivity along with our studies of how structural parameters such as metal identity, chain structure (linear vs. zig-zag), and metal/ligand oxidation state can influence the overall electrical and magnetic properties of the resulting chain.

The alpine zone (i.e., area above treeline) is Washington's most pristine habitat due to minimal human disturbance. However, plant species richness and the distribution of those species (collectively floristics) are poorly documented by herbarium specimens due to sheer spatial scale, physically challenging access, and a short (10-12-week) growing season. The goal of this project is to generate baseline data for future studies examining climate change impacts on alpine plant communities, improving distribution data for alpine species in Washington, and locating new populations of rare plant species. In 2021 we initiated a 5-year project to conduct comprehensive botanical surveys on 50 alpine peaks in the Cascades Range between the Goat Rocks Wilderness and the Canadian border in Washington. Peaks were selected on the basis of accessibility (i.e., do not require technical climbing to access the summit), elevation (summit area is exposed), latitude (dispersed between the north and south ends of the sampling area) and position relative to the Cascades Range crest (representation of both East Cascades and West Cascades peaks). In 2021 we sampled 11 peaks. Species lists for each peak were generated from historic herbarium specimens, collecting specimens during field work, and generating lists on the basis of sight identification of specimens in the field. We collected at least one specimen of each species, subspecies, or variety of plants encountered in the field. Here I present my analysis of the data examining the frequency distribution of species across sampled peaks, dispersal mechanisms, and flower color, and report notable species range extensions and new rare plant populations.  This analysis can be used as a comparison against future collections in new sites or collections at the same sites in the future to observe the effects of climate change on alpine plant populations.

Chronic pain -- pain that continues beyond expected healing time that may or may not be linked with tissue damage -- is a common condition among older (≥65 years) adults, often caused by osteoarthritis (OA) in weight-bearing joints, such as the knee. Older adults with knee OA often experience movement-evoked pain that is associated with reduced mobility, activity avoidance, and social relationship disruption. Knee OA is also associated with increased falls risk, which is a leading cause of injury and mortality among older adults. However, the role of movement-evoked pain in knee OA and falls risk is unclear. It is conceivable that movement-evoked pain contributes to falls risk via impaired neuromuscular function and knee buckling. Thus, better understanding movement-evoked pain might help identify rehabilitation targets to reduce falls risk. While literature demonstrates differences between movement-evoked pain and pain-at-rest, current assessments cannot accurately discern which surveyed pain levels are movement-evoked. Example methodologies that correlate activity and pain levels include pain diaries, accelerometers, or ecological momentary assessment (EMA) surveys. Such methodologies are often dependent on a participant’s ability to recall and differentiate pain experienced in relation to movement, and do not directly assess what movements evoke specific pain levels. The proposed pilot project: (1) develops a protocol for novel trigger-based smartphone EMAs using the Move 4 accelerometer, (2) evaluates the protocol’s feasibility when implemented in older adults with knee OA, and (3) analyzes relationships between movement-evoked pain, knee buckling, and falls. We anticipate that EMA collection will be acceptable for most participants, but are concerned about the accelerometer. The non-invasive Move 4 is capable of triggering EMAs when specific movement thresholds are achieved in real time. Thus, the proposed project addresses a major shortcoming in the field that currently relies on participant recall and does not capture pain levels immediately following specific movement patterns.

 In humans, gait changes with age; changes that have been associated with the onset of disease. We also see age-related changes in the fruit fly, Drosophila melanogaster, which shows a decrease in the ability to climb vertically. However, the effects of age on walking patterns of flies on a flat surface, which more closely mimics human walking, have not been fully characterized. In my research, I use D. melanogaster as a model to investigate such effects. During the past year, I followed cohorts of D. melanogaster over their lifespans and recorded videos of them walking in an enclosed arena. A wide-field camera captured the entire arena while a higher resolution camera captured the leg movements of individual flies. I analyzed the trajectories of each fly from the wide-field videos to evaluate walking velocity and duration. Based on my preliminary analysis, I have discovered that flies walk less frequently and at slower average speeds with increasing age. As a next step, I am analyzing the high resolution videos to investigate the possibility that changes in gait might explain the slower walking velocities at older age. To do this, I trained a neural network using multi-pose animal estimation software to track the movement of individual legs in relation to the fly’s thorax. This will allow me to look at gait (step length, swing duration, stance duration) as well as coordination. I expect to see age-related changes in gait and a loss of limb coordination over fly lifespan, which might explain why flies walk slower with increasing age. With the findings from my study, I hope to establish a foundation for how gait changes with age in D. melanogaster.

The COVID-19 pandemic caused many college courses to shift to emergency remote instruction and instructors had to alter their in-person teaching methods for an online format. For example, some active-learning instructors opted to embed questions in recorded lectures, asking students to pause the video and attempt to answer on their own, and then continue playing the video to hear the answer. We hypothesize that if a professor has a video “autopause” before the video provides the answer (instead of asking the students to pause a video themselves), then students will be more likely to try the question on their own. Further we hypothesize that if students generate their own answer, they will develop a stronger understanding of the material. We investigated these hypotheses in Introductory Biology III. Students (n=550) completed “lecture follow alongs (LFAs)” assignments as they watched the video. Half of the students were randomly assigned to the control group, in which they were asked to pause lecture videos and answer the questions. The other half were part of the treatment group, in which lecture videos autopaused, and the students had to positively affirm they answered the question before the video continued. We are investigating differences in exam performance, LFA responses, interview transcripts, Panopto video data, and pre- and post-surveys. Preliminary statistical analyses show that autopause did not impact student exam performance. However, preliminary analyses of LFAs indicate that autopausing can decrease copying behavior and increase the chance that students try to answer a question on their own. Through further analysis, our results can give insight into the effectiveness of autopause questions in reducing copying, which can help encourage independent student thinking. At the same time, our results will teach us how to adapt autopause questions, so they might improve student performance, in addition to encouraging student thinking.

The Washington Department of Natural Resources (DNR) publishes statewide predictions of various forest characteristics—diameter, density, height, age, species composition—in publicly-available spatial datasets called RS-FRIS (Remote Sensing Forest Resource Inventory System). Digital aerial photogrammetry is used to produce this model because of its relative ease of acquisition as compared to active remote sensing (LiDAR). However, the point clouds extracted with this method provide less detail and may be less representative of forest conditions in areas with more vertical variation. Additionally, the RS-FRIS model is an empirical model best suited to predict forest conditions reflected in field plots used in model development. I investigate how forest metrics reported by RS-FRIS compared to ground data in the Long-Term Ecosystem Productivity (LTEP) experiment in Sappho, WA. The purpose of my comparison is to evaluate if the RS-FRIS data can make reasonably accurate predictions of treatments in the LTEP experiment site and the feasibility of using this in other research sites with novel silviculture treatments. I expected the RS-FRIS to work well on the areas of our site that are likely represented in the DNR’s field plots, and my results do show more agreement between the datasets in metrics for tree diameters greater than 15.2 cm (6”) and broadly in the Douglas-fir treatment and unthinned control. I find that RS-FRIS does differ from ground measurements for predicted basal area, trees per acre, and species, particularly in the thinned treatments and where predictions account for hardwood. These results suggest that RS-FRIS is less suitable for inventory of areas that may not be well represented in the DNR’s land base. Thus, for remote sensing to be used in monitoring, additional models must be developed.

Several age-related neurodegenerative diseases include abnormal protein deposition of tau, amyloid-β (Aβ), and TDP-43. The most common dementia, Alzheimer’s Disease (AD), is tauopathic, meaning it is mainly characterized by tau deposition. In neurons, tau normally functions to bind and stabilize microtubules (MTs), proteins made of tubulin dimers that provide neuronal structure and assist in molecular transport along axons. However, in AD, tau becomes hyperphosphorylated, resulting in disassociation from MTs and aggregation in the cell. Previous genetic screening identified several new mutations in genes encoding tubulin proteins that ameliorate the effects of tau toxicity in tauopathy models of Caenorhabditis elegans; however, the various genes confer suppression to varying levels. Given that tubulin genes are expressed differentially in neurons, I hypothesized that the level of gene expression may correlate with the level of tau toxicity suppression for a given gene. To test this, we constructed transgenic C. elegans strains overexpressing mutant tubulin at differing levels. I characterized age-matched worms for tau-induced motility defects and assessed tubulin transgene expression by qPCR, observing that higher levels of mutant tubulin gene expression correlated with higher levels of toxicity suppression. Additionally, since previous work showed that tubulin mutations could confer suppression in a tau-Aβ copathology model, I sought to elucidate whether this result is generalizable to a copathology model with tau and TDP-43. To test this, I generated strains of worms expressing human tau and TDP-43 as well as mutant tubulin, and assessed them for motility defects. I expect that suppression will be conferred in these animals, although not to the same extent as in a model without copathology. These experiments will help increase our understanding of the molecular mechanisms underlying mutant tubulin mediated tau suppression. Additionally, we will gain knowledge about the mechanisms behind tauopathic disease progression in models of pure tauopathy and copathology.

The events of May 1968 in France are overwhelmingly understood as a period of civil unrest punctuated by demonstrations, general strikes, and the widespread occupation of universities as well as factories. As the capital city of France, Parisian narratives dominate the academic field of French studies. The current understanding of May '68 is based predominantly upon the events that occurred in Paris, which has colored how stakeholders understand French politics, creating serious implications for foreign policy. Although the violent protests that occurred in the Latin Quarter of Paris are integral to understanding 1968 as a whole, they are not representative of the entire country. This study investigates the ways in which the events of May '68 unfolded differently in Brest – a port city in Bretagne (a region in northwestern France) that is exemplary of a province standing outside of the aforementioned interpretive grid. As the lead researcher, I conducted interviews with individuals involved in the events of '68 in Brest with the aim of highlighting and qualitatively analyzing their lived experiences. Additionally, a focused literature review was performed on local newspapers to gain insights into the visual record of '68 in Paris and Brest respectively. Analysis focuses on the difference of trends found in the language used for protest slogans, chants, graffiti and physical posters. May '68 in Brest was a more moderate revolution – less explicitly tied to Communist party politics and far more influenced by a uniquely Breton political ideology of progressive Catholicism. Making both a historiographical and a methodological intervention brings more visibility to 68 events outside of the Parisian narrative –a step towards bringing nuance into the living memory of 1968. This rethinking of binary understandings of protests and politics in France leads to a significant reframing of our collective understanding of French leftist politics.

Hypoxia ischemia (HI) is one of the leading injuries in term neonates and can lead to death or long-term disability. The current standard of care is therapeutic hypothermia (TH). Our lab has developed a model for HI injury in rats which is used to test the efficacy of potential treatments. At postnatal day (P) 10, which is the equivalent of a full-term human neonate, rats undergo unilateral carotid artery ligation followed by hypoxia. The animals are given a 30-minute rest period before rectal temperatures are gauged to determine the post-surgical (time 0) temperatures. Animals are then placed in warm water baths for a period of normothermia (NT). Every 15-30 minutes rectal temperatures are recorded, and water baths are adjusted in order for the rats to maintain the goal temperature of 36.5°C (± 0.5°C). After 6 hours of NT, a randomized subset of animals undergo 3 hours of TH to reach 32°C (± 0.5°C). On P43 the rats are euthanized and perfused transcardially for brain removal and formalin fixation. The brains are assigned an injury severity score between 0 and 4, and % injury by volume is detected by MRI. The correlation between time 0 temperature and injury score will be calculated, as well as the correlation between the coefficient of variation of the temperatures for each animal throughout NT and the injury score. Further statistical analysis will be conducted throughout this experiment as the project is currently ongoing, including analysis by treatment group (NT vs TH). This project aims to determine if time 0 temperatures and variation of temperature are indicative of injury severity. Based on a similar study observing the relationship between the time 0 temperatures of ferrets and HI injury severity score, we expect to see that rats with lower time 0 temperature and greater temperature variation will have more severe brain injuries.

In 2020, the COVID-19 pandemic caused students living in rural areas to experience exacerbated educational disparities. This included familial financial stresses, which also pushed many migrant students living in rural communities to prioritize work over school. The pandemic shed light on educational disparities featured in rural public-school education systems. The purpose of the study was to examine how the education trajectory of students in rural communities had been affected by the social and economic impacts of COVID-19. To accomplish this purpose, we examined the extent to which familial needs impacted students’ post-high school educational plans, how financial strain influenced their post-graduation choices, and how students practiced resourcefulness and resilience despite experiences of economic hardship. In this community-based qualitative research project, we conducted semi-structured interviews with Eastern Washington high school seniors who are 18 years of age or older and used a phenomenological thematic analysis to gather themes related to our research questions. As part of the research, we collaborated with a community advisory committee composed of teachers and recent high school graduates from Eastern Washington communities to develop the project’s research methods and to ensure the analyses and interpretation of interviews are reflective of the students’ experiences. We predicted that students will plan to alter their post-high school paths to accommodate their families’ needs. Anti-racist, strength-based frameworks were used to make academic support recommendations for students in rural communities. Ultimately, our study can help inform collaboration with community members to find solutions so we can best support students and encourage them as they navigate pathways after high school graduations.

Herbivory is a concern for agriculture because it results in loss of crops. Plants have innate immune systems that allow them to defend against pests that once better understood, can be utilized to help mitigate this loss. In Steinbrenner et al.’s 2020 work, the authors identified an herbivore-specific immune receptor termed INR in Vigna unguiculata (cowpea) which induces plant defenses upon perception of the protein inceptin, found in the oral secretions of caterpillars. To further investigate the herbivory defense signaling pathway, we acquired EMS (ethyl methanesulfonate) mutagenized seeds, which contain mutations called single nucleotide polymorphisms (SNPs). These F1 parent seeds were allowed to self-propagate, producing M2 seeds. We did this because the F1 seeds likely have a SNP in only one allele of their chromosome, thus being heterozygous at a particular loci where the SNP occurred. When the F1 seeds self-propagate, we expect to get a ratio of 1:2:1 of homozygous for the wildtype allele (HH), heterozygous (H*H) and homozygous for the mutated allele (H*H*). This would give us a more varied phenotypic response when screened for sensitivity to inceptin. Individuals that exhibited a compromised inceptin response will be further investigated to determine causative mutations. We identified a number of families with abnormal immune responses, rescreened those families and verified specific families. Next we will self-propagate individuals from verified families, and expect phenotypes to segregate in a 3:1 ratio if a single dominant mutation is causative. Then bulked segregant resequencing will be used to determine which mutations are co-associated with phenotype. After knowing the genotype, we can begin to determine the molecular mechanisms behind the phenotype. This would further our understanding of plant immune responses, which we can harness to better develop more resilient crops, thus mitigating crop loss due to herbivory.
 

Postpartum depression (PPD) for new mothers has exceptionally increased since the start of COVID-19. The pandemic has created grave health and economic risks for new mothers. Additionally, the pandemic is found to have higher contraction rates and worse outcomes for those in racial and ethnic minority groups. In this research project, I will be conducting a secondary analysis to ask, “What are the unique barriers to mental health services during the COVID-19 pandemic that have affected women in Washington State that identify in racial and ethnic minority groups, clients with low socioeconomic status, and those with limited English proficiency?” This study aims to document the barriers and best practices in mitigating mental health care services for new mothers of color. I will be using a mix-method approach with data from the 2020 U.S national cross-sectional sample of women who gave birth in the last 12 months. I will be identifying themes in the client population (e.g., primarily White clients vs. mostly non-White clients) and how they differ in response to services. Early findings show that women from historically marginalized communities are at a higher risk of lower mental health services. Race and ethnicity play a pivotal role in determining who is affected and when care is accessible. By adopting the participatory action research model, I can inform and educate society on the barriers women face after pregnancy while providing suggestions for equitable care for all women and their children.

With continued population growth in urban areas, salmon populations along the coastal margins of the United States are being increasingly impacted by stormwater runoff. Urban stormwater has been linked to pre-spawn mortality events in Coho salmon (Oncorhynchus kisutch), killing them just hours after exposure. This phenomenon is known as urban stormwater mortality syndrome (USMS) and has been shown to affect large portions of Coho populations in heavily developed watersheds (≥ 60% of entire run). This study is a continuation of research that started in 2017, focusing on the incidence of USMS in Swan Creek (Tacoma, WA), which is a tributary stream that feeds into the lower Puyallup River. Live salmon and carcasses were counted weekly from October 1-December 15 and used for estimating annual spawner abundance in 2017-2021. Female Coho carcasses were examined for evidence of USMS (≥50% egg retention). Additionally, this research included community engagement efforts in collaboration with The Pierce Conservation District to educate local middle school students on the salmon life cycle, water quality, and human impact on ecosystem health. The results of this study have not yet been analyzed but will be presented visually via poster during oral presentation. Little is known about the incidence of USMS in the Puyallup River watershed, this research will support future projections for Coho populations and contribute to better understanding the implications of USMS.

Standard diagnostic tests for common bacterial infections such as strep throat are invasive and uncomfortable, especially for children. Difficulty performing these tests may delay or prevent diagnosis, which leads to more severe consequences, such as secondary conditions or organ damage. To address these barriers to care, we developed the CandyCollect, a novel saliva collection platform that is child-friendly, effective at pathogen collection, and suitable in at-home and clinical settings.The purpose of this study is to test the feasibility of our device’s bacteria capture for home use in healthy people. Staphylococcus aureus and Streptococcus mutans are common oral bacteria that are targeted in this study to assess our device’s ability to capture and detect bacteria. The findings in this study will inform our ability to use the CandyCollect for the bacterial pathogen responsible for strep throat, Streptococcus pyogenes, in future studies. We initally recruited 5 participants (>18 years) nationwide to test our CandyCollect device against oral swabs and spit tubes to compare the efficacy and user feedback of CandyCollect with traditional methods of saliva collection. The study is continuing with additional participants. qPCR analysis and fluorescent imaging were used to detect the target oral bacteria in collected samples. Responses from the user feedback survey indicated that the majority of the participants were satisfied with the design and were optimistic about implementing this device for children. Future applications include quantitative determination of bacteria concentration as well as targeting S. pyogenes. We aim to expand this human subjects study to recruit younger participants, especially children (> 3 years), which will help us achieve the ultimate goal of delivering a comfortable saliva collection platform to pediatric patients.

This research aims to explore the link between mRNA translation and cancer growth specifically in bladder. Using genetically engineered mouse models, this work demonstrates that protein synthesis is essential for efficient urothelial cancer formation and growth but dispensable for bladder homeostasis. Through a candidate gene analysis for translation regulators implicated in this dependency, we discovered that phosphorylation of the translation initiation factor eIF4E at serine 209 is increased in both murine and human bladder cancer, and this phosphorylation corresponds with an increase in de novo protein synthesis. Employing an eIF4E serine 209 to alanine knock-in mutant mouse model (in vivo), we show that this single posttranslational modification is critical for bladder cancer initiation and progression, despite having no impact on normal bladder tissue maintenance. Using murine and human models of advanced bladder cancer and immunostaining techniques, we demonstrate that only tumors with high levels of eIF4E phosphorylation are therapeutically vulnerable to eFT508, the first clinical-grade inhibitor of MNK1 and MNK2, the upstream kinases of eIF4E. Our results show that phospho-eIF4E plays an important role in bladder cancer pathogenesis, and targeting its upstream kinases could be an effective therapeutic option for bladder cancer patients with high levels of eIF4E phosphorylation.

Preserving the stories of Holocaust survivors is vital to remembering the inhumane and harrowing events that transpired during the Nazi Era as well as preventing them from ever occurring again. This project aimed to translate a collection of narratives from Holocaust survivors from Mizoch, a village in Ukraine. These stories consisted of personal memories, folklore, short tales about cultures and customs, written accounts of the genocide, images, and more. These Hebrew texts were published in 1961 and needed to be translated into English to increase access to these stories. One of the greatest challenges we faced was finding a balance between maintaining the literal integrity of the text and preserving the poetic and stylistic nature of the writing(s). Cultural, religious, and generational differences in language prevented direct verbatim equivalences. Was a literal translation with choppy flow better or worse than an English version that lost some of the original interpretation? In working together, we were able to combine our varying linguistic skills and understandings to solidify our writing. We found that the most powerful and complete translations both retained the tone of the original text and allowed for relatively easy reading. Our work has been published online and is accessible to all for free. This aligns with our goal to help disseminate these Mizoch narratives. Additionally, our translations have been used by a Ukrainian university scholar to create Ukrainian versions that have also been published online. As these English texts continue to serve as sources for future translations, protecting the authenticity of these stories is critical.

Hot and cold spring travertine (calcite mineral) deposits record integrated histories of climate and tectonism. Describing the evolution of these features informs our understanding of the driving forces behind major changes in local hydrology. Dixie Valley, Nevada, hosts abundant active hot springs, fossil spring deposits, once extensive lakes, and records of large earthquakes. We present results from a geologic and geochemical investigation of Cottonwood Travertine, a large, enigmatic, mid-Pleistocene (<200 ka) deposit recording some of the oldest spring activity in the region. Of particular interest is the temperature of spring water that precipitated the Cottonwood Travertine; determining whether the spring was hot or cold has implications for the relative effect of hydrothermal versus climatic processes influencing the formation and cessation of this massive deposit. Cottonwood Travertine covers 24 acres of a steep canyon wall 1.5-2 km upcanyon from range front faults, the modern hydrothermal field, and the shores of ancestral Lake Dixie. Several generations of calcite-filled veins and faults are exposed at the base. Three samples from distinct fracture or vein systems were analyzed: centimeter wide subvertical veins with fibrous calcite in altered rocks below the travertine, low angle veins with banded calcite below thick travertine sheets, and a subvertical, decimeters-wide fault vein. Average isotopic values of ð18O_calcite(VPDB) are -29.42‰, -14.45‰, and -14.88‰. Assuming ð18O_water(VSMOW) values of spring fluids similar to values measured in modern surface and spring water, the higher ð18O_calcite values suggest deposition between 10-12^oC. Analysis of fibrous, sub-travertine veins suggests deposition at 99^oC; these veins are likely older and unrelated to the deposit. Ongoing work will provide independent constrains on Cottonwood Travertine temperature and fluid composition with clumped isotope thermometry. Now extinct, this massive cold spring deposit suggests changes in local hydrology were driven by climate change rather than migration of the modern high temperature hydrothermal system.

High-Level Synthesis for Machine Learning (HLS4ML) has created an open-source library to optimize the resources and time needed to run machine learning inference on hardware devices. This library has converted common machine learning algorithms into C files and subsequently into Verilog code that can be uploaded onto these devices. This process works on creating efficient techniques to produce neural networks and convolutional neural networks that can be mapped to Field-Programmable Gate Arrays (FPGAs). Our group seeks to analyze the strengths and weaknesses of this process through exploring the performance, resources, time, and other criteria of models derived from a high-level neural network library, Keras. We have chosen to explore the KERAS1_Layer and KERAS_conv2d models. To evaluate the efficiency of the low level code produced by HLS4ML, we implemented our own SystemVerilog versions that mimic the machine learning algorithms. In doing so, we are able to understand what HLS4ML is doing that is replicable in just Verilog, and what is not. We are currently working on matching HLS functionality, but preliminary results show that our KERAS_1Layer implementation has faster clock speeds while lagging behind in terms of resource usage and latency. This is due to a higher number of pipeline stages and we are working on rectifying this. We expect to see similar results for the KERAS_conv2d model.

The mechanistic target of rapamycin (mTOR) is a protein kinase that is closely linked to growth and nutrient control in a multitude of organisms. Inhibiting TOR with the drug rapamycin has been shown to increase lifespan in many species. In the fruit fly, Drosophila melanogaster, rapamycin slows development, an outcome that is perhaps closely related to its effect on lifespan. Recent work in the Promislow lab on larval development has shown that the effect of rapamycin varies greatly across different genotypes, from no impact in the time of development to a nearly doubling of development time. However, it has not yet been determined which of the three larval stages is most sensitive to rapamycin. My project attempts to answer this question. I tested the delay in larval development of larvae treated with rapamycin across six different fly genotypes, four that are known to be sensitive to rapamycin treatment, and two that are resistant. After transferring eggs to food containing rapamycin or control food, I collected larvae over three days and staged them based on specific characteristics of each stage. The data were compared between treatments and genotypes to see if there was a delay in specific larval stage development that resulted in the overall delay seen in previous experiments. These data were analyzed using R, and the results indicate that there is a significant delay in development of the first instar larvae of the sensitive strains, and no delay in the resistant strains. Based on these results, I will next use single cell sequencing of first instar larvae raised on rapamycin-treated or normal food, with the goal of better understanding the specific mechanisms by which rapamycin leads to a decrease in larval development time, and the genetic basis of variation in the response to this treatment.

The Wnt signaling pathway plays a critical role in mammalian cell development and regulates cell growth and differentiation. Two central proteins in this pathway are glycogen synthase kinase 3β (GSK3β) and β-catenin. Another protein, Axin, is responsible for holding these two proteins in close proximity in order to promote the reaction between them. The kinetic mechanism for the Axin-mediated reaction is well understood, but the relationship between this function and Axin’s structure is poorly characterized. I am currently generating a cryogenic electron microscopy (Cryo-EM) structure of Axin bound to GSK3β and β-catenin in order to investigate this mechanism. Cryo-EM is a technique for determining the structures of proteins that is especially suitable for large protein complexes, such as the one I have produced. Inital results indicate that the stoichiometry of the complex is much more complicated than initially assumed and does not follow the predicted 1:1:1 complex. Elucidating this structure will provide insight into how the structure of Axin promotes the reaction between GSK3β and β-catenin as well as how it provides Wnt signaling specificity. This structure will also be important for understanding and intervening in diseases such as cancer where Wnt signaling is dysregulated.

One year prior to the 1968 presidential election, CBS News hosted a town hall debate between rising political stars Robert Kennedy and Ronald Reagan that captured their opposing beliefs and styles as partisan leaders, especially on racial issues. In post-civil rights America, no tool shaped race relations as effectively as the two types of populistic rhetoric distinctly epitomized by Reagan and Kennedy. The reactionary style and colorblind rhetoric of the former was used by the Right to promote white backlash and racial hierarchy. Conversely, Kennedy’s inclusive style and rhetoric was used by the Left to break down hierarchy and expand political representation. This research explores the rhetoric and policy proposals in the campaigns of Reagan in 1966 and Kennedy in 1968 on the issues of government size, welfare, and law and order. In a macroscopic view, it also looks at partisan trends that followed the loss of the inclusive style, with Kennedy's death, and the adoption of the reactionary style throughout the Republican Party. I argue it was these compounding processes in the mid to late 1960s that made white backlash mainstream and pushed the country to the Right where it has largely remained since. This research utilizes primary source material that reflects Reagan’s and Kennedy’s campaigns, identities, and beliefs, namely newspaper reports, opinion pieces, and contemporary political analyses. This body of material is supplemented with secondary biographies that have shaped understandings of the campaigns in historical scholarship. Altogether, the chosen sources enable analysis of the populist messages Reagan and Kennedy promoted in their campaigns and brings to light refined explanations of the twentieth century party realignment, race’s primary role in the process, and America’s current conservative tendencies and beliefs.

According to the World Health Organization, 10% of pregnant women and 13% of recent mothers suffer from adverse mental health, particularly depression. The pregnancy process is an already tumultuous experience as mothers undergo many physical and psychological changes, so the addition of other prenatal stressors like limited resources that disproportionately affect low-income mothers can further negatively impact maternal mental health, birth outcomes, and an infant's cognitive and emotional development. The purpose of the study is to examine the impact of maternal mental health on low-income mothers and children and how mindfulness can be a potential tool of intervention. I carried out a literature review using preliminary outcomes of an ongoing, longitudinal study examining maternal well-being following a mindfulness-based intervention among low-income new mothers with data acquired from the New Moms Connect Lab. Early findings show prenatal mindfulness resulted in lower depression and lower anxiety caused by pregnancy, however its effects were not long-term. Postnatal mindfulness did not yield conclusive results due to lower attendance. Social- Emotional Attunement (SEACAP) resulted in reduced depression in comparison to all three. Overall, mindfulness practices reduced maternal stress more than the control group. Research is still ongoing and more data is still being collected regarding the long term effects of postnatal mindfulness. This research is important as it provides data on the maternal health of low-income mothers who have been grossly underrepresented in previous similar research. These findings also indicate the potential benefits of mindfulness programs geared towards serving low-income communities.

Advances in molecular techniques have allowed for genome-wide studies to examine which genes are associated with many different human diseases, including those of the skeletal system. In one such study, mutations in the human SOST gene were linked to the skeletal disease Sclerosteosis 1, which presents as hyperostosis in the skull and long bones of the axial skeleton. Functional SOST genes influence the production of sclerostin, a protein which inhibits osteoblastic bone formation. The homologous gene in zebrafish is hypothesized to have a similar function based on ongoing work examining the axial skeleton in mutant zebrafish. Our goal for the current study is to test how the zebrafish cranial skeleton is affected by the SOST gene. I used the open-source 3DSlicer software to place landmarks on micro-CT scans of 27 zebrafish (9 SOST mutants, 9 SOST heterozygotes, and 9 wildtype fish) from the same clutch. I also used 3DSlicer to generate digital models of the cranial skeleton as well as to place 308 pseudolandmark points on the models. From here, I used geometric morphometric methods implemented in R to analyze the complex shape differences between the three groups. Preliminary results suggest that SOST mutants have narrower posterior cranial skeletons than heterozygous or wildtype fish, and that groups may vary in their degree of cranial asymmetry. In addition to quantifying the effect of SOST on zebrafish cranial morphology, this study is part of a larger project to establish a baseline craniofacial analysis method, and create a screening tool for examining genotype-phenotype relationships in genes associated with human skeletal diseases within zebrafish models.

Perfluorinated carboxylic acids (PFCAs) such as perfluorooctanic acid (PFOA), perfluoronanoic acid (PFNA), and perfluorodecanoic acid (PFDA) are worldwide environmental pollutants that are used in many consumer products due to their thermal and chemical stability. They are commonly found in products that resist sticking, water, stains, and grease (e.g. stain-resistant carpet, fire-proof products, paper, and cardboard packaging). The goal of the study was to determine how these PFCA compounds regulate the hepatic transcriptome and especially transporters that are involved in the absorption, disposition, and excretion of drugs, environmental toxicants, and endogenous nutrients. Fully differentiated human liver-cancer-derived HepaRG cells were incubated with 45 μM in the following PFCAs: PFOA (perfluorooctanoic acid), PFNA (perfluorononanoic acid), and PFDA (perfluorodecanoic acid) for 24 hours. The advantage of using HepaRG cells as compared to other common immortalized human hepatocyte cell lines is that HepaRG cells retain more characteristics of human livers and express most drug-metabolizing enzymes, thus serving as an excellent tool to recapitulate toxicological responses in vivo where the research is done within the organism. Total RNA was isolated from vehicle and PFCA-treated HepaRG cells (n=3 per group) and was subjected to RNA sequencing. Pathway analysis showed that PFCAs with increasing carbon chain lengths up-regulated the mRNAs of many amino acid transporters, which are essential for protein synthesis. PFCA compounds with longer carbon chain lengths also down-regulated several uptake transporters for xenobiotics and bile acids, but up-regulated several efflux transporters - likely as a compensatory mechanism to limit further toxic exposures and pump out toxic substances, respectively. Overall, our study is significant because the regulatory responses may function as biomarkers of PFCA exposures in the liver even at low doses; this can help prevent liver injury such as inflammation and possibly liver cancer.

Between 1946 and 1958, the United States conducted 67 nuclear tests in the Marshall Islands, a country in Oceania composed of volcanic islands and coral atolls. The Burke Museum holds biological specimens collected from the Marshall Islands as part of a project sponsored by the Atomic Energy Commission. I am locating, contextualing, and digitizing these specimens in order to democratize access to this biological and cultural information. I am locating as many specimens as possible in the Burke Museum collected during and after this nuclear testing period, beginning in the Herbarium. I am then digitizing these specimens by imaging, databasing, and georeferencing them. There was also an opportunity to test these samples for radiation. I am collaborating with colleagues to contextualize the specimens culturally, and bring attention to the University’s involvement in this part of our nation’s history. Overall, we aim to make this information freely available online so that communities have access to knowledge and resources to inform their decisions. Early investigation suggests that samples exist in at least the ornithological, invertebrate, and botanical (Herbarium) collections, but the total number of specimens remains unknown. This is an opportunity to unite multiple departments in the Burke Museum as we make connections across cultural and biological collections. Ultimately, this research addresses the ethical implications of the University’s scientific past and makes the specimen data of the Burke Museum more accessible.

It has been observed that the degree of mesoscale convective clustering is correlated to more intense total precipitation. We seek to examine this observed correlation in the Model for Prediction Across Scales [MPAS] model, developed at the National Center for Atmospheric Research. Following a recent study by Angulo-Umana and Kim (2022), which examined this relationship in data from the Tropical Rainfall Measuring Mission [TRMM] satellite, we i) collect various mesoscale snapshots of tropical precipitation, ii) stratify them by saturation fraction (how much water vapor the area contains divided by how much it is capable of holding) and convective area fraction (how much of the area is comprised of convective clouds), and iii) examine quantitative differences in precipitation between strongly clustered and weakly clustered snapshots. Work thus far has primarily consisted of gaining familiarity with and adapting code to the MPAS model, which has significant differences to the TRMM dataset in output variables. Additional literature research has been performed to adapt the stratiform vs. convective precipitation distinction to the MPAS model, which outputs 3D data as opposed to TRMM’s radar data. Our investigation into the MPAS model’s ability to capture the relationship between convective clustering and intense tropical precipitation will assess its overall ability to forecast tropical precipitation accurately.

The Rubber Hand Illusion (RHI) is a perceptual illusion used to examine the sense of body ownership. Traditionally, the illusion is induced by stroking a visible rubber hand synchronously to a participant’s own hand, which is placed out of view. Our research is aimed at developing a virtual reality (VR) RHI task to more closely investigate how body ownership is experienced by participants in a fully immersive setting. Following the implementation of our prototype, we will conduct a signals analysis of either an EEG or invasive electrode dataset, depending on clinical patient availability. The results of this analysis will provide greater insight into the neural correlates of body ownership, as well as provide the foundation for future studies related to perception and integration of stimulation in VR environments. The VR simulation is created using Unity 3D and the SteamVR SDK. The experiment is split into four parts: (1) initial proprioceptive drift measurement, (2) induction of the illusion, (3) final proprioceptive drift measurement, (4) movement to “break” the illusion, repeat. During the proprioceptive drift measurements, participants are asked to point and click on a virtual ruler that is placed in front of them. For the illusion phase, we render a virtual hand and colocated touch of the virtual hand in synchrony with touch of the subject’s real hand -- the induction is thus accomplished in “real-time” (no pre-recorded animation and sync up), which requires precise and accurate tracking of the hand, fingers, and touch object. To achieve the precision and accuracy needed, we will use Hi5 VR gloves alongside SteamVR’s pose system. While more difficult to design, real-time induction will allow us to more flexibly explore novel ways to induce the illusion, opening new pathways for future RHI studies.

Plants have a variety of pathways and receptors to defend against herbivores, pathogens, and damage. While receptors involved in damage and pathogen perception are well studied, receptors involved in defense against herbivorous pests are not well understood. The first known receptor involved in plant-herbivore interactions is the inceptin receptor (INR). INR recognizes inceptin, a short 11 amino acid polypeptide found in the oral secretions of caterpillars. When inceptin binds INR, the plant produces ethylene, peroxidases, and reactive oxygen species to reduce herbivory. However, only some legume species express INR. To confer novel resistance against herbivores to other plant species, we designed plasmids containing INR as well as its coreceptors to reconstruct the cell surface signaling pathway. These plasmids enable transgenic expression of the INR cell surface signaling network in the model organism Arabidopsis thaliana. After constructing these plasmids, I transformed them into Arabidopsis thaliana via Agrobacterium-mediated transformation, which allows for assessment of immune activation in response to inceptin treatment by measuring ethylene and reactive oxygen species production. If this strategy successfully transfers INR function into Arabidopsis thaliana, then this same approach may be used to transfer INR into crops. Higher resistance to herbivory would limit pesticide use while increasing crop yield, reducing the amount of land and water needed to sustain populations.

Throughout our lives, we generally base our idea of age upon someone’s ‘chronological age’, or how many years they’ve been alive. This, however, is not always the best indicator of aging, as people reach social and biological milestones at different ages. As an alternative, someone’s ‘biological age’ can be more representative of their progression through life. As such, research has focused on identifying biomarkers of biological age to help us better understand aging. Recent work in our lab has sought to determine the impact of several metabolites - biomolecules used for metabolism - on the biological age of the fruit fly, Drosophila melanogaster. Among the metabolites studied, histamine - a neurotransmitter involved in wakefulness and visual processing - had one of the strongest correlations with lifespan, suggesting that it plays a role in aging. In this study, we attempted to manipulate the biological age of female D. melanogaster by altering either their metabolic levels of histamine, or their perception of histamine. To do this, flies were given food supplemented with histamine or with the antihistamine hydroxyzine, a competitive inhibitor of histamine receptors. These experimental conditions were compared to control food that lacked additives. Treatment was administered continuously starting at 4 weeks and the lifespans of flies in each condition were measured. Based on our previous results, we expected to see a negative effect of added histamine on lifespan and an increase in lifespan in response to antihistamine. Our study could highlight histamine’s role in aging and lay the foundation for demonstrating that biological age can be influenced by a single metabolite.

One in seven children will experience child abuse in their lifetime. While places like schools and day care facilities were once able to detect signs of abuse, the move to online school and closing down of childcare facilities have made it more difficult for children to seek out help during the COVID-19 pandemic. The purpose of this literature review is to review current papers that investigate how cases of child abuse have been reported over the past two years. This literature review asks the question: How has the COVID-19 pandemic impacted the reporting of child abuse cases? Using sources from Google Scholar and UW Libraries, I found readings from the U.S, France, Uganda, and Canada that included keywords such as “child abuse”, “COVID” and “underreporting”. Findings include some countries finding higher rates of reporting which may be accounted for due to the widespread domestic violence campaign that brought awareness to, other countries reporting lower cases which may be accounted for due to the lack of contact with education personnel who are critical reporters, and the potential for online predators to seek out children who are mostly online all day. Overall, this research is important for childcare providers, teachers, and social workers who have been working during the pandemic to provide resources for their students who they may not be seeing in person and to further a conversation of continuing research on the impact of remote learning on child welfare.

Plastic pollution is a growing concern in the microecology of the oceans. Studying bacteria colonization rates on plastic provides one way of understanding of how toxic debris can move through the food chain through ingestion. This process of toxins moving through the food chain is called biomagnification and can eventually reach humans. To evaluate bacterial colonization rates, I collected seawater in the coastal waters of Hawaii and near the Pacific garbage patch (GPGP). Seawater was intermixed with 5 different kinds of clean plastics then timed to determine how long it took bacteria to colonize the plastic surfaces. Bacteria on the plastic were counted under an epifluorescence microscope then divided by the time of colonization to determine the rate. Alongside the colonization rate, surface microplastics were collected with a manta net; then sized and classified with a dissection microscope. Seawater was collected from a Niskin bottle attached to a CTD (Conductivity, Temperature, and Density sensors) rosette to calculate bacterial abundance with the use of a Guava flow cytometer. The findings of the research displayed little to no correlation between surface bacterial abundance and plastic density, with an R2 value of 0.1072. Bacteria were found to colonize plastics at 48 and 96 hours in the waters near the Pacific garbage patch with a rate of 7.4E+04 cells/mm. The colonization rates and plastic abundance support evidence of plastics being integrated into the ocean ecology.
 

Patterns in three dimensional artefact orientation can provide insights into the environmental and cultural processes that form archaeological deposits. A range of post-depositional processes can be inferred from non-random patterns of artefact orientations. We apply statistical and graphical techniques to artefact orientation data from Madjedbebe, a site in northern Australia with evidence of human activity at 65 thousand years ago. The three-dimensional positions of artefacts were recorded in situ using a total station (survey equipment), and analysed following McPherron’s (2018) methods. Through this I investigate how accurate the site's dates are through analyzing deposition processes and site formation processes. I compare the orientations of the archaeological artefacts to orientations of experimentally trampled artefacts, and to orientations from simulated slopes and terrains. I found that the archaeological orientations are significantly different from the trampled artefacts, which have a strongly linear orientation. The orientations of archaeological artefacts are most similar to simulated orientations from sloped and irregular surfaces. The upper phases at Madjedbebe present more isotropic orientations than the lowest phases, which are strongly planar. These patterns suggest minimal post-depositional disturbance in the deposits containing the earliest artefacts at Madjedbebe, which provides support for a stable stratigraphy, which provides added confidence for the association of stone artefacts and the dates of 65 kya. This methodology can be used to reinforce conclusions about site dates in other sites with questionable stratigraphy.

STAND (Supporting Teens Autonomy Daily) is an evidence-based practice modular therapy that helps ADHD (Attention deficit hyperactivity disorder) teens build academic and interpersonal skills. STAND has been shown to be cost-effective in promoting evidence-based practices compared to the usual-care psychological services and medication for ADHD. The purpose of this study is to assess the utilization and efficacy of online resources and training tools for therapists and supervisors implementing STAND. Six therapists and four supervisors administered STAND with the use of LYSSN, an AI (Artificial Intelligence) based tool that provides feedback on therapy audio tapes, and Care4, a dashboard that provides supervisors with a library of resources. I analyze 49 therapist sessions and 3 supervision sessions to determine the attitudes and engagement of the therapists and clients with STAND activities by calculating the average of scores provided by therapists and supervisors from Care4. Therapists login onto the Care4 system after a therapy session and answer questions about feedback they received from LYSSN and the preparation for their session. Supervisors used the data collected from the therapist’s LYSSN score and formulate their feedback for therapists. Therapists spent an average time of 11 to 30 minutes reading through the STAND session description and preparing client worksheets and an average of 1 to 10 minutes reviewing LYSSN feedback, watching STAND demonstration videos, seeking consultation from the STAND research team, and reviewing feedback from their supervisor. Therapists found LYSSN to be “occasionally” accurate and credible. Collection of this research will provide insight about the implementation of STAND through an online format and about the general appeal of the use of AI, the workload of supervisors and therapists could be reduced and give feedback more efficiently to them with regards to STAND treatment and other evidence-based treatments.

Target of rapamycin (TOR) signaling is a nutrient-sensitive pathway that plays a role in cell growth and aging. Caffeine is a factor that inhibits TOR signaling and growth, compromising overall cellular fitness. The budding yeast S. cerevisiae is an ideal organism to study TOR signaling in relation to caffeine tolerance as there are many resources available to study yeast genetics, and yeast shares many basic biological properties with other eukaryotic cells. Additionally, yeast allows us to study TOR signaling using experimental evolution, where under a defined selective pressure - here, caffeine - we observe what rare beneficial mutations arise and increase in frequency. While many inputs to the TOR signaling pathway are known, others are yet to be identified. Furthermore, how other pathways compensate for TOR signaling inhibition is not completely understood. Our goal is to identify factors involved in TOR signaling by growing yeast in inhibitory concentrations of caffeine to select for better growing mutants with increased resistance. We will then sequence the resistant strains’ genomes and study the resultant mutations to determine how they connect to TOR signaling and caffeine tolerance. We evolved yeast for 5-10 weeks in increased doses of caffeine, and sequenced clones from the evolved populations. Our yeast evolved increased caffeine tolerance, and mutations arose in drug-response pathways including in the Pdr1 transcription factor. We also observed mutations in processes regulated by TOR, such as nutrient sensing. With further genome sequencing of more evolved populations we aim to identify novel mutations and factors involved in caffeine tolerance and TOR signaling. Ultimately, these findings increase our understanding of how caffeine impacts TOR signaling and how other cellular processes are regulated by TOR signaling. More broadly, this research can aid in the continued development of how cell signaling pathways are related to nutrient response, aging, and growth.

Campus safety issues are not typically included in institutional diversity efforts. However, it's important to understand how students with diverse ranges of social identities perceive campus safety; promoting safety should be prioritized. To understand marginalized student perceptions of campus safety and policy, I analyzed focus group transcripts (31 participants) from PNW LSAMP (Pacific Northwest Louis Stokes Alliance for Minority Participation) students and recent alumni from the University of Washington, Washington State University, Oregon State University, Portland State University and Boise State University. Data was coded deductively in NVivo by a pre-determined coding scheme guided by the research question but allowing for inductive coding as other themes emerged. My findings indicate that gender identity has a high impact on perceived safety. Women and non binary individuals tended to discuss feeling uncomfortable in campus areas at night and mentioned taking preventative safety measures like carrying tasers. Participants identified emergency blue lights and safe rides as initiatives which maintained or improved safety feelings. This indicates that on some level, universities are already protecting marginalized students. Little discussion on policy improvement emerged, but instead how universities can use clarity and police presence (or absence) to show students they value their safety. Namely, several Black/African American participants expressed preference for more detailed notifications on how to take action when safety threats are alerted to the university community. Participants generally expressed that police did not improve their safety perceptions or that police presence could detract from other students’ safety depending on their race. These findings suggest that universities are protecting marginalized student safety to some degree. Next steps include clarifying safety notifications, expanding safe ride and emergency blue light operations, and conducting broader surveys on perceptions of campus and municipal police, given police brutality incidents and mixed literature.

A comparison of rates of osteoarthritis in the spine by occupation allows us to understand the impact of different jobs on spine health. This study will compare the rates of spinal osteoarthritis between non-manual labor workers (such as retail/service and information technology workers) and heavy-lifting occupations (such as agricultural, building, and manufacturing laborers). We will be evaluating the spines of decedent individuals with known occupation categories, available through the New Mexico Decedent Image Database, for spinal disk space narrowing and osteophytosis with a grading scheme of 0-3. For disk degeneration, 0 = normal disk spacing, 1 = mild disk space narrowing, 2 = moderate disk space narrowing, 3 = severe degeneration of the disk space. To assess osteophytosis, we will grade the anterior side of each vertebral body for the presence of osteophytes on the 0-3 scheme (0 = no osteophytes, 1 = mild osteophytosis, 2 = moderate osteophytosis, 3 = severe osteophytosis) with a score for each vertebral body and intervertebral space. We will calculate the prevalence of osteoarthritis and perform regression analyses between the occupation categories to determine the relationship between occupation and spinal osteoarthritis. As osteoarthritis is positively correlated with age, age categories will be established for the analyses. In addition, we anticipate that different trends in the development, location, and prevalence of osteoarthritis within the spinal column may exist between the occupation groups due to differences in biomechanical loading and areas of "wear and tear." We will perform regression analyses to determine if any such pattern is present in our sample. The results of this study will increase our understanding of the prevalence of osteoarthritis by occupation, aiding health care providers in developing preventative care and treatment plans for their patients and occupational health officials find effective ways to protect their employees.

Dilated cardiomyopathy (DCM) is one of the most common causes of heart failure and is often associated with mutations in sarcomeric proteins. The sarcomere is the basic contractile unit of the heart and depends on Ca²⁺ for force generation. Two sarcomeric protein mutations, D230N in tropomyosin and I61Q in cardiac troponin C, have been shown to decrease force production and Ca²⁺ sensitivity of the sarcomere, leading to DCM. We are investigating if the small molecule, Danicamtiv, is effective at improving Ca²⁺ sensitivity in D230N and I61Q mouse models. Danicamtiv binds to myosin and has been previously shown to enhance force production capacity in cardiac muscles. Using demembranated D230N and I61Q tissue positioned between a force transducer and length controlling motor, we move the tissue between different solutions of varying Ca²⁺ concentrations in the presence and absence of Danicamtiv. We experimentally determine Ca²⁺ sensitivity for each solution by calculating the calcium concentration (pCa = -log[Ca²⁺]) required for half maximal force (pCa50). Additionally, we measure the rate at which the tissue is able to redevelop force (ktr). By exposing each tissue preparation to physiological solutions both with and without Danicamtiv, we can determine how maximal force, pCa50, and ktr are affected. Preliminary results suggest that Danicamtiv increases pCa50 in both D230N and I61Q tissues, and also decreases ktr by about half. Data gathered in this project will contribute to our understanding of the mechanism by which Danicamtiv improves cardiac function. In the future, these data will also help build and strengthen computer models of the sarcomere that can predict the effects of different mutations or small molecules on force production and contractile kinetics.

Test questions are commonly written with the intent to assess a student's higher level thinking skills, but the phrasing of a question may not align with this intention. One way to remedy this is to assess the cognitive complexity of existing test questions, and use these findings to reform instructional materials and subsequent examinations. We used Marzano’s Taxonomy to characterize a bank of multiple-choice exam questions from general chemistry by the level of cognitive complexity required of students' thinking. Marzano’s Taxonomy has been applied to introductory physics questions to characterize quantitative thinking and problem-solving skills (Teodorescu et al., 2013), and to general chemistry curricula to guide course and assessment development (Toledo & Dubas, 2016). Here we will describe our implementation of Marzano's Taxonomy in the context of large-lecture general chemistry, and our development of a rubric that chemistry instructors can utilize to evaluate their own exam questions. The development team included an undergraduate biology major and former general chemistry peer mentor; a chemistry graduate student and experienced general chemistry teaching assistant; and a chemistry faculty member. When applied to multiple choice questions, we found that the cognitive level of most questions was lower than expected, even for lengthy questions that are normally considered challenging by students and instructors alike. We will discuss ways to increase the cognitive complexity of problems to allow instructors to elicit the intended level of thinking from the student, and to align the cognitive levels of assessment and instruction. This can also improve the clarity of expectations of the cognitive complexity that is required of the student and promote higher-order thinking.

In Southeast Asia, it is common for there to be a lack of clear typological categories in stone artefact archeology, but this is typically informally observed rather than empirically demonstrated. In Northern Vietnam, Mau A, an open air site located near the Red River in Yen Bai Province, Northern Vietnam, stone artefact flakes were excavated and analyzed in order to answer these questions: What variation exists within the assemblage of flakes, and are there distinct typological groupings corresponding to different tool categories? 1058 stone artefacts were excavated from four square meters using hand tools. The artefacts were measured using calipers, and these measurements were converted into outlines that we analysed using geometric morphometry and Principal Components Analysis (PCA). The PCA plot we made revealed that there was a great amount of overlap between reduction categories with a lack of typological groupings distinct from one another, although there was high variation in the amount of dorsal scarring. Primary flakes (the flakes that have been worked upon the least) have the least variability in terms of shape, with tertiary flakes having the most variability. We also created additional boxplots which revealed a lack of shape and material variation as well as confirming the PCA's results that dorsal scarring impacts flake variation. Our results suggest that flake shape is sensitive to assemblage reduction intensity, and may give useful comparative insights when other attributes show little variation. These results are important for understanding stone artefact assemblages from Southeast Asia which often yield little variation when analysed with traditional approaches.

GNE myopathy (GNEM) is an autosomal recessive inherited disease that causes progressive muscle weakness starting around ages 20 - 40, which leads to physical disability. GNE myopathy is caused by a mutation in the GNE gene, which encodes the enzyme glucosamine (UDP-N-acetyl)-2-epimerase/N-acetylmannosamine kinase. This enzyme is responsible for sialic acid (SA) production. SA is a sugar that is required by all cells, including muscle cells, to produce energy. SA is also an important component of the cell membrane. Thus, lacking SA causes muscle wasting. Interestingly, sialic acid produced in the liver could be used by skeletal muscle. Thus, our lab is developing new promoters for GNEM gene therapy that combine liver and muscle expression. By using PCR and Hifi-assembly, I cloned three control promoters--one muscle-specific enhancer/promoter (Ck8e), one liver-specific enhancer/promoter (ApoE-hAAT), and one constitutive promoter (CMV)--into three reporter plasmids ahead of firefly luciferase. These reporter plasmids also have CMV-renilla luciferase as an internal control. Now I am transfecting six tandem expression cassettes plasmids (already cloned) and the three plasmids that I cloned into the culture of liver cells (HepG2) to observe the expression of the genes based on the ratio of firefly and renilla luciferase. In this way, we can compare expression in liver cells from the novel promoters and the control promoters. Another member of the lab will be transfecting the plasmids into muscle cells (C2C12), so we will be able to compare our results. This research is important to figure out the most optimal promoter to express the GNE gene in both muscle and liver cells. Thus, after treatment, the cells could produce SA by themselves to maintain their functions and prevent muscle wasting for patients with GNEM.

A high-throughput yeast two-hybrid method has recently been developed within the Seelig Lab at the University of Washington to measure thousands to millions of pairwise protein-proteins in a single tube. In each yeast cell, only one pair of proteins is expressed from the same single copy plasmid. Currently, the plasmid in each cell is assembled from 5 double stranded DNA fragments using yeast homologous recombination machinery upon yeast transformation. Particularly, 2 fragments encode for a pair of proteins and the other 3 fragments form the backbone of the single copy plasmid. To optimize the protocol and reduce the number of transformed DNA fragments, 3 of the aforementioned 4 fragments that form the backbone can first be assembled together into a separate accessory plasmid that is subsequently linearized using a restriction enzyme. This reduces the number of transformed fragments that assemble into the single copy plasmid to just 3, which increases the efficiency of the overall transformation. In my research with other members of the Seelig Lab, I have already assembled the accessory plasmid, and am now running the optimized version of the experimental protocol to confirm that this strategy leads to the same results as the established protocol when applied to a control set of coiled coil proteins. Upon completion of this optimization step, I will further try to optimize the protocol by investigating whether the two double stranded DNA fragments that encode for a pair of proteins as well as assemble the single copy plasmid, can be made from a pool of single stranded oligonucleotides rather than ordered as two separate fragments from a DNA synthesis company. This optimization should further reduce the labor and the cost of materials while performing the protocol, and will allow a broader range of individuals to successfully complete yeast transfections with limited resources.

 For 9 months, a human fetus is fully dependent on the placenta which provides oxygen and nutrients, regulates waste transport, and acts as an immunological barrier. These 9 months are critical as the placenta has been linked to various health outcomes later in life. Glial Cells Missing Transcription Factor 1 (GCM1) is a transcription factor (TF) that plays a crucial role in placental development. We hypothesize that reducing the expression of GCM1 will affect placenta-specific genes such as the placenta growth factor gene (PlGF), and pathways that control embryonic development such as the sonic hedgehog. Studying how GCM1 alters gene expression in the placenta will reveal specific biological pathways that are affected by GCM1. Primary villous trophoblast cells were collected from 4 male and 5 female placentas following routine c-sections and a siRNA knockdown was conducted to decrease GCM1 expression. High-quality RNA has been isolated and sequenced from these samples. I am using the RNA sequencing data to compare gene expression between two groups: normal placental cells and cells where GCM1 has been knocked down. Using standardized differential gene expression approaches in R, I have identified genes with statistically significant differences between the two groups. Preliminary results indicate that 15 genes are decreased and 2 are increased. A sex-stratified analysis has been performed to determine if fetal sex plays a role in the different placental expression levels between the two conditions. I also performed a pathway enrichment analysis to determine if certain biological pathways are enriched for genes whose expression was associated with a reduction in GCM1. These results will help us better understand the molecular changes that occur due to a reduction of GCM1 and potentially how that contributes to maternal health conditions such as preeclampsia which harms both the mother and fetus.

The human brain is highly sophisticated and its functions are influenced by a multitude of factors, many of which play a role in the complex aging process. Certain individuals appear to possess more resilience to environmental and biological stressors as they age compared to others. However, why they are more resilient is not understood. Resilience refers to an individual's capacity to respond to stress (physically, psychologically, emotionally) by resisting damage and bouncing back. In my research in the Promislow lab, I use the fruit fly, Drosophila melanogaster, to explore the intricate process of aging. In this experiment, I applied a biological stressor on the flies halfway through their lives and examined mortality and motor function as measures of health to study resilience throughout the fly’s lifespan. I stressed the flies with a sublethal dose of paraquat, a neurotoxin that causes oxidative stress and mitochondrial dysfunction upon acute exposure. If the stressed flies return to the mortality and motor function levels of the control flies, this tells us the flies are resilient. I hypothesize that all of the flies that receive the paraquat dosage will experience an increase in mortality and a decrease in motor function when compared to the control flies. While I think the majority of the flies will fail to recover from these stressed levels, I hypothesize a small number of flies will return to the mortality and motor function of the control flies, demonstrating resilience. In my project, I aim to understand how flies can recover from biological stressors, and how their ability to recover changes throughout their lives. My long-term goal is to understand how exposure to biological stressors affects the aging process, and in particular, how and why resilience varies with age.

When mainstream media and off-line spaces fail to provide representation and community, many trans fans move to online fan spaces. When centering trans fans’ voices, common themes emerge surrounding their experiences within these spaces that may otherwise go unnoticed. It is then vital to hear what fans, particularly trans fans, have to say about the fan spaces in which they participate. My project uses qualitative and quantitative data collected from a survey I created and distributed across online fandom spaces. From the survey results, I isolated data from trans fans and conducted a case study on their experiences within fandom and with fan fiction and fan creations. My project centers their voices as I examine their experiences holistically to recognize how the intersection of multiple facets of their identity inform their experiences in online fandom spaces. In the scope of my project, it is seen that trans fans clearly benefit from fan communities and fan creations with 94.93% of respondents viewing fan communities as beneficial. Yet, these benefits are not always easily gained and can be accompanied by negative incidents as evidenced by 82.43% of respondents having such experiences. These results give rise to further lines of inquiry about the circumstances of trans fans' participation in online fan spaces and highlight the importance of centering trans fans' voices in future research.

 In vivo recordings of neural ensemble activity in non-human primates (NHPs) have contributed to the understanding of how neural activity relates to motor function and intent. Traditionally, neural recording has been conducted in constrained environments, such as head-fixed experiments using bulky wired equipment to achieve high data throughput and reduce measurement noise. To collect more natural data from free-moving NHPs requires a high speed, low power wireless uplink of the neural data. However, wireless communication inside a metal NHP cage suffers from dense multipath interference, due to multiple signal bounces from the cage walls, which decreases communication reliability. We explored an approach called "Electronic Mode Stirring" to obtain better communication reliability. We found that by adding 4 switchable reflecting antennas to the roof of the cage to perform mode-stirring we were able to improve the mean one-way path loss and improved the worst-case signal loss across 126 measured positions in the primate cage. We expect that the reduced signal loss with the mode stirring system will lead to improved wireless communication reliability inside the cage.

In the lungs, conventional dendritic cells (cDCs) survey the tissue for foreign antigens and initiate immune responses. A subset of lung cDCs, known as cDC1s, are phenotypically distinct from cDCs and critical for T cell activation in diseases such as lung cancer and respiratory viral infection. Lung-specific cDC1 generation in vitro has not yet been achieved, and performing mechanistic studies in vivo remains difficult because cDC1s exist in small numbers in the lung. I aim to identify an in vitro culture system to develop sufficient numbers of murine (mouse) lung-specific cDC1s. I examined how the addition of transforming growth factor-beta (TGF-β) to bone marrow from mice affects cDC1 differentiation. TGF-β is a factor of interest because it is an abundant lung cytokine during development that upregulates genes associated with DC development, but its effect on cDC1 differentiation has not been documented. I first harvested mouse bone marrow, cultured these cells in FMS-like tyrosine kinase 3 ligand (FLT3L), and subsequently added either recombinant granulocyte-macrophage stimulating factor (rGM-CSF) or recombinant TGF-β (rTGF-β) individually or together. I then used flow cytometry to assess the phenotype of generated cDCs in these cultures. Preliminary results indicate a combination of FLT3L, rGM-CSF, and rTGF-β increases the expression of the epithelial adhesion molecule (EpCAM), which our lab has newly identified to be expressed in subsets of lung cDCs. Additionally, GM-CSF suppresses XCR1 expression, another marker of cDC1s, while FLT3L induces XCR1 expression. Based on these findings, I will continue to improve the culture conditions by tracking the expression of preDC markers in only FLT3L to determine the best day to add GM-CSF and TGF-β. Understanding the development of in vitro murine cDCs would allow us to replicate similar in vitro studies for human bone marrow-derived lung cDC1s and use them for developing cell-based therapies against lung diseases.

The adolescent age is ideal for Attention-Deficit/Hyperactivity Disorder (ADHD) treatments, such as Supporting Teen's Autonomy Daily (STAND) which was developed to make therapy accessible to disadvantaged youth with ADHD, since these treatments may be carried from the teenage years into adulthood. To increase the quality of care for these adolescents and improve their outcomes, six therapists and four supervisors have received STAND training. This training uses technology, such as the Artificial Intelligence (AI) tool that gives them feedback: Lyssn, is used to enhance provider engagement and delivery. The purpose of this study is to examine whether this STAND training and its technology are effective in improving therapists' STAND delivery methods to patients. Through 64 previously coded audio recordings of provider sessions that took place in Spring and Summer 2021, I assess the effectiveness of this AI training for therapists and their supervisors on their performance and delivery of STAND treatments. To do this, I analyze the Motivational Interviewing Treatment Integrity (MITI) coding system scores that the therapists received as feedback from Lyssn, when the therapy sessions occured, when those sessions were uploaded, and when therapists received feedback. I also examine questionnaires given to therapists regarding their thoughts on Lyssn before and after their sessions. Though data is preliminary and lacks a control group, results could inform us of therapists' positive engagement with Lyssn. Using AI feedback to train therapists on new practices or more efficient ways to deliver ADHD therapy sessions could potentially produce significant benefits to disadvantaged adolescents with ADHD so that they receive optimal care and see positive outcomes.

Misinformation is one of the defining issues of the twenty-first century. Prior research has pointed to the significant role that a handful of influential accounts can play in spreading misinformation on social media. This project focuses specifically on misinformation originating from small accounts but amplified by major influencers, to understand further the role such influencers play in the spread of online misinformation. In a case study of Twitter posts during the early stages of the Covid-19 pandemic, I quantitatively analyze the effects of strategic amplification by high-profile influencers on the amplified user’s behavior, including how their behavior on the platform changes and the persistence of engagement with new audience members gained after being amplified. Additionally, I conduct a qualitative analysis of topics posted about by users, in order to see whether users shift discussion topics after amplification, particularly whether being amplified for posting misleading information increases the likelihood of posting misinformation in the future. I anticipate to find that users increase their engagement with the platform, and that audience engagement with amplified users reduces from peak levels when going viral, but remains higher than pre-amplification levels of engagement. I also anticipate finding that users are more inclined to post content similar to what was amplified, including having a higher likelihood of spreading misinformation. This research will further inform discussions about the role of social media platforms in the spread of misinformation, and how inter-user interactions on social media platforms impact this spread.

Over the past decade, we have seen major forest loss due to events like deforestation and tree die off. Previous studies have examined ways in which the distribution of plant types and how they function impact local and global climate. Local climate can be impacted because plants alter fluxes of water, energy, and momentum between the land surface and the atmosphere. Global climate can be impacted because local changes influence atmospheric response in clouds, humidity, and gradients in energy which drive changes in circulation. This project aims to identify if observed forest loss has a measurable effect on the noisy climate system. We have compiled spatial data of actual forest loss derived from satellite observations and test the climate impact of forest loss in simulation experiments using an Earth system model. We will assess differences between simulations with and without forest loss to identify how forest loss impacted the atmosphere and surface climate over land. We use the identified impacts as hypotheses for the expected climate response to forest loss and will subsequently analyze if these patterns can be seen in observed environmental conditions following forest loss. We expect to see some effect in the climate due to the observed forest loss. This project serves to advance our understanding of the effect of forest loss on global climate, atmospheric circulation, and energy balance, and thus will help to coordinate efforts to mitigate climate change by identifying potential unwanted impacts of forest change due to human actions.

Alzheimer’s Disease (AD), despite its prevalence, remains a much-researched yet incurable condition. Most prominently characterized by cognitive decline with advanced age, AD is associated with beta-amyloid plaques, tau protein tangles, and neurodegeneration in the brain. Previous research has shown that a relevant contributor of AD is neuroinflammation in the brain and spinal cord. Neuroinflammation is driven by cells such as microglia, brain immune cells, and astrocytes, which regulate the blood-brain barrier and homeostasis. However, our understanding of cell-type specific changes in their gene expression is lacking. We hypothesize alterations in gene expression in microglia that result in a more inflammatory phenotype in AD compared to controls. We also expect to observe alterations in the prevalence and inflammation of astrocytes in AD brain. Single-nucleus RNA sequencing (snRNAseq) provides a more detailed analysis of cells, allowing us to examine expression differences between cells in the same brain region. Applying this technology to compare phenotypic differences in specific cell types of AD patients and non-affected controls has yielded a more comprehensive look at differences in cell types that influence AD. This study examines a snRNAseq dataset generated by a cohort of 20 individuals, using brain tissue acquired post-mortem. This tissue is separated into individual cells, which are analysed using our lab's sequencing pipeline in R, allowing us to determine gene expression for each cell. My role in the project involves piloting this pipeline and analysing the various outputs to identify differentially expressed genes and how they are characterized between groups (control vs AD) for specific cell types such as microglia. I have also begun to validate these findings with protein expression assays such as Western blots, as well as in cultured microglia and astrocytes. This study enables us to determine and characterize the alterations in gene expression in AD, and hopefully identify potential therapeutic targets in the future.

From their ability to perch perfectly on nearly any surface to their capability of balancing on any perch, birds are phenomenal animals. Avian anatomy includes mysterious features like the lumbosacral organ (LSO) within the spinal cord. Research surrounding the LSO has led to the discovery of a mysterious network of nerve fibers that seem to cross the midline of the spinal cord only in the LSO. The goal of my research was to identify the neuron cell bodies giving rise to these axons and to track their targets. To do so, I used immunohistochemistry (IHC) and tracing. A tracer was injected into the glycogen body, where the surrounding neurons took up the tracer and transported it both retrogradely, toward the cell body, and in the anterograde direction, toward the axon endings. Tracing revealed the structure of the circuit. IHC was subsequently used to test if proteins indicative of sensory nerves were present in the glycogen body, such as CGRP and substance P. These methods, thus far, have indicated the presence of CGRP and substance P in nerve fibers across the glycogen body. These findings suggest the mysterious nerve network in the LSO is a network of sensory neurons abundant in substance P and CGRP, also implicating the glycogen body as a facilitator for the passing of these nerve fibers.

Age-related macular degeneration (AMD) is the leading cause of vision loss for people aged 50 years and older. Variants in the complement factor H (CFH) gene are associated with an increased risk of developing AMD, making it one of the main drivers for disease progression. Rare mutations that affect the expression of CFH protein and its isoform - Factor H-like protein 1 (FHL-1), have been linked to early-onset macular drusen (EOMD), an inherited degenerative disease with similar clinical characteristics to AMD. Our lab recently found a novel CFH variant of two family members with EOMD. This single nucleotide polymorphism (SNP) in the conserved splice site of intron 3 resulted in a frameshift with premature stop codon and no translation, causing ~50%reduction in CFH/FHL-1 production. Since CFH/FHL-1 are cofactors for C3b cleavage in the alternative pathway of the complement system, this haploinsufficiency will likely affect complement regulation in retinal pigmented epithelial (RPE) cells, hence contributing to the development of AMD-like pathology. Using patient specific iPSC-derived RPE cells, this study aims to investigate the role of CFH/FHL-1 in maintaining complement homeostasis. I will examine the expression levels of several active complement components and multiple regulators of complement activation (RCA). ELISA and Western blots will be used to quantify protein expression, and protein localization was revealed through immunostaining. RPE cells will also be exposed to normal human serum and stained for the membrane attack complex (MAC) to assess the susceptibility of cells to complement-mediated damage. I will also examine reversal of phenotype in EOMD RPE supplemented with exogenous CFH or FHL-1, as well as in Crispr-corrected EOMD iPSC-RPE cells. Results from this study will provide insights into the role CFH/FHL-1 plays in regulating players of the complement pathway RPE cells and its contribution towards progression of AMD.

Inherited retinal diseases (IRDs) are a diverse family of disorders which cause progressive vision loss and retinal degeneration. IRDs impact over two million people globally, with an estimated prevalence of 1 in 2000 individuals. With more than 280 genes currently implicated in IRD phenotypes, a retinal gene therapy approach offers hope for resolving the underlying genetic causes of IRDs. In 2017, the first FDA-approved retinal gene therapy for Leber congenital amaurosis, an IRD causing severe vision loss at birth, served as an encouraging landmark for their efficacy in therapeutic application. The therapy employs adeno-associated virus (AAV) vectors to replace mutant RPE65 genes. However, AAVs only have a carrying capacity of approximately 4.7kb, significantly limiting their application to other target genes. We hypothesize that lipid nanoparticle (LNP)-mediated delivery of messenger RNA (mRNA) may be a viable alternative to existing retinal gene therapy methods. We characterized cell type specificity of enhanced Green Fluorescent Protein (EGFP) expression in retinas after intravitreal injection of mRNA LNPs to anesthetized male, CD-1 mice. Retinas were subsequently prepared for cross section and flat mount confocal imaging 48 hours after injections. Immunohistochemical staining with SOX9, GFAP, and RPE65 indicated EGFP expression in retinal pigment epithelial cells (RPE) and Müller Glia (MG). These neuronal cell types are impacted in various IRDs and other degenerative disorders, making them promising targets for retinal gene therapy. In future work, we will quantify the intensity and duration of EGFP expression in mouse retinas after intravitreal injection with EGFP mRNA using flow cytometry analysis at timepoints 6 hours to 14 days post-injection. Additionally, we aim to deliver therapeutic replacement mRNAs to improve IRD phenotypes. The results of our investigation may demonstrate the therapeutic prospects of LNP-mediated gene therapy in mouse retinas.

H. pylori is a gram-negative, helical shaped bacteria that infects more than 50% of the world's population. The shape of bacteria is determined by the peptidoglycan cell wall, which is a macromolecule composed of glycan strands cross-linked by short D-amino acid containing peptide stems. The Salama Lab has identified multiple cell-shape-determining proteins that are required for the helical cell shape of H. pylori, including Csd5. Csd5 is a transmembrane protein and loss of Csd5 causes cells to be straight rods instead of helical. The C-terminal bacterial SH3 domain of Csd5 binds directly to the peptidoglycan cell wall. This project aims to identify whether the SH3 domain of Csd5 binds to a specific feature of the peptidoglycan cell wall. We hypothesize that the SH3 domain binds specifically to tetrapeptides in the peptidoglycan cell wall. To investigate whether the SH3 domain binds to tetrapeptides, we performed a pull-down experiment with purified SH3 domain and purified peptidoglycan from mutants that have varying tetrapeptide content. From this we confirmed that peptidoglycan with higher tetrapeptide content pulls down purified SH3 domain from Csd5 at a higher level than peptidoglycan with lower tetrapeptide content, suggesting that the SH3 domain of Csd5 preferentially binds to the tetrapeptides in the sacculus. In addition, we are visualizing where tetrapeptides are localized in H. pylori cells and identifying whether purified SH3 domain incubated with purified cell walls localizes to the same location by 3D microscopy. By investigating how the SH3 domain of Csd5 interacts with the peptidoglycan cell wall, we will learn more about how Csd5 controls the helical cell shape of H. pylori cells.

Why do identical twins have different lifespans? Beyond genes, what else might influence the aging process? Variation in any phenotype is due to the combined effects of genetic variation and environmental variation. In fact, there are two types of environmental variation—one is extrinsic environmental variation, such as food, temperature, etc., and the other is intrinsic environmental variation, which can lead to subtle differences in behavior, such as how much an individual eats, how long it sleeps, etc. We hypothesize that these differences can be predicted by an individual’s underlying metabolism. There are thousands of molecules that make up the structural and functional building blocks of all organisms, a domain known as the metabolome. Previously, many studies have shown that genotypes vary in lifespan, but even within a single genotype there is enormous variation in lifespan. Here we address how intrinsic environmental variation influences aging by controlling the genetic and extrinsic environmental variation under lab conditions. We designed an experiment using Drosophila melanogaster, and since Drosophila has a natural tendency to climb upwards against gravity, and climbing ability of flies decreases with age, we hypothesized that we might use climbing ability as a biomarker of future mortality risk. Using mid-life climbing ability, we separated genetically-identical flies and then analyzed each group’s lifespan. We found that within a genotype, strong climbers had a longer lifespan than non-climbers. Finding strong support for this hypothesis led us to propose that the metabolome between climbers and non-climbers might be different. Our goal is to understand the role of intrinsic variation in aging. If we can find metabolites that associate with climbing ability, and as we have shown, climbing ability is associated with aging, we might be a step closer to explaining how intrinsic environmental variation influences aging.

The gut microbiota, or the microorganisms that live in the gastrointestinal tract (GI), play a significant role in both gut and overall health. In LMICs (Low-Middle Income Countries), diarrheal diseases are among the leading causes of death in children under five. The ongoing EcoMiD (Enteropatógenos, Crecimiento, Microbioma, y Diarrea) project aims to study the interaction between infants' gut microbiomes and viral, bacterial, and parasitic enteric pathogens along a rural-urban gradient in Ecuador through a cohort study of 600 mother-child dyads. The objective of this portion of the study was to validate a method for assaying multiple enteric pathogens in stool samples simultaneously using TaqMan Array Cards (TAC). Before samples can be analyzed using TAC, each of the 56 pathogenic targets of interest must be validated to determine the limit of detection, limit of quantification, inhibition effect of the stool matrix on detection, repeatability and reproducibility. The TAC Positive Control validation process initially involves growing up each of the organisms of interest, extracting its' total nucleic acids (TNA) and quantifying this through (RT)-qPCR and Qubit. Analysis of this data enables seeding the positive controls at known concentrations to quantify target TNA for TAC. Of the 56 targets, we have completed this work with 35 targets (26 reference organisms, 9 gBlocks). For targets that are time/cost intensive to work with or not culturable, gBlocks are utilized in place of reference organisms. gBlocks are synthetic gene fragments with a known sequence. Next steps of this process includes validation of the 21 remaining pathogens, some of which involves working with anaerobic organisms. Analysis of this data will inform seeding concentrations to validate the developed TAC and enable quantification of these targets in stool samples from Ecuador. 

Historically, one’s social and geographical position can result in disproportionate levels of exposure to harmful environmental toxins and contaminants. The field of Environmental Justice aims at addressing this inequality by redistributing both the environmental burdens and benefits among all members of society. My research focuses on access to “blue spaces,” or public spaces built around bodies of water that confer health benefits and promote active lifestyles, as an environmental justice issue. Given that Minnesota prides itself with being the land of “10,000 lakes” on every license plate, my research aims to see if the state slogan may be misleading as these lakes may only be accessible to some members of my community. Exploratory in nature, my research intends to reveal possible inequalities regarding access to blue space—even if not clearly visible in the everyday landscape. Through the formulation of a Blue Space Equality Index, which includes several proxy variables for accessibility such as transportation routes, water quality, sidewalks and bench availability, and recreational usages, I will examine three blue space regions in Minneapolis. Using this index in conjunction with demographic and property values data, I intend to represent the local environment surrounding these spaces. A closer investigation of Minneapolis is relevant and timely considering the several high-profile cases of systematic racism in the city’s recent past which have made global headlines; Minneapolis is just a snapshot of what is occurring across the country and that closer examination of the environment is needed to better understand the forces underlying environmental and socioeconomic disparities in Minneapolis. My investigation of blue space accessibility will shed light on how the residents of Minneapolis connect with the environment around them and potentially expose the harsh realities for some community members who call the Twin Cities home.

Age-related macular degeneration (AMD) is a multifactorial eye disease characterized by drusen formation and thickening of the Bruch’s membrane (BrM). The BrM is a extracellular matrix (ECM) located between the retinal pigment epithelium (RPE) and choriocapillaris, which play roles in structural support, transport of nutrients and interactions with RPE. Changes to the ECM composition and architecture, contributed partly by accumulation of basal deposits during AMD could interfere with nutrient diffusion and transport. Moreover, RPE cells perceive changes that occur in the ECM, therefore age-related changes that occur in the Bruch’s membrane can affect RPE pathology in AMD cells. Early onset macular drusen (EOMD) is an inherited retinal degeneration with similar clinical phenotype to AMD. Patients with EOMD have rare genetic variants in the complement factor H (CFH) gene that affects both CFH and factor H-like protein 1 (FHL-1) expression. We recently described a novel mutation in a EOMD family, which resulted in CFH and FHL-1 haploinsufficiency. Peripheral blood mononuclear cell were isolated from EOMD patients, reprogrammed into induced pluripotent stem cells (iPSC), and differentiated into RPE cells. Preliminary results indicated the cells to have phenotypes consistent with early AMD, making it a beneficial in vitro model for the study of the disease. The ECM composition has not yet been explored in the EOMD RPE cells, therefore the purpose of this project is to examine the expression of and staining pattern of ECM components, as well as several common drusen components. RPE seeded on transwell filters were collected, sectioned and immunostaining was performed to visualize the ECM components. Expression was further validated by western blot. Results from this study will determine if CFH and FHL-1 haploinsuffiency leads to alteration of ECM, which could play a contributing role towards pathology of the RPE.

Mycobacterium abscessus (MABSC) is a species of rapidly growing nontuberculous mycobacteria (NTM) that is most frequently encountered in human NTM infections and is very difficult to treat. Active MABSC disease commonly emerges from pulmonary infections, to which populations with underlying lung disease and depressed immune systems are most susceptible. There is no official standard of care for MABSC infections and current treatment regimens lack efficacy and are met by challenges of intrinsic and acquired resistance mechanisms. Studies evaluating pulmonary disease outcomes report unsatisfactory treatment success rates of approximately 45%. Thus, there is an urgent clinical need for novel antibacterial agents and drug combinations to efficiently and effectively cure MABSC infections. Repurposing FDA-approved drugs can help us economically discover new treatment options with a shorter drug development time, which is critical for antibiotics as the emergence of resistance often outpaces drug development. We approached the repurposing of approved drugs to treat MABSC infections through the whole-cell screening of a drug library of about 2400 FDA-approved compounds and clinical molecules, followed by the selection and characterization of the activity of the most promising hits. Our data revealed a novel antiemetic compound, netupitant, that exhibited a potent anti-MABSC activity (MIC 4-16 µg/ml) and was able to interact synergistically with standard first-line MABSC therapeutics. Netupitant has a good safety profile and accumulates preferentially in lung tissues, making it suitable for treating pulmonary MABSC infections. Additionally, our screen revealed two promising antibiotic drug combinations: eravacycline/clarithromycin and bedaquiline/amikacin that were able to exhibit potent synergistic interactions against clinical MABSC isolates, as determined by checkerboard microdilution assays. Further mechanistic and in vivo studies are needed to evaluate these hits as potential treatment options to improve the current clinical outcomes for MABSC patients.

Global climate change is here and it is already affecting exposure to heat across Washington state. Heat exposure can exacerbate climate-sensitive health threats by increasing risks for groups who are more susceptible--particularly in non-acclimatized areas. The World Health Organization has predicted that diabetes will be the 7th leading cause of death by 2030. It is critical to define the interactions between heat and diabetes to prevent excess risk; having diabetes leads to reduced thermoregulation (body's ability to regulate heat) which affects other organs critical for sustaining life. My research focuses on the intersection of health impacts of heat on people with diabetes and how these relationships change with increasing temperatures. It also considers how mitigation and adaptation can protect the 1 in 8 people in Washington who have diabetes as part of the overall goal of managing the health risks of climate change. I consider the relationship between heat and diabetes by performing a literature review of peer-reviewed, original research in different academic journals. Then, outcomes are compared and synthesized to determine if there is a causal relationship where temperature affects the odds of diabetes morbidity and mortality. My research found that those who have diabetes have higher odds of negative health outcomes and death as temperatures increase. Through a variety of mitigation and adaptation strategies, we can reduce the odds of and prevent excess morbidity and mortality. Taking this research into account, Washington can be more conscious in its public health interventions during periods of increased heat to prevent excess diabetes-related morbidity and mortality. More research is needed on the effectiveness of specific interventions for Washington on reducing the odds of morbidity and mortality for people with diabetes.

Undergraduate nursing students—the next generation of health care providers—play a pivotal role in caring for people living with dementia (PLWD) as their numbers continue to rise. Unfortunately, new graduates may not be adequately prepared by their educational programs to care for PLWD. This study aimed to capture students’ current attitudes about and barriers to working with older adults and PLWD post-graduation, especially related to their desire for receiving additional clinical and didactic curricular content. We also evaluated students’ interest in enrolling in the elective dementia and Long-Term Care (LTC) Externship. An initial survey was distributed to first and second-year bachelor of nursing (BSN) students at the University of Washington School of Nursing (SoN) in April 2021. The survey consisted of 20 questions modified from the Dementia Attitude Scale (DAS) and 8 questions related to respondents’ health care experience and attitudes about nursing care of older adults. The survey was completed by 76 first and second-year BSN students. The survey focused on students’ comfort level working with PLWD and their willingness to expand geriatric and dementia care skills. To gather more illustrative qualitative data, we conducted a focus group with 8 BSN students. The survey results indicated low self-reported knowledge of nursing care for PLWD as well as a lack of desire to work in the discipline immediately following graduation. However, the majority of focus group members (N= 6) expressed interest in acquiring additional knowledge and hands-on learning opportunities in care for older adults and PLWD. Key components that would attract students to geriatrics training and a LTC externship included: structured seminar; access to interactive, relevant pre-study materials developed by professionals; and 4-6 hour clinicals of hands-on skills. Conclusions: The findings from these surveys have informed the development of a LTC Externship, which is currently being piloted and evaluated.

Polar cyclones are extremely common, but their relationship to sea ice melt and growth is not always clear. They survive longer than midlatitude cyclones and thus have significant impacts on the seasonal variability of sea ice. Sea ice can have major impacts on ocean circulation, local ecosystems, and is sensitive to important feedback systems that affect global climate. No comparison has been made between the two hemispheres of sea ice response to cyclones. I use data from a combination of ERA5 reanalysis for cyclone tracking and observed sea ice concentrations to select intense cyclones and compare their effects on sea ice. Atmospheric data is ‘regridded’ so that Antarctic cyclones are central to the area of analysis. This allows for further study into atmospheric features that make up cyclone structure and spatial patterns in the sea ice response. Atmospheric features show similar spatial patterns in both Antarctic and Arctic cyclones, however differences in temperature, water vapor, and cloud cover are much greater in Antarctic cyclones. Typically, Antarctic cyclones are considered to be more intense than Arctic cyclones, however I show that when differences in latitude and typical background sea level pressure values are accounted for, cyclone intensity is similar in both hemispheres. In both hemispheres, sea ice growth is found to the west of cyclones and loss is found to the east, however greater east-west differences are found in the Antarctic. Understanding the impact of polar cyclones can improve short-term sea ice forecasting and paint a clearer picture of the variables that control the climatology and variability of the polar atmosphere.

Current research in Anthropology of children and their agency is generally lacking as they are often viewed as passive appendages to adults. This study seeks to study the moral agency of children from a gender perspective in a conventionally patrilineal, patriarchal cultural context. To investigate the ways in which gender interacts with and exert influence on children's moral development, we examine an unpublished archive of interviews and observations of rural Taiwanese children compiled by the late anthropologist Arthur P. Wolf in the 1950s using modern computational methods. For this research, I apply techniques such as statistical modeling and social network analysis to explore children’s interpersonal behaviors. Our preliminary findings show that although boys and girls exhibit different behavioral patterns, gender does not seem to be a factor that affects a child’s centrality in the network based on co-occurrence. My future analysis will focus on children's conflict and cooperation, the dark and bright side of moral development, by studying the general behavioral patterns within and across gender and/or age group, but will also highlight representative or epicentic individual observation entries. By looking into the social life of school-age and pre school-age children of rural Taiwanese children in the 1950s, this study also sheds light on the influence of gender on children's moral development in a non-Western historical and cultural context.

Thor: Ragnarok hit theaters in 2017 in the midst of the mid-2010s surge in the popularity of the Marvel Cinematic Universe. As media conglomerates continue to grow and subsume smaller artists and studios, attending to the types of narratives they are producing and how those narratives serve or complicate the aims of the overall corporation becomes increasingly important. Furthermore, Disney’s global reach as an American media franchise can be considered a product and extension of historical colonial violence, even as some of its products purport to critique such forms of oppression. In this project, I examine how Thor: Ragnarok, directed by Māori filmmaker Taika Waititi, reframes the glorified civilization of Asgard as a violent colonial empire, a fascinating recontextualization that potentially critiques not only sanitized narratives of colonialism but Disney’s utilization of them. The movie was commercially successful, raising further questions on how the film simultaneously critiques and benefits Disney. I question how Thor: Ragnarok constructs its critique of its fantasy-colonialism and the parameters of that engagement. To accomplish this, I perform a close reading of the film alongside contextual materials such as previews, interviews, and previous Marvel media from which it draws. I argue that Thor: Ragnarok reframes and subverts the previous Thor movies to critique historicized and overtly violent colonialism, and constructs a simultaneous critique of ongoing legacies of colonialism and systems of oppression. Its critique of ongoing oppression is couched in humor, avoiding being provocative in a way that would alienate audiences accustomed to the previous whitewashed depictions of power and empire or fully implicate Disney’s participation in those narratives. I trace how this critique’s possibilities and limitations coexist alongside each other in the film, creating an amalgamation that reenacts cultural tensions and discourses rather than producing a monolithic narrative.

Ewing sarcoma (ES) is a bone and soft tissue tumor that primarily occurs in children and young adults. The tumor is driven by an oncogenic fusion gene that fuses the EWSR1 gene located on chromosome 22 to FLI1, an ETS family transcription factor located on chromosome 11. EWS-FLI1 promotes tumorigenesis through transcriptional and epigenetic dysregulation. Despite maximally intensive cytotoxic therapy, the outcome for patients with metastatic ES remains poor, thus the need to identify new therapeutic agents. Given ES’s epigenetic dependencies, there is strong rationale to investigate epigenetic modifying drugs. Bromodomain and extra terminal domain (BET) proteins function as epigenetic readers that facilitate transcription and are upregulated in many tumors. I have shown that BET inhibition slows the growth of ES cells in vitro and in mice. However, BET inhibitors (BETi) will not be successful as a single agent. I hypothesize that combining BETi with other biologically targeted agents will lead to synergistic effects and tumor regression. Based on established literature, RNA-seq data of BETi-treated ES cells, and in silico drug screen, we selected Copanlisib (PI3K inhibitor), GSK-126 (EZH2 inhibitor), PP121 (Src, mTOR, VEGFR2, PI3K inhibitor), Danusertib (Aurora Kinase inhibitor) and Infigratinib (FGFR inhibitor) to test in combination with BETi (BMS-986158). Using standard in vitro cytotoxic assays and calculating synergy using the Chou-Talalay method, my preliminary results showed strong synergy between Copanlisib and BMS-986158. I detected no synergy between BMS-986158 and GSK-126. Our initial results from our in silico drug screen predict that Danusertib, Infigratinib and PP121 may be synergistic with BETi. Ongoing in vitro studies are testing this. Promising combinations will be tested in vivo xenograft models. It is our goal to identify drug combinations from the drug screen that will enhance the cytotoxic effects of BETi in vitro and lead to tumor regression in vivo.

Madjedbebe is the earliest known site of human activity in Australia with artifacts dating to 65,000 years ago. However, the presence of termite mounds in the landscape of the site have led some to question this notable date. Termites have the potential to affect the stratigraphic integrity of archaeological sites, their subterranean movements mixing sediments from deposits of different ages. Such activity may have led to the overestimation of the age of artifacts at Madjedbebe. Our study investigates the extent to which termites disturbed Madjedbebe’s archaeological deposits, thereby providing key insight into the reliability of the ages of the site’s sediments and artifacts. We used micro X-ray fluorescence (μXRF) to quantify and compare elemental concentrations in blocks of resin-impregnated archaeological sediments and sediments collected from termite mounds. We analyzed the μXRF data using log ratio analysis. Our research seeks to identify whether chemical traces of termite activity exist within sediments at Madjedbebe. Our results indicate that termite-impacted sediments have distinctive chemical signatures that distinguish them from archaeological deposits; generally, this signature is not present in any of the Madjedbebe samples. There is therefore little evidence for termite activity and mixing in the archaeological deposits. Our findings are indicative of high stratigraphic integrity at the site, suggesting that the proposed 65,000-year-old ages of the Madjedbebe artifacts are sound. This is important for understanding the timing of the arrival of humans in Australia, the earliest movement of humans out of Africa, and the chronology of human interactions with Denisovans and Neanderthals. Our research also demonstrates the applicability of material science methods to archaeological questions.

Congenital Heart Defects (CHDs) are the most common birth defect within humans and are characterized by having multiple structural problems with the heart, often leading to problems with blood circulation. Our lab has identified a set of genes potentially involved in birth defects that are also expressed during heart development, thus identifying candidate genes for congenital heart defects. Amongst the gene candidates is a gene encoding proteasome maturation protein (POMP). Proteasomes are the main system for protein degradation, but the role of proteasome factors in CHD development is not known. To establish the importance of POMP and the proteasome in proper heart development, I examined zebrafish embryos that are mutant for the POMP gene and the effects of chemical proteasome inhibitors on zebrafish heart development. Zebrafish (Danio rerio) are commonly used to study human development and birth defects because their genes and organs strongly resemble those of humans, and the zebrafish heart develops in only 3-4 days. Using fish that have been tagged with green fluorescent proteins to visualize heart development, I tracked the developing hearts of POMP mutants and proteasome for signs of heart malformation. I expect to find structural similarities between the developing hearts of the POMP mutants and those with inhibited proteasome function, thus indicating the proteasome system as having a prominent role in CHDs. Gaining insight into the molecular mechanisms and genes behind CHDs aids in the development of human patient diagnoses, targeted therapeutic treatments, and an increased understanding of complex diseases.

Microfluidics is the technology of systems in which microscale channels are used to manipulate small quantities of fluids (microliter to picoliter or less). Open microfluidics provides a platform to control the movement of microscale volumes of liquid in open space, making every position more accessible than conventional closed microfluidics. In many applications–including cell culture, chemical synthesis, and high throughput screening–merging droplets is essential for the experimental workflow. For example, merging droplets containing reagents can initiate a chemical reaction, or adding a drug to a cell culture can stimulate cells. In conventional closed droplet-based microfluidics, merging techniques often rely on external components such as merging by electrofusion using electrodes. In other cases, like merging using hydrophilic strips in a channel, the merging method is built into the device and the location of merging must be determined in advance. Therefore, the ability to easily initiate droplet merging in an open microfluidic system would be beneficial to researchers and offer flexibility in experimental design and applications. In this study, I developed an easily accessible droplet fusion technique with the prick of a needle at the liquid interface; my method fuses multiple droplets simultaneously (2-15 droplets). Importantly, this fusion method can be used on-demand at any point in the open microfluidic device and does not require external equipment nor features built into the device. This presentation characterizes the experimental parameters required for successful and controlled droplet fusion and explores the physics behind this phenomenon.