Throughout the United States, children and adults are forced to engage in sexual acts and use illicit drugs against their will, in what many would call modern day slavery, commercial sexual exploitation, or sex trafficking. Victims are controlled through coercion, force, fraud, physical and sexual violence and they endure many emotional and physical consequences. Approximately 80% of victims access healthcare during their time of exploitation for health reasons such as acute injuries, sexually transmitted infections, and mental illnesses, such as depression, anxiety, trauma, and suicidal ideation. The emergency department (ED) is the frontline to these patients. To date, little is known about nurses' confidence or competence in identifying or caring for this vulnerable population in Seattle, WA. The aim of this study is to examine what current practices exist at emergency departments in three, large referral hospitals in Seattle to identify victims, and to assess facility and nurse readiness for improved identification. Nine ED nurses in Seattle were interviewed to gain insight into how they currently identify trafficking victims, their opinion of the acceptability of existing screening questions (compiled from nationwide piloted studies) in their ED setting, and whether they felt such a standardized screening tool may be effective in identifying victims and improving subsequent care. Through transcription of audio interviews, nurses’ responses were analyzed and coded for common themes. Improved identification of trafficking victims by nurses and other frontline health workers in emergency settings may lead to more efficient and effective linking with vital support services and resources to assist them in safely exiting the sex trafficking industry. Results from this study will be shared with the participating hospitals, as well as public health officials and stakeholder organizations in order to improve awareness of sex trafficking, victims, and provision of necessary care and resources to support this vulnerable population.

WMlife Network Technology Company Ltd builds shared bookcases located in kindergartens to rent picture books at a low annual fee. WMlife builds bookcases of different sizes based on the number of students in the kindergartens. For all existing kindergartens in the system, WMlife has been choosing the same set of books for every kindergarten. However, now they are considering supplying different book sets to different kindergartens and exchanging books between them each year to provide new books while reducing total cost. To minimize the time investment by WMlife, we modeled the problem as a periodic traveling salesman problem and used a modified Prim’s algorithm and brute-force to find the route with the shortest traveling time. We also constructed a feasible book exchanging algorithm. With our new strategy, WMlife could get a dynamic instruction for book exchanging instantly with kindergarten locations and starting locations as input. WMlife can exchange books easily according to our instructions and significantly reduce cost by 30% in book reallocation approximately. As WMlife expand their services in more cities in China or adding more new kindergartens to the existing city, they could have a smart start of purchasing the book for future cost saving. The system not only solves a one-time problem but also can support WMlife to provide better books and more book types for kindergarten children, help them to form reading habits, widen their horizon and raise Chinese average reading rate.

Traumatic fracture of the bone surrounding the eye (the orbit) can result in functional impairment of vision. Double vision, diplopia, is one such sequelae. The cause of diplopia in orbital fracture patients has most commonly been attributed to loss of orbital volume caused by an open fracture that allows the contents of the orbit to shift. However, in cases where the orbit is surgically repaired and orbital volume is restored, some continue to have diplopia. Our hypothesis in these cases is the anatomy of extraocular muscle pulleys is affected by the trauma. The aim of this study is to test this research question by analyzing computed tomography (CT) scans. Patients were selected from a previous study of orbital fracture patients in which all patients had CT scans available for review and consented to be part of a research study on orbital fractures. Locations of the extraocular muscle pulleys were measured relative to the center of the orbit in the lateral, anterior and superior directions. Pulley location was determined by a tangent plane drawn where the optic nerve meets the orbit and crosses the muscle. The measurements were input into Orbit, simulation software, to produce a prediction of visual changes in Lancaster plots. The Lancaster plots were compared to the data from the patient’s clinical visual testing to determine the accuracy of the prediction at each point in the visual field. Our results showed that the accuracy of our prediction varied, as the location of the pulleys is not the only factor that contributes to diplopia. Other factors that may contribute include weakened muscle, entrapment and damage that is not visible on CT. More research is needed to determine how the muscle pulley system is affected in orbital fracture, and to determine how clinical interventions including surgery may help improve diplopia.

Autism Spectrum Disorder (ASD) is a developmental disorder in which individuals experience challenges with their behavior, social skills, and communication. There is a significant disparity between the numbers of males and females diagnosed with ASD; males are 4x more likely to receive a diagnosis, and it has been suggested that females with ASD are less likely to receive a diagnosis because they may “mask” their symptoms. Females with equally-high levels of autistic traits are less likely than males to be diagnosed with ASD, and females with ASD demonstrate enhanced social skills and higher levels of social motivation compared to males. Additionally, research by Parish-Morris shows that school-aged ASD females had stronger pragmatic language ability than ASD males. Pragmatic language is an individual’s ability to communicate verbally and nonverbally, understand and reciprocate social cues, and express appropriate social behavior. If females with ASD have stronger pragmatic language skills, it may serve as a linguistic camouflage and contribute to their ability to conform. The goal of this project is to examine sex differences in pragmatic language in children with ASD and explore what factors are related social communication differences between males and females with ASD. 15 male and 15 female participants with a confirmed diagnosis of ASD between the ages of 8 and 17 participated in this study. All participants completed a recorded conversational interview with a clinician. Videotapes were then rated by coders using the Pragmatic Rating Scale-Modified (PRS-M). First, we evaluated sex differences in PRS-M scores. We predicted that females with ASD will be rated as having better pragmatic language than males with ASD. Second, we investigated whether IQ, Age, and Autism Severity predicts PRS scores, and if Sex moderates these relationships. The results of this study shed light on sex differences in social communication in autistic children.

Orbital fractures are one of the most common facial fractures and often result in functional deficits if not treated appropriately. No prior reports have assessed dynamic visual acuity (DVA) in these patients, which is critical in visually tracking objects during head movement. Our research seeks to determine the effect of orbital fractures on DVA and assess whether DVA will change over time in those who undergo surgical reconstruction of the orbit. We hypothesize that DVA is adversely affected in orbital fracture patients and improves after surgical reconstruction. We designed a prospective pilot study at a Level I trauma center. Adult patients who presented with an orbital fracture between November 2017-January 2019 without prior ocular history were eligible. All enrolled subjects underwent static visual acuity (SVA) and DVA testing twice: once within one month post-injury before surgery, and another at least ten days following surgery. SVA was assessed using a mobile Snellen eye chart. DVA was measured by having the subject rotate their head at a standard frequency while again reading a Snellen chart. DVA is defined as abnormal if acuity falls more than two lines on the Snellen chart compared to SVA. Eleven patients have been enrolled and tested. Four(36%) patients exhibited abnormal DVA prior to surgery, compared to two(18%) after surgery. For all patients, the mean change in visual acuity (DVA logMAR minus SVA logMAR per subject) before surgery was 0.17 (SD=0.09). Following surgery, the mean difference in visual acuity across all patients was 0.07 logMAR (SD=0.07). Our data shows that, surprisingly, the majority of orbital fractures patients tested did not have abnormal DVA. In patients that did demonstrate abnormal DVA, half improved after surgery. The mean difference in visual acuity of all patients also improved after surgery. A larger study is needed to determine the prevalence of abnormal DVA in this patient population.

Modern cells discriminate among the amino acids chosen to be included in protein synthesis: some, like leucine, serine, glycine, and alanine appear in most proteins across most cells, whereas other amino acids, like (g)-aminobutyric acid and aminoisobutyric acid do not appear. This raises the question of how selectivity among amino acids arose – does the selectivity rely on modern protein machinery or could it have arisen in the first protocells of the Early Earth? For example, could certain amino acids have, by some mechanism, associated more strongly with protocell membranes, increasing their chances of being integrated into the first peptide chains? To test this hypothesis, our group assembles rudimentary protocells from molecules that would have been present on the early-Earth: decanoic acid (a fatty acid), sodium mono-phosphate, salt, and water. Vesicles of the decanoic acid spontaneously form. We then add different amino acids to the solutions and measure their turbidity to determine whether each amino acid causes the number of lamellae in the vesicles to increase or decrease. Increased lamellarity correlates with a sturdier vesicle. If certain amino acids increase lamellarity of protocells, that could serve as a method of selection for certain amino acids rather than others. Our results are that particular amino acids (most notably serine, glycine, and alanine) do in fact increase the lamellarity of fatty acid vesicles significantly, whereas other, less common, amino acids do not. We are currently exploring the plausibility of a mechanism for this occurrence involving ease of rotation around the alpha carbon of the amino acids, and we are investigating other ways in which interactions between amino acids and fatty acid membranes might be manifested, for example by a shift in the solution’s critical vesicle concentration. Our results will fit into the overarching goal of understanding peptide formation and protocell stability in order to gain insight into the origins of life on Earth.

Alzheimer’s disease (AD) is a neurodegenerative illness affecting millions of individuals in the United States and represents the leading cause of dementia worldwide. AD is pathologically defined by abnormal accumulation of interneuronal plaques composed of amyloid beta protein and intraneuronal tangles composed of tau protein. The focus of our current research involves understanding how tau protein homeostasis is restored using the model organism C. elegans. One process by which protein homeostasis is modulated is through the endoplasmic reticulum unfolded protein response (UPRER), which is composed of three signaling branches. Initially, the UPRER acts to restore normal protein folding, but if restoration is impossible, it triggers apoptosis. Our lab became interested in this signaling pathway after we identified the UPRER master transcription factor XBP-1s as a modifier of tau pathology (tauopathy). Specifically, XBP-1s overexpression protects against tauopathy in our C. elegans model. Previous RNA sequencing (RNAseq) analysis identified several downstream target genes upregulated with XBP-1s overexpression in our pathological tau C. elegans model. To follow up, I am working on understanding the molecular basis of XBP-1s-mediated tauopathy suppression by generating C. elegans models containing target genes of interest. Assessment of the effect on tauopathy phenotypes includes behavioral analysis by performing a liquid thrashing assay and tau protein analysis via the Western blot technique. Using the microinjection technique, I have generated a transgenic C. elegans model overexpressing hsp-4, an upregulated XBP-1s target gene identified by RNAseq that potentially modulates tauopathy. We are currently determining whether overexpression of hsp-4 can ameliorate tauopathy similar to XBP-1s overexpression. Gaining further information about the role of these XBP-1s target genes in tauopathy using the model organism C. elegans is valuable for future translational studies in mammals, along with being integral to research for neuroprotective therapies for neurodegenerative diseases such as AD.

Computer vision models are used to help analyze biomedical images for diagnosis and treatment through looking for differences between images by a comparison to a template image. For instance, optical coherence tomography (OCT) is used to diagnose and treat retinal issues. When looking at the brain, injury, cellular uptake and characteristic features vary across regions, therefore images are often segmented into established brain regions to determine how the brain is impacted in a particular study. Current models fail to work in segmenting brain regions because each brain has variation in local microstructure, making it difficult to compare one brain to another. Furthermore, when brains are sliced, the exact location within the brain can be difficult to pinpoint, particularly in regard to depth, because the regions vary slice to slice. Therefore, my research addresses the increasing need for a method of analysis to align and compare images from brain regions across slices from a single brain, and from brain to brain. Using scikit-image analysis tools, I extracted information from cell images and videos of nanoparticles obtained in brain slices and determined trends within various regions. My program extracted cell density, shape, and death, then analyzed the uptake of nanoparticles to determine where a small segment of an image is most likely located within the brain. Iterating over the entire image generated a rough map of the regions within the brain which is refined using mapping descriptions detailed in literature. This research resulted in a systematic program that uses image analysis tools to extract features of defined brain regions. This program allows for quick, accurate and consistent analysis of regional differences of cellular features, nanoparticle distribution, toxicity, and other important measures.

Archaeologists increasingly use three-dimensional modeling to analyze artifact attributes and document in situ relationships. Less attention has been paid to modeling’s potential in strengthening connections between descendant communities and their cultural heritage. In collaboration with the Grand Ronde Historic Preservation Office, we created three-dimensional models of excavation units and belongings associated with the Molalla Encampment, a late nineteenth and early twentieth century settlement area on the Grand Ronde Reservation in northwestern Oregon. This work proceeded in two steps. First, we established a modeling methodology applicable to a variety of excavation contexts and artifact types. This required defining proper artifact handling techniques and modeling procedures, including optimal camera settings and software workflow. Second, we used this methodology to capture hundreds of artifacts photos in order to build each model. Photogrammetry provides an alternative approach to heritage curation, allowing us to share interactive, three-dimensional models of historic belongings within the Grand Ronde community. This not only lends additional transparency to our research process, it can also initiate conversations with tribal members about the functions and meanings of belongings in historic reservation lifeways. Photogrammetry can thus play an important role in the development of community-based research practices.

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.

Analysis of faunal remains provides crucial insight into the subsistence patterns of past groups. This is particularly important for the Grand Ronde community in northwestern Oregon, where past and present populations have balanced traditional practices and lifeways with imposed colonial assimilation pressures and economies. This research examines faunal remains found at the Molalla Encampment and Grand Ronde Agency Schoolhouse, two nineteenth and early twentieth century archaeological sites on the Grand Ronde Reservation. Our research focuses on documenting and analyzing faunal remain attributes such as degree of fragmentation, taxonomic presence, and bone modification at the two sites. In addition to comparative faunal analysis, our research takes a step further by looking into the implications of observed bone modifications. Aided by archival and primary sources, we reconstructed the taphonomic pathways of these remains, determining whether they have been altered by either humans or animals. For example, our analysis of cut marks revealed if meat was butchered commercially at the site, what meat cuts were consumed, and meat prices at Grande Ronde during the early 1900s. Comparing these data by site and stratigraphic level paints a better picture of life at Grand Ronde across space and time. This research provides insight into subsistence patterns within the Grand Ronde community and how they navigated the challenges and opportunities of life on the reservation.

Black bears are the most abundant bear in North America and Washington, but present knowledge is dominated by telemetry studies on home range size or vegetation class preference. Camera trapping data indicating black bear presence were used to build an occupancy model for the two study sites in Washington, one in Okanogan county and one in the northeast. The camera traps are intended for a predator-prey study focused on wolves, cougars, and ungulates, but detected a large number of black bears, suggesting the importance of black bears in these ecosystems. Black bear distribution may be driven by land-use patterns including variables such as elevation, vegetation and habitat type, approximate percent canopy cover, slope, aspect, and land management type. The constructed single season occupancy models of black bear presence can be used to better understand habitat selection by black bears in different regions of the state. This study seeks to fill in gaps of knowledge about black bears in Washington and provides a framework for future occupancy studies on black bear habitat use.

Analysis of plant foods and medicines is a growing field within archaeological research. However, few scholars have examined plant foods among Native American communities in colonial settings. This project combined archival and archaeological data to better understand plant use within the Grand Ronde community during the late nineteenth and early twentieth centuries. Field Methods in Indigenous Archaeology (FMIA), a community-based research partnership between UW and the Confederated Tribes of Grand Ronde Historic Preservation Office, has collected macrobotanical remains from the Grand Ronde Reservation in northwestern Oregon. I explored how and why these remains were used within the community. Knowing this information will provide a better sense of local environmental contexts and cultural lifeways. To study this question, I selected ten plants that have been recovered during FMIA’s excavations and examined ethnographic, archaeological, and documentary sources from the region to understand each plant’s cultural context. I found that most plants found on the reservation were used as food sources, though some also had medicinal and spiritual significance. This work sheds new light on the relationships between Native communities and plants in the Pacific Northwest over the past two centuries, helping us understand how people navigated the challenges and opportunities introduced by colonialism.

Through rapid industrialization and development, much of the Green River Valley landscape and townships from the earlier periods of settlement have been obscured. However, several historical cemeteries remain largely preserved. Cemeteries can have an equalizing power when it comes to representation of past populations, while preserving societal ideas of hierarchy. While cemeteries cannot tell the full history and lives of those interred, they do provide names, ages, and the placement of the individual in conversation with other deceased people. A cemetery is not a passive location in a town, but rather a place created with the ability to reflect societal values regarding death and social integration and spiritual significance for the living. My research focuses on what relationships can be rediscovered in the development of the Green River Valley’s townships, as these relationships may speak to the social organization of the Valley townships in development, the role death had in society, and how race, gender, religion or other factors were presented. These relationships will be investigated following three lines of inquiry: a) relationship among the dead in cemeteries, b) relationship between cemeteries and the living in modern and historical times, and c) relationship of the deceased’s identity and their position within the cemetery landscape.  Though each cemetery has been researched and written about separately, conducting a survey of several cemeteries in the region and analyzing them together will allow for a bigger picture of health discrepancies between men and women, white and non-white residents, and differences of land quality broadly associated with members of different social classes, building a more comprehensive history of the Green River Valley.

Self-efficacy has been shown to be associated with fewer problem behaviors in adolescence. However, the research investigating this association has been primarily based on self-report measurements of delinquency. The purpose of this study is to see if this association is maintained with a more objective measure of delinquency by examining juvenile offenders’ self-efficacy, and its predictive relationship with probation violations. Using self-report survey data of juvenile offenders’ self-efficacy along with probation violation data from Pierce County Juvenile Court, we conducted a statistical analysis of self-efficacy measurements and official probation violations. The self-reported self-efficacy measures address feelings in desire of academic and career success, prosocial behavior, and a reduction in the desire to express aggressive behaviors. If self-efficacy is shown to be a good predictor of fewer probation violations, further research can investigate determinants of high versus low self-efficacy, or how to improve self-efficacy, in order to promote probation adherence of juvenile offenders.

The influenza virus is known for its rapid evolution, or the ability to fix many mutations over a short period of time. Some of these mutations lead to amino acid substitutions in regions of the virus targeted by the immune system. Such changes are often selected for because they confer a fitness advantage by allowing the virus to "escape" immune response. This pattern of repeated immune escape is a detriment to public health because it necessitates an annual update to the influenza vaccine. Therefore, identifying sites on the influenza virus which are targeted by the immune system could help predict which influenza strain will circulate in the future, inform vaccine design, and help understand basic evolutionary questions. Using molecular phylogenetic techniques, we can identify sites potentially targeted by the immune system by looking for "positive selection", a phenomenon which manifests as a higher than expected rate of evolution. To identify sites evolving faster than expected, we defined a null expectation of the evolutionary rate of influenza in the absence of immune pressure. This null model is defined using empirical measurements from a high-throughput functional assay known as deep mutational scanning. This null model differs from traditional phylogenetic models in that it describes the constraints on influenza on a site-specific basis and, as a result, has been shown to be a more accurate and powerful null model. I have implemented an empirical Bayes approach to identify sites which deviate from the null model by an unexpectedly high evolutionary rate, suggesting positive selection. Preliminary results show that my method outperforms other methods for identifying sites under positive selection. Next, I will apply these methods to the influenza virus surface protein, hemagglutinin, which is a major target of the immune system.

The #MeToo movement has sparked a national conversation about what constitutes sexual consent, and a series of debates have erupted over the actions of famous men like Brett Kavanaugh, Louis C.K., and Aziz Ansari. Popular hits like Robin Thicke’s song “Blurred Lines” have also come under fire for promoting sexual violence by celebrating confusion around sexual consent. At this moment, when norms and expectations around sexual interactions seem to be changing quickly, it is especially important that we have comprehensive and systematic data on how people define sexual consent and interpret ambiguous sexual experiences. Existing studies have shown characteristics of an individual matter when it comes how they conceptualize appropriateness. However, we still do not know to what extent these characteristics matter. This study addresses this gap by answering the question: How do people of varying groups define appropriate sexual behaviors and boundaries? A Q-Method study of over 200 people will reveal how individuals of different characteristics sort scenarios of various sexual interactions into categories ranging from “most appropriate” to “most inappropriate.” Q method is similar to a rank-order techinque and allows for representation or reconstruction of perspectives concerning any given topic of interest. In addition to providing insight about how different people make sense of sexual behaviors, this study shows how Q-sorts can provide a more detailed look at sexual norms than the survey and vignette studies currently used in the field.

Autism Spectrum Disorder (ASD) is a collection of neurodevelopmental disorders characterized by social-communicative and behavioral impairments that affects approximately 1 in 59 children in the US. Children with ASD, on average, exhibit increased rates of externalizing behaviors compared to their typically developing peers. Studies have shown that children with relatively low familial risk for ASD are more likely to develop ASD if they have a history of pregnancy complications and that presence of pregnancy or birth complication can exacerbate the severity of externalizing behaviors. In typical development, pregnancy complications are associated with a number of child psychiatric disorders and externalizing behaviors and males have increased levels of externalizing behaviors compared to females. In a sample of children with ASD, this study (1) investigated the relationship between pregnancy complications and externalizing behaviors and (2) compared sex differences in externalizing behaviors. The study involved 200 children with and without pregnancy complications, between the ages of 8 and 17 years. Pregnancy complications were provided by the parent during a medical history interview and refer to a wide range of complications including, among other things, health concerns for the mother or child, difficult birth, or coexisting pregnancy diagnosis such as gestational diabetes or preeclampsia. To assess for externalizing problems, parents of participants completed the Behavior Rating Inventory of Executive Functioning (BRIEF) questionnaire about their child. We predict that ASD children (1) with pregnancy complications (compared to those without complications) will demonstrate more externalizing behavior problems and (2) males with ASD will exhibit more externalizing behavior problems than females in the subscale mean scores. The results of this study provide insight to the connection between pregnancy complication and externalizing behaviors in children with ASD.

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. 

Microbial communities are essential to numerous biological systems from aquatic and land ecosystems to the human gut. Our understanding of interactions among bacterial strains can help us understand how these communities form and function. Such interactions may depend on the species themselves but can also be sensitive to the abiotic environment. Harsh environments, where mortality is increased, can reverse the outcome of a competition between species. Simple models predict that increasing mortality favors faster growers, and past experiments repeatedly confirmed this prediction. In these pairwise competition experiments, slow growers excluded fast growers in low-mortality environments, but increasing mortality caused fast growers to dominate. A necessary condition for observing the reversal is that slow growers are good competitors in low-mortality, high-density environments. While this tradeoff between growth rate and competitive ability was common in well-characterized lab strains of bacteria, it was unclear if it might exist elsewhere. In this project, we tested for the existence of a growth/competition tradeoff in a natural community of bacteria collected from MIT’s Killian Court. Contrary to past results and some evidence from the literature, we found a weak positive correlation between competitive ability and growth rate, which does not support the idea of a tradeoff.

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.