Hydrogen sulfide (H₂S) is a common cause of workplace injuries and deaths for industrial workers. In our project, we use Caenorhabditis elegans (C. elegans) as a model organism for investigating how cells behave under an environmental stressor and the long-lasting effects of that behavior. Previous work in our lab has shown that early exposure to low H₂S (50 ppm) enable C. elegans adults to survive a much higher subsequent exposure by forming a cellular memory known as a “bookmark.” Bookmarked animals survive at high H₂S (150 ppm), while animals without previous exposure do not. In a genetic screen, we identified various epigenetic factors that are involved in this process; however, it is still unclear when in the “life” of the bookmark and where in the animal these factors are required. The required bookmarking gene swsn-4 is part of the SWI/SNF complex, a group of proteins that regulate compaction of DNA and thus the accessibility of genes. We are interested in assessing the spatial requirements for swsn-4 by rescuing mutant animals that lack this chromatin-remodeling factor. For the first part of the project, we use Gateway recombination cloning technology to enable tissue-specific expression of swsn-4. In the next part of the project, we test whether introducing swsn-4 in specific tissues rescues bookmark retention. A recent study identified hif-1, a transcription factor, to be broadly needed to rescue animals exposed to both low and high H₂S, suggesting that the response is needed in most cells to ensure survival of the animal. Because swsn-4 is also present broadly in the body of C. elegans, we predict it will be needed in a similar way to hif-1. We hope that our investigation would lead us to discovering methods in which we can utilize the properties of H₂S as a chemical messenger to help patients.

Organic photovoltaic (OPV) cells are an emerging technology that is experiencing continued breakthroughs such as reaching a power conversion efficiency (PCE) of 17.3% in August, 2018. OPVs have the potential to become a major source of energy in our future and a more sustainable energy option than traditional solar cells. In addition to contributing a lower environmental impact than common silicon-based solar cells, OPV cells can be made to be flexible, lightweight, and are comparably inexpensive to fabricate. They are also quite customizable via molecular engineering providing the opportunity for much novel architecture. Our research team focuses on innovating a modular processing system for OPV cells in the form of multi-component fibers by continuously coating device layers onto wires and winding the fiber with a secondary electrode. Using a small, user-friendly system allows us to focus on the most important factors that affect the morphology and PCE of the resulting OPV fiber. After characterizing the fibers we are able to consider what changes need to be made to the modular system, allowing us to better advise on the design of a larger-scale manufacturing process for organic photovoltaic fibers.

Pre-hospital patient protocols standardize medical procedures for EMTs and increase the quality of care for patients. In Washington state, these protocols vary by county and often contain a section for psychiatric emergencies with variant restraint protocols. The immediate use of restraints in these situations can be seen as dismissive to the patient and can lead to agitation and violence. Verbal de-escalation can enhance provider-patient relations and decrease likelihood of restraints, seclusion, and hospital admissions.  The goal of this project was to explore restraint and de-escalation methods used in WA state county-level EMS pre-hospital patient protocols. To complete the project, a codebook was created with binary variables with definitions that determined whether the procedures listed in the protocol satisfied the definition. Using the codebook, each protocol was independently coded by 2 people and disagreements were reviewed by a third person. Finally, we created descriptive statistics from the restraint protocols and stratified based on rurality. Of the 39 counties in WA state, 77% of counties mentioned verbal de-escalation methods in their pre-hospital patient protocol. The de-escalation method with the highest proportion in all counties recommended reassuring the patient that the providers care for them (28%). The second highest proportion in all counties was to mention the use of de-escalation methods without explicit instructions (23%). Counties that do not have specific instructions and just mention the use of de-escalation methods can be confusing if the provider does not have de-escalation training or has not trained recently. These findings highlight the need for future research concerning the adherence of EMTs to these protocols and to see the what the outcomes are of the different protocols. More research can be done through contacting individual EMS agencies to see if internal protocols exist outside of county protocols.

This research involves the design of a system for an unmanned aerial system (UAS) to operate and navigate in an environment devoid of a Global Navigation Satellite System (GNSS) such as the Global Positioning System (GPS). The system operates by interrogating an aviation transponder (either mode C or S) that is carried by the UAS and measuring the time elapsed for the response to multiple, ground-based antennas and using triangulation (multilateration) to locate the transponder and by association, the UAS. The ground-based system then routes this position information back to the UAS via the UAS’s data telemetry link. The autopilot then utilizes this position information for navigation in much the same way it would utilize a GPS-based position report. Our research focused on the system architecture to enable a UAS to operate in a GPS-denied environment. Flight test results are presented utilizing a customized version of the popular Pixhawk/ArduPlane avionics platform and demonstrate that the system is capable of guiding a UAS through a series of waypoints in the absence of GPS signals. Furthermore, the customized controller that was designed to consume this alternate source of position information performed well in highly unfavorable environmental conditions. This success illustrates the feasibility of the system as a practical alternative to GPS.

All over the globe, an escalating number of people are developing distressing sensitivities to our environment’s everyday intoxicants. Multiple chemical sensitivity (MCS)—also termed idiopathic environmental intolerances (IEI), among other names—is an acquired disorder characterized by a wide range of symptoms and reactions to low-level chemical exposures. Studies conducted in Japan, USA, Sweden, and Denmark found that 12% of the population has been diagnosed with chemical hypersensitivity, while total underreported prevalence is estimated between 15% and 27%. More importantly, research states that prevalence has increased by 300% within the past decade. People diagnosed with MCS experience a range of symptoms, examples including asthma, heart irregularities, headaches, dizziness, confusion, skin irritation, and fatigue. Abundant in our everyday environment, commonly reported triggers include pesticides, building or renovation materials, car exhaust and chemical smog, smoke, perfumes and fragrances, and cleaning supplies, among others. With a myriad of proposed titles, contrasting research perspectives and approaches, and a staggering dissensus on physiology and diagnosis, it is critical that we compare and contrast the existing literature by geographical region. Three primary research hubs generate the majority of contrasting MCS research; these regions include the United States and Canada, Scandinavia, and Japan. The problematic inconsistencies include (1) a lack of consensus definition or diagnosis for the illness, (2) varying perceptions on pathophysiology, and therefore (3) contrasting treatment strategies. This research aims to examine the global, contrasting research approaches, proposals for treatment or action, and subsequent healthcare experiences within the geographical regions of focus. Preliminary results suggest that inconsistencies in the medical diagnosis of MCS lead to deficiencies in research and ineffective healthcare. By any given title, the escalating number of chemically hypersensitive patients is a growing global health concern. Integrating the globally contrasting approaches and diagnosis criterion is the first step to developing efficient healthcare strategies.

My research is on the intersectionalities of racism and sexism as they apply to the healthcare of women of color, and specifically that of African-American women’s maternal mortality rate. African-American women die of after-birth complications at a rate that is three times higher than that of White women. I am using Critical Race theory and Intersectional Feminism to analyze how racism and sexism are interconnected, and how they together implicitly bias many healthcare professionals. I am working from the viewpoint that racism is widespread throughout American society, and highly influences the way that people of color, and white people, live their lives. However, I also believe that an intersectional approach is necessary to fully understand the influences on women’s healthcare, and that examining the sexism inherent in women’s care is vital to understanding the full extent of the bias society holds against Black women. I am creating a literature study on the history of bias in the medical system with a focus on historical methods of medicalizing racism, researching the explanation for Black women’s historical distrust for the white healthcare system, and a focus on current research on implicit bias among healthcare professionals. I am hoping to discover whether the underlining differences in Black women’s maternal mortality rate, due to their societal positionality, is being addressed by their doctors in their healthcare strategies. I am hoping to show that a healthcare system which addresses bias and the biological and psychological effects of racism and sexism rather than ignoring them and treating every woman the same is vital to ensuring every woman’s optimal health outcome.

Detection of uncomplicated mild traumatic brain injury (mTBI) is difficult because there are no visible brain lesions that are often associated with more severe forms of TBI. New biomarkers would allow doctors to more sensitively screen for mTBI using neuroimaging methods. One promising biomarker technology is resting-state functional connectivity, which is brain activity measured at rest using functional magnetic resonance imaging. One particularly salient resting-state network is the Default Mode Network (DMN). Our research focused on identifying differences in resting-state functional connectivity between individuals diagnosed with mTBI and healthy controls. We examined mTBI in 254 U.S. military personnel deployed to a combat theatre in the Middle East from 2010-2013. Each subject underwent initial magnetic resonance imaging and screening for TBI following medical evacuation to Landstuhl Regional Medical Center (LRMC), the primary triage center for all evacuated combat casualties, up to 30 days post-injury. We used four distinct groups for our analysis; Blast/Non-Blast (n=79, 44) TBI, and Blast/Non-Blast Control (n=35, 96) while covarying for age and gender. We hypothesized that resting-state networks will be disrupted in TBI and blast populations when compared to controls. We used two different methodologies; the first was a seed-based analysis examining group differences in the correlations from the Posterior Cingulate Cortex (PCC, a key hub within the DMN) to the whole brain. The second analysis used the Yeo Seven Network parcellation to compute correlations between all seven networks to the DMN. We were unable to distinguish any group from controls, suggesting that early differences in functional connectivity are not a robust biomarker of injury.

Epithelial-mesenchymal transition (EMT) refers to a biologic process that allows a polarized epithelial cell, which normally functions in the basement membrane of a cell, to undergo biochemical changes that makes it express as a mesenchymal cell phenotype. This mesenchymal phenotype allows the cell to have enhanced migratory capacity, invasiveness, elevated resistance to apoptosis, and increased production of extracellular matrix (ECM) elements. The process of EMT is considered completed once the underlying basement membrane breaks down, and the mesenchymal cell becomes migratory. Another component that proves EMT is the loss of e-cadherin. E-cadherin refers to cell-to-cell adhesion and the degradation of e-cadherin levels are a hallmark of EMT happening. There are three distinct types of EMTs; I will be focusing on type II EMT. Type II EMTs are associated with inflammation/wound repair but usually stops once inflammation subsides. However, in the context of organ fibrosis, type II EMTs can continue to over-respond to a persisting inflammation and can lead to organ death. In my experiment, I hypothesize that in HEK 293 human cells, SNAP -Δ1-91 alpha-1D adrenergic receptors undergo type II EMT. SNAP -Δ1-91 alpha-1D adrenergic receptors are a truncation of the extracellular portion of the receptor. Certain receptors undergo this truncation to increase its expression. It is shown that in SNAP – Full Length alpha-1D adrenergic receptors  (wild type receptors) do not undergo EMT. I will be able to observe the process of type II EMT through imaging the breakdown of the cell membrane in SNAP- Δ1-91 alpha-1D adrenergic receptors and the measuring of e-cadherin levels. The purpose of this research would be to potentially influence future therapeutic interventions that target wild type receptors to induce would repair. 

Individuals with hearing loss have a much greater difficulty understanding speech in noisy environments than their normal hearing counterparts, leading to reduced participation in communicative activities and a lower quality of life. Audibility explains some of the difficulty this population experiences, but even with audibility recovered by hearing aid amplification, impairments exist in spectral and temporal processing. The extent of spectral and temporal processing impairment is captured in detection of spectro-temporal modulations, which is closely related to speech understanding in noise. This study explores the potential effect age has on STM sensitivity, while controlling for severity of hearing loss. In order to investigate, we measured STM sensitivity in participants with a hearing loss across a wide age range. In the STM test, they are asked to listen to two sounds, a broadband noise and the same broadband noise with spectro-temporal modulation applied, and identify which sound contained modulations. The modulation applied will adaptively change based on response accuracy until the listener can no longer discriminate between the two sounds. A multiple linear regression model is used to analyze the data, with predictors of hearing loss severity and age. We expect to see a decline in STM threshold with age, independent of hearing loss status. The results are discussed in terms of potential for future clinical application and the ability to better the quality of hearing aided listening.

Short-term exposure to diesel exhaust particulate matter can cause headaches, dizziness, and irritation of the nose, throat, and eyes. Prolonged exposure has been shown to increase the risk of developing cardiovascular disease, respiratory disease and lung cancer. Because heavy machinery is often fueled by diesel, occupations such as coal miners, truck drivers, railroad workers, and construction workers are at high risk of exposure. Non-occupational exposures are also of concern, especially in locations impacted by high volumes of vehicle traffic. A potential way to determine diesel exhaust exposure is by measuring the amount of nitrated polycyclic aromatic hydrocarbons (NPAHs) in household dust. There is no current method for measuring NPAHs in dust. We conducted a literature review of methods for measuring related chemicals in household dust and measuring NPAHs in other matrices. We tested three different methods before establishing the optimized sample preparation and cleanup process using silica gel solid phase extraction followed by analysis using high performance liquid chromatohraphy with tandem mass spectrometry detection (HPLC/MS/MS). To evaluate method performance we analyzed replicates of spiked and unspiked household dust and silica gel. The method was used to analyze 42 household dust samples collected in two communities – one with expected high and one with expected low traffic-related air pollution. Future research should include comparisons between dust NPAH measurements and other measures of diesel exhaust exposure, including NP (nitropyrene) metabolites in urine, air filter NP, or a priori predicted exposure based on home location.

Cranial elaboration, in the form of domes, horns, and bony bosses, has evolved multiple times in vertebrate history, including in non-mammalian synapsids, the extinct ancestors of mammals. The lack of similarly thickened skulls in living animals has meant that very little can be inferred about how these features developed or why they evolved. Here we use comparative bone histology to describe the microanatomy of Oudenodon, a 255-million-year-old synapsid with distinctive nasal bosses. We compared thin-sections from the thickened portions of Oudenodon to the non-thickened skull of its relative, Diictodon, in order to better understand how cranial elaboration developed in synapsids. We took thin sections from the thick nasal bosses as well as non-thickened regions of Oudenodon to capture changes in tissue microstructure. Then, we compared these thin-sections to similar regions of the skull in Diictodon, which lacks cranial elaboration. In the bone tissue of both Oudenodon and Diictodon, we observed a cancellous, spongey interior sandwiched between two layers of compact cortical bone. This pattern of bone is typical in many modern skulls but the thickness of the cancellous region in Oudenodon is strikingly thicker, even in regions away from the boss. We hypothesize that increased cranial thickness in Oudenodon is due to this middle expansion of spongey, cancellous tissue. As a result of the increased bone thickness, sutures in Oudenodon are deep, wide, and convoluted when compared to the relatively simple interdigitated sutures in Diictodon. These and other histologic comparisons with distantly related taxa allow us to interpret the growth and construction of cranial elaboration in synapsids, adding to our understanding of how these thickened skulls evolved. Future work can also explore the possible soft tissue covering of these bosses and domes, which may have been used for intraspecific display or combat. 

Drepanosauromorpha is an extinct group of reptiles known from the Middle to Late Triassic (237–212 MA). The clade currently includes seven genera (Avicranium, Dolabrosaurus, Drepanosaurus, Hypuronector, Kyrgzsaurus, Megalancosaurus, and Vallesaurus) that are known from fossils collected in Europe (Italy, UK), North America (Arizona, New Mexico, New Jersey), and Asia (Kyrgyzstan). The first described drepanosauromorph, Drepanosaurus unguicaudatus, was based on a flattened holotype preserving most of a complete skeleton. Subsequently described drepanosauromorphs display the following diagnostic features: the length of the chevrons (ventral spines below the tail vertebrae) is substantially longer than corresponding tail neural spines, the cervical (neck) vertebrae are heterocoelous (saddle-shaped articular surface), the cervical ribs are absent as distinct ossifications, and the chevrons are fused to their respective centra. In recent years, both three-dimensionally preserved partial skeletons and isolated material of drepanosauromorphs have been found across both Europe and North America. These discoveries have helped shape our understanding of the biology and diversity of drepanosauromorphs. However, comparing isolated, three dimensionally preserved specimens to the more complete, yet two dimensionally preserved articulated specimens is difficult due to differences in preservation. Here, we describe a new drepanosauromorph species from the Chinle Formation in Petrified Forest National Park, Arizona based on the left second manual ungual (claw) . Some of the characteristics that distinguish this claw from those of most drepanosauromorphs is its size. It differs significantly from all known Drepanosaurus specimens (like the Italian holotype and the Hayden Quarry Drepanosaurus) because of the ventral placement of the cotyle (articulation surface), the height of the claw, the lack of compression along the pre-axial/post-axial plane, and a furrow along the midline. This new taxon not only highlights unsuspected morphological variation within Drepanosauromorpha, but also helps sheds light on the evolutionary history of smaller-bodied reptiles within Late Triassic ecosystems.

Myocardial infarction (MI) is the leading cause of death globally. Methods to regenerate cardiac tissue after MI has focused on inducing proliferation in adult cardiomyocytes near infarcted tissue or injecting stem cell-derived cardiomyocytes with proliferative capacity into the infarcted tissue. However, optimal regeneration has not been achieved with these methods, as the mechanism behind adult cardiomyocyte proliferation is not well understood and proliferative stem cell-derived cardiomyocytes are phenotypically and functionally immature. Exploration of the mechanism of cardiomyocyte proliferation is therefore necessary to enable optimal regeneration of cardiac tissue and function and MI. We hypothesize that the sarcomere structure, the basic muscle unit of the cardiomyocyte, is the limiting factor in proliferation of cardiomyocytes. To investigate this hypothesis, we have performed a thorough characterization and comparison of stem cell-derived wild type cardiomyocytes (WTC-CMs) and troponin I double knock out cardiomyocytes (TNNIDKO-CMs) which have an incomplete sarcomere structure due to the lack of troponin I. After confirming TNNIDKO-CMs and WT-CMs only vary in their sarcomere structure, we developed a coculture platform to demonstrate the mechanical weakness of TNNIDKO-CMs sarcomere structure. We then performed proliferation assays utilizing multiple proliferation markers to observe if proliferation was higher in the TNNIDKO-CMs with the incomplete sarcomere structures. Preliminary results have shown that TNNIDKO-CMs are more proliferative than WTC-CMs, thus implicating that sarcomere structure plays a role in controlling cardiomyocyte proliferation. Successful characterization of TNNDKIO-CMs and their increased proliferative capacity will elucidate the sarcomere structure’s role in proliferation as well as develop a more comprehensive understanding of the underlying mechanism behind proliferation to help progress therapies for regeneration of cardiac tissue after MI.

Reinforcement Learning (RL) is a subcategory of machine learning, in which an agent (the decision maker) observes its environment and executes the best course of actions to maximize rewards. This is similar to teaching a pet to perform tricks using treats as positive reinforcement. Our research compares different RL methods on low-performance devices like a Raspberry Pi in real-time, real-world environments. RL has gained popularity recently with breakthroughs from DeepMind’s paper, Playing Atari with Deep Reinforcement Learning, where an agent learns to play Atari games from raw pixels and from DeepMind’s AlphaGo (DeepMind, https://deepmind.com/research/alphago) program that was the first computer program to beat a world champion Go player. RL projects like AlphaGo have utilized big data, powerful computing resources, and simulated environments that do not require real-time interaction to train the machine learning models. Our group compares the effectiveness of different RL methods on an accessible level of computing power on offline devices that an average consumer could acquire. The team constructs a physical environment for the robot to navigate, creates an OpenAI Gym environment that our agents will use to control the robot and get feedback from the environment. We train our agents using different RL methods to optimally navigate the environment and avoid collisions. We then compare the performance of the different methods in our physical real-time environment. Reinforcement Learning in small, offline devices could pave the way for a variety of devices that learn over time without being connected to a network. Imagine a small Mars rover that learns to navigate its environment efficiently over time.

Solar energy has unmatched potential as the energy source of the future and semiconducting polymers (SP) offer a unique set of properties that can address many of the current barriers that restrict solar technology. SP are exciting because they have untapped potential for improvements in efficiency and they offer a cheap, energy-efficient alternative to silicon due to the ability to scale their production to industrial applications via film deposition techniques, like roll-to-roll printing. Solution processing via roll-to-roll printing is transformative, allowing for low-energy, high-throughput manufacturing of flexible devices. Previous work by Tatum and Resing investigated crystallinity in SP film microstructures through the self-assembly of highly ordered nanowires. This project expands upon this by utilizing a Python classification program to generalize relationships between morphology, optoelectronic properties and processing conditions of organic photovoltaics (OPV). Films of these materials will eventually enable stretchable and deformable electronic devices, but the nano- and microstructures are currently stochastic and inconsistent in their morphologies and resulting properties because processing and chemical conditions influence the domain size of the components and the distribution of those domains throughout the film. Using atomic force microscopy (AFM), a relatively cheap and quick technique, the active layer domains have been spatially resolved based on differences in their mechanical properties. These properties are strongly correlated to electronic performance factors such as fill-factor, short-circuit current and open-circuit current. For this project, OPV with an active layer of Poly(3-hexylthiophene):Phenyl-C61-butyric acid methyl ester has been fabricated with systematically varied processing conditions. A library of data has been established, containing AFM images, the device morphology and OPV performance data. This experimental data set of unprecedented compositional resolution aids in the evaluation of cutting edge simulation techniques, creating a more accurate computerized simulation model for OPV. 

Mental illness is a national concern; for rural residents, the availability of mental health services magnifies the problem. The 2017 National Survey on Drug Use and Health found close to 1.5 million non-urban residents had psychiatric illness or serious thoughts of suicide during 2017. Hypothesis: In Washington State, there is less access for outpatient mental health services in rural areas than in urban areas. The 2018 Washington State Directory of Certified Mental Health, Substance Use Disorder, and Problem & Pathological Gambling Services contains 503 entries of mental health facilities. Using the Office of Community Health Systems Series on Rural-Urban Disparities, facilities with available information were categorized as urban or rural. Statistics were calculated to characterize the availability of individual therapy, and crisis intervention information via public-facing websites for included clinics. The association between rural-urban status and the availability of outpatient psychotherapy and crisis intervention contact information was tested via the chi-squared test of independence. Approximately half (n=131, 50.6%) of clinics with available information regarding outpatient psychotherapy indicated that these services were available. Similarly, nearly half (n=120, 46.1%) of clinics with available information had crisis lifeline information presented via their public-facing website. There was no significant association between urban-rural status and availability of outpatient psychotherapy services (_X²=0.21, p=.647), nor between urban-rural status and publicly available crisis lifeline information on mental health clinic websites (_X²=0.80, p=.371). Washington State rural clinics are not significantly different from urban clinics with respect to saying they provide outpatient psychotherapy and crisis lifeline information. Limitations include methods relying on public reporting of services and limited scope, not allowing the study of other factors that might affect rural residents, such as distance traveled to obtain care. Rural counties in Washington State do not differ from urban counties with respect to the presence of clinics providing outpatient psychotherapy.

2D cell models have traditionally been used in labs to test the effects of new drugs on certain cell types due to the ease and convenience of use. While 2D methods are great, they often simplify the cell-to-cell interactions and may not accurately represent cell systems in humans. 3D methods show the complex cell communication systems and better simulate actual organ systems. Research comparing these two methods can inform scientists on the benefits of 3D models which can help efficiency in creating new drugs. Our lab looked into various 3D models to determine their effectiveness and reliability and looked into the differences in perceived cell mechanics and functionality between 2D and 3D methods. We tried Corning Matrigel and Corning 3D Spheroid microplates for 3D cell modeling using HEK293 cells, which are human embryonic kidney cells that were grown in lab. They are known for being easy to grow and transfect. We used SNAP-Gels, which are protein assays that show the protein levels in the cells, to ensure that the protein levels were similar between the 2D and 3D systems. We then did florescent imaging to determine cell localization and EPIC dynamic mass redistribution (DMR) to determine cell functionality. We found Matrigel to have inconsistent results, so we focused on using the spheroid microplates. Based on our initial results, we saw increased functionality and expression levels for full-length protein cells compared to cells with a truncated N-terminal protein in the 3D method. This increase in functionality and expression levels was not seen in the 2D method. Our results show that 3D modeling methods can be reliable, and do show results that differ from 2D models. This is important for future studies that require cell modeling because 3D models can provide a more accurate and reliable modeling system to create novel therapeutics.

G-protein coupled receptors (GPCRs) - characterized by seven transmembrane alpha helical domains - are the largest family of membrane proteins, constituting ~1% of the human genome. The α1D-adrenergic receptor (A1DAR) is a GPCR that regulates function of the cardiovascular, urinary, and central nervous systems. Dysfunction of this receptor can lead to various diseases including schizophrenia, benign prostate hypertrophy, hypertension, and PTSD. Prazosin, a non-specific α1-antagonist is the first line treatment for PTSD, however, chronic use has deleterious side effects including orthostatic hypotension and potentially fatal reflex tachycardia due to interactions with off-target related receptors. Thus, understanding how A1DARs are regulated will allow for the development of targeted therapeutics. To this end, the Hague Lab has previously discovered that A1DAR undergoes an endogenous cleavage of its extracellular N-terminal domain, affecting its membrane localization and response to agonist stimulation. Located within the N-terminal domain of A1DAR are two glycosylation sites at amino acids 65 and 82. Currently, how glycosylation of these sites regulates the cleavage event remains unknown. To characterize this phenomena, I used molecular cloning to mutate the glycosylation sites of A1DAR in the pSNAP vector for expression in Human Embryonic Kidney 293 (HEK293) cells. Near Infrared PAGE analysis revealed that glycosylation of both amino acids is required for cleavage and proper expression of A1DAR. Sucrose density gradient and dynamic mass redistribution further showed that glycosylation controls function and trafficking of A1DAR to the membrane. These results allow for the development of targeted medications specific to the N-terminal glycosylation sites of A1DAR, further reducing the potential side effects experienced by patients.

G protein-coupled receptors (GPCRs) contain seven transmembrane domains and are the largest family of cell surface receptors, making up ~1% of the human genome. GPCRs can interact with a variety of ligands, such as odors, pheromones, hormones, and neurotransmitters. At least 30 human GPCRs contain a C-terminal Type-I PDZ ligand that allows for interactions with adapter proteins which can regulate receptor trafficking, stability, and signaling. The Hague Lab has previously found that the α1D-adrenergic receptor, which contains a C-terminal Type-I PDZ ligand, also undergoes an endogenous N-terminal cleavage event, which enhances receptor function and may play a role in which PDZ domain containing proteins interact with this receptor. We propose that this unique observation of the α1D-adrenergic receptor may be prototypical of a new class of GPCRs which contain a Type-I PDZ ligand and undergo an N-terminal cleavage. CysLT2, MAS1, and NPFFR2 are understudied GPCRs and potential members of this family with distinct PDZ ligands, though it remains unknown if their extracellular N-terminal domains regulates receptor function. Thus, I have cloned wildtype and N-terminal truncation mutants of these three GPCRs into the pSNAP vector to create fusion proteins with N-terminal SNAP tags. These constructs were transfected into HEK293 cells and subjected to near infrared PAGE analysis to elucidate the presence of N-terminal processing. Furthermore, dynamic mass redistribution revealed how the N-termini modulate receptor signaling. The combination of biochemical and pharmacological techniques allows me to determine if these receptors belong to this new subfamily of GPCRs. These results increase our understanding of how GPCRs are regulated within the cell, which can lead to the development of more efficient drugs.

Imidacloprid (IMI), a neonicotinoid insecticide, is being sought by shellfish growers to control burrowing shrimp (ghost shrimp, Neotropea californiensis) in Willapa Bay and Grays Harbor, Washington. The shrimp destabilize sediments resulting in poor survival and low yields of the commercially harvested Pacific oyster (Crassostrea gigas), threatening the local shellfish industry. A permit for the use of IMI has been denied by the State, Pacific County has declared an economic emergency, and the outcome of an appeal by the growers remains uncertain. We have undertaken studies to determine if un-iodized table salt may be an alternative to IMI, specifically targeting juvenile shrimp (recruits) inhabiting the upper 10-15 cm of the sediment. Studies in 2018 indicated that a 2-3-fold increase in salinity resulted in 100% mortality when juveniles were exposed in artificial seawater. In 2019, we exposed juveniles (3 replicates, 5 shrimp each) within 10 cm of native sediment to five different salt solutions to achieve sediment pore water salinities of 25 (ambient, control), 35, 50, 70, and 100 ppt. Salt solutions were prepared with native seawater, added on top of the sediment (depth = 2 cm), and allowed to percolate through the sediment column for 6 h (low tide). At 6 h, 2 cm of ambient seawater (25 ppt) were added to simulate tidal inundation and allowed to remain on the surface for 12 h (low-high + high low tide) with two subsequent drawdowns and tidal inundations at 25 ppt (total test duration = 48 h). The sediment was then sieved to remove the shrimp and determine mortality. Average survival of controls (25 ppt) was 73.3% whereas none of the shrimp exposed to elevated salinities survived. Additional tests are underway to examine different exposure scenarios. Results to date suggest table salt may be a viable and greener alternative to IMI.

Spinal cord injury is known to cause an inflammatory immune response in the central nervous system (CNS), activating local immune cells microglia and astrocytes as well as recruiting circulating macrophages. Previous studies have shown that distinct populations of macrophages can exhibit either neurotoxic or reparative effects, depending on the cytokines activating them. Injury to the CNS also results in enzymatic breakdown of extracellular matrix (ECM) molecule hyaluronic acid (HA) into lower-molecular weight glycoproteins. It is known that after injury, many of these cleaved glycoproteins from the spinal cord extracellular matrix act as damage-associated molecular patterns to modulate immune response. However, it is still unclear which facets of the inflammatory response are subject to control by HA, a key ECM component. In this study, we seek to determine whether the breakdown of HA causes proliferation and activation of microglia and astrocytes. Introducing the enzyme hyaluronidase that degrades HA to an uninjured spinal cord decreases the levels of HA in the local area. Subsequently, we used double immunohistochemistry with markers for either microglia or astrocytes in combination with a marker for cell proliferation, to analyze cell activation and proliferation after treatment with either active or heat-inactivated hyaluronidase. We expect hyase treatment to result in both increased activation and proliferation of microglia and astrocytes. The expected data would indicate that the breakdown of ECM HA leads to activation of microglia and astrocytes. Understanding these cells’ activation could lead to future treatments for spinal cord injury focused on modifying the ECM to induce beneficial rather than neurotoxic inflammation.

Growing concern regarding brain injury in football has encouraged research on head impact exposure (HIE), incidence of concussion, and strategies to reduce injury risk. Reducing HIE in games has presented challenges due to the physical nature of the sport. However, limiting the amount of full-contact in practice may present a means for both decreasing HIE and minimizing concussion risk. In the past few years, guidelines limiting contact have been proposed, including decreasing the intensity of contact (limiting "two-a-days"), decreasing the number of full-contact practices, and eliminating practice drills associated with greater contact (such as the "Oklahama drill"). A few states have even modified their existing concussion legislation to include provisions limiting contact. The goal of our study is to review the literature regarding limitations on contact as a means for improving safety in football and to evaluate whether evidence supports this approach as a means for mitigating risk. We have identified studies assessing the impact of limitations on contact for either reducing HIE or concussive risk in football. We have then categorized the resulting studies by level of evidence, and summarized our findings. We concluded that reduced contact should result in a decrease in HIE and thereby a decrease in injury rate. 

The study of firn is integral to determining past climate from ice cores and calculating present and future melt runoff from ice sheets. The Herron and Langway model is a semi-emperical firn densification model. While the model's simplicity makes it easy to use, it assumes constant temperatures and accumulation rates, but Earth's climate is changing. The goal was to recalibrate the model by adding data from new ice cores to a gap in the model's dataset to create more accurate depth-density curves. Preliminary results show the recalibration is a better fit for 57 percent of depth-density profiles. The recalibration could help determine melt runoff, informing sea level rise forecasts. The recalibration could also increase the precision of timing between past CO2 and air temperature changes.

Thermoregulation, the maintenance of core body temperature in a constantly changing enviroment, is a critical aspect of homeostasis. Despite its importance, the neural mechanism by which thermoregulatory processes occur is not very well understood at the circuit level. Afferent skin temperature information travels through the spinal cord to the parabrachial nucleus (PBN), where it passes on to the preoptic area of the hypothalamus (POA). A subset of prodynorphin (Pdyn)-expressing neurons in the PBN (PdynPBN neurons) are activated when mice are exposed to warm environments, and 80% of these neurons project to the POA. The exact role of PdynPBN neurons has not been characterized, however, and their full projection profile is not established. Using genetic and viral techniques, we inserted a Cre-dependent designer receptor exclusively activated by designer drugs (DREADD) into mouse PdynPBN neurons and labeled their synaptic projections with GFP-bound synaptophysin, an abundant synaptic vesicle protein used for neurotransmitter trafficking. The use of Cre-dependent DREADD and synaptophysin-GFP allowed us to specifically label and activate PdynPBN neurons. We found that activation of these cells increases tail-skin temperature with a concurrent drop in core-body temperature. These data suggest that PdynPBN neurons may convey environmental temperature information that is sufficient to activate heat-defense responses. Establishing the genetic identity of neurons in a circuit that helps to maintain constant core body temperature will allow for the elucidation of downstream nodes in this circuit.