Research suggests that parental alcohol use increases alcohol use in their children. Longitudinal studies also consistently suggest that alcohol expectancies for young people are influenced by ads in the mass media, and exposure to commercials is associated with increased likelihood to initiate drinking. The present study investigates the relation between family history of alcoholism, exposure to alcohol related media and alcohol use among high school students. High school aged participants throughout the state of Washington (n = 2,497) completed a baseline online survey as part of a larger trial that assessed the alcohol related negative consequences via the Young Adult Alcohol Problems Screening Test score in the past three months, the incidence of alcoholism in the family via the family history risk score and the ratings of average alcohol ad exposure. Regression analyses revealed a significant positive main effect of ad exposure and family history of alcoholism onto alcohol related negative consequences (p < .05). A significant moderating effect of ad exposure onto the relationship between family history and alcohol use (β = -.10, F (3, 2497) = 15.88, p < .05) was found, such that those with higher family history of alcoholism and higher ad exposure have higher alcohol related negative consequences. These results suggest that individuals who are exposed to alcohol at both a familial and cultural level have the most negative alcohol related consequences. Future research should continue to investigate the relationship between familial and cultural exposure to alcohol.

Organisms rely on numerous proteins that carry out all of life’s essential functions. Although each protein has the potential to have many functions, we researchers tend to over-simplify and associate a protein as having one primary function without having performed the due-diligence to prove this. I am interested in determining the multiple functionalities of the previously uncharacterized protein, Yil166c. A suppressor screen recently performed in the Dunham Lab has implicated missense alleles of YIL166C to encode low affinity sulfate transporters. However, many lines of evidence – including sequence comparison to the canonical S. cerevisiae sulfate transporters, Sul1 and Sul2, and phylogenetic classification – suggest that native wt Yil166c may have other functions within the cell. Currently, I am in the process of testing whether wt Yil166c is a sulfate transporter by performing growth curves in media containing a range of concentrations of ammonium sulfate. If Yil166c is a sulfate transporter, I would expect to find concentrations at which a strain missing only the canonical sulfate transporters (sul1∆ sul2∆) will grow and a strain missing all three (sul1∆ sul2∆ yil166c∆) will not. In the future, I aim to characterize the growth of wt and yil166cΔ strains in various standard growth media including media containing suspected substrates and a range of basic nitrogen, phosphorus, sulfate, and carbon sources to discover if the protein imports any of these substrates. I also plan to study tradeoffs in transporter specificity caused by the mutations generated in the suppressor screen by isolating the specific mutant clones and characterizing their growth in media containing a range of ammonium sulfate and other nutrient concentrations. In doing this, I hope to get a glimpse of how a transporter of a given substrate can change preference – an extremely important aspect of evolution for which few examples are characterized.

Despite recent successes in reducing the total number of new HIV infections and deaths due to AIDS, a vast disparity persists in the diagnostic and treatment capabilities between high- and low-resource settings. This can be attributed partly to the fact that many common diagnostic procedures are too expensive and technically complex to be practical in low-resource settings. The Lutz Lab is working to address this challenge through the development of paper-based diagnostics for HIV that are inexpensive, rapid, sensitive, specific, and easily operated. My research focuses on the twin challenges of diagnosing acute HIV infection and diagnosing drug-resistant HIV. One promising method for diagnosing acute infection is the detection of the p24 antigen, a viral protein that can be detected two weeks after HIV infection. I have demonstrated sensitivity levels in a paper-based p24 antigen assay near that of the equivalent lab-based assay. Furthermore, I am working in collaboration with the Lai Lab to investigate the use of stimuli-responsive polymers to improve the sensitivity of this assay. I have gathered initial data that suggests that this coupling can enable a significant improvement in the performance of the assay. In order to diagnose drug-resistant HIV, we must switch from targeting proteins to targeting the nucleic acid of the virus. This will be accomplished by using an oligonucleotide ligation assay (OLA) to probe for point mutations indicative of drug resistance. Prior work in the lab has indicated that we are able to successfully detect the OLA product in a paper-format. However, current hapten-antibody capture methods limit testing to two mutations per strip. I am developing a DNA hybridization approach that, if successful, will enable us to scale up this level of multiplexing. The completion of this project will bring us one step closer to increasing access to these tests in low-resource settings.

Duchenne Muscular Dystrophy (DMD) is a debilitating disease caused by a genetic mutation in the dystrophin gene, located on the human X chromosome, that leads to muscle degradation and ultimately death. Induced Pluripotent Stem Cells (iPSCs) can be derived by reprogramming patient-specific somatic cells. Therefore patient-specific iPSCs provide unprecedented in vitro models to study pathological mechanisms with disease-specific genetic backgrounds. Recently it has been shown that urine stem cells (USCs) can be easily and efficiently reprogrammed into iPSCs. As USCs do not require invasive procedures to collect parental cells for reprogramming, USC-derived iPSCs are an ideal in vitro model for study of human disease.The overall goal of this project is to characterize the epigenetic landscape of USCs and USC-derived iPSCs to understand how the stem cell state of USCs contributes to efficient reprogramming, and to eventually investigate the role of epigenetic mechanisms in the pathology of DMD using iPSC-derived cardiomyocytes. We now know that epigenetic modifications regulate gene expression and, in turn, type and function of the cell. The goal of my project is to analyze the epigenetic profiles of USC-derived iPSCs obtained from two healthy subjects and one DMD patient. I will analyze chromatin immunoprecipitation with massively parallel DNA sequencing data generated for both activating (H3K4me1, H3K4me3, H3K27ac) and repressive (H3K27me3) histone modifications. Histone modifications are class of epigenetic modification in which molecules added to a DNA bound protein regulate transcription. Using tools from the HOMER bioinformatics suite to annotate locations of histone modifications and preforming comparative analyses in R, I will determine epigenetic mechanisms that define the urine stem cell state, efficacy of reprogramming the epigenome, and if significant condition-specific variation occurs in iPSCs derived from DMD patients.

Single base pair changes, SNPs (Single Nucleotide Polymorphism), in bitter taste receptor T2R38 influence the sensitivity to 6-n-propylthiouracil (PROP) and have been associated with caries risk/protection. However, no study has been reported on the role of T2R38 in the oral innate immune responses to bacteria. The goal of this study is to investigate if T2R38 regulates oral innate immunity and if this regulation is genotype-specific. Primary gingival epithelial cells carrying three common genotypes, PAV/PAV (PROP super-taster), AVI/PAV (intermediate) and AVI/AVI (non-taster) were stimulated with four bacteria: tooth decay causing Streptococcus mutans, periodontal pathogen Porphyromonas gingivalis or non-pathogens Fusobacterium nucleatum and Streptococcus gordonii. QRT-PCR analyzed T2R38 mRNA, and T2R38-specific siRNA and ELISA were utilized to evaluate induction of IL-1α, IL-8, and hBD-2, associated with regulating the immune system, in various donor-lines. Experiments were set up in duplicate and repeated three times. T2R38 mRNA induction in response to S. mutans was highest in PAV/PAV (4.3-fold above the unstimulated controls; p<0.05), while lowest in AVI/AVI (1.2-fold). IL-1α secretion level was higher in PAV/PAV compared to AVI/PAV or AVI/AVI with S. mutans stimulation, but was reduced by half when T2R38 was silenced (p<0.05). In contrast, only AVI/AVI showed 4.4-fold increase (p<0.05) in T2R38 expression in response to P. gingivalis, whereas the levels in PAV/PAV and AVI/PAV remained close to that of the controls. When T2R38 is silenced, the level of hBD-2 secretion in response to S. mutans was significantly decreased in PAV/PAV. Our data suggest that T2R38 plays a role in the regulation of gingival innate immunity and that these responses are T2R38 genotype-dependent. We envision the knowledge gained from this study will provide a broader understanding of new ways to control caries and oral inflammatory responses. 

Horizontal ocean currents are well-documented, but vertical movement of seawater is an important physical aspect of the ocean that is difficult to measure directly.  Accurate vertical water velocity (W_w) data provides better quantitative understanding of internal waves and ocean circulation—both of which have important roles on Earth’s climate system.  To shed more light on these phenomena, I researched the ranges of W_w that occur in the ocean around Drake Passage using data from five EM-APEX floats—robotic instruments that move vertically by adjusting their density relative to that of seawater.  I investigated the float parameters that optimize W_w measurements, the distribution of optimized vertical velocity values, and the internal wave frequencies of W_w.  I used pressure and time data from the floats to compute float vertical velocities (W_f), modeled still-water vertical float velocities (W_s) using an equation balancing drag and buoyancy, and calculated W_w by subtracting W_s from W_f.  I then tuned the W_s model by numerically optimizing the float compressibility and piston expansion coefficients.  I found that W_w values were normally distributed about 1.1 mm s^-1 downward on average with a mean spread of approximately 1.1 cm s^-1 across all floats, which agrees with past results.  The reason(s) behind the negative bias in W_w values require further investigation, and could be due to potential float ballasting procedure errors or to a signature of gradual downward motion in large-scale oceanic currents.  I also determined that the frequencies of W_w were very close to N, the maximum internal wave frequency.  The similar W_w frequencies to N confirm that the internal waves responsible for the vertical velocities oscillate primarily in the vertical direction, as expected.  Vertical ocean velocities can be further studied by applying a different buoyancy model and integrating ocean data from elsewhere in the world.

Chronically homeless individuals in the United States have higher rates of medical, psychiatric, and substance use disorders than the general population, which puts them at higher risk for suicide. Although a handful of studies have explored suicidality in the general homeless population, there are no studies to date that describe suicidality among chronically homeless individuals in the US. The current study aims to fill that gap in the literature by providing descriptive data on various aspects of suicidality (i.e., current and lifetime suicidal ideation, plans and access to means) in this population. Participants (N = 134) were chronically homeless individuals with severe alcohol problems who were frequent utilizers of publicly funded criminal justice and health-care services in King County, Washington. During in-person interviews, participants were administered the Screen for Suicide Risk, a modification of the Screen for Youth Suicide Risk. Findings indicated that nearly half of participants had experienced suicidal ideation in their lifetime, and 33.8% reported having attempted suicide. Alcohol was involved in 79.5% of these prior lifetime attempts. Considering current suicidality, 9.3% of participants reported having suicidal ideation in the past two weeks, with 6.6% having a plan and 20.0% having access to means to die by suicide. Findings indicate a high level of suicide risk in this multimorbid population and signal a need among providers to be aware of this risk and address it effectively in providing treatment and services.

Prednisone is a medication that reduces inflammation and inhibits activity of the immune system. Many conditions can warrant its use in pregnant and lactating women, so understanding the pharmacokinetics of prednisone, how the body handles the drug, during pregnancy and lactation is important for choosing the best dose. Therefore, the objective of this study was to evaluate the pharmacokinetics of prednisone and its active metabolite, prednisolone, in pregnant and lactating women. Nineteen women received oral doses of prednisone ranging from 2 to 40 mg per day in early- (n=3), mid- (n=9), and late-pregnancy (n=13), as well as postpartum with (n=2) and without (n=5) lactation. Blood and urine samples were collected over one dosing interval from all participants, and breast milk was collected from those who were lactating. Concentrations of prednisone and prednisolone were used to calculate pharmacokinetic parameters. The apparent oral clearance, the rate the body clears oral drug, of both prednisone and prednisolone significantly increased between the 5 and 20 mg doses. Higher concentrations of prednisone and prednisolone resulted in higher percent unbound in plasma. These data are consistent with dose- and concentration-dependent pharmacokinetics of prednisone and prednisolone due to concentration-dependent plasma protein binding. Unbound prednisolone’s terminal half-life, the time it takes to clear half of the drug, was prolonged in pregnancy compared to postpartum. Other parameters were unchanged, however, suggesting that dose adjustments may not be necessary during pregnancy. The ratio of breast milk to plasma drug exposure ranged from 0.49 to 0.57 for prednisone and 0.019 to 0.030 for prednisolone. Infant exposure to prednisone and prednisolone was 0.35 to 0.53% and 0.09 to 0.18% of the mother’s weight-adjusted dose, respectively. This suggests that oral prednisone administration is compatible with breastfeeding.

Over the last fifty years, crows have become more prevalent in urban and suburban areas. Crow roosts can consist of thousands of birds, which generate copious amounts of nitrogenous waste. It is critical to understand how this waste can impact urban greenspaces, and especially ecological restoration projects. In the Seattle area, there are at least two wetland restoration sites that have recently become inhabited by roosting American Crow (Corvus brachyrhynchos); North Creek Wetlands in Bothell and Springbrook Creek Wetlands in Renton. Previous research showed increased soil nitrate levels and lower pH on roost versus non-roost sites in the North Creek Wetlands. We were able to confirm that a 21-day biweekly application of crow waste to non-roost soil was enough to increase nitrate levels. In addition, we investigated whether crow-induced increases in nitrate levels could affect symbiotic relationships between plants and nitrogen-fixing bacteria. We tested this by adding crow waste to inoculated garden pea plants (Pisum sativum) and observed that these plants had more leaves, fewer nodules, and shorter taproots than controls after 21 days of biweekly waste applications. This is significant, as red alder (Alnus rubra), which form a symbiotic relationship with the nitrogen-fixing bacteria Frankia spp., appear to be becoming dominant on the North Creek Wetlands roost. Additionally, we tested soil from the Springbrook Creek Wetland restoration roost to confirm crow-induced increases to nitrate levels and decreases to pH. We hope these studies will help with future management efforts by providing a better understanding of how large crow roosts, which are increasingly common urban disturbances, are affecting the ecological viability of urban restorations.

Gene regulation, a crucial mechanism in living cells, has been applied in fields ranging from biofuels to targeted delivery of pharmaceuticals. RNA, in particular, acts as one of the main sources of gene regulation in cells.  sRNA (small RNA) is an important part of gene regulation in prokaryotes. Current research in synthetic biology has turned to the use of sRNA for engineered genetic regulation in place of, or in tandem with, conventional gene knockdown --reduction of gene expression by design -- strategies. sRNA regulated gene expression is an attractive possibility due to its ease of implementation compared to conventional methods. By incorporating the concepts of feedback system modeling, we are investigating sRNA's efficacy as a regulatory device in a two-plasmid system. Using kinetic rate-based computational modeling, we will determine the impact of intracellular regulation using sRNA to downregulate expression of GFP (green fluorescent protein) of a target gene. Sampling of the computational space will allow us to determine the most robust constructs to implement in vitro. The system is expected to show lowered expression of GFP in the test model when compared to the control model which will have no sRNA present. Characterization of novel pathways of control using sRNA will add to the synthetic biology suite of techniques. Synthetic Biology's aim is to use fine tuning techniques like this and others to engineer systems that provide some purpose. In characterizing this technique, we hope to aid in furthering the field of Synthetic Biology as a whole.

Traditional computing is based around manipulating 1s and 0s, or bits, through logic gates. However, quantum computers have been proposed which replace classical bits with quantum bits or qubits. As these qubits can be in a superposition state, that is both 1 and 0 at the same time, this opens up a wide range of computational possibilities. Our research focuses on trapping ions, which can be used as qubits and potentially scaled up. However, in order to reliably operate on trapped ions, they must be very cold. Traditionally, this cooling is done with lasers that exploit the Doppler effect to slow down ions. My work has focused on deploying a 370 nm laser, which can be used to cool ytterbium ions. For our usage, some novel technical solutions were required. To bring the peak wavelength down to 370 nm, the laser diode needed to be cooled to sub-zero temperatures and placed in a dry nitrogen environment to avoid condensation. Work will continue as we begin to cool ytterbium ions, and ultimately create cold chains of barium and ytterbium.

The Kuril Islands form an archipelago that stretches between the northern tip of Hokkaido (Japan) and the south of the Kamchatka Peninsula (Russia). Although relatively unknown outside of Russia and Japan, these sub arctic islands provide an ideal setting for studying hunter-gatherer population structure and resilience in harsh environments. Archaeology has been conducted in the islands for about 100 years, with a particular resurgence in the last 15 years with the International Kuril Island Project (IKIP) and Kuril Biocomplexity Project (KBP), the goal of which is to study human environment interaction, hunter-gatherer maritime adaptation and biogeographic and ecological sensitivity in the marine subarctic. This project examines the formal stone tools of the Kurils collected by the KBP during their 2006, 2007, and 2008 field seasons through photographs of the full collection. The goal of this research is to explore the degree to which adaptations in hunter-gatherer subsistence strategies affect and maintain human resilience in harsh or extreme environments. Particular attention is paid to evaluating the relationship between geographic insularity– or the relative isolation of a population– and technology. Tool kit composition and variation through the islands should reflect the terms of this relationship. Quantity and quality of tools between the remote (central) and proximal (nearer mainland) islands is expected to be limited by access to and availability of raw material.

Energy transfer between the atmosphere and snow surface is one of the primary drivers of snowmelt. With more than 50% of the global population relying on snow for a significant portion of its water resources, understanding how the local climate impacts snow in this way is essential. Accurately measuring atmospheric turbulence near the snow surface can be difficult, and quantifying the amount of energy exchanged is even more complicated. Current methods include expensive and specialized instruments called sonic anemometers used to measure 3D wind speed, but these instruments do not function well during precipitation or freezing events, which account for much of the heat exchange and are common in the Pacific Northwest mountains and other alpine regions. This project focuses on deploying a vertical array of temperature and wind sensors on a meteorological tower at Snoqualmie Pass, Washington to create a vertical data profile for use in an energy transfer comparison with the existing sonic anemometer. This type of array utilizes simple, low-energy instruments and requires very little maintenance once established, making it a promising substitute for the multi-thousand dollar and maintenance-intensive sonic anemometers. I hope to determine whether this relatively inexpensive alternative can provide continuous quality data for turbulent flux (heat exchange) during inclement weather events in the mountain environment. The outcome of this project will have implications for decisions to expand comprehensive snow station networks around the world and will contribute to the global effort to measure the impact of climate change on water resources. Although low snowpack this season has been an issue for some mountain hydrology research, it is still possible to compare turbulence profiles above the ground without snow cover; establishing, troubleshooting, and calibrating the system through this year will lead to more accurate measurements and better calculations of energy transfer in future winters.

Spinal cord injuries (SCI) are a devastating condition, and incomplete injuries of the cervical spinal cord are the most common among patients. In incomplete injuries, recovery of function can be partially regained through sprouting and/or rewiring of spared neural tissue. Our group has created a more permissive environment for these effects through the use of Chondroitinase ABC (ChABC), which enzymatically removes inhibitory Chondroitin Sulfate Proteoglycans (CSPGs) from the lesion site, as well as perineuronal nets. We paired ChABC injections with therapeutic open-loop intraspinal microstimulation (ISMS) in order to further promote plasticity. While previous studies have shown that each of these strategies can help promote recovery of function alone, our preliminary data suggests that much greater effects are seen when they are combined. A major symptom of SCIs is muscle weakness and fatigue. This is caused, in part, by wide-scale transitions of muscle fiber types from slow (Myosin type I) to fast-twitch (Type IIa, IIb, IId/x) following loss of motor neuron input. After treatment with ChABC and therapeutic ISMS, we completed an immunohistochemical analysis to characterize their effects on slowing this transition of fiber type in the triceps muscles of rats with incomplete unilateral spinal contusion injuries (n=7). After isolating specific myosin isoforms, we quantified the number of each muscle fiber type in each triceps, as well as characteristics of the fibers themselves, such as cross sectional area. We hope that these results will lead to a new understanding of injury-related fiber type changes and strategies to combat them after central nervous system damage.

Bruton’s Tyrosine Kinase (BTK) is a non-receptor tyrosine kinase that is crucial for B-cell maturation and proliferation. BTK is linked to the development and maintenance of B-cell malignancies. A small molecule inhibitor of BTK, ibrutinib, shows impressive response rate in B-cell malignancies, chronic lymphocytic leukemia and mantle cell lymphoma. Early results from clinical trials identified only the C481S resistance mutation; however, other mutations in BTK could also confer resistance to ibrutinib. In a leukemia-causing oncogene BCR-Abl, there are 15 known mutations that can cause inhibitor resistance. Understanding the full spectrum of BTK resistance mutations could have profound clinical applications, including personalizing treatment by combining several different inhibitors to make a drug cocktail that reduces the chance of resistance. I will use deep mutational scanning to quantify the level of resistance of all possible single mutations in BTK. In a deep mutational scan, a library of BTK variants is created, functional variants are selected with a functional assay, and the frequency of each variant both before and after selection is quantified using high-throughput DNA sequencing. I propose to select for ibrutinib-resistant BTK variants using a modified version of the Yeast 2 Hybrid system (Y2H), a system used to test protein-protein interaction. Here, I will link the expression of a reporter gene to the phosphate-dependent interaction between a BTK substrate, PLCγ2 and an SH2 domain. Under these conditions, yeast will only grow when PLCγ2 has been phosphorylated by BTK, enabling the interaction with the SH2 domain. The effect of each variant on resistance will be proportional to the growth rate of yeast expressing the variant in the presence of inhibitor. Variants that increase in frequency during the selection have a higher level of inhibitor resistance. Profiling of BTK inhibitor resistance is critical for future research and therapies targeting BTK.

Burnout is a negative emotional state characterized by exhaustion and cynicism due to prolonged exposure in stressful work environments. Burnout is frequently associated with professional staff working in human services in response to intense emotional engagement with clients who experience stressful life events. Maslach and Jackson advanced the research with empirical examination and incorporated a tridimensional model including emotional exhaustion, depersonalization, and reduced personal accomplishment which led to the development of the Maslach Burnout Inventory (MBI). Research done on mental health professionals has examined various factors that potentially cause burnout. Although research on burnout is plentiful in mental health professionals, there has been limited examination of those implementing Dialectical Behavioral Therapy (DBT) as a method of treatment. The intensive and often demanding training that is required of therapists prior to practicing DBT, accompanied by the nature of symptoms DBT is often used to treat, may lead to increased occupational stress. More recently, a new assessment tool for burnout was developed by Kristensen et al. – the Copenhagen Burnout Inventory (CBI). Research has pointed out numerous advantages of the CBI over the MBI. The CBI has been demonstrated to have high internal validity and its scales are able to discriminate between different professions. Less research has been done using the CBI. In this study, we measure the potential factors that cause burnout in mental health professionals using the CBI. Data in this study are collected from questionnaires filled out by mental health professionals participating in Dialectical Behavioral Therapy intensive trainings (N=411). Result for this study by employing the CBI will provide us with an opportunity to compare the findings to those made with the MBI. It will also be able to shed light on the characteristics of burnout in DBT practitioners.

ATLAS is a particle detector located at the Large Hadron Collider that is used to perform experiments in high-energy physics. In 2014, the Insertable B-Layer (IBL) was implemented into the center of the ATLAS detector to protect detector layers against radiation damage and provide additional particle tracking accuracy. During the years leading up to the implementation of the IBL, the ATLAS scientific community had asked for computer visualizations of the detector that were updated to show the addition of the IBL. Using the computer graphics software 3DS Max, we implemented the IBL in the detector images to produce a new, up-to-date generation of ATLAS detector images. Following this graphical update of the detector itself, we are planning on developing a new set of 3D computer animations that visualize the physical concepts behind the ATLAS experiments, including the LHC’s role in searching for discoveries related to dark matter, origins of the universe, and the Higgs boson. These animations will be used in scientific education as a useful way to visualize important concepts in particle physics.

Providing students high quality educational opportunities is important, especially in schools where there are high poverty rates and students who have been historically underrepresented. The Washington STEM project was a collaboration between researchers in UW’s College of Education and a community of educators at a local elementary school that was consistently underperforming in mathematics achievement. School leaders and university researchers worked together to develop a model called “math labs” to help improve the experience of learning mathematics for both the students and teachers. The purpose of this project is to study whether or not the use of math labs were statistically significant in contributing to the students’ growth in understanding mathematics. We drew on four consecutive years of student achievement on state’s benchmark mathematics assessment for grades 3-5 to assess our hypothesis. We hypothesized that math labs and other professional learning for teachers resulted in a growth that is statistically significant in improving the students’ academic performance in mathematics. Thus, we expect an increase in the benchmark scores for the students. To analyze possible growth, with my mentors, I ran multiple linear regression analysis for each sequential year to examine if there was a positive correlation between each year of the students’ benchmark scores. I also ran t-tests to examine if any of the suggested growth were statistically significant. There are multiple variables that attributes to a students’ academic growth, one important variable is the assigned teacher. To examine this variable, we ran a hierarchical linear models (HLM) to analyze cohorts of students nested within their teachers, necessary for our multi-level data set. The project is to provide evidence that the professional development model provides a positive growth for teachers’ instructional practices and students’ academic achievement.

In a marine environment, attachment to substrate is vital to the success of many invertebrates, particularly those in the intertidal. Wave action, tidal flow, and predation are challenges that these organisms combat through a variety of mechanisms. Among these are attachment mechanisms, which help the animals stay in place. Chitons belong to the phylum Mollusca, which often use a mixture of suction and glue to attach to a substrate. In 1911 and 1916, G.H. Parker laid the groundwork for the study of chiton attachment forces. He made the assumption that chiton attachment “depends almost exclusively upon suction”. However, his observations did not involve formal testing, which left the questions of how and with how much force chitons attach to a substrate virtually unexplored. In this study we measured the attachment forces of the chiton species Mopalia muscosa. Specimens were allowed to attach to substrates with and without perforation. Perforation in the substrate inhibited the chiton’s use of suction. In order to also investigate the impact of surface roughness on chiton attachment, we performed the same experiment using substrates of three different types of roughness: smooth, grain size of 0.267mm, and grain size of 1-2mm. We measured a significant decrease in the attachment tenacity for all roughness type, which displayed between 30% and 50% of the attachment tenacity upon solid substrates. On solid smooth and 0.267mm grain substrates, M. muscosa attached with tenacities of 21 and 22 kPa, respectively, while attachment tenacity upon the roughest surface was much lower (10kPa). Our results show that suction plays a significant role in chiton adhesion to substrates of varying roughness. However, other attachment mechanisms such as glue clearly play an important role as well

The computer science (CS) field lacks female and minority diversity, and this is due in part to the female and minority students’ negative stereotypes of the tech field. Previous research suggests that these students decide that they do not belong in computer science during or before high school. To recruit these populations into computer science, various programs have been implemented to educational programming, namely STEM specialized high school curriculums, federally funded CS initiatives, and accessible support guides. This study examined the role of these computer science programs or support systems on women and minority students’ perceptions of computer science. We predicted that the underrepresented students who participate in a special program would identify more favorably with computer science. Students completed surveys measuring cultural stereotypes, sense of belonging, identification with computer science, and whether they had participated in a special CS program. The sample population included students from a two-year public college and a four-year public university who were at least eighteen years of age. Data collection is currently ongoing. Follow-up research should measure students’ identification and sense of belonging in CS prior to enrollment in a diversity program and again after they participate. If these programs can help increase underrepresented students’ sense of identification and belonging in computer science, then more colleges and universities across the country should adopt these CS diversity programs to help address the long-standing recruitment problem.

Bridging between the innate and adaptive immunity, dendritic cells (DCs) are the most potent antigen-presenting cells. To develop vaccines against sexually transmitted diseases (STDs) like human immunodeficiency virus (HIV), DCs that locate in and beneath the epithelium of the female vaginal mucosal tissue are of particular interest. Unlike the vaginal submucosal DCs, which tend to induce aggressive adaptive immunity against viral infection, the vaginal epithelium DCs generally induce immunosuppressive adaptive immunity. The ultimate goal of this project is to develop nanoparticles that can be delivered as a vaccine via the female genital tract to enhance protective immunity of the vaginal mucosal tissue. We designed our DC transfection nanoparticles by encapsulating antigen-encoded DNA plasmids for antigen presentation, as well as a double stranded RNA adjuvant, poly(I:C), for provoking DC maturation. The combination of these two types of nucleic acids is condensed by a biocompatible nucleoprotein called protamine, which is known for condensing DNA in sperm due to its rich positive charge. We hypothesize that this adjuvant-encapsulated design will have an advantage over administration of extracellular adjuvant as used in conventional vaccine. This is because extracellular adjuvant promptly induces DC maturation, which generally decreases cell uptake, resulting in less antigen-encoded plasmid internalization by DCs. Alternatively, if we induce DC maturation via the intracellular pathway using poly(I:C), by encapsulating the maturation-inducing adjuvant along with the antigen DNA, more antigen-encoded DNA can be internalized before DC maturation is induced. As for future work, we will extend our DC-targeting intravaginal immunization system to mouse models on which the impact of the nanoparticles will be evaluated.

Lithic curation practices as denoted by the amount of retouch, or reworking, of the edges has been shown to be related to the degree of sedentism and by proxy to a transition from hunting and gathering to agriculture. In order to provide insight into when an agricultural transition may have occurred in Indonesia I have analyzed retouch at two Sulawesi sites, Talimbue and Gua Mo’o hono, in order to see if this evidence matches with the small amount of faunal and other evidence from nearby sites. I calculated a perimeter retouch ratio (PRR) by analyzing images of the artifacts using Gimp and ImageJ software to compare the determined retouched length of the perimeter to the total perimeter length. The retouch intensity denoted by PRR was then plotted over time using dates obtained from radiocarbon dating material associated with the stone tools. My results indicate that during the time period from 500-3400 BP retouch was significantly high at both sites. PRR values were at their highest at Talimbue during this period and showed a peak at Gua Mo’o hono as well. This would support the notion that a fundamental shift was occurring, quite possibly associated with the beginning of agriculture in the region. More corroborating evidence and better temporal resolution would help elucidate this connection further. Being able to pinpoint and find converging evidence for when the agricultural transition occurred in Sulawesi has implications for tying the region into a larger network of migration patterns and cultural exchange.

Botulinum neurotoxin A (Botox), a frequent treatment for maxillofacial disorders, has been shown to cause bone loss. A previous study in which rabbits were terminated 1-3m after Botox injection into one masseter muscle (final age 6.5-8.5mo) observed less bone in the injected-side mandibular condyle (upper jawbone). To understand how repeat injections (clinically common) would affect bone, a second group of rabbits received 3 injections with 3mo recovery after each. Bone loss was again seen, but the morphology of the condyles differed so much that previous analytic methods could not be used, perhaps because the repeat-injection rabbits were older. My goal was to test the hypothesis that repeat-injection results in greater bone loss and to investigate age-effects on rabbit condyles by re-analyzing the single-injection condyles using uniform methodology to the repeat-injection experiment. Group A rabbits (14.5mo) received 3 unilateral injections of either Botox or saline (control) into one masseter every 12 weeks whereas Group B received single injection and were sacrificed after 1 (6.5mo) or 3mo (8.5mo). All condyles were frontally sectioned and stained for histology. Each slide was imaged, and all non-bone tissue digitally removed. Cross-sectional area was measured and the proportion of bone pixels (bone areal density) was calculated. Saline animals all had symmetrical condyles (p>0.1). Botox animals had less dense condyles on the injected versus uninjected side (p≤0.03 all groups) but were not smaller except in younger rabbits. The side difference in areal density was 8.6% at 8.5mo and 7.6% at 14.5mo. Thus, repeat injections did not result in greater bone density loss than a single injection. With age, area decreased in all condyles (p≤0.004, ANOVA), but areal density decreased significantly only in the Botox-injected side condyles (p=0.02), suggesting Botox exacerbated natural density loss. Our findings have clinical implications for administration of Botox and age-specialized care in humans.

In HIV-1 infection, dendritic cells (DCs) are among the first cells thought to encounter the virus and contribute in shaping the course of the disease. However, we know little about how DCs detect HIV-1 or how innate activation of DCs is regulated. DCs play an active role in cell-to-cell transmission of HIV-1 to CD4 T cells. Loss of CD4 T cells is the hallmark of AIDS, but only a subset of dying CD4 T cells are productively infected with HIV-1. Recent evidence indicates host sensor, interferon gamma inducible protein 16 (IFI16), can detect HIV-1 in CD4 T cells. IFI16 then mediates pyroptosis – an intensively inflammatory form of cell death. Dead CD4 T cells release many pro-inflammatory cytokines, which act to attract and cause even more death of these CD4 T cells. Moreover, HIV-infected DCs are shown to produce type-I IFN, display hyper-inflammatory characteristics, and die at a high frequency, but it is unclear by which mechanisms these processes occur. In this project, we aim to exploit a novel gene-editing program identified in bacteria - CRISPR/Cas9 system (clustered regularly interspaced short palindromic repeats) - to perform unbiased genome wide screens for new regulators of HIV-mediated IFN production and HIV-induced cell-death in DCs. Monocytes isolated from whole blood of healthy donors first get transduced with lentiviral vectors of GecKO library. DCs derived from these monocytes then get infected with reporter virus (HIV-GFP). These cells are collected at different time points and analyzed for infectivity, activation level, and death using flow cytometry. Final step of deep sequencing helps identify potential gene targets. These new discoveries might be a promising avenue for the development of future therapeutics.

Differential scanning calorimetry (DSC) tests are being conducted to determine the melting enthalpy (heat flow) values of high-oil content wax used as the binder in synthetic granular composite materials. An important application of these granular composites are the surfaces of synthetic horse racetracks. The thermal behavior of the binder is important as it affects the mechanical properties of the track in terms of shear strength and hardness. Test temperatures span a range from -30°C (-22°F) to 94°C (201°F) and include the thermal transition regions where predominant melting of the wax occurs. These temperatures are within the range many racetracks are exposed to throughout the year in various parts of the country. Thermal transition ranges and peak temperatures were taken from the second heating run and plotted with thermal endotherms pointing downwards by the standards for DSC wax testing and ASTM D4419. The 1st transition peaks typically indicates the predominant melting regions showing enthalpies of the wax samples as heat is absorbed. Our results from an ongoing seven year trial indicate a decrease in crystallinity over time as the wax degrades from environmental and mechanical surface interactions.

This study explores the connection between foraminiferal assemblages and environmental pollution in Commencement Bay near Tacoma. Foraminifera are microscopic single-celled organisms that produce an external shell made of either calcium carbonate or agglutinated (organically cemented) sediment. These marine organisms are sensitive to environmental change, and are thus useful tools in monitoring ecosystems. Some species, such as the opportunistic Eggarella advena, are highly pollution tolerant. A strong presence of these species would provide evidence of water degradation. In this project, foraminifera are utilized as indicators of pollution, which is an immediate problem in Commencement Bay. Since the mid-1800s, the bay has seen the development of sawmills, smelting plants, refineries, and shipyards. Extensive industrial pollution has resulted in designation of 12 square miles of the area as a Superfund site. Assessing the environmental conditions of the site is one aspect of a larger effort to restore its ecosystem. The research documented here helps provide the EPA with such information. The twenty-five samples used in this study were collected in 1997, 2008, and 2014, allowing analysis of changes over an extended time. More than 1,500 foraminifera were identified and calculations of foraminiferal density and species diversity were performed. Fifteen species were recorded, including Cribroelphidium excavatum and Elphidiella hannai which are the most common species in Commencement Bay. A decrease in agglutinate foraminifera and an increase in species diversity was observed from 1999 to 2014. Overall low species density and diversity per sample, along with evidence of dissolution of calcareous shells, indicate that Commencement Bay is still an environmentally stressed ecosystem.

Serotonin (5-HT) is a monoamine neurotransmitter that facilitates behavioral arousal and regulates movements by affecting synaptic receptors on spinal neurons. Serotonergic neurons project directly to motoneurons in the brain stem and ventro-medial part of the spinal cord that innervate extrafusal muscle fibers directly responsible for muscle contraction, as well as other secondary motor structures. Damage to the serotonergic neurons that facilitate movement is therefore, of great relevance to research of motor impairment and motor recovery after central nervous system injury. Our research on promoting neuroplasticity with electrical stimulation has demonstrated improved recovery following surgically induced spinal cord lesions, presumably by strengthening synaptic connections. This encouraged further research into the anatomical changes in the neural circuits that follow spinal cord injury and stimulation therapy. Using immunohistochemistry techniques, we studied changes to serotonergic innervation of the spinal cord and its correlation to functional recovery. In addition, we investigated perineuronal nets, specialized extracellular matrix structures composed of chondroitin sulfate proteoglycans (CSPGs), which are critical to neural development and synaptic plasticity. Past studies have shown that in adulthood CSPGs in glial scar tissue inhibit axon growth and regeneration following damage to the nervous system.

The objective of this project is to identify how concussion mechanism of injury varies between males and females, and to determine if this variation influences response to post-concussion Sub-Threshold Exercise Program (STEP). It has been estimated that 1.6-3.8 million children sustain concussions each year and that female athletes are at greater risk of injury compared to males. Over the past decade, the number of youth female athletes has risen to 41% and the leading cause of sport concussions for female athletes is now soccer. Given this information, mechanism of injury and its variation between sexes has become an interesting topic of study which may inform our approach to treatment and prevention. Thus far, there has been a good amount of research done on rates of concussions between males and females but not on differences in mechanism of injury, or on response to exercise treatment programs. With this study, a retrospective chart review of 55 patients with concussions to determine differences in mechanism of injury. We are currently in the midst of completing the chart review, but preliminary analysis suggests that males more commonly had concussions resulting from person to person contact while females more commonly collided with the ground or another object. In conducting this research alongside the larger STEP evaluation, we hope to get a better understanding of the ways in which youth athletes of different sexes are commonly injured and how these mechanisms of injury affect overall recovery. Moreover, we hope this knowledge assists in creating treatment programs that are better targeted to addressing common issues that each sex faces as well as prevention programs that address the common mechanisms of injury for each sex. 

It is now understood that a multitude of clinically-relevant antibiotic resistance genes (ARGs) are widespread in environmental systems. Previous research has confirmed that municipal wastewater can be a particularly important route for ARG discharge into the environment from such sources as hospitals, medical clinics, and nursing homes. Conventional biological wastewater treatment processes can help to attenuate levels of ARGs. However, significant ARG levels often remain in treated wastewater within viable antibiotic resistant bacteria (ARB) or within cell debris associated with inactive biomass. Therefore, additional research is needed to evaluate the effect of water disinfection methods – frequently applied following biological treatment – on decreasing the prevalence of ARGs in wastewater prior to discharge into the environment. This work focuses on quantifying the effectiveness of the disinfectant chlorine in degrading ARGs in wastewater. Chlorine – one of the most effective and widely used disinfectants in wastewater treatment plants (WWTPs) – is well known for its ability to inactivate live bacterial cells, including ARB cells. However, little is known about its efficacy in directly degrading ARGs. In this project, methods are being developed to enable DNA extraction and quantitative real-time PCR (qPCR) analysis of the ARGs tetA, vanA, mecA, ampC, NDM-1, and KPC within municipal wastewater. A non-native ARG (blt) present within cells of a Bacillus subtilis strain spiked to wastewater samples is being used as an internal standard to normalize for DNA recovery during sample processing. Samples of secondary wastewater effluent will be collected before and after chlorination from King County’s West Point WWTP. Using the methods indicated above, the six monitored ARGs will be quantified in each sample to assess the effect of chlorination on ARG copy numbers. Measured ARG losses will be compared to predictions based on rate constants for ARG degradation determined in parallel studies undertaken in the same lab.

Playing sports is a setting in which traditional gender norms are often policed and reproduced. In addition, sports are often strictly sex-segregated. Because of these factors, I hypothesize that transgender individuals may feel alienated from the world of sports and are less likely to participate. This study aims to identify what aspects of sports-related settings discourage or restrict the participation of transgender people, and what solutions may help to remove those barriers. To reach this goal, in-depth interviews of five transgender athletes were conducted, as well as a survey of 66 transgender people from varying backgrounds. Results indicate a majority of transgender people have felt unwelcome playing sports because of both their gender expression and sex, and identify the need for private changing areas in locker rooms, statements of explicit inclusion of transgender athletes, and opportunities to play sports in non-gender-segregated settings. Interview responses also indicate that public discourse debating the genders of athletes causes harm to transgender athletes and causes them to feel unwelcome playing sports. The findings of this study provide implications of some ways to make sports more accessible to transgender people, which is greatly needed given this demographic’s high rates of depression, social isolation, and suicide. Future research is needed on the impact of implementing these proposed changes on both transgender and cisgender athletes.

The use of fermentation to create alcoholic beverages is one of mankind’s first known biological engineering tasks. Beer, one such product, is made by the saccharification of starch and the fermentation of the resulting sugar to ethanol by yeast. We are exploring the genetic factors that allow yeast to adapt to the selective pressures of this fermentation process, as well as which genes shape the resulting byproduct.  To study S. cerevisiae’s genomic evolution during fermentation, we have competed thousands of barcoded gene deletion and amplification strains within a medium composed of malted cereal grains (wort). As the beer underwent this fermentation process, samples were collected over time and the frequency of each mutant was tracked using unique barcodes through high-throughput sequencing. The barcode abundance over time was then used to estimate the fitness impact of each mutation. Throughout the experiment we also recorded such things as the glucose, maltose, and ethanol concentrations, cell viability, and plasmid retention. This data allows us to identify the genetic components that control growth in wort.  In the future we plan to try several other variations of the experiment, including the use of different medium/wort compositions and different temperature conditions for lager vs. ale strains. We will also be performing a competition using a heterozygous deletion collection in a hybrid similar to common lager strains, as well as with a mutant collection in Saccharomyces uvarum, another species commonly found in winemaking and cider fermentations. There is also the potential to compete strains within a bread dough starter.  This project will expand the potentials of microbial biotechnology, as new targets for genetic engineering have the potential to be identified, and are thus relevant to emerging technological and industrial applications.

Positive outward remodeling of arteries in response to arterial injury is highly dependent on the migration of smooth muscle cells (SMCs) and their ability to degrade extracellular matrix (ECM) around existing vessels. The p27 protein is a cyclin-dependent kinase inhibitor that stalls the cell cycle when in the nucleus. It affects cell migration and interaction with the extracellular matrix when in the cytoplasm. We have previously demonstrated that mice lacking p27 have improved collateral artery enlargement in response to ischemia. We hypothesized that decreased expression of p27 improves SMC migration and ability to degrade ECM. To test this hypothesis, we performed several in vitro assays using aortic SMC isolated from p27 knockout (KO) and wildtype (WT) mice. A wound assay, performed under growth arrest, was employed to assess SMC migration in the absence of a chemoattractant, and a modified Boyden Chamber assay was used to assess migration in response to a SMC attractant platelet derived growth factor-BB (PDGF-BB). Both assays were then repeated with the addition of a matrix metalloproteinase (MMP) inhibitor to assess migration in the absence of ECM degradation. Wound assay data shows that p27 KO SMCs exhibit a higher degree of migration. Addition of an MMP inhibitor, BB94, to the wound assay abolished cell migration for both WT and KO cells. Preliminary Boyden Chamber results showed that p27 KO cells migrate more than WT cells in response to PDGF-BB. Preliminary experiments involving a more specific MMP2 inhibitor are now in progress. A better understanding of p27’s role in arterial remodeling may lead to improved therapies stimulating arterial growth to treat ischemia resulting from arterial occlusion or inhibited arterial growth to treat cancer.

Prochlorococcus are abundant photosynthetic bacteria in the ocean, but Prochlorococcus in ocean oxygen minimum zones are understudied. We are examining the viruses that infect Prochlorococcus in these anoxic waters. To do so, we look at three genes: a gene essential for photosynthesis (psbA), the viral capsid assembly protein gene g20, and a viral DNA polymerase gene which is used in viral replication. We use phylogenetic trees to find relationships between oxygen minimum zone viral sequences and previously studied organisms. We also assembled our short sample sequences (150 base pairs) into longer, more complete gene sequences. These longer assembly sequences were incorporated into our reference tree to include organisms relevant to the oxygen minimum zone. Both Prochlorococcus and Synechococcus carry the psbA gene. Some of their viruses also carry the psbA gene to help the host remain alive long enough for the virus to reproduce. In the oxygen minimum zone, we see many fewer psbA reads in general compared to the oxic waters. Most of our psbA reads in the oxygen minimum zone are the lowlight IV Prochlorococcus instead of viral. Viral psbA reads decreased with depth. We included the g20 and DNA polymerase genes in our analysis, so we could see a larger cyanophage community since psbA isn’t found in all cyanophage. From our research on the viral DNA polymerase gene, we found an abundance of myovirus sequences in our assembly data. Results from psbA are consistent with myoviruses being the dominant cyanophage in our oxygen minimum zone samples. The g20 gene is only found in myoviruses. Results from our g20 gene analysis informed us that cyanophage viral sequences decreased as Prochlorococcus decreased. These results imply that there are at least some cyanophage present in the oxygen minimum zone without the psbA gene.

In the yeast genome, there are two copies of nearly all genes that encode the proteins that make up the ribosome, the cellular structures that are responsible for assembling proteins. When one copy of the gene encoding one of these ribosomal proteins, RPL20A, is deleted, the cells are sick and grow slower; however, eventually variants arise that are able to grow faster. Most variants acquire a second copy of the remaining gene as part of an adjacent, direct, tandemly duplicated chromosomal segment through unequal recombination at flanking transposable elements. However, one strain acquired two additional copies, and the ends of the amplified segment did not map to the expected transposable elements. I tested the hypothesis that the two extra copies were structured as an inverted triplicon (all three copies side-by-side with the middle copy in the opposite orientation). Our lab had proposed a new model for how these inverted amplifications arise and I wanted to see whether this gene amplification event was an example. I used a CHEF gel to compare the sizes of the parental and evolved chromosomes and determined that all three copies reside on the same chromosome. By cutting within the amplified sequence with restriction enzymes and performing Southern blots, I tested whether the additional sequences generated an inverted triplicon, direct repeats, or were scattered throughout the chromosome. I also performed a snapback assay to see whether the ends of the amplified sequences had an inverted structure. My results indicated that this unusual amplicon had a direct repeat structure that had occurred at a different pair of repeated sequence. Using yeast as a model organism to study the mechanisms of gene amplification helps us understand how gene amplification occurs and has implications for diseases in humans, including cancer.

Recent studies have shown that within tumors of many cancer types exist a sub-population of cells, termed cancer stem cells (CSCs). These cells are responsible for tumor relapse and the development of drug resistant tumors. Because of their role in tumor biology and their inherent chemotherapy-resistance, targeting CSCs has become an active area of cancer research. We hypothesize that one way to sensitize CSCs to chemotherapy treatment would be to inhibit signaling pathways in CSCs. For this study I am working with stem and non-stem cancer cells isolated from an ovarian tumor. CSCs can be isolated by the presence of the cell surface marker CD133. We used RNA-seq to identify genes differentially expressed between these tumor cell types. We found that 107 phosphatases and kinases are upregulated in ovarian cancer stem cells compared to the non-stem cancer cells. These up-regulated proteins are potentially important in ovarian CSC signaling pathways, and could be key to their chemotherapy resistant phenotype. Based on the upregulation in CSCs, I used small molecule inhibitors to target a subset of these molecules. After inhibition, I exposed the cells to Cisplatin—a common chemotherapy drug. I compared the inhibited cells against an untreated control to identify differences in cell survival. I measured cell response by FACS sorting, using the cancer stem cell marker CD133 and a cell viability fluorescent dye. This information on cell survival and composition will provide valuable information on the sensitivity of the cells to chemotherapy drugs after signaling pathway inhibition. It will also give us key insights into how kinase and phosphotase activity affects the CSC phenotype. We expect that CSC's targeted with small molecule inhibitors will exhibit a greater sensitivity to Cisplatin. This could have a significant impact in the treatment of ovarian cancer through enhancing existing chemotherapy treatment with small molecule inhibitors to reduce tumor resistance and recurrence. Small molecule inhibitors could give physicians a way to target the CSC population previously resistant to traditional chemotherapy treatment.

Studies suggest that excessive alcohol consumption is a leading cause of premature mortality in the United States, beginning in late adolescence and peaking in college. Specifically, impulsivity has been linked to alcohol consumption in young adults. Additionally, numerous studies suggest that alcohol consumption is often precipitated by negative feelings. Thus, impulsivity and mental health seem to be important factors in young adult drinking behaviors. The present study investigates the relations among impulsivity, mental health and alcohol consumption in young adults. High school seniors from Washington state and Sweden (n=3175) completed a series of online surveys assessing alcohol related negative problems via the Young Adult Alcohol Problems Screening Test, mental health as measured via the 8-item dichotomized stress symptoms checklist (SCL-8 D), and impulsivity as measured via the sensation seeking scale. 
Regression analyses revealed a significant main effect of impulsivity on alcohol related negative consequences (p < .05). A significant moderating effect of mental health on the relationship between impulsivity and alcohol related negative consequences (β = .01, F (3, 3174) = 104.50, p < .05) was found, such that those with higher impulsivity and higher mental health symptoms have higher alcohol related negative consequences. These results suggest that individuals who are both higher in impulsivity and experiencing mental health symptoms have the most negative alcohol related consequences.

Volcanoes are among the most violent and unpredictable forces on earth. As population grows and increasing numbers of people live and work in direct proximity to volcanoes, it is important to develop useful models of the behavior of volcanic magma and eruptions. La Palma is the most volcanically active of the seven Canary Islands, a chain of ocean island volcanoes off the northwest coast of Africa. Records of historic eruptions date from 1480 to 1971. All of the historic eruptions on the volcanic ridge of La Palma have a Volcanic Explosivity Index rating of two, which indicates that the eruptions have to this point been relatively non-explosive. However, the presence of water-bearing amphibole crystals in the 1480 (Quemada) lava flow suggests that a steam-driven explosive eruption is possible. It is unknown whether the amphibole crystallized from the Quemada lava or an older unrelated rock. My study will consist of three tests: microscopic evaluation of sample thin section, microprobe analysis of amphibole mineral composition, and lead isotope analysis of mineral and host magma. This data will provide chemical signatures that I can use to track the origin of the amphiboles. If the amphiboles did not crystallize from the host lava, it is possible they were incorporated into the magma from an older lava flow. However, if the amphibole crystals and host lava have the same Pb isotope ratios then we can infer that they originated from the same magma, which indicates that the explosive potential of the volcano is high.

Engineered in-vitro human tissue models for evaluating drug-effects harbor great potential. Establishment of competent models will greatly reduce the number of laboratory animals expended in the drug discovery process, enable higher-throughput than is possible with animal models, and yield more humanly accurate responses. Still, high-throughput electrophysiological assessment remains elusive. Microelectrode-arrays (MEAs) hold promise for in-vitro cardiac drug testing because they can rapidly acquire tissue-level electrophysiological data on key metrics of cardiac function. These include the conduction velocity, QT interval of the tissue, and other attributes that pharmaceuticals can impact for better or worse. Furthermore, one can integrate several MEAs into one multiwell-plate. MEAs are typically imprinted on flat substrates that don’t accurately mimic the topography of the native myocardium. Consequently, cardiomyocytes atop MEAs exhibit a functionally and morphologically immature phenotype distinct from the phenotypes of native cardiomyocytes. Accordingly, MEA-based cardiac tissue models exhibit physiologically inaccurate responses to compounds with well-understood electrophysiological impacts. This limits the utility of MEAs when assessing unexplored compounds. We hypothesize that an MEA-based cardiac tissue model incorporating biomimetic topography will yield responses to well-explored drugs closer to responses observed from mature cardiac tissue. To investigate this, we test several thoroughly understood compounds on rodent and then human stem cell-derived cardiac tissues cultured atop a biomimetic MEA platform and compare the responses to those of native cardiac tissue. Our device consists of a flat MEA below an anisotropic nano-fabricated uniaxial grooved surface, which promote overall cardiomyocyte maturation and help to organize the structure of the engineered cardiac tissues, giving them an aligned and more biomimetic architecture. Alignment causes these tissues to conduct electrical signals in a directed and efficient fashion representative of native myocardial tissue. Should our model prove effective, it will accelerate the pace of drug discovery and improve the safety of current and future pharmaceuticals. 

 Food sovereignty is defined as a universal right to have control over the source and content of one’s sustenance. Within Canadian First Nations and Native American tribes, food sovereignty is integral to health, wellness, economic stability, and environmental consciousness. Cross-cultural knowledge sharing has been instrumental across Native and Non-Native communities in defining food sovereignty. My question is twofold: how has the discourse and practice of food sovereignty influenced the efforts of Native communities to reestablish their traditional food systems, and what is the role of cross-cultural, participatory engagement and political alliance within the Food Sovereignty movement? I argue that food sovereignty is a spatial and epistemological "contact zone" with points of commonality and contention. Through cross-cultural ethnographic accounts of scholars engaged with food sovereignty in Washington and Canada, I have compiled case studies of allyship and mentorship within specific communities. I develop guiding principles for relational thinking across boundaries of culture and ways of knowing. This response provides opportunities for productive and meaningful partnerships as well as ways to redefine and decolonize the language of Food Sovereignty.

I examine, in an art historical context, two TV advertising spots for Lindt brand chocolates. Using critical analysis of the gendered and erotic images depicted in these commercials, I study the manner in which Lindt's ad campaign appropriates sexuality (specifically, the female orgasm) in order to heighten the allure of their product. Several strands of art historical context are useful in establishing the heredity of the campaign's sensibilities. I treat, firstly, the long-standing cultural conflation of chocolate with sexuality (due to its supposed properties as an aphrodisiac), then examine prevailing cultural attitudes about the  female orgasm as it pertains to male/female sexual relations, and finally look as far into the past as the 16th century to analyze the factors that Titian's languishing Venus and Manet's courtesan "Olympia" have in common with the women in Lindt's commercials. I examine how the codified image of the sexually available woman is present in Lindt's ad campaign, and hope as my research develops to assess the effectiveness of the campaign itself due to the imagery involved. In this, I hope to broaden the scope of art criticism and context awareness to include everyday images (like advertisments). Though it may seem dubiously relevant to compare a modern commercial to works by Titian and Manet, I aim to show the linkage, visual and substantive, between these manifestations of the western visual tradition.

Alcohol-use disorders are more prevalent among people who are homeless than in the general population. Thus, homeless individuals experience disproportionately high levels of alcohol-related problems and resulting publicly funded criminal justice and healthcare system utilization. Available treatment services, however, are not effective at engaging and treating this population. To better tailor treatment services to their needs, it is important to understand this population’s perceptions of their alcohol use. The aim of this study was to provide qualitative descriptions and relative rankings of the advantages and disadvantages of alcohol use from the perspectives of this population. Participants (N = 44) were people with lived experiences of alcohol-use disorders and chronic homelessness who were recruited from community-based agencies in Seattle, Washington. Open-ended prompts were used to assess the perceived role of alcohol in participants’ lives, including participants’ perceptions of the advantages and disadvantages of their current drinking, and a conventional content analysis was conducted. The most commonly mentioned advantages of drinking included positively and negatively reinforcing psychological reasons, perceived control over drinking, and social benefits. Physical effects, concerns about dependence on alcohol, and health problems were the most commonly mentioned disadvantages. By documenting the perceived advantages and disadvantages of drinking among people with the lived experience of homelessness and alcohol-use disorders, this study supplied information providers may use to better tailor treatment services to this multimorbid, high-utilizing population’s needs and interests.

As the use of antiretroviral drugs for HIV treatment becomes more widespread in developing countries, the prevalence of drug resistant virus populations is beginning to rise. Drug resistant strains heavily reduce the success of antiretroviral therapy and also pose an increased threat to unaffected individuals via transmission. Our group is currently engineering a new test for HIV that utilizes an Oligonucleotide Ligation Assay (OLA) to diagnose HIV infection and drug-resistance. OLAs are more specific and sensitive than gold standard antibody tests but require equipment and personnel that low-resource areas often lack. To improve the feasibility of using OLAs in these settings, the goal of my project is to investigate methods for long-term preservation of ampligase ligase, a crucial enzyme used in the ligation process of OLAs. First, we will use a trehalose-dextran solution, which has been reported as an effective protein stabilizer, to desiccate the enzyme. After being subjected to a range of storage conditions (37-72°C, 50-70% relative humidity) for up to 30 days, the performance of the dehydrated enzyme will be compared to that of fresh ligase. Once a successful enzyme preservation method has been established, further work will focus on engineering a technique to anchor the enzyme onto a matrix. Effective enzyme stabilization and anchoring will eliminate the need for refrigeration in shipping and storage and can incorporated into the lateral-flow HIV detection device that our group previously developed. This simplification of the diagnostic process will ultimately reduce costs, detection time, and lab use, thereby allowing the test to be better suited for application in low-resource settings. Furthermore, improved access to disease and drug resistance diagnostics for HIV will prevent rampant transmission and enable proper treatment for infected individuals.

Kosrae is a developing island approximately 111.3 km² incorporated in the Federated States of Micronesia. Within the past 50 years, increased conversion and degradation of coastline has drastically reduced the area of mangrove habitat as the once secluded country has opened itself to tourism and higher quality of living for residents. Unsustainable methods of wood harvest threaten mangrove diversity and the benthic faunal community. Mangroves are the only species of plant able to survive in brackish environments and are the only terrestrial flora present in intertidal zones. They are essential figures in tropical ecosystems controlling erosion, acting as global carbon sinks, and housing a multitude of unique flora and fauna. While abroad in Kosrae, I took core samples of sediment from below stands of four prominent mangrove species: Bruguiera gymnorrhiza, Sonneratia alba, Rhizophora stylosa, and R. mucronata. Utilizing Shannon Wiener Diversity Indexes and sample t-tests for the difference in means, I discovered the Kosraean benthic macro-invertebrate community to be evenly distributed and low in species richness and abundance. There was little evidence to conclude a statistically significant relationship between the species of mangrove and its faunal composition. Abiotic components such as substrate type and salinity were determined to have the highest likely influence on phylum distribution and abundance. Although low in macro-invertebrate diversity and abundance, mangrove forests are increasingly recognized as key elements in habitat conservation vital to the protection of the island’s terrestrial and marine organisms. Local government organizations are working to promote and protect the mangrove forests, ensuring sustainable harvest and educating the public about their significance both ecologically and economically.

Phononic Crystals (PCs) are periodic structures that control the behavior of elastic and acoustic waves. Promising and mostly unexplored types of PCs are woodpile structures consisting of a chain of cylindrical rods. This system possesses a considerable number of parameters, including rod length, contact angle and added load, which can tune the wave dynamics of the system. The purpose of this project is to understand the effects of local resonance (i.e. low frequency vibration of individual particles in the chain) on wave propagation in the woodpile structure and assess the feasibility of potential applications. Focusing on the applications side, the main goal is to create a system that converts pulses that aren't linear or periodic in localized vibrations from which electrical energy can be extracted. Currently, we are numerically investigating the wave propagation in woodpile structures having various local resonances at different locations under impact excitation with a Discrete Element Model (DEM). We are also experimentally measuring the wave propagation and amount of energy trapped in the system using a Laser Doppler Vibrometer coupled with post-processing software and verifying the wave dynamics in the woodpile structure. The understanding obtained from this study will enable my team and me to focus on deriving an analytical expression that describes this system's behavior and developing a new type of energy harvesting technology. If successful, this project would represent a significant contribution in mathematics and nonlinear dynamics; woodpile PCs could become the building blocks of highly tunable devices used for energy harvesting, impact mitigation, and elastic/acoustic wave filtering. 

This research focuses on University of Washington (UW) student-athletes and how the principle of sleep affects their success in sports and academics. Do student-athletes experience sleep deprivation because of their schedules, and if so, do they think sleep deprivation impacts their education or athletic success? I found out that fluctuation in sleep could affect the academics and sport success of student-athletes which could later affect them in their future careers. I hypothesize that academic curriculum conflicted with sport practices leading to sleep deficiency in sport athletes, thereby affecting their achievement in both areas. To research this subject further, I did one-to-one interviews, focus groups interview and discourse analysis on UW student-athletes and took some data on their sleeping patterns and performance in both sports and academics. I hope that through my research I could show the importance of sleep and its effect on the progress of student-athlete's sport and academics. This research matters to student-athletes because their inability to do well in both sport and academics could limit their future employment.

Using bathymetric imagery and current velocity readings from a Teledyne R.D. Instruments Workhorse Mariner 600KHz ADCP, the influence on benthic currents exerted by a set of dunes in the southern half of Skagit Bay, a part of the Puget Sound estuary, was examined. A strong correlation was found between theoretical bathymetric influenced currents and actual currents measured by the Acoustic Doppler Current Profiler (ADCP). Taking the horizontal current flow and slope of the dunes, and working with the assumption that vertical currents can be heavily influenced by horizontal flow being constrained by the slope of the seafloor, a bathymetric driven vertical velocity was calculated. The theoretical vertical velocity was calculated using the observed vertical velocity at various depths as well as the calculated bathymetric driven vertical velocity. Findings strongly suggest a strong influence of the bathymetry on tidal currents lower in the water column. These findings can lead to a better understanding of the currents in an around the Pudget Sound allowing for better understanding of forces actinve on ecosystems and pottentially allowing for a better informed descision about locations for tidal energy stations.

Homeless individuals are at a greater risk of suicide than the general population. Chronically homeless individuals experience even greater severities of psychiatric and substance use disorders than the general homeless population. No studies to date, however, have documented means of suicide in this particularly high-risk group. The present study assesses means of suicide among chronically homeless individuals with severe alcohol problems. Participants were chronically homeless individuals (N= 134) who were high utilizers of publicly funded services in King County, Washington. Suicidality variables were measured during one-on-one interviews using the Screen for Suicide Risk (SSR) questionnaire, a modification of the Screen for Youth Suicide Risk (Thompson, Eggert, & Herting, 1993). A conventional content analysis was conducted to categorize means of lifetime suicide attempts and current suicide plan. Findings showed that cutting was the most commonly reported means of previous suicide attempts; however, cutting was not named by any participants as currently planned means. Overdose was the most commonly reported means for current planned attempts. A noticeable difference existed between reported means of suicide for past attempts and current plans, which may be due to the age differences at the times of suicidal ideation within participants. Understanding aspects of this population’s self-reported means of suicide may help providers reduce suicide risk through restricting access to means.

Endocrine disrupting compounds (EDCs) in wastewater treatment plant effluents are a surface water quality concern. Bioaugmentation of Sphingobium sp. BiD32, a bacterium capable of degrading the EDC bisphenol A (BPA), is a new technology for enhancing BPA removal. Degradation declines over time after augmentation due to endogenous respiration and predation by protozoa. However, recent laboratory reactor studies have shown that the DNA of the bioaugmented bacteria persists even as their activity declines, suggesting that the measured persistence may be associated with inactive cells. The aim of this project was to develop a method to distinguish between DNA from live and dead cells, and to apply this method to determine how predation affects the activity and concentrations of bioaugmented bacteria. The current method for monitoring bacterial concentrations, quantitative PCR (qPCR), measures the 16S rRNA gene of Sphingobium sp. BiD32, but cannot distinguish between DNA from viable and dead cells. Propidium monoazide (PMA) is a cell membrane-impermeable dye that reacts with DNA from nonviable cells and interferes with PCR amplification, so only DNA from viable cells will be amplified and quantified. Testing is underway to determine the best conditions for use of PMA-qPCR with activated sludge samples. Batch reactors will be set up in duplicate to determine bacterial death rates through concentration measurements of Sphingobium sp. BiD32. To determine endogenous respiration rates, batch experiments will be conducted in which the predators have been deactivated by either physical (rapid blending) or chemical (nystatin) means. To determine predation rates, controls will include two reactors with untreated sludge containing predators and one reactor with only mineral media so that the bacteria can survive but not grow. This work will develop a new method for quantifying viable bioaugmented bacterial concentrations in activated sludge and determine the rates at which predators are removing the bacteria.

Autism Spectrum Disorder (ASD) is the fastest growing developmental disorder in the U.S. and affects 1 in 68 children. It is a disorder characterized by deficits in social interaction and the presence of repetitive and restricted behaviors. Despite the wide variability in clinical presentation, a unifying deficit in social motivation has been proposed to account for the behavioral heterogeneity. Advances in genetics indicate nearly 1000 genetic risk factors for ASD. To determine if variability in social motivation may be due to these distinct etiologies, this study’s objective was to examine differences in social motivation in individuals with ASD with two distinct genetic events: 16p11.2 and 1q21.1 copy number variation (CNV). We utilized data collected in the Simons VIP study, a study applying a genetics first approach to understanding ASD. The sample consisted of 33 (22 males; 11 females; mean age: 150.5 months; range 20 to 281) individuals with ASD with 16p11.2 deletion or duplication and 7 (7 males; mean age: 131 months; range 46 to 216) individuals with ASD and 1q21.1 duplication and deletion. We used two measures of social motivation: the “social motivation subscale” from the Social Responsiveness Scale (SRS), a 65-item parent based measure of symptoms associated with ASD and the “social overtures item” from the Autism Diagnostic Observation Schedule (ADOS), a semi-structured clinician administered assessment of ASD symptoms. We will conduct two independent sample t-tests comparing both genetic groups on social motivation metrics. Given clinical reports that individuals with 16p11.2 CNVs make a greater number of social overtures, we hypothesize that individuals with a 16p11.2 CNV would score more highly on parent and clinician measures of social motivation relative to individuals in the 1q21.1 group. Potential implications from this study include symptomatic predictive power based on etiologically defined subtypes, which can aid in clinical management of ASD.

The neutrino remains the only standard model particle with an unknown mass. The neutrino mass could be inferred by means of precise energy measurements of tritium beta decay electrons. Project 8 is an experiment to do this using cyclotron radiation emission spectroscopy (CRES). We are currently evaluating the energy resolution of this technique by trapping and measuring Krypton 83m conversion electrons. Observing these trapped electrons can also provide information about the residual gas in the trap. I will present the results of identifying and analyzing electron-gas scattering events and speculate on how this can be used to improve future designs of the Project 8 experiment.

Xerostomia (dry mouth) is a condition that can lead to plaque accumulation, cavities, eating problems, and overall oral discomfort. Research over the years has shown that the prevalence of xerostomia is correlated with gender and age, however these studies have only been conducted on adult populations. Adolescents with psychiatric conditions take medications that might predispose them to xerostomia. In this pilot study, we estimated the prevalence of xerostomia from a convenience sample of adolescents hospitalized in an inpatient psychiatric clinic (N=25) and examined whether: 1) gender and age were associated with xerostomia; and 2) if xerostomia was associated with self-reported dental caries (ever having had a cavity). Adolescents were asked to complete the Xerostomia Index to assess self-reported xerostomia (no/yes). Participants were between the age of 9 and 14, of whom 72% were male and 28% female. 60% of those studied reported having xerostomia. My findings show that there is no statistically significant relationship between xerostomia prevalence and the subject’s gender (P=.99), age (P=.659), and self-reported ever having had a cavity (P=.26). Thus, gender and age are not associated with xerostomia prevalence and adolescents with xerostomia did not appear to have higher rates of dental caries. Other factors may be associated with xerostomia in adolescent populations. Future research focusing on xerostomia in adolescents should be conducted.

Duchenne muscular dystrophy (DMD) is a disease caused by a mutation in the dystrophin gene which results in muscle degeneration and progressively leads to death. Currently, there is no cure for DMD and patients are dependent on palliative care. Cardiomyopathy has become the major cause of mortality in these patients. Thus it is important to better understand the pathophysiology of DMD cardiomyocytes. Recently, dystrophin deficient cardiomyocytes have been differentiated from urine-derived induced pluripotent stem cells (iPSC-CMs). These cardiomyocytes retain the DMD mutation and are being used for disease modeling and drug discovery. The dystrophin protein connects muscle fibers to the surrounding extracellular matrix, the contractile properties of dystrophin-deficient iPSC-CMs could also be affected. Traction force microscopy (TFM) has been used to quantify cellular traction forces on soft substrates with fluorescent beads covalently bonded to the surface. By tracking the displacement of the beads embedded near the surface of the substrate and using the known elastic properties of the gel, the cell traction field can be calculated using Fourier transform traction cytometry. TFM techniques have previously been used to quantify contractile forces of micropatterned cardiomyocytes. Here we comparatively analyze the contractile forces of dystrophin-deficient iPSC-CMs and wild type iPSC-CMs. Contractile forces of IPSC-CMs on polyacrylamide substrates were measured from high resolution time lapsed microscopy of cellular contractions using the custom made ImageJ scripts. Our results show that dystrophin-deficient DMD iPSC-CMs exhibit less contractile force than wild type iPSC-CMs. Our quantitative analysis of contractile forces can be used to further characterize DMD iPSC-CMs for advanced disease modeling.

The impact of cultural differences on non-expert’s decision making styles with risk information will be investigated. In everyday life, people are required to make difficult decisions that involve uncertainty in areas in which they are not experts such as financial investment and provisions against natural disaster. Several factors such as the manner in which uncertainty is expressed are known to influence people’ preferences. However, empirical studies have also shown that aspects of cultures impact people’s perception of riskiness. In this study, a cultural aspect called uncertainty avoidance from Hofstede’s classification model (2001) is used to determine whether cultural differences in risk perception can be detected and if so, whether they are related to decision making under risk. For this study, students from high uncertainty avoidance culture such as Korea and low uncertainty avoidance culture such as U.S. participate in a computer-run experiment, in which they take the part of an agricultural consultant making decisions about which crop to plant in drought susceptible areas. Participants choose between crop A, with a higher potential of profit if no drought occurs but no profit if it does, and crop B with a lower but guaranteed profit, regardless of drought. It is predicted that students from high uncertain avoidance culture will choose more conservative option, Crop B, compared to people from low uncertainty avoidance culture, in respect to the same probabilistic information. The number of risk averse errors, defined as choosing a risk-free option when the expected value of the risky option is higher, will be measured. Finally, statistical analysis will be conducted to assess the significance of differences between cultural groups.

The purpose of this study was to use foraminifera to assess the bottom water quality of the San Juan Islands. Foraminifera are microscopic shelled single-celled organisms that live in marine environments. The foraminifera used in this study are benthic and live within or on top of sediments. These organisms form shells of calcium carbonate (calcareous) or agglutinated sand grains, and different species assemblages can be used to determine water and sediment quality. The San Juan Islands have fewer pollution sources than other areas in the Puget Sound. However, the city of Victoria, British Columbia releases raw sewage into Northern Puget Sound, which could affect water quality in the islands. In this study, 26 species of foraminifera were recovered from 20 sediment samples collected by the Washington Department of Ecology in 2002, 2003, and 2012. In each sample, the density of individuals, species richness (number of species in each sample) and number of calcareous specimens showing signs of dissolution (from acidification) were determined. The species richness varied dramatically between samples. Some shallow embayments, including Prevost Harbor, Westcott Bay, and Reid Harbor, contained very few foraminifera. Conversely, several samples within Roche Harbor had a particularly high diversity with up to 19 different species. The samples within the shallow embayment of Roche Harbor contained a high number of partially dissolved calcareous individuals, especially in two species, Elphidiella hannaii (54% dissolved) and Islandiella limbata (up to 25% dissolved). The high species richness in Roche Harbor is an indication of better water quality, however the high number of dissolved individuals implies that the water has a low pH. Acidification may be caused by human recreation in Roche Harbor or by acidity coming from the Pacific Ocean.

Existing literature concerning mental health of the elderly indicates that late-onset major depression or a subset of its symptoms occurs following experiences of later-life events including bereavement, loneliness, new onset disease, and loss of physical ability. Prior lifetime depression can influence presence and severity of later-life depression/depressive symptoms but no work has been done to determine the precise relationship between prior lifetime depression and later depression/depressive symptoms. We will examine adjustable health factors including obesity, smoking status, and level of physical activity and their effects on depressive symptom trajectory, using linear regression to determine significance when compared to trajectory of those of the out-groups of health factors of interest. We aim to utilize a data set of Medicare managed care beneficiaries to determine the role of these health factors on depression trajectory across a two-year period. Preliminary results suggest that adjustable health factors can play a significant role in depression trajectory. These results will inform existing literature pertaining to the elderly of when later-life depression/depressive symptoms occur and how they may be prevented and managed.

Possession Sound is located in the northeast arm of Whidbey Basin in Puget Sound, and is part of the larger estuary system of Puget Sound, with freshwater influence from the Snohomish River. The Snohomish River is the second largest river in the Puget Sound watershed, at 1,730 square miles (USGS). It has many anthropogenic influences, including agricultural inputs from local farms, as well as influence from urban population centers along its banks. It has the second largest wastewater treatment plant-WWPT- input in the region (USGS.) Parameters such as temperature, salinity, dissolved oxygen (DO), chlorophyll, turbidity, nutrient, and plankton abundance would all fluctuate spatially in relation to the Snohomish River. Initial questions about how the Snohomish River influences water chemistry, nutrients, and plankton abundance in Possession Sound led to data collection from March-May, 2015. It was predicted that with proximity to the river, temperature and salinity would decrease, while dissolved oxygen, nutrients, turbidity, chlorophyll, and plankton abundance would all increase. Temperature, salinity, DO, chlorophyll, turbidity, nutrient, and plankton samples were taken along a horizontal transect line from the mouth of the Snohomish River to Whidbey Island. A Conductivity/Temperature/Depth Instrument, or CTD, was used to determine the temperature, salinity, DO, chlorophyll, and turbidity data with depth. A Niskin bottle was deployed at the halocline to collect nutrient concentrations and plankton abundances. In an increasingly urbanized ecosystem, discovering the effect of a river influenced by agricultural and urban practices on water chemistry, nutrient loads, and plankton abundance, could help to determine how these parameters will change in the future, and could fuel further studies on how these changes will affect the ecosystem as a whole.

Prolyl 3-hydroxylation is a rare post-translation modification unique to collagens. The molecular sites and sequence motifs of the modified proline residues, 3-hydroxyproline, have been thoroughly characterized in the major fibril-forming collagens (types I, II, III and V). No function is yet known for the modification although defective prolyl 3-hydroxylation has been linked to certain subtypes of osteogensis imperfecta and more recently high myopia. We investigated a role for 3-hydroxyproline in the proteolytic protection of collagen fibrils against matrix metalloproteinase-1 (MMP1), a major protease involved in the breakdown of the extracellular matrix. Preliminary data from our laboratory supported potentially altered MMP1 cleavage patterns in type I collagen of mouse tail tendon as compared with mouse skin. To eliminate tissue-specific variability, type I collagens were isolated from mouse tail tendons of control mice (P3h2c/c) and genetically engineered mice (P3h2n/n), which are known to have reduced levels of 3-hydroxyproline. Mass spectrometry was used to identify proteolytic cleavage sites and peptide fragments resulting from MMP1 digestion. Under the conditions tested no significant differences were observed between the MMP1 cleavage patterns of type I collagen from P3h2n/n and P3h2c/c mice. These findings suggest that the 3-hydroxyproline alone does not play a major role in protecting collagen fibrils against MMP1 degradation. Further research is needed to understand the functions for the prolyl 3-hydroxylases in regulating connective tissue growth and remodelling, chronic fibrotic disorders, inhibiting tumor growth and metastasis, and the resistance of polymeric collagens in general to proteolysis.

Circadian rhythms are daily oscillations in behavior and physiology that are fundamental to maintaining proper health. Research has shown that these endogenous circadian rhythms can be “entrained” by different cues, such as the light-dark cycle. However, it is unknown whether they can be entrained by fearful stimuli. In this project we test the hypothesis that cyclic fearful stimuli during the night, when nocturnal mice typically forage and feed, can act as an entraining signal to the circadian system. We use a mouse cage divided between a nesting area and a foraging area where the animal needs to retrieve its food from. Within this foraging area mice can receive footshocks, which act as aversive fearful stimuli. We are testing whether footshocks delivered during the nighttime are sufficient to induce mice to shift from their natural nocturnal pattern of foraging and feeding to a diurnal pattern in these behaviors. If feeding and foraging shift to the daytime, as it is the case for rats under similar conditions, we will also test whether this behavioral shift represents the output of a circadian clock entrained by cyclic fear by releasing the animals into constant environmental conditions. Our experiments in mice will open the gate to new experiments to unmask the genetic and neural pathways by which fear and anxiety may lead to changes in circadian activity patterns.

American crows (Corvus brachyrhynchos) are intelligent, communal animals that show a large vocabulary of calls. In this study, we compared crow communication on post-roost aggregations, where crows gather as they leave their roost in the morning, with that on diurnal activity centers, where groups of crows forage during the day. We recorded audio and video on post-roost aggregations and diurnal activity centers, and analyzed the total number of crow calls and the intra-call variation within two call types, short caw calls and assembly calls, using an audio analysis program, Raven Pro 1.5. Intra-call variation was analyzed by measuring the length of syllables, discrete sounds emitted by individual crows within calls, and of gaps, breaks between syllables within calls. We observed variation in the length and number of syllables, and the duration of gaps, in assembly calls and in calls using short caws. Crows made more calls in post-roost aggregations than on diurnal activity centers. Gap lengths in assembly calls were shorter on post-roost aggregations as compared to diurnal activity centers. These findings are consistent with the hypothesis that crows are sharing different types of information in these two contexts, and add to a body of pre-existing literature that suggests that crows use complex vocalizations in a context-dependent manner.

Environmental critical area (ECA) is a precautionary land use term that raises red flags for most land developers in Seattle for good reason. However, ECA sites often present opportunities for community leaders to enhance wildlife habitat and create recreational public space. Nearly all of the undeveloped parcels of land in Seattle have numerous ECA designations due to the glacier carved topography of the Puget Sound region. Many local developers have been improving the means and methods for integrating construction designs with the naturally occurring landscape and these breakthroughs have made it possible to build on ECA sites in a reasonable timeframe at an affordable cost. As the project manager of the Yesler Swamp Bird Blind project I have researched and found the means necessary for completing a feasibility study for the construction of an elevated boardwalk and viewing platform on the shoreline of Lake Washington. My case study research and literature review summarizes fundamentals of low environmental impact design and outlines steps for protecting vegetation, soil, and wildlife habitat during and after a construction project. The primary methods for my case study research involved job shadowing construction managers at the Yesler Swamp and meeting with experienced consultants who specialize in environmental protection. During the case study I uncovered three critical site conditions that must be carefully assessed before designing and permitting a low impact construction project in a riparian habitat. These three conditions include underground peat settlement, surface soil drainage characteristics, and native plant composition.

In natural environments, and symbiotic systems, bacterial horizontal gene transfer is an important means for acquiring new genes. Natural transformation is one method bacteria use to take up free DNA from the environment and incorporate it into their genomes. Some bacteria only take up free DNA that contains specific sequences known as DNA uptake sequences (DUS). In this project, we aim to test the specificity of DNA uptake by Verminephrobacter species, a symbiotic bacterial genus in earthworms. To evaluate the specificity of Verminephrobacter DNA uptake, we will incubate V. eiseniae with free DNA from other Verminephrobacter species with marker genes (antibiotic resistance) incorporated through Tn5 transposon mutagenesis, and test for acquisition of antibiotic resistance. The presence of the new gene in the genome will be confirmed with PCR. A second series of experiments will test DNA uptake from more distantly related free-living Acidovorax species that take up specific DNA from the environment. We hypothesize that V. eiseniae will be able to take in DNA from other Verminephrobacter species but not from the closely related environmental genera (Acidovorax), since transformation is sequence specific and closely related species have more similar DNA. If uptake is detected, as a follow up study the sequences of the DNA around the incorporated genes will be sequenced to discover the possible specific DNA uptake sequences.

Bacterial vaginosis (BV) is the most common vaginal infection among women of reproductive age and is associated with increased risk of premature labor, pelvic inflammatory disease, and both acquisition and transmission of HIV. BV is characterized by a change in the vaginal microbiota: most notably, a decrease in dominant Lactobacillus species coupled with an increase of overall species diversity and increased number and amount of anaerobic bacterial populations. Much of the pathogenesis of BV still remains unclear. For example, administration of antibiotic therapies generally results in alleviated BV symptoms, but recurrence within six months affects 50% of women treated. Our study attempts to understand such recurrence by characterizing a possible extravaginal reservoir for bacterial vaginosis-associated bacteria (BVAB) in the oral cavities of male sexual partners through a cross-sectional and longitudinal study. Gingival swabs were obtained from 23 men whose partners were assessed for BV at baseline. Gingival swabs were also obtained on daily basis from another cohort of men who kept diaries outlining their sexual activity. Our study used species-specific quantitative PCR on bacterial DNA extracted from gingival swabs to test for oral BVAB colonization in male partners. From these assays, we found prevalent oral colonization of BVAB in men and that such colonization is highly dynamic over time. Establishing the oral cavity in male partners as an extravaginal reservoir for BVAB would have implications for BV treatment and may suggest cunnilingus as a possible mode of BV recurrence.

Attention-Deficit Hyperactivity Disorder (ADHD) affects approximately 5-7% of children worldwide. ADHD is associated with traits such as hyperactivity, impulsivity, and a lack of focus, and appears to be influenced by various genetic and environmental factors. Among these factors, ADHD has a significant genetic contribution with heritability estimates suggested to reach ~75%. One gene commonly associated with ADHD and ADHD-like traits is the dopamine receptor D4 gene (DRD4); with the 7R allele of the 48 bp repeat polymorphism in DRD4 associated with ADHD patients. In addition, DRD4 7R alleles are highest in frequency in human populations that have experienced long-term and short-term migration. Although the 7R allele in DRD4 is associated with ADHD and migration, the role of ADHD, and particularly the genetic signatures of ADHD, on human migration patterns is currently unknown. We hypothesized that ADHD-related alleles are commonly associated with highly migratory populations. Firstly, macro-migration measures, distances in kilometers populations have migrated from an ancestral origin, and micro-migration measures, whether a population is nomadic or sedentary, were determined for 52 worldwide populations from the Human Genome Diversity Project (HGDP). Then, ADHD genes were prioritized from ADHDgene, a database that compiles all genetic association studies on ADHD. Once these ADHD genes were identified, genotypes for these 52 worldwide populations were obtained from publicly available sources of genome-wide data and allele frequencies for each population were computed. To test for the association between the ADHD-related alleles and migration, a linear regression was performed while controlling for neutral genetic population substructure. We expected ADHD-related alleles to be highly correlated with migratory distance and nomadic populations. This research provides insight into the maintenance of ADHD in worldwide human populations and the influence of neurodevelopmental traits on populations that have undergone rapid migration throughout human history.

I researched the plausibility of using remote sensing to find evidence of sub-surface processes occurring in the Columbia River estuary. An estuary is a place where fresh water from rivers mixes with saltwater from the ocean. It acts as a bioreactor, transforming organic matter and nutrients before they enter the ocean. The Columbia River estuary is a salt-wedge estuary, meaning that as the river empties into the Pacific Ocean – and because fresh water is less dense than ocean water – the fresh water floats on top of the salt wedge that tidal forces are pushing into the river. My research aimed to find remote sensing data, ranging from satellite images to camera images from the experiments that showed surface manifestations of the salt wedge pushing into the river. Using data collected by the autonomous underwater vehicles (AUVs) used in the experiments conducted in the last five years, I aimed to find periods of overlap between AUV data and the remote sensing data. Comparison of these data then allowed me to check whether the remote sensing results were supported by the AUV results. These data can be used to improve numerical models which will assist in plotting where the salt wedge is occurring in the estuary. Understanding the interactions between the salt wedge and fresh water is important not only for the marine ecosystem – as many species rely on these estuaries for food and as places to nest and breed – but also for the human community as people rely on these estuaries for recreation, food, and jobs. Comprehending the physics of why and how these salt wedges occur can allow insight into how the nutrients and organic matter in the buffer zone are processed, which in turn will affect the marine life feeding on those resources and thus human communities.

Heart failure (HF), the inability of the heart to keep up with its work load, affects about 5.1 million people in the U.S. and only half of people diagnosed with HF survive more than five years. Past studies of successfully transplanting human embryonic stem cell derived cardiomyocytes into damaged myocardium of monkeys have demonstrated it as a viable approach to improve survival in HF. Despite decreasing the scar size and making new cardiac muscle cells, there was an increase in arrhythmias possibly due to immaturity of the injected cells. Therefore, success of transplanting cells into damaged myocardium depends on the ability to produce more mature cardiomyocytes. I am currently investigating the ability of 2-deoxyadenosine triphosphate (dATP) to reduce maturation times in induced pluripotent stem cell derived cardiomyocytes (iPSC-CM). The rate-limiting step in dATP production is enzyme ribonucleotide reductase (R1R2). Past studies on the overexpression of the subunits encoding for R1R2 resulted in a 40% increase in shortening magnitude and 80 % increase in the shortening rate in adult rat cardiomyocytes. I am testing the hypothesis that the use of an adenoviral vector that overexpresses-R1R2 will decrease maturation time in human iPSC-CMs. The following maturation factors are being used to determine decreased maturation time: cell size-mature cells are larger and more elongated; switch from fetal slow skeletal troponin I (ssTnI) to adult isoform of troponin I (cTnI); alignment of myofibrils, magnitude of contraction, and mitochondrial number. We have shown that treatment of iPSC-CM with viral vector that overexpresses R1R2 increases contractility. Now, we are assessing the ability of this treatment to decrease maturation time. By demonstrating dATP as a successful agent in decreasing maturation time, the HAMM Lab hopes to be able to use transplantation of more mature cells as an effective means to improve morbidity and mortality in heart failure.

Vocal communication among birds has been studied across many species. American Crows (Corvus brachyrhynchos) may employ “information centers” to collect information about foraging sites, yet how the information is gathered, actively or passively, is not well understood. To explore the information center hypothesis, we collected audio and video recordings of American Crows in the field over a three-month period, in 8 different locations across two distinct behavior contexts: post-roost aggregations (potential information centers) and diurnal activity centers (foraging sites). Recordings were analyzed using Raven Pro 1.5, a sound analysis program, and the types of calls used in each location type were compared. There was no significant difference in the average number of call types used in post-roost aggregations versus diurnal activity centers, but there were significant differences in use of the alert, assembly, and rattle calls, all calls that were previously categorized in literature. We also saw significant differences in calls not previously named or categorized, which we named the short caw and extended caw. All of these calls were shown to be used more at post-roost aggregations than diurnal activity centers. These data are consistent with the hypothesis that American Crows may use post-roost aggregations as information centers.

Of continuing interest in the field of synthetic biology is the nature of inter-protein interactions and the optimization of protein-protein interfaces. To better understand these phenomena, we combined directed evolution with computational design to optimize the dimerization of an engineered ligand-binding protein, DIG 10.3. Our goal was to couple the stability of DIG 10.3 to the binding of digoxigenin. DIG 10.3 binds digoxigenin and exists natively as a stable homo-dimer. We used unstable mutants of DIG 10.3 and identified pairs of proteins that are dependent on the presence of digoxigenin for dimerization. These mutant proteins are unstable and rapidly degraded. However, when digoxigenin is present, the DIG 10.3 mutants bind to this small molecule and are stabilized, forming dimers. Additionally, mutations to the ligand-binding site have been identified that change the ligand-binding specificity of DIG 10.3 from digoxigenin to progesterone. Using these variants, we created a system to combine digoxigenin- and progesterone-binding proteins in order to identify hetero-dimer pairs optimized for response to digoxigenin, progesterone, or both. This approach is theoretically modular and could be applied to downstream applications like the construction of biosensors and gene circuits.

The human pathogen Helicobacter pylori's only natural niche is the human stomach, and chronic infection with H. pylori is associated with the development of gastric ulcers and cancer. The bacterium has a characteristic helical cell shape that has been shown to be essential for flagellar-based motility through the gastric mucus and subsequent colonization of the underlying gastric cells. In our study, we are interested in a particular non-enzymatic protein Csd5, that is a part of a complex helical shape-defining machinery. When the Csd5 protein is not expressed in H. pylori, we observe a complete loss of helical morphology i.e. straight, rod-shaped cells. Within this Csd5 protein is a small protein domain called the SH3 domain that is ubiquitous throughout prokaryotic and eukaryotic domains of life, where it commonly mediates interactions between proteins. A specific region within the SH3 protein domain called the RT-loop, contains a highly invariable or conserved amino acid reside arginine (R), and a moderately conserved amino acid residue threonine (T). We hypothesized that the two conserved amino acid residues within the RT-loop are important for the shape defining function of the Csd5. By replacing these two conserved amino acid residues using site directed mutagenesis, we observed two distinct changes to the helical cell morphology of H. pylori. From these preliminary results, we conclude that the two conserved amino acid residues are important for the normal function of the Csd5 protein in vivo, and using a combination of biochemical and molecular approaches, we will continue to study the mechanistic roles of the RT-loop in the Csd5 SH3 domain. 

There has long been an interest among climate scientists in glacier trends as indicators of climate change. In particular, glacial length is a commonly reported measurement because it is easy to measure and provides a summary of a glacier's response to climate perturbations. However previous research has shown that kilometer scale changes in length can be expected as a result of random climate variability even when there is no significant climate trend (Roe & Oneal 2005, Roe 2011). In light of this, we have elected to study mass balance data as a more direct measure of the interaction between glaciers and climate. We have applied a number of conventional statistical measures to every available glacier mass balance record longer than 5 years that we are aware of. In particular, we focus on characterizing the variability and trends observed in the mass balance record. Furthermore, we investigate the significance of mass balance trends and constrain confidence intervals on our statistical results. We report all of our statistics as a resource for future research.

Our research investigates the visual system of rhesus macaques. One of the experimental approaches we use is to deliver genetic material into visual neurons using modified viruses. A significant obstacle to this approach is that not all viruses work equally well in all animals. My project is designed to determine the relative infectivity of each virus as it applies to individual animals and thereby guide virus selection. We use nine different forms (or serotypes) of adeno-associated virus (AAV) in our experiments. One factor that likely affects the infectivity of each viral serotype is host immunity conferred by circulating antibodies. Different animals in the lab have been exposed to different antigens through the natural course of events or by experimental manipulation. Consequently, different animals express different antibodies, rendering them immune to some viruses but susceptible to others. To determine the infectivity of each type of virus, we are producing the nine serotypes of AAV, each containing a gene that encodes for a green fluorescent protein. We intend to infect cultured cells with these viruses in the presence and absence of antibody-containing serum extracted from monkey blood. By comparing the levels of fluorescence in the presence and absence of serum, we can determine the relative infectivity of each type of virus for a particular animal. After having determined the serotype of virus which is best suited for each animal, we can use that serotype for future experiments and thereby improve our ability to manipulate the visual system.

The American Crow (Corvus brachyrhynchos) is a highly intelligent bird that uses complex vocalizations as a means of social communication within different contexts. We compared how these birds communicated in two distinct behavioral contexts, pre-roost aggregations and diurnal activity centers. Pre-roost aggregations are large groups that crows form prior to flying to their roost at night, while diurnal activity centers consist of foraging groups. Here, we located four sites for each behavioral context and collected a total of ten video and audio recordings of American Crows near Bothell, WA USA. The videos were used as a control for counts obtained in the field and the audio recordings were analyzed for number of calls and number of types of calls using Raven Pro 1.5. Crows made more total calls and more types of calls in pre-roost aggregations than diurnal activity centers. These findings remained significant after controlling for the number of crows present within each context. It may be possible that pre-roost aggregations are used as information centers where crows communicate about the locations of good foraging areas discovered while at diurnal activity centers. Assessing the structure of vocal communication in the American Crow will not only help us better understand complex animal communication, it will also give us insight into the nature of intelligence across species.

The reduction of cardiac function can be caused by many different factors including myocardial infarction (heart attack) and genetic cardimyopathies. There are limited pharmacological treatments to improve heart performance in these conditions. However, the Heart and Muscle Mechanics (HAMM) laboratory is investigating a novel gene therapy, which can improve cardiac performance. Studies have shown that dATP (2-deoxyadenosine triosphosphate), a naturally present nucleotide used by the cell during DNA synthesis, works as a better energy substrate for muscle contraction than ATP. dATP allows for an increase in contractile capacity due to faster acto-myosin cross-bridge turnover. The dATP is produced by the enzyme Ribonucleotide Reductase (RNR) during DNA synthesis and cell division. By overexpressing RNR, more dATP can be produced, allowing for contractions with more force. Previous studies have shown that increasing the expression of RNR in cardiac cells increases contraction both in animals and in cell culture. In the current study, we are investigating if increased expression of RNR can improve cardiac function in a genetic model of dilated cardiomyopathy caused by mutation in tropomyosin (D230N). Mice with dilatd cardiomyopathy will receive a systemic injection of an adveno-associated virus serotype 6(AAV6) that restricts the expression of RNR to cardiac cells. In vivo and ex vivo cardiac function will be tested in these mice in weeks following injection. Tissue samples will later be harvested for DNA and protein analysis to determine uptake of the virus. Through qPCR and western blotting, levels of RNR present in the mice before and after being treated with the virus will be compared to determine the effectiveness of uptake of the vector. Successful uptake of the virus and overexpression of RNR in mice models could lead to the development of a new gene therapeutic treatment for those suffering from dilated cardiomyopathy. 

Myocardial infarction (MI), more commonly known as a heart attack, results in a temporary loss of blood flow to the heart which leads to tissue ischemia and necrosis. An event such as this commonly leads to further complications such as heart failure and arrhythmias that can severely affect quality of life or cause death. An MI occurs most commonly due to an occlusion of vascular branches that supply oxygen and nutrients to various parts of the heart, causing tissue death and adverse remolding. Current methods of regenerating cardiac tissue using stem cells lack proper vascularization. Improving this aspect of stem cell treatment has important clinical implications that can lead to a more sustainable solution. Here, we developed a novel in vitro microvascular platform that mimics luminal vascular architecture and function. This platform is generated from a biocompatible polymer, polydimethylsiloxane, using a soft lithographic method. A single perfusable channel is formed by first placing an acupuncture needle through the well of the platform and subsequently lining the well with type I collagen, an extracellular matrix component. Optimization was performed to provide consistent cell seeding of the channel by controlling flow rate and improving the design of the apparatus. Human umbilical vein endothelial cells (HUVECs) were seeded in the channel and cultured for 3 days. We observed retention of structural integrity of the channel and cell survival. On-going studies include evaluating the robustness of the HUVEC lined channel with a cardiomyocyte embedded extracellular matrix, as well as cell-cell interaction under flow. The study is expected to not only provide a base platform to develop preventative or therapeutic medicine for those affected by MI, but also as a tool for disease modeling of other pathologic phenomena.

Most cases of human bacterial infection begin with the immobilization of free-floating planktonic bacteria to host cells—such as human endothelial cells. In this biological context, flow is often pulsatile rather than constant due to normal cardiac rhythm and peristalsis. Many types of bacteria bind in pulsatile flow environments, yet the effect of pulsatile flow on shear-enhanced bacterial adhesion is still not understood and has not been thoroughly investigated. Realizing the effect of pulsatile flow on bacterial adhesion could potentially inform the structure of medical biomaterials or anti-adhesive therapy. Using E. coli expressing the adhesin FimH as a model organism, this research aims to develop and validate a dynamic simulation elucidating how the velocity and frequency of pulsatile volumetric flow in a bacterial microenvironment modulates binding. Current data suggest that pulsatile flow mediates robust bacterial adhesion in situations where high constant flow would be too harsh for bacteria to adhere. Additional data demonstrates that increasing the frequency of low-to-high flow alternation results in increased adhesion, providing diminishing returns past one hertz. Emergent properties of the simulation were tested by microfluidic flow chamber experiments, whereby the predictive properties of the model are validated. Because FimH-expressing E. coli display a unique binding pathway that begins with surface rolling followed by immobilization, attempts are being made to understand whether the simulation can be applied to other clinically relevant bacteria that bind in the same way. A generalized and validated computational explanation of this adhesive behavior could enable researchers and clinicians to better understand how pulsatile flow environments might contribute to the initiation of various types of endothelial infection—such as endocarditis.

Over 90% of cancer-related deaths are caused by the migration of invasive cells along bundles of extracellular matrix (ECM) fibers. The ECM itself is an important part of a human cell and composed of many proteins, such as collagens and fibronectins. Two factors affect the migratory behaviors of cells: the stiffness of the ECM and the activity of the PI3K pathway. These two factors are essential in our quest to understand the process of cancer cell migration. First, as the stiffness of a cell's ECM increases, many biochemical changes take place, such as increased cell division and stronger anchorage to neighboring cells. Most importantly, cell migration is substantially increased. Second, the PI3K pathway plays a crucial role in cell migration. Two prominent cell lines in the pathway are the PTEN knockout and PIK3CA knockin breast epithelial cell lines, which are known to decrease and increase the activity of PI3K respectively. In order to explore these relationships we employed novel nanoengineered platforms with defined topographies and breast epithelial cells with or without a cancer-inducing mutation of the PI3K pathway. Ultraviolet light assisted capillary force lithography was used to fabricate nanoengineered cell culture platforms with arrays of grooves-ridges. Additionally, polydimethylsiloxane was used to vary the stiffness (100 kPa to 800 kPa)  of these platforms. The migration velocities of each cell line on substrates of varying stiffnesses were quantified with ImageJ and MATLAB. Results show that there is statistical significance between the migration velocities on patterned and unpatterned substrates of low stiffness (100 kPa), indicating that the stiffness of a cell's ECM does indeed play a role in cell migration. 

Salt wedges form when an underlying layer of saline water intrudes up an estuary due to the density difference between the incoming freshwater and the saltwater body it is being discharged into. Salt wedges are an important avenue of research in environmental fluid mechanics because they have the ability to profoundly influence estuarine dynamics. One important consequence of salt wedge formations is that they force faster moving freshwater away from the riverbed, shielding the sediments under the salt wedge from being uplifted and exiting the estuary. Simultaneously, the freshwater forms a thin, buoyant plume that extends into the saltwater basin. While previous research has tackled the dynamics of both the salt wedge and the plume separately, taking their interface at the mouth of the estuary as constant, this project examines the interrelation of the two in a combined system. In order to explore how these two formations modify the fundamental dynamics, a simple numerical model was setup and calibrated. This model was used to investigate the effects of input parameters, such as estuary slope and discharge, on the salt wedge and thus establish relationships that can better predict its behavior. A greater understanding of salt wedge behavior will enable engineers and policy makers to make more informed decisions regarding salt wedge estuaries, such as the Duwamish River Estuary, where predictions of salt wedge intrusion would allow for a more targeted dredging scheme.

As the demands of health care services increases, the use of biomedical mixers also increases. However, using a traditional vortex mixer (electric motor runs with a cupped rubber piece that oscillates rapidly) can damage biomedical samples because of the violent vibration. My purpose is to investigate a method that mixes biomedical samples gently without damaging cells, corpuscle, etc. As a result, less samples are required from patients and more accurate test results can be achieved. In my research, there are two main components: 1) using permanent magnets to deflect artificial cilia, creating a vortex in the mixing channel, and 2) using the theory of chaotic mixing to improve performance and guide the design process. Cilia can be found on cell’s surface and beat in coordinated waves in human body. My artificial cilia were made from a silicone elastomer with iron microparticles. I used additive printing to build a specific wheel from polylactic acid which can hold permanent magnets. To power the system, I programmed a motor using LabView. As the wheel rotates, the cilia deflect and generate vortices in the channel. The performance depends on many factors such as the length of the cilia and size of the channel. When both cilia beat together, stagnation is observed in the channel. Using an alternating pattern motivated by chaos theory, the stagnation points can be eliminated.

I am conducting the groundwork research to propose a design intervention for a shoreline street end at 12th and Elmgrove Street, located along the Duwamish Waterway in the South Park Neighborhood. South Park is plagued by environmental justice concerns including pollution and major industrial zoning which conflicts with the residential areas. It is in great need of additional and improved green space, which is a key step in improving the quality of life for its residents and workers. Shoreline street ends are community assets that have been designated by the City of Seattle as special rights-of-way that should be preserved and improved for public use. Being on the waterway, this street end is critical for enhancing ecological function. It can be re-designed to filter stormwater before it goes into the river and enhance habitat for migrating salmon. Through my design studio with my partner Colleen Brennan this past winter quarter, we have come up with a potential design that responds to a key community concern: providing educational opportunities. The educational theme for this site is green storm water infrastructure. On this site, water movement plays an important role in showing and explaining different processes such as water filtration and estuarine habitat. My research involves the collection of both primary and secondary data. The primary research includes site analysis that consists of careful experiential and physical observations, photo documentation, and sketching. I have been looking at the materiality of the site, tidal changes, human circulation, opportunity for views, topography, water drainage, and neighborhood context. Other primary research methods include community meetings with neighborhood activists and residents. I will continue to refine this design proposal in response to the community’s needs with the goal that it will be a demonstration project for the other seven street ends along the Duwamish Waterway.

Honeybees (Apis mellifera) use a caste system to organize individuals into different roles. The queen is the only bee in the hive to ingest royal jelly and the only fertile female. Many sterile workers perform different jobs in their short lives, moving from larvae-nurse to keeping the hive clean to foraging for food. At each stage the worker bees learn a new task. Such different phenotypes and behavior would indicate a difference in gene expression in the brain. In a population of organisms that are genetically similar, these changes in gene expression must be explained in chemical modifications of the genome, or the epigenome. Thus far, much attention has been given to DNA methylation as an epigenetic feature known to be associated with variation in gene expression. Royal jelly, however, contains butyrate, a histone deacetylase inhibitor, which suggests that histone tail modifications could be a major mechanism for regulating gene expression in queen bees. To further investigate the role of histone modification in determining honeybee caste division, we are performing chromatin immunoprecipitation with massively parallel DNA sequencing (ChIP-seq) to profile key chromatin modifications: H3K4me1, H3K4me3, H3K27ac, and H3K27me3. H3K27ac and H3K27me3 are histone modifications that have opposing effects; where H3K27ac marks active regions of the genome, and H3K27me3 is indicative of repressive chromatin. H3K4me1 is associated with enhancer regions and H3K4me3 is associated with promoters. We anticipate when using ChIP-seq to map chromatin modification in the brains of queen and worker bees, we will observe key differences in genome regulation and be able to identify genes important to honeybee behavior. As an initial step toward achieving our goal, we describe the chromatin state of brains from forager honeybees.

This project comprises of a series of sketches each detailing aspects of a specific streetscape in the many neighborhoods of Seattle. In examining the quality of streetscapes, different parts of neighborhoods and even whole neighborhoods in general can vary significantly due to differing needs addressed. For instance, a good streetscape for pedestrians cannot be replicated everywhere if the primary goal is to move as much traffic as possible in an industrial area. This project documents these contrasts in design philosophies expressed throughout the city. Also, major changes are happening in the city, primarily in those walkable places, whose ideals have again come into fashion. This project was conducted on foot, each sketch informed by a walking person’s analysis. Drawing from the work of Victor Steinbrueck, who sketched parts of Seattle in seminal works both in 1962 and 1973, this project seeks to document this Seattle’s time of change, consistency, improvement, and loss.

Viruses are the most abundant microorganisms found in the ocean and are important in maintaining marine food webs. Viral diversity is immense. Within this diverse group of viruses are the single-stranded DNA (ssDNA) viruses, many of which infect marine eukaryotic organisms. Unfortunately, very little is known about the environmental distribution and diversity of these phytoplankton pathogens. To help understand the diversity and distribution of the ssDNA viruses, I constructed phylogenetic trees using the capsid and replication-associated genes for the ssDNA viruses. These phylogenetic trees were used to identify viruses in the marine environment. I looked at virus communities from four locations off of the Washington coast. I hypothesize that the ssDNA virus communities will vary between these location because of differing host communities. Ultimately, this will help to understand the diversity of ssDNA viruses and their distribution in the ocean.

Promoting increased contraction in cardiomyocytes is a potential therapy for diminished heart systolic function due to problems such as myocardial infarction or genetic cardimyopathiese. The primary biological substrate used to initiate actin-myosin mediated contraction of sarcomeres within cardiac muscle cells is the molecule adenosine triphosphate (ATP). It has been shown that an alternative substrate 2-deoxyadenosine triphosphate (dATP) can significantly increase cardiomyocyte contractility. Overexpression of the enzyme ribonucleotide reductase (R1R2), the rate-limiting step in de-novo dNTP synthesis, increases the intracellular dATP levels. dATP is also able to diffuse across gap junctions and increase contraction in neighboring cells. The next step in this research is to engineer human embryonic stem cell cardiomyocytes (hESC-CMs) that globally overexpress this enzyme. These cells can be transplanted into diseased hearts to provide a source for increased dATP. This was done initially by engineering an adenoviral vector containing the gene for overexpresssion of R1R2 and infecting the cells. The virus was initially characterized in human embryonic kidney cells. Virus uptake was confirmed using fluorescence. Western blot analysis indicated successful overexpression of the R1R2 enzyme with a 16 fold increase of R1 and and 28 fold increase of R2 expression in nucleofected cells. In the future, hESC-CMs will be infected to characterize how the overexpression is affecting the cardiomyocyte contractility. I then plan to engineer and characterize an immortal line that constitutively overexpresses R1R2. These cells would then be grafted into injured myocardium and aid the tissue to regain normal function. This research advances these therapies one step closer to the clinical setting.

At the Ocean Research College Academy (ORCA), an early college program at Everett Community College, students engage in boat-based research utilizing data that is collected monthly at five locations incorporating chemical, physical, geological and biological parameters. Previous studies demonstrate the presence of heavy metals in sediment and marine invertebrates. Human derived activities contribute heavy metals, including zinc, lead, copper, cadmium, mercury, and arsenic to the estuary and the greater sound. From previous work in the spring and summer of 2014, heavy metal concentration in benthic marine invertebrates was investigated. Based on the background sediment data, the levels in marine invertebrates, specifically ghost shrimp, Neotrypaea californiensis, and various crab species, showed a trend of bioaccumulation of heavy metals. Copper levels in the sediment were under 15 mg/Kg while the ghost shrimp levels ranged from 200-400 mg/Kg. Zinc levels in the background sediment were under 50mg/Kg while ghost shrimp levels were above 80mg/Kg. Arsenic also bioaccumulated in the ghost shrimp tissue to 10 mg/Kg while the sediment levels were under 5 mg/Kg. In order to better understand the impact and significance of the bioaccumulation, further testing of specific internal organs from two invertebrates species was conducted. For both the crab and shrimp, the focus was on testing the hepatopancreas, an organ that regulates heavy metal levels in crustaceans, for evidence of elevated levels of heavy metals. Tissue from the hepatopancreas from both organisms was analyzed under a microscope to visualize any damage to the tissue and compared to pictures of unaffected tissue. Crabs exoskeleton and muscle tissue were also isolated and tested separately.

Communities at risk of natural hazards may misinterpret the risk information they receive. Consequently, such misunderstandings may influence a community’s estimated likelihood of a natural hazard and their willingness to take precautionary action. For example, past research has found that flood-vulnerable communities fail to recognize that the return period term, “100-year flood”, refers to a flood that has a one percent chance of occurring in any year (Ludy & Kondolf, 2012). Instead, they interpret it as a deterministic forecast for a single flood over that time period. This study examines two factors that may affect people’s estimated likelihood of a flood occurring in the near future and their interpretation of risk expressions: 1) the way risk information is communicated and 2) the recency of a previous flood hazard. In a computer-simulation task, participants were given flood risk information and were tasked with minimizing a flood-vulnerable firm’s losses over forty years by purchasing flood insurance. Participants were told that the area experienced either “10-year floods” or floods at a “10% chance per year”. They were also told that the most recent flood happened either last year or ten years ago. We anticipated that participants who received the “10% chance per year” term would have estimated flood likelihoods more congruent to the flood risk term (10%) than participants who received the “10-year flood” term. We also anticipated that participants in the return period condition who were told a flood happened recently would estimate lower flood likelihoods than participants who were told a flood happened some time ago. These results suggest that flood risk communication should avoid terms such as “100-year floods”. Additionally, this study offers insight into the factors that influence how communities estimate the likelihood of future natural hazards. These results can better inform future natural hazard risk communication.

Adolescence is a critical period of risk for the emergence of both internalizing psychopathology and risky behavior. This period of development is also characterized by the differing developmental trajectories of the brain regions associated with self-regulation. Throughout adolescence, brain regions associated with emotions and motivation, including subcortical and limbic structures, develop at a faster rate than the region primarily associated with self-control, the prefrontal cortex. The greater levels of risk-taking observed in adolescence may be driven by enhanced motivation for immediate reward coupled with delayed ability to regulate behavior in accordance with long-term goals. Accordingly, this study will examine the influence of individual differences in reward sensitivity on levels of risky behavior and internalizing behavior. In addition, gender will be examined as a possible moderator of the influence of reward sensitivity on internalizing symptoms and behavior problems, as gender differences have been observed in the development of risky behavior and internalizing symptoms across adolescence. We hypothesize that both risky behavior and anxiety symptoms will be associated with heightened reward sensitivity, and that poor inhibitory control will differentiate adolescents who engage in risky behavior from those with anxiety problems. In contrast, we expect that greater levels of depressive symptoms will be correlated with blunted response to reward. We investigate these questions in two cross-sectional samples: a multisite urban sample (N=287) of 16 and 17 year olds as well as a local community sample of (N=60) 13-16 year olds. Participants completed self-report questionnaires assessing internalizing symptoms and behaviors such as substance use, risky driving, and risky sexual behavior. Two tasks were used to assess behavioral response to reward, the delayed discounting task and the piñata task. Results may provide critical evidence on the role of reward sensitivity in the development of psychopathology in adolescence.

Axions are a type of particle that could make up the mysterious dark matter in our universe. The Axion Dark Matter Experiment (ADMX) at the University of Washington (UW) is searching for axions with masses around 3 ueV using a microwave cavity, but higher mass axions could be dark matter as well. We constructed an axion experiment called "Orpheus" using a novel technique involving a Fabry-Perot resonator in spatially varying magnetic field to search for axions at higher masses. We were able to show a sensitivity to dark matter axions competitive with large laser experiments and have a path to upgrade the experiment to explore new dark matter axion parameter space.

Collectivism is defined as a set of feelings, beliefs, and behaviors related to the concern for others (Hui, 1988). Previous studies indicate a negative correlation between collectivism and drinking, suggesting collectivism serves a protective function against drinking (Foster, 2014). Furthermore, collectivism is associated with attitudes that favor family, elders, and community over personal pursuits (Foster, 2014). Such community-oriented attitudes are likely to contradict with heavy drinking, a generally self-indulgent behavior. The theory of planned behavior postulates that attitudes about a behavior are influential in predicting the occurrence of that behavior (Ajzen, 1991). The current study examines whether attitudes toward risky drinking mediate the relationship between collectivism and drinking among high school seniors. We hypothesized lower levels of collectivism to correlate with increased drinking and related consequences. We predicted collectivism level would be related to drinking behavior through attitudes, indicating mediation. Four individual measures: the Individualism and Collectivism Scale (Triandis et al., 1986), four items assessing attitudes toward drinking (Baer, 1994), the Daily Drinking Questionnaire (Collins, Parks, & Marlatt, 1985), and the Rutgers Alcohol Problem Index (White & Labouvie, 1989) were taken from 1095 high school seniors. We examined correlations among collectivism, attitudes toward drinking, number of drinks per week, and alcohol-related consequences. All correlations were significant in the expected directions, except for finding no correlation between collectivism and alcohol-related consequences. Multiple regression analyses and a Sobel test were performed to test whether attitudes serve as a mediator of the relationship between collectivism and drinking. Results indicated that the association between collectivism and drinks per week was partially mediated by one’s attitudes toward drinking (Z = -3.53, p<0.001). These findings suggest that level of collectivism influences attitudes, which, in turn affects drinking behavior. Interventions aiming to moderate student’s attitudes toward alcohol may strengthen the protective function of collectivism on drinking behavior.

Cardiovascular disease (CVD) is currently the leading cause of morbidity and mortality across the world. Although much research has been conducted in the treatment of CVDs in the past decades, there are still numerous problems that need to be addressed, including the development of effective therapeutic drugs and accurate disease models. Current screening methods, however, simply utilize two-dimensional culture of cardiac cells. These environments do not accurately simulate the complex, three-dimensional structure of cardiac tissue. Cardiac muscle function is dependent on the highly anisotropic nature of its structure. Recent efforts have shown to produce engineered muscle tissue possessing a well-defined anisotropy that is capable of being independent within each layer. This accomplishment was made possible through the use of thermoresponsive nanofabricated substrata (TNFS) that contain an array of ridges allowing for cells to linearize themselves in a single, uniform direction. Subsequently, layering multiple single cell sheets using TNFS allows for the cells of each layer to maintain their initial anisotropy and be minimally influenced by direct contact with other sheets. This research seeks to further develop the fabrication of tissues in a high-throughput manner with greater reproducibility. These tissue samples can then be applied to the study of disease modeling, cardiotoxicity, and the effects of drugs on the three-dimensional structure of cardiac tissue. This researcher utilized 3D printing, C2C12 mouse myoblast cells, and magnetic nano-particles to develop a method for the creation of multi layered, uniformly aligned, tissue for disease modeling. The nanoparticles non-specifically bind to cell membranes allowing for cells sheets to be transferred and stacked in a high throughput manner using a magnetic drive. This method will then be utilized to develop a multi-layered, uniformly aligned, tissue for disease modeling of DMD cardiomyopathy.

Recent developments in vaccine research for the human immunodeficiency virus (HIV) have demonstrated the importance of the mucosal immune system to protect against HIV infection. The vaginal tract is the only route of mucosal vaccination that induces both local and systemic immunity against HIV infection. However, current vaccine delivery systems are unable to overcome the mucosal barrier and promote cellular uptake of vaccines. The proposed project will develop a novel multifunctional delivery system of vaccines (E-PS) by coating an amine-functionalized polystyrene nanoparticle (NP) (200 nm) with a pH-sensitive polymer, Eudragit S100. Eudragit S100 is unique polymer that will dissolve in physiological pH but will remain precipitated in vaginal pH of 4. E-PS is hypothesized to penetrate the mucosal layer and upon dissolution of Eudragit, E-PS will be uptaken by immune cells. A modified emulsion method was used to fabricate E-PS. Dynamic light scattering assessed the zeta potential and hydrodynamic diameter. E-PS exhibits a neutral zeta potential of +7.13 mV in pH 4 for mucus-penetration, whereas uncoated polystyrene NP exhibits +40 mV. Mucus-penetration of E-PS will be assessed using NanoSight Tracking Analysis. E-PS is anticipated to have a diffusion coefficient for mucus penetration at least 1000-fold greater than NPs. Upon removal of the Eudragit, the positive (+40 mV) zeta potential of the NP was exposed for improved cellular uptake. Cellular uptake studies will be conducted using a dendritic cell line and vaginal epithelium cell line. Flow cytometry and microscopy will be used to examine cellular uptake and cytotoxicity. Finally, the functionality of E-PS will be assessed in vivo using a mouse model previously established. This work would represent the first of a mucus-penetrating nanoparticle with improved cellular uptake for mucosal HIV vaccine delivery. This project will further demonstrate the feasibility of a mucosal HIV vaccine by combining the core NP with vaccines.

Anemia is a blood disorder where the body fails to circulate an adequate amount of oxygen-rich blood, which can lead to a range of health complications. One of the main causes of anemia is the lack of red blood cells, which is produced in a process called erythropoiesis. Erythropoiesis occurs within a bone marrow niche that is supported by the bone marrow vascular system. Even though the process of erythropoiesis has been studied extensively, very little is known about how erythroid progenitors interact with the bone marrow vascular system, and some studies have shown that endothelial cells play a role in stimulating erythropoiesis in vitro. I aim to investigate the effect of erythroid progenitors on the vascular endothelium by co-culturing them in a 3D microvessel model. The erythroid progenitors are expanded from whole peripheral blood mononuclear cells (PBMC). They are then embedded in a collagen matrix, which is moulded into a device containing microfluidic channels lined with endothelial cells to form a structure that resembles the microvasculature. This system mimics the in vivo arrangement within the bone marrow, where the cells can interact via both adhesive proteins and soluble factors. The behaviours of the cell populations in this system are quantified via confocal microscopy to determine the effect of one on the other during erythropoiesis, as well as in the presence of different cytokines or disturbances. The proposed 3D culture system is the first step in the construction of an in vitro model for studying anemia pathogenesis and a screening platform for anemia treatment. Additionally, it could also be used to investigate methods to improve in vitro production of red blood cells.

Throughout the corpus of extant Greek literature, particularly in poetry, a number of references to feet and ankles appear in the context of abduction, rape, and heroic death in battle.  In my project, I sought to determine the reason for, and meaning of, such occurences.  I began by seeking out specific instances of the terminology involved in such references, then analyzed them in context of the surrounding passages, taking into account both the narrative content of each passage and the meanings and connotations of the words composing it.  By so doing, I sought to piece together the connotations that such mentions of feet and ankles would convey to the original Greek-speaking audience.  In the end, a nuanced picture began to take form: each term, such as an adjective meaning "slender-ankled," had a distinct set of connotations, and the author selected which term to employ based on factors such as the gender and circumstances of the character that he or she was describing.  I concluded by demonstrating that, depending on these factors and on the specific term employed, such terminology could convey acute vulnerability, relative safety, or an ambiguous mixture of the two to the original audience.

Affordable 3D-printers have created a new toolset that may help improve the fabrication and function of a variety of commercially available orthoses that are designed to help improve movement for individuals with disabilities. The purpose of this study is to improve the design of wrist-driven orthoses (WDO) by reducing their complexity and fabrication time and increasing their availability through the use of 3D-printing technology and open-source designs. WDOs are prescribed for patients with incomplete spinal cord injury (SCI) at the 6th cervical (C-6) level who exhibit strong muscle activity in the wrist extensor muscles and little to no mobility in the fingers. By utilizing wrist motion, the device assists in opening and closing the hand and imitates a grasp-and-release motion, which consequently enhances performance of tasks of daily living. The study consists of two phases, which are still in progress. The focus of the first phase is to evaluate the potential of 3D-printing technology for orthotic fabrication from the perspective of practitioners and students who currently fabricate orthoses, including orthotists, therapists, and physicians. Feedback from practitioners will be used to further improve design and fabrication methods. The second phase focuses on evaluating the device from the perspective of end users. The specific aims of this phase are to receive feedback from end users and test the effectiveness of the device in improving hand function by using a variety of functional tests, electromyography, and ultrasound.

This study aims to examine the relationship between children’s emerging morphological knowledge, as evidenced in their generation of novel “invented” morphological forms and “morphological awareness” among 5th and 8th grade students. “Morphological awareness” is the understanding of units of meaning that can be removed or added to root words to change their meaning. “Invented” morphology, as defined by McCutchen and Stull (2015), are errors in forming derivations of morphologically complex words that result from adding inappropriate affixes to roots. Tyler and Nagy (1989) observed that acceptance of morphological inventions during reading seemed to increase until 6th grade and taper off by college, the next age examined by Tyler and Nagy. The present study aims to extend the Tyler and Nagy study by examining inventions generated by students and the relationship of such inventions with a measure of students’ own writing. The data were previously collected as part of a larger study looking at the relationship of morphological awareness to writing skill. Participants included 146 5th grade students, and 132 8th grade students from the 2012-13 school year, and 86 5th grade students, and 58 8th grade students from the 2013-14 school year, with 52% female. We hypothesize that grade, gender, and whether a student produced morphological inventions will predict morphological control in a more extended sentence-writing task. We plan to examine this effect by using a 2 (Grade: 5th vs. 8th) x 2 (Inventor: no inventions vs. one or more inventions) x 2 (Gender: girl vs. boy) ANOVA design with successful, accurate morphological changes in a sentence-combining task as the dependent variable. The results of this study may provide a more nuanced understanding of students’ emerging morphological knowledge as it relates to their writing than previous research.

This poster project will present a diachronic study of French ethnography, literature, and theatre which examines French conceptions of the landscape and living space of Native Americans from early colonial times to the Romantic era. In particular, this project will show how French ethnographers and writers make use of a stereotypical, mythologized image of Native Americans and their relationship to their environment in order to conceive of Native American society as a counter-example which serves as a philosophical or pedagogical tool to contrast with French society. The finished poster will diachronically convey important historical phases in which distinct French conceptions of the Native American landscape emerged, showing a progression of the French imaginary’s construction of Native American ecological identities. The poster’s visual organization will convey this diachronic structure, beginning with an examination of how French Catholics qualified the Native American wilderness as a place of sin and inferior social and economic development in the early colonial period. The poster will proceed with an analysis of how secular, anti-monarchical French writers and artists during the Enlightenment reimagine the Native American landscape in order to engage in social critique, contrasting the uninhibited, edenic landscape of the Native Americans with the artificially-constructed, highly cultivated environment of the French courts. The end of the study will show how French Romanticism responded to this Enlightenment conception of the Native American landscape by reasserting the importance of Christianity and reinterpreting the Enlightenment’s positive conception of the uninhibited, natural quality of this landscape as emerging from a profound, spiritual unity with nature. By examining the subjective claims and values of literary, artistic, and ethnographic texts, the project will show how collectively shared ecological values contribute to shifting essentialist notions of race in the context of French and Native American colonial encounters.

What role do Black women have in American Politics? How is this different from the role of Caucasian women? Is there a necessity for Black Feminism? How has Black Feminism and its application transformed over time? Traditional Feminism recognizes the economic, social, and political oppression of women throughout society based off of gender. Contrarily Black Feminism recognizes the intersectionality of race and gender, which influence the disenfranchisement of women throughout the economic, social and political spheres. Using a historical analysis to describe the characteristics of Black Feminism, I will illustrate a differentiation from Traditional Feminism while analyzing pioneers of Black Feminism and contemporary Black Feminists. A historical analysis will contribute to the interrogation of predisposed notions with regard to Black Feminism, while reasserting the necessity of Black Feminism over time. The double minority status which recgonizes the heightned discrimination of Black women because of their gender and race within the U.S provides additional explanations regarding changes in the degrees by which Black Feminism has been applied throughout time. The double minority status also supports the necessity of Black Feminism due to the egalitarian values instilled within Traditional Feminism which dismiss the participation of Black Women. The experiences of oppression based off of race and gender is central in the understanding of Black Feminism historically. This analysis of Black Feminism will create a foundation to explain the conceptualization and relevance of Black Feminism chronologically.

The ENCODE Consortium is an NIH-funded effort, consisting of around 50 groups across the country, whose objective is to find all the functional elements (genes, enhancers, promoters, etc.) in the human genome. This is done by performing genome-wide assays on hundred of human cells. The Noble lab at the UW has produced a software tool called “Segway” that performs unsupervised machine learning analysis to aid in human interpretation of the resulting data. The goal of my project is to migrate our Segway into the DNAnexus cloud computing platform and then use the software to annotate the active genomic elements in a diverse set of cell types. The current analytical challenge is to correctly model the different types of genomic data to produce an accurate annotation. We aim to model the genomic elements as a mixture of Gaussians. Segway takes as input real-valued data tracks defined across the genome and outputs a partition of the genome, where each segment receives an integer label such that genomic segments with the same labels have similar values in the data tracks and, hence, correspond to the same biological function. For our analysis, we choose cell types that have at least two of three distinct types of assays (DNase-seq, histone modification ChIP-seq and transcription factor ChIP-seq). These assays were chosen because they describe the regulation of genes. The project will produce functional annotations of 92 cell types.  These annotations will be made publicly available on the UCSC Genome Browser.  In addition, the project will migrate the Segway annotation software to the DNAnexus cloud computing platform.

I am interested in exploring the link between perceived access to youth sports and collegiate athletic career choices. Does student athletes’ perceived spatial and economic access to youth sports within respective native neighborhoods and communities affect which collegiate sport is chosen to pursue? If so, how? This research is important because the psychological and social benefits of youth sports should be available to all children, although such opportunity for participation may not exist for youth from underserved communities. Youth sports are becoming increasingly privatized, inaccessible and unaffordable for low-income students which has a long-term effect on youth’s pursuit of sports at higher levels, collegiate and beyond. I will use the methods of interviews, voluntary surveys and local sports facility data analysis to analyze my question. Through the intersection of qualitative and quantitative research, I hope to capture the systemic impacts on individuals within the community concerning youth sports opportunities. I anticipate my research results will indicate that students from underserved communities will perceive less spatial and economic access to particular sports (golf, gymnastics, tennis) and more access to other sports (soccer, basketball), impacting student athlete’s choice of sport at the collegiate level. If such results are found, I would urge for the implementation of affordable and accessible youth sport programs within underserved communities in order to provide the opportunity to participate for all children, regardless of socio-economic status.

Current stab resistant materials are heavy, bulky, and expensive. My teammates and I set out to create a lightweight, cost-effective material that would give a higher degree of flexibility while protecting against stabs typical of hostile prison and mental institution environments. Cloth failure due to puncture does not represent the cleavage of the individual fibers, but rather the windowing, or separation, of fibers to allow penetration. We investigated impregnating nylon with a non newtonian shear thickening fluid (STF) in order to prevent windowing. The STF is a suspension of nano-sized silica particles in polyethylene glycol (PEG). The fluid, by itself, reacts to an imposed pressure by transitioning from a colloidal suspension to a state of flocculation resulting in an instantaneous separation of colloid and medium. This phenomenon is demonstrated by imagining the difference between walking on dry sand vs. wet sand. By impregnating the cloth with a silica based STF the shear force of a spike attempting to separate the nylon fibers will be met with a thickening reaction of the STF. This thickening will prevent the fibers from spreading and allow them to absorb more of the impact energy. We tested our STF fabric by creating a spike drop apparatus, allowing us to mimic typical stab conditions and measure the total energy dissipation. Through trial and error we were able to determine a 1:1 ratio of silica to PEG provided the highest energy dissipation to the lowest weight gain. The perfection of this lightweight, affordable stab resistant material can compete with top shelf STF Kevlar and will prove to have applications in more areas than just body amour.

There is a farm worker labor movement taking place in the state of Washington led by Triqui, Mixtec and Mexican activists as well as other farm workers in the Skagit Valley. These activists are changing the face of field work by confronting the Western belief of who works in the fields and who has the capability of standing up for themselves to secure suitable and just labor treatment. Drawing on Marshall Ganz’ work on California farmworkers, I focus on the significance of leadership, organization and strategy to explain the success of the contemporary movement in the Skagit Valley. Additionally, I argue that Indigeneity has played a critical role in the movement’s ability to gain visibility and exert pressure on powerful economic interests. I will also explore what this farm worker movement means to the campesinos (farm workers) all over the state.

The Snohomish River estuary is located on the Everett, Washington waterfront, at an interface between freshwater originating from Cascade mountain range runoff and saltwater from the Pacific Ocean. There, marine environments are blended with industry and suburban living along the Snohomish River and the banks of Possession Sound. These anthropogenic influences introduce harmful heavy metals into the ecosystem, including: copper, a common building material; arsenic and cadmium, both of which are found in diesel fuel. Studies demonstrate that long-term heavy metal exposure can be harmful to both marine organisms and humans. These heavy metals are toxic to marine organisms and inhibit genes that metabolize carcinogens in humans. The Ocean Research College Academy (ORCA) has been collecting benthic sediment samples and analyzing heavy metal content in Possession Sound since 2009. 10 sites with varying anthropogenic influence were tested for these heavy metals. Sites along the Everett waterfront were focused on due to the high influence of industry, such as the Burlington Northern and Santa Fe Railway (BNSF) tracks, and a high population on the bluffs above the waterfront tracks. It is hypothesized that copper, arsenic and cadmium will be found in higher concentrations at sites with the closest proximity to shoreline due to anthropogenic influence.

Disorders that impair memory affect millions, but our physiological understanding of memory processes is limited. Research designed to uncover the neurological bases of different memory types contributes to the broader goal of addressing these impairments. Over the past several decades, a delayed match to sample (DMS) task has been used to observe short term working memory because its simple, versatile design allows for use across species (e.g. rats, mice, primates). Image-based DMS experiments in primates reward monkeys for holding a sample picture ‘in mind’ and signaling by releasing a touchbar when the sample picture is repeated within a sequence of pictures, ignoring other stimulus repetitions. A recent publication by Wittig and Richmond questioned the strategy that monkeys use in performing this task. Using a novel training procedure and DMS task design, they argue that monkeys do not selectively hold the sample stimulus in memory, but instead nonselectively memorize all stimuli and use a recency rule to respond to stimuli that match those presented early in the sequence. They also argue that monkeys do not respond only to stimuli that match the sample, but also respond indiscriminately to repeated non-match stimuli. The Wittig and Richmond results challenge previous use of DMS to assess working memory. Here, we examined results from 9 monkeys performing the traditional version of the DMS task. The data show that while some passive, non-selective strategy was employed in early learning stages of DMS, as the task difficulty increased (by including more intervening non-match distractors), monkeys used this strategy less in favor of selective memory. In these later sessions, we identified a decrease in error-rate in trials with repeated non-match stimuli. We suggest that any of several adaptations made by Wittig and Richmond to the standard DMS task might have caused the monkeys to adopt non-selective memory strategies.

Our research focuses on developing new methods of producing composite structures, namely locally-resonant granular metamaterials and phononic crystals, particularly at the microscale. Phononic crystals require strict periodicity, a trait which necessitates precise structure and organization between the individual component spheres. This required precision is difficult to attain through methods of self-assembly, as these methods result in many defects in the crystal structure. This restricts some of the possibilities for testing the samples, as the imperfections in periodicity produce scattering effects and make reaching meaningful conclusions more difficult. The alternative method of syringe printing allows for the placement of individual spheres into specific configurations. Here we explore how to utilize syringe printing to produce ‘perfectly’ periodic granular crystals. The process of syringe printing is achieved through the use of a syringe pump paired with a syringe of a very small volume (we use a 1 microliter syringe). This makes it possible to dispense spheres individually, while a two-axis motion stage beneath the syringe tip allows for microsphere positioning. This setup allows for a considerably more refined ability to create granular crystals on a microscale, introducing the possibility of future studies of the dynamics of locally resonant metamaterials containing particles of multiple materials. Thus far, we have verified that we can control the dispensing of several spheres through the syringe; going forward, we hope to be able to isolate a single microsphere and are currently developing methods for precise spatial control of sphere placement.

North American river otters (Lontra canadensis) are an apex predator in the Snohomish River estuarine system, eating fish, crab, other small crustaceans, birds, and occasionally ingesting anthropogenic products. Students at Ocean Research College Academy (ORCA), an early college high school academy through Everett Community College, have been conducting undergraduate research on the local river otter population for the past three years. Since the river otter study began in 2012, over two-hundred non-invasive fecal samples from three latrine sites near the Snohomish River, a contributing inlet of the Possession Sound, have been collected and dissected, supplying information on the river otter’s diet, diseases, genetics, and behavior. Through the dissections of our scat samples, students are able to create observational hypotheses on the diet of the river otters, such as spatial and temporal studies on the fish and crustacean consumption. Recently, anthropogenic products, such as Styrofoam, have been noted and further studied to pinpoint where upriver these plastics are being sourced. The scat samples also supply information on potential diseases that the river otters may have. For this branch of the research, samples are sent to local veterinarians and analyzed for diseases such as Amebiasis, Coccidiosis, and Giardiasis, ailments that are found in the scat of domestic animals upriver. It is also through our scat samples that Polymerase Chain Reaction (PCR) tests are conducted to determine the genetic diversity and gender of the local population. Finally, camera traps record river otter’s presence and behavior. These data can be connected to tide stages, time, and water chemistry. All aspects of this research project are imperative because little exploration has been done on this population of Snohomish River otters and an unawareness of their diseases and potential anthropogenic impacts could endanger the population.

The perception of color by the brain depends on the nature of light reaching the retina and on the effects of contextual stimuli on that light. Bright and dark objects are assigned color differentially by the brain, characterized by long-wavelength lights that appear yellow when brighter than their surroundings but that change to a different hue, brown, when they are made darker than their surroundings. To explore the temporal and spatial aspects of this context effect, we asked how quickly the change from yellow to brown occurs once the surroundings brighten and whether the latency depends on the proximity of surround stimuli to the target. Nine observers viewed a 2° diameter foveal target disk surrounded by a large, bright (124 cd/m2 CIE 1964) annular surround with a gap of 0°, 0.25° or 1° between target and surround. Both surround and target stimuli were presented for either 1 s or 30 ms. Observers adjusted the luminance of the target to the highest light level at which the target appeared pure brown, with no trace of yellow: the higher this “brown boundary”, the more effective the surround is at inducing brown. Observers also performed this same task but for the “yellow boundary,” adjusting the luminance to the lowest light level at which the target appeared pure yellow with no trace of brown. We found that bright surround stimuli raise boundaries for both brown and yellow. These induction effects occur rapidly (within 30 ms) but generally strengthen slightly with longer-duration (1 s) stimuli. The degree of proximity effects across all conditions varied considerably among observers but without any clear differences across conditions. We hypothesize that the brightness of the target, determined by stimulus duration surround stimuli configuration, has a similar effect on the light levels at which both brown and yellow appear.

In eukaryotes, the ribosomal RNA genes (rDNA)—encoding the RNA component of ribosomes—are present as tandem repeats in the genome. The yeast Saccharomyces cerevisiae carries around 150 copies of the rDNA genes located on chromosome XII. rDNA has been shown to be an important contributor to genome replication and replicative lifespan. Previously we performed a screen in the Brewer/ Raghuraman lab to investigate genetic regulators of rDNA copy number using a single gene deletion collection. However, chromosome XII size phenotypes of deletion mutants from this genetic screen failed to match the phenotypes of the de novo knockouts of the same genes. Additionally, previous experiments from our lab also showed that when wild type cells are taken through the lithium acetate transformation process they frequently show chromosome XII size variation. We therefore hypothesize that the transformation process itself induces changes in the number of rDNA tandem repeats. I have successfully replicated our lab’s previous findings that cells taken through the transformation protocol have changes in their rDNA size. I am testing which specific process in the protocol causes the variation. I have harvested cells at each step of transformation (i.e., heatshock and freezing) to see which particular action is causing the variation. Moreover, preliminary data shows that the A364a background strain tends to have more variation in rDNA size than the S288c background strain leading me to question whether the magnitude of change in rDNA size is dependent on the strain background. Intriguingly, I also found that transformation processes can change the sizes of other chromosomes along with chromosome XII. Therefore, we are interested to ask if transformation is affecting overall genetic stability as well. From the results of this study, I hope to gain insights on the causes of change in rDNA copy number and potentially genome instability.

Although previous studies have found that commuting increases our salivary cortisol (stress) levels, little is known about how it affects salivary testosterone, the sex hormone associated with aggression. Even less is known about the impacts that different modes of transportation may have on these hormone levels. My project therefore aims to 1) identify how commuting influences both salivary cortisol and testosterone patterns and 2) compare the salivary cortisol and testosterone levels of commuters when they drive to when they take the bus to determine how different modes of transportation affect these hormone levels. I predict that 1) commuting (in general) will have a noticeable impact on salivary testosterone and cortisol levels, 2) that testosterone levels will be highest shortly after commuting for both busing and driving, and 3) that salivary cortisol and testosterone levels will be higher when study participants drive than when they take the bus. Twenty male and female participants will select one day to commute home via bus and in the same week one day to commute home via driving their own car or vehicle. Shortly before and after both of these commutes, each participant will complete mood surveys and stress scales and collect 2 mL of saliva specimen. Cortisol and testosterone will be assayed in the specimens using enzyme immunoassays. Bus commute day hormone levels will be compared to car commute day hormone levels. Findings from this research project could have implications for future research on stress and transportation which may encourage commuters to use one mode of transportation instead of another since long-term exposure to elevated stress levels has been shown to be bad for one’s health.

This research aims to investigate whether Escherichia coli bacteria isolated from water and crow droppings from the wetlands of Bothell Campus demonstrate antibiotic resistance. For investigational purpose, 18 feces and 10 water samples were collected from the wetlands over a period of two months. The feces samples were plated on Eosin Methylene Blue (EMB) agar and colonies with blue green sheen were isolated as putative E. coli. Water samples were filtered and the E. coli isolated on m-ColiBlue as blue colonies, were verified on EMB agar. Colonies from EMB agar were isolated as pure cultures on Tryptic Soy agar for use in antibiotic testing by the Kirby Bauer method, and for DNA extraction for genotyping. In total, 8 antibiotics were examined. Preliminary results with 36 feces isolates have shown that 11/36 (31.6%), 10/36 (27.8%), 4/36(11.12%) and 3/36(8.3%) were resistant to streptomycin, ampicillin, tetracycline and nalidixic acid, respectively, while 4 of 24 (16.66%) water isolates, respectively, were resistant to streptomycin and ampicillin. 15 of 60 (25%) isolates from water and feces samples were resistant to multiple antibiotics. All isolates were sensitive to ciprofloxacin and enrofloxacin. Interestingly, 5 isolates from different feces samples shared the exact same pattern of antibiotic resistance with 6 water isolates, suggesting, some of the contamination of the wetlands by E.coli may be due to crow excretions. These isolates will be targets for source tracking studies based on the sequence of the malate dehydrogenase (mdh) gene. This gene is a suitable candidate for such studies because it is not prone to horizontal gene transfer. It consists of a highly variable region flanked by conserved regions and thus contains sufficient polymorphism for host specific differentiation of E.coli strains. The results from this study will inform students and staff who engage in various activities in the wetland about its safety.

When we step into a dark room, it takes time for our eyes to adapt. This adaptation process increases the range of light levels our retinas are able to encode. However, two different photoreceptor types (rods and cones) are responsible for light transduction, and they transmit signals through distinct pathways that converge at the output of the retina. The goal of my work is to characterize the independence of post-receptoral adaptation between these two pathways, i.e. exploring whether adaptation of rod signals affects cone adaptation, and vice versa. To this end, I have designed a psychophysical experiment to probe perceptual sensitivity in human observers. Observers perform a task to perceptually match the intensity of two flashes of light presented in their periphery. A persistent light called an adaptive field is then shown in the region where one of these flashes occurs to adapt the retinal circuitry at that location; the intensity of light that corresponds to a perceptual match for an observer thus gives a measure of the gain suppression brought about by the adaptive field. By designing an adaptive field that selectively activates cones, and then another adaptive field that activates cones to the same degree but additionally activates rods, I am investigating how adaptation in one pathway affects the sensitivity of the other pathway; truly independent cone adaptation should be unaffected by additional rod content in the adaptive field. By corroborating my results with electrophysiology recordings from primate retina, my work helps to elucidate the precise locations of post-receptoral adaptation, offers insight into biological mechanisms of parallel processing, and draws connections between neuronal mechanisms and perception.

Arterial hemostasis, or blood clotting, is mediated through the large plasma protein von Willebrand Factor (VWF). The A1 domain of VWF facilitates platelet adhesion in blood vessels through binding to the glycoprotein (GPIb) domain of platelets, a unique process characterized by activation of VWF by high levels of shear stress. This project focuses on how mutations in VWF A1 domain affect function in other parts of VWF, in particular those causing increased enzyme-directed proteolysis at the A2 domain. Our method uses molecular force spectroscopy instrumentation to stretch recombinant VWF, requiring an anchor strong enough to immobilize VWF under high force conditions. I have expressed VWF with a specific covalent bond-forming protein called HaloTag, yielding results that strongly support ligand-receptor binding. A1 optimized with the covalent tether can then be bound to a tag on the other end, and in the next steps I will begin collecting data from A2 cleavage in the stretching experiments. Results gained from these experiments can suggest a genesis for the VWF fragments observed in patients with the bleeding disorder Type 2B Von Willebrand Disease (VWD), which is caused by mutations in VWF that counterintuitively can increase VWF-platelet binding. Since different mutations have different effects on A2 cleavage, protocols developed in this research could potentially be used clinically to classify VWF mutations in Type 2B patients.

Recent reforms in the collection of eyewitness evidence have included the suggestion that the investigator administering identification procedures should not know which individual in the photoarray is the suspect. This recommendation for a “blind” administrator stems from the belief that administrators with this knowledge may implicitly or explicitly influence the witness’s decision. As an alternative to this “blind” administration, policy-makers have proposed a “blinded” method in which a knowledgeable administrator is unable to know which photos are being shown to a witness at any given time. In our study, we have evaluated how this “blinded” method compares with “blind” and “non-blind” administration. In addition, we have compared the interactive effects of these procedures within sequential (each photograph shown one-at-a-time) and simultaneous (all photographs shown at the same time) presentations. Participants viewed individuals on a computer and then interacted with a blind, blinded, or non-blind administrator. During this interaction, participants viewed 4 simultaneous or sequential photoarrays testing their memory for the individuals they viewed on the computer. To date, data has been collected from 256 participants. Analyses examined the relative influence of blind, blinded, and non-blind administration on eyewitness accuracy and choosing rates in simultaneous and sequential lineups. Using ROC (Receiver Operating Characteristics) analysis, we assessed the extent to which administrator knowledge and line-up presentation type influences the witnesses’ discrimination accuracy and response bias (i.e., willingness to make an identification). Consistent with past research, we hypothesized that blind administration will reduce the likelihood of administrator influence on witness decisions. Furthermore, we believe that the blinded administration produces similar outcomes resulting in improved discrimination accuracy for blinded administration compared with non-blind administration. Results of this research will inform policy decisions regarding what procedures are best for collecting eyewitness identification evidence.

Surface EMG (sEMG) signals have been used frequently in clinical applications involving neuromuscular disorders. Recently, sEMG sensors have emerged as a viable, non-invasive human-machine interface for control of active prosthetic devices. In particular, it is starting to be used widely in robotics control. The goals of this research project are to i) develop an sEMG-based activation signal whose amplitude can be controlled by the intensity of the captured sEMG signal, and ii) develop a robotic hand in order to investigate how the activation signal can be used to achieve simple robotic hand motions, e.g. grasping. The development of the activation signal is based on the fact that an EMG sensor can detect the intensity of a muscle contraction. With this fact, users will be able to deliver their intention by varying the muscle intensity. First, the raw EMG signal was obtained from the user’s muscle activity, filtered by an electrical circuit, and then amplified with a LabView program; the resultant signal was obtained as a DC voltage. Second, the robotic hand was constructed using publicly available part models, which were 3D printed and assembled. It was driven by servo motors powered with a simulated Pulse Width Modulation (PWM) output, capable of performing the required grasping motion. The next step will be to generate a PWM output using the EMG signal, which will then control the motion of the robotic hand. In future work, hand orientation and location will be considered as one of the control variables.

Trapped ion quantum computing uses laser cooled trapped ions as qubits (a quantum equivalent of classical bits) to store quantum information. The combination of barium and ytterbium ions is a promising candidate for this experiment because of their similar atomic masses which allows for efficient sympathetic cooling, and the convenient structure of those ions’ atomic energy levels. Each ion species needs different energy to change their electron state. The goal of my project is to improve ion trapping mechanism by building a stable power supply for the ovens that heat metallic barium and ytterbium to create neutral atoms which are later ionized and trapped. Stable oven temperature is needed to controllably evaporate these metals while maintaining a stable pressure of the system and minimizing light radiated from the oven itself. Therefore, I designed a circuit to make a stable dual channel total power regulating supply for the system which accommodates changes in the oven’s resistance. The main circuitry provides up to 20W to each oven. An additional digital interface allows the experiment control system to switch the ovens on or off as needed. A display of power supply’s current, voltage and power is included in the circuit to provide more information about the heating mechanism. This low cost, yet stable power supply is expected to increases the efficiency of the experiment and enhance the overall ion trapping process.

Forty percent of the world’s population relies on mountain rivers for fresh water. A very important metric in understanding and accurately quantifying these water flows is snow water equivalent (SWE). Snow water equivalent is a measure of the water content in a given snowpack. In snow hydrology research, snow depth and air temperature are more easily and reliably measured than SWE. Accurate estimates of SWE can help forecast how much water will flow through mountain streams during a given time period, which in turn helps estimate how much water an area will have throughout the dry season. I have developed an empirical model that estimates SWE as a function of air temperature and snow depth through time. The model was designed for maritime mountainous climates, as data collected from Olympic National Park and Snoqualmie Pass was used in developing the model. When snow falls, it has a snow density that can be estimated as a function of air temperature, which in turn can be used to estimate SWE from observed snow depth. In general, colder air temperatures result in lighter and less dense new snow. After falling, the new snow undergoes a phase of compaction, where snow settles over time, resulting in increased snow density. Over the snowfall events of a given winter, there will be both compacted snow and new snow at the same time. This model estimates SWE by predicting the SWE of new snow based on conditions when the snowfall occurred before adding it to the SWE of snow that has already been compacted. Preliminary results at one snow study site indicate accurate real-time SWE values for any point throughout the year.

We have designed and 3D printed quasi-1D and 3D periodic lattices with negative Poisson’s ratios (NPR). NPR materials contract laterally under compression. The lattice is based on the conventional 2D NPR re-entrant hexagon. The 3D version, like its 2D counterpart is anisotropic, and can be constructed from a unit column tiled along a hexagonal grid. Both periodic structures and those with NPR have been shown to rapidly and selectively attenuate certain frequency ranges of acoustic vibrations, corroborating our preliminary simulations. This makes periodic and NPR structures particularly adept at filtering or damping vibrations in solids and structures. The lattices will be excited by an actuator on one end, and the frequency response will be measured at the opposite end by a force sensor. We expect that the frequency response will be highly dependent on the lattice geometry: relative strut lengths, unit cell size, and characteristic angles, with specific frequencies rapidly attenuating where the structure shows local resonance outside the axial direction. Achieving the ability to predict these dependencies will allow us to produce a 3D acoustic filter, selectively suppressing acoustic frequencies. This technology has application in sensitive electronics and mechanical devices where vibration can cause output signal noise, or in transportation, where excessive vibration can cause discomfort or damage. 

This study develops methods to determine the pathogenicity of bacteria isolated from the wetland waters and crows feces within the UW Bothell campus. This emanated from the need to determine whether the American crow (Corvus brachyrhyncos) has a role in the epidemiology of diarrheal disease along with its significant contribution to the high level of fecal coliforms in the stream water that runs through the crow roosting area. Modified from previous studies, we developed protocols to culture Escherichia coli and Campylobacter, both are known to be pathogenic and present in crow feces, isolate DNA from cultures or samples, and perform PCR/qPCR to detect virulence genes. We found that the virulence genes eae and rfb that are necessary to cause diarrhea were absent in a representative number of E. coli strains isolated from the water samples and the fecal samples. The virulence genes flaA and cad in Campylobacter species were detected in fecal samples (77.8% and 73%, respectively) and in water samples (75% each). Our results suggest that the Campylobacter isolated from wetland water and crow feces are potentially pathogenic. However, our results are not conclusive and more sample and virulence genes specific to extraintestinal pathogenic E. coli, need to be screened in order to accurately assess the pathogenicity of these bacteria.

The collection of eyewitness evidence by police is a process that must strike equilibrium with the complex processes of human behavior, all in the pursuit of seeking truth from memory. Our research investigates the influence that the administrators of these identification procedures may have over witnesses, and how that affects their accuracy and decision confidence. Jurors rely on decision confidence when assessing a witness’s credibility, and considering past research that suggests how confidence can be manipulated by the actions of an administrator, we are researching how administrator knowledge (across blind, blinded, and non-blind administrators) might influence a witness. Recently, in an effort to reduce the potential influence of administrator knowledge on witness decisions, there has been push to have “blind” administrators that do not know the identity of the suspect administer the identification procedure. However, due to concerns of law enforcement officials over utilizing blind administrators, this recommendation has not been widely embraced. Thus, policy-makers have suggested the use of a “blinded” method, in which a non-blind administrator conducts the procedure, whilst their knowledge of the suspect’s identity has been forcibly reduced by the constraints of the procedure. In our research, we seek to test this method empirically. To date, data has been collected from 256 participants who viewed individuals on a computer, and were later tested for their memory of those individuals with four simultaneous (all members presented at one time) or sequential (members are presented one at a time) photo arrays. We used Juslin, Olsson, and Winman’s (1996) calibration technique to evaluate how administrator knowledge influences the confidence accuracy association, and if blinded administration reduces that influence. Results will inform our understanding of the efficacy of blinded administration as a method of preventing administrator influence.

To cope with the forces of wave activity, organisms living in the intertidal have developed a variety of mechanisms to stay in place. Metridium farcimen (white plumose sea anemone) are capable of withstanding wave action by strongly attaching to solid substrates. Through observation it was thought that M. farcimen attach themselves to substrate by secreting an adhesive glue from the basal foot. Limpets, another intertidal invertebrate has been documented attaching to substrate by utilizing the force of suction. This research explored the possibility of suction and glue working in unison to attach M. farcimen to substrate. Given that the force of suction is reliant on the formation of an area of sub-ambient pressure, suction cannot occur on a surface with holes. Therefore our experiment had two groups. One group was comprised of M. farcimen attached to acrylic glass with holes, and the remaining group was comprised of M. farcimen attached to acrylic glass without holes (control). The specimens were removed from the substrates using a clamp attached to a mechanical testing machine. The machine recorded the pull off force (N) required to remove the anemone from the substrate. This measurement and the area of each basal foot was used to calculate tenacity (kPa). Specimens attached to holed substrates averaged tenacities of 5.48 kPa ± 2.31 whereas the mean tenacities of specimens attached to solid substrates was 9.36 kPa ± 5.35. These data imply that suction is a component of M. farcimen attachment. M. farcimen attachment site selection behaviors were also examined. When given the option of attaching to acrylic glass substrates with and without holes, M. farcimen showed no preference for either substrate. This research provides insight on the diversity of intertidal invertebrate attachment mechanisms and how these mechanisms influence behavior. 

Ice sheets are formed in areas with a net accumulation of snowfall leading to a compaction of buried snow, turning the snow into something glaciologists call firn and eventually ice. The pathways connecting the pores within the firn eventually close off during this process as more snow accrue. As a result, gases that were otherwise diffusing in these pathways become trapped in air bubbles. Ice cores in dry snow zones can provide a unique record of Earth’s past climate history and atmospheric composition for up to 800,000 years. However, in order to interpret the ice-core data accurately and efficiently, scientists must take many other variables into consideration. The firn physics group has developed an open source Community Firn Model at the University of Washington to analyze firn densification and gas transportation processes. As an application for this model, we have calculated the spatial distribution of depth, age, and porosity of the firn at the close off zone (where all air bubbles are effectively closed off). To achieve this, we used (Jason Box)’s datasets for the average temperature of Greenland over a 173 year period and we further looked at the snowfall accumulation rate to process these data through the firn model. The results are mapped for straightforward interpretations and are critical to a better understanding of past climatic changes and future predictions. Knowing the depth-integrated porosity will allow mass balance studies to be done more accurately, and knowing the close-off depth and age will provide guidelines for future core sites for the investigation of pre-industrial levels of trace gases.

Project 8 is an experiment using a new technique, Cyclotron Radiation Emission Spectroscopy (CRES), to measure the mass scale of the neutrino. Currently, the setup is focused on demonstrating the energy resolution of this method by studying single-electron radiation from Krypton 83m. This experiment requires the coordinated and simultaneous control of many instruments using a slow control system. I will discuss the design, deployment, and operation of this modular, cross-platform system for the Project 8 experiment, how it is used in calibrating and characterizing the resolution of the CRES technique, and the future opportunities for controlling a full-scale neutrino mass experiment.

The model organism Caenorhabditis elegans experimentally exhibits several well-defined behaviors when put into a controlled environment. Using a biophysical model of neuro-sensory integration built on experimental findings of neuron conducatances and the resolved connectome developed by Kunert et. al., we explore the bifurcation structures that drive a behavior known as an "omega turn." We start by simulating the circuit presented in Gray et al. to see what kind of low dimensional bifurcation structures exist within the model. Using the known structure of forward motion we can project this new behavior into the reduced, known, space and see that this new structure behaves consistently with how an omega turn should be represented. We found a unique bifurcation structure within this circuit, a bi-stable point when attempting to activate the neurons experimentally related to inducing omega turns. Running simulations with various perturbations from this condidtion, we map out the separatrix to give us a better understanding of what drives the system from this bi-stability into one of the two stable solutions. This is significant because one question about the numerical model was whether or not it could display behavior that in experiments is induced by chaning the environment of C. elegans. We see that even without inducing these environmental conditions, the model is robust in characterizing these behaviors with only stimulus of the key neruons in the relevant circuit. 

The sequencing of the human genome revealed that there is substantial variation between the DNA sequences of different individuals. On average, two human individuals differ at approximately one in one thousand nucleotides. However, the consequences of that genetic variation are still largely unknown. The advent of high-throughput sequencing allows the characterization of phenotypic effects at the molecular level, ranging from quantifying mRNA expression levels to characterizing DNA-protein interactions. We focus on the molecular phenotype of chromatin accessibility. Because most DNA is tightly packed around proteins called histones, DNA must be unwound from the histones in order to interact with the cellular machinery. The regions of DNA that are unwound from histones are called accessible, and these regions can be identified using a high-throughput DNA sequencing experiment. We looked at how regions of accessible chromatin vary across 70 individuals of African ancestry by using publicly available DNase I hypersensitivity data. We used a cross-validation-like approach to maximize our sensitivity to regions of open chromatin while minimizing false positives across individuals. Using the patterns of variation seen, we hope to gain insight into how natural selection shapes patterns of accessible chromatin. Moreover, we will characterize how variability in chromatin accessibility correlates with functional features of the genome, such as distance from genes, and the impact of DNA sequence variation on chromatin state variation.

A positive sense of control is vital to an individual’s psychological well-being. A negative sense of control or lacking control has been shown to induce stress, depression, impairments to performance, hostility, and perceptual changes. It has been suggested that when an individual is in a state of lacking control they will first try to generate control objectively and if that fails they will try to regain control perceptually. This perceptual control comes in the form of illusory pattern perceptions in the immediate environment. Illusory pattern perception is defined as the identification of a coherent and meaningful interrelationship among a set of random or unrelated stimuli. Past studies have shown that individuals who have an induced sense of lack-of-control will see more illusory patterns in a range of modalities including: pictures, natural objects, and stock market information. In our experiment we induced a sense of lack-of-control in subjects and tested the notion of increased visual pattern perception using digitized static pictures and also tested for increased illusory numeric pattern perception using a series of numbers. We found no significant difference between groups for illusory pattern perception in either modality. We also tested to find if detection rates (Hit/False Alarm) changed among groups for real pattern recognition. We found that those in the lack-of-control group were not significantly better at detecting real patterns than the control group for numeric stimuli. The significantly high hit rate and the significant interaction between stimuli and hit suggests that the two tasks have different levels of difficulties.

It is known that representations of electromagnetic plane waves (EMW) are difficult for students to understand; examples of electromagnetic waves are radio waves, visible light, and x-rays. Student responses to a question that focuses on using only an EMW and wires to light a lightbulb, no batteries or outlets involved, where examined. The question requires students to understand the 2D representation of an EMW as a 3D image, and know how the representation changes over time. In studying student responses to the question, one type of logic was identified in student reasoning that causes confusion based on the given representation. This poster will discuss implications from student responses to this question and provide an alternate phrasing that might impact student responses and understanding of electromagnetic plane waves.

Antibodies play a key role in imaging, diagnostics and therapeutics. Antibodies are large molecules, often proteins, produced by our immune system that recognize and bind to specific targets. It is our goal to create an antibody-like switchable recognition protein that can be triggered to bind and release on command. This protein can be created by mutating and/or adding parts to an existing protein that would act as a scaffold for our design. FimH, found on the end of bacteria fimbria, is an ideal protein scaffold because it can be triggered by force to have a significant allosteric conformational change that increases its binding affinity to its ligand. In recent experiments, it has been found that inserting a sterically straining peptide between the two domains of FimH also activates high affinity binding. We propose that inserting a small peptide into the region between the two domains that can bind to a larger externally added compound would allow for regulation of FimH binding by pushing FimH into high affinity binding. We have chosen two promising peptides in which to focus our efforts because they meet the requirements of being small and binding to a larger binding protein. Using a crystal violet based assay that I have refined, we can express the switchable recognition proteins on the outside of bacteria and test for binding, expression, and activation. The results suggest that the peptide alone does not activate the protein, indicating that they could be used for the protein switch. When testing peptide binding to the external protein, there was an issue in expressing the peptide correctly and assessing if the external binding protein activates FimH binding. We predict that our end technology could improve the specificity of diagnostics and imaging by exacting control through triggering binding and unbinding of targets in these systems.

Possession Sound as a part of the larger ecosystem of Puget Sound is significantly influenced by both freshwater and nutrient input from the Snohomish River as well as tidal exchange that input of saline water from the Pacific Ocean. It was hypothesized from the years 2010 to 2015, the site closest to the Snohomish River in Everett will have the largest fluctuations in salinity, dissolved oxygen, temperature, chlorophyll and a higher level of nutrients than in comparison to the site in the middle of Possession Sound. Furthermore, the location closest to the city of Mukilteo will express similar characteristics as Everett, but to lesser extremes. The hypothesis was supported in terms of salinity, dissolved oxygen and temperature. The Everett location had the largest range of salinity, between 7 ppt to 26 ppt, temperature from 6◦C to 18◦C, and dissolved oxygen from 5 mg/L to 13 mg/L. However, higher chlorophyll levels and nutrients were not supported at Everett. The location with the greatest chlorophyll range was at in the middle of the Sound, at 0.2 RFU to 2.1 RFU and nutrient levels were at the greatest range at the location near the Snohomish River with 0.1 µM to 27.6 µM in nitrates, 5.43 µM to 75.7 µM in silicate, and 0.245 µM to 3.6 µM in ammonium. However the location in the middle of the sound had the greatest range in terms of phosphates from 0.09 µM to 3.14 µM as well as nitrites from 0 µM to 0.6 µM.

A sediment core collected from Barora Lake in the Solomon Islands can be used to analyze the recent precipitation patterns of the South Pacific Convergence Zone (SPCZ), the Southern Hemisphere’s most prominent weather feature. Barora Lake is a shallow freshwater lake located at 8.722°S 157.600°E and its lack of oxygen one meter below the water’s surface has left its sediment in a uniquely undisturbed state. Because of this, and a high accumulation rate, the sediment in this lake may provide a detailed view of the rainfall in the Solomon Islands over the last millennium. Dinosterol is a sterol produced almost exclusively by certain species of dinoflagellate microalgae, and for this reason it can function as a “molecular fossil,” or biomarker. The goal of this project is to purify dinosterol using column chromatography and High Performance Liquid Chromatography, and evaluate its concentration over time. Gas Chromatography-Mass Spectrometry and Gas Chromatography-Flame Ionization Detection will be used to identify and quantify dinosterol found in the core samples. If there is more than 1.5 micrograms of dinosterol present in these samples, we will be able to determine deuterium/hydrogen (D/H) ratio of dinosterol, which can then be used to determine the precipitation changes in the SPCZ region over time. The D/H ratio in dinosterol tracks the D/H ratio in lake water, which is determined by the amount of rainfall and evaporation that takes place. Since this core provides such a high-resolution view of recent years, we may be able to see the effects that El Niño or La Niña has had on the SPCZ.

Freedom of press allows journalists to cover news without fear of government restraint. During the pro-democracy protests in Hong Kong in late September 2014, approximately 100 activists across China were detained for their support of the Hong Kong protests. Thirty-one remained in detention at the end of the year. Hong Kong and China have a close relationship based on the “one country, two systems” principle adopted by China for governance in Hong Kong in 1984. The Chinese government released a white paper in 2014 that states Beijing holds “comprehensive jurisdiction” over Hong Kong. Under “one country, two systems,” is freedom of press in Hong Kong being suppressed by the Chinese government? My research examines legal powers the Chinese government uses to limit press freedom in Hong Kong, and asks to what extent the Chinese government interferes with Hong Kong news publications, including denying public access to certain news stories through the Golden Shield Project--the Chinese government’s internet censorship program. The research looks closely at China’s censorship program, the laws in the People’s Republic of China, Hong Kong Basic Law, annual reports from Amnesty International and Human Rights Watch, international agreements signed by China related to freedom of speech, and legal and journalism scholars’ research on media relationships between Hong Kong and China. Through observing two major issues that received huge media attention--the 2014 pro-democracy protests in Hong Kong and the Yili Milk Scandal in 2008 that killed six babies and made 300,000 ill--I show how the Chinese government responds to negative media coverage in Hong Kong.

HIV remains a major global health problem due to a high level of transmission and an increased emergence of drug resistant strains. In 2013, sub-Saharan Africa had the most widespread occurrence of HIV in the world, with 1 in every 20 adults infected, accounting for 70% of people living with HIV in the world. A high number of circulating drug resistant HIV remains undetected due to the lack of affordable HIV genotyping methods. Drug resistant HIV is associated with point mutations, therefore screening for specific sets of drug-resistant mutations will be more effective than complete-genome sequencing, which is complex and expensive. The Oligonucleotide ligation assay (OLA) is a point mutation assay that has been used to detect drug resistant HIV. The high specificity of OLA is given by its ligation step, which currently requires a lab instrument (thermo cycler) to cycle between two reaction temperatures: 95 °C allowing oligonucleotide strands to dehybridize and 37 °C allowing ligation to occur. This presents a challenge to adopting OLA in low resource settings where sophisticated laboratory technology, such as thermo cyclers, may not be available. Developing a ligation step that runs at a constant temperature (isothermal ligation) would eliminate the need for expensive equipment. We propose to generate isothermal ligation by using the helicase enzyme, which unwinds DNA naturally in the human body, to dehybridize oligonucleotide strands at a fixed temperature (helicase dependent ligation). With the development of isothermal ligation, the OLA can be translated to a paper device (like a pregnancy test) with potential for widespread use in low resource settings. This will allow HIV infected individuals to be properly diagnosed and receive the proper line of HIV drugs, controlling transmission rates and reducing the evolution of drug resistant HIV within entire populations of people.

Travel time reliability is a key measure of a freeway system’s performance. Traffic incidents are one of the more important factors affecting travel time reliability, since they reduce the capacity of a freeway segment and generate a temporary bottleneck. In this study, an empirical travel time reliability analysis was conducted to compare the performance of general purpose lanes with HOV lanes. This study used four years of travel time and incident data collected on Interstate 5 and Interstate 405 in the Seattle metropolitan area. Three incident types (i.e., shoulder, single lane, and multiple lane incidents) are considered in the data analysis. Travel time reliability was analyzed through several measures, including travel time variability, buffer index, and the probability of freeway segment traffic breakdown. The results for both general purpose and HOV lanes show that incidents result in higher values for all these measures. Specifically, multiple lane incidents induce the highest buffer index and variability in freeway route travel times, while shoulder incidents induce the lowest buffer index and variability. Compared to normal conditions, shoulder incidents can significantly increase the probability of freeway segment traffic breakdown. In general, incidents can significantly reduce travel time reliability. The findings in this study can help shape incident mitigation and management policies for different incident types, especially when aiming to improve freeway travel time reliability.

Automobiles encompass many different integrated systems that require the driver to interface visually, manually, as well as auditory. Automakers are designing cars that allow connectivity to embedded and cloud-based systems. However, some users find it easier to use their hand-held device regardless of whether the vehicle can sync the device to the car interface. A contextual interview was conducted in Seattle, WA and Washington, DC to identify what types of commands users typically execute using voice while driving. The study was conducted on-road with 64 participants, where they were asked to perform voice control tasks for communication, navigation, accessing information, and entertainment. The interviews were recorded using video and paper sheets were used to log in the types of errors made. Cognitive workload while conducting the tasks was assessed using the NASA TLX ratings. The study showed that drivers use several types of voice control devices including smartphones, car embedded systems and a combination of both. The types of errors made and the level of workload perceived varied among the different types of tasks, indicating that varying levels of distraction can occur with voice interfaces.

The Hidden Valley is a new theory that extends the Standard Model, our current best description of particles and forces that make up our universe. The Hidden Valley theory, along with others, extends the Standard Model in an attempt to describe phenomena like dark matter not yet included in the Standard Model. Hidden Valley particles do not interact with Standard Model particles, and are therefore impossible to directly detect. But by using their predicted decay products we maybe be able to prove their existence. We are running simulations to try to give us an idea of what we should be looking for when we use data from the Large Haddon Collider at CERN. Features of the model and the search strategy will be presented.

Peripheral artery disease (PAD) is the narrowing of the peripheral arteries in most commonly the legs. Atherosclerosis causes this disease where plaque buildup inside the arteries causes blockage of oxygen-rich blood flow. A preferential treatment of PAD in the legs is vein bypass surgery as techniques such as angioplasty and stenting have not been useful. However, there is a 30% chance of vein graft failure within the first year due to the thickening and contraction of the vein. The lab has been testing the hypothesis that differences in healing result from differences in the vein wall as opposed to blood-borne elements. They showed growth differences in the vein cells of patients with and without graft failure. My project is to determine if novel genes recently found to be associated with vein graft failure regulate venous cell migration and proliferation, processes that contribute to graft failure. A network analysis of global gene expression by cells obtained from vein tissue at the time of bypass surgery produced a group of genes that was associated with graft failure as a group but not as individual genes. The primary gene of this group is called SCARA5 for which nothing is known regarding vascular function. I first confirm the presence of human SCARA5. I used antibodies to human and mouse SCARA5 from a Scandinavian collaborator that created a SCARA5 knockout mouse. They identified mouse SCARA5 as a 100kD protein. Using the human SCARA5 antibody, I have consistently found a ~70kD protein in human cells which disagrees with the commercial values of 50kD or 100kD. It is not clear whether these differences result from different antibodies or different cells being tested. The next step is to do immunostaining for SCARA5 in hopes of seeing differences and then relate that information to their migration and proliferation data.

The biocompatibility and biodegradability of Bombyx mori silk fibroin (SF) make it an ideal biomaterial for tissue engineering applications. We have chemically modified SF with a thin layer of polypyrrole (PPy) to impart electroconductive properties for engineering electrically-excitable tissues. We also nanopatterned this electroconductive SF-PPy to mimic the extracellular matrix topography of native muscle tissues; such biomimicry has been shown to enhance differentiation and maturation of cultured muscle cells. By combining the beneficial aspects of electroconductivity and nanopatterning, we aim to generate implantable grafts of mature cardiac and skeletal muscle to be used to treat chronic diseases such as myocardial infarction and muscular dystrophy. Aqueous SF solution was produced from B. mori cocoons by dissolving in lithium bromide and then dialyzing against water. Using capillary force lithography, we fabricated large area SF substrates with well-defined nanotopographies by casting SF solution onto nanopatterned molds. To increase substrate hydrophilicity, we modified the tyrosine side chains in the SF protein with sulfonic acid groups using a diazonium coupling reaction. Substrates were made electroconductive by immersion in an aqueous solution containing pyrrole monomers that were subsequently polymerized with FeCl₃. We seeded human embryonic stem cell derived-cardiomyocytes and primary murine muscle cells expressing the GCaMP reporter onto flat and nanopatterned substrates both with and without PPy, and found rapid initial cell attachment and alignment on acid modified SF scaffolds. Furthermore, we enhanced cell retention by adsorbing fibronectin or gelatin onto substrates prior to cell seeding. Finally, we assessed cellular morphology and organization using immunofluorescent staining, and cellular functionality by fluorescently imaging calcium ion transients in response to cellular contraction. Cells cultured on nanopatterned electroconductive substrates appeared more mature than on other conditions tested.

The Human Powered Submarine (HPS) is a student-run research project within the Mechanical and Electrical Engineering departments at the University of Washington. The are several teams comprised of mechanical, aerospace, electrical, and industrial engineering students. Each academic year, the club designs and constructs a submarine that is entered in an international collegiate competition, the International Submarine Races (ISR), which are held each summer. As the title of the club suggests, each submarine is run solely on human power. One or two operators sit or lie inside of the submarine and manually provide power to propel the submarine through water. This is be primarily done by providng power to a drive train via pedals (similar to those used on a bicycle). The drive train transfers power to a thrusting propeller located at the rear of the submarine. Additionally, the operator must wear SCUBA equipment while in the submarine, as the competition rules require that all vehicles be flooded with water. Today, we are presenting our submarine which will be participating in ISR this summer. 

Scientists have been using strontium/calcium ratio (Sr/Ca), magnesium/calcium ratio (Mg/Ca), δ¹³C and δ¹⁸O as proxies for SST. Because each of these proxies has their weaknesses under certain environmental conditions, it is imperative to search for new proxies to complement existing proxies. My project investigates the relationship between SST and magnesium (Mg) isotope fractionation in corals as a proxy for paleo temperature. Samples were collected from each annual growth-ring and then powdered before dissolved in concentrated acid. The Mg in the samples was separated by cation exchange chromatography. The values of ²⁶Mg/²⁴Mg and ²⁵Mg/²⁴Mg for each sample will be measured using a multi collector inductively coupled plasma mass spectrometer (MC-ICP-MS). If δ²⁶Mg in coral is found to have a direct correlation with SST, then fossil coral could be analyzed to provide a long term record of paleo temperature. Having yet another tool in the paleo thermometer toolbox would help scientists improve on global SST records and better model past and future changes in the climate. 

Individuals who wish to save for retirement by participating in defined contribution savings plans are responsible for making their own asset allocation decisions. It is commonly assumed that allocation of resources should satisfy the risk preferences of the individual. However, previous research suggests that the decision to invest in specific assets can also be influenced by the context in which the decision is made. Therefore, in this study, I am using the asymmetric dominance effect (ADE) paradigm to investigate the role of risk perception in determining choice behavior for asset allocation decisions in different choice option contexts. The ADE is a context effect that is well established in consumer behavior research but has not been widely studied in research involving long-term financial decisions. ADE suggests that people’s preferences between two options of equal expected value but different risk levels can shift when presented with a third option (dominated option) that is inferior in all respects to one of the first two options (dominating option) but is only partially inferior to the other option. Participants completed a risk-assessment survey and then 10 trials in which they selected between investments of varying risks and returns. Participants were randomly assigned to a condition that either includes or excludes the dominated option. If individuals’ risk perception is susceptible to ADE there will be a higher percentage of risk-averse individuals preferring the risky investment (dominating option) in the presence of the dominated option relative to when it is absent, and vice versa. This effect will suggest that the presence of the dominated option increases the attractiveness of the dominating option, but does not influence the attractiveness of the other option. I hope the results of this study will provide further support for the influence of context effects on long-term financial decisions.

Age is a major risk factor for cancer, heart disease, and diabetes, and changes in lipid metabolism are increasingly being linked to these diseases. However, the changes that occur in lipid metabolism during aging and contribute to disease are still unclear. Using the yeast, Saccharomyces cerevisiae, as a model system for cellular aging, I identified age-associated changes that occur in conserved lipid metabolism pathways. Specifically, I tagged conserved enzymes of lipid metabolism endogenously with Green Fluorescent Protein (GFP) or mCherry, and I then used microscopy to analyze the distribution and levels of the fluorescent enzymes in aging cells. I found that most enzymes remained the same during aging, but some enzymes accumulated on increasingly more lipid droplets (LDs). LDs are conserved across eukaryotes and function in the storage of energy used during cellular stresses (e.g., nutrient deprivation). Importantly, LDs are known to accumulate in specific tissues during obesity and aging and in many diseased cells. Thus, an age-associated accumulation of LDs could reflect changes in lipid and energy metabolism. To begin to understand how LDs accumulate during aging, I next asked which genes were required for this accumulation. I targeted conserved enzymes of lipid metabolism for deletion, and screened these deletion strains for decreased LD accumulation as above. Using this approach, I identified three genes that are required for LD accumulation with age and that function in neutral lipid synthesis, sphingolipid synthesis, and membrane phospholipid remodeling. These data indicate that age-dependent changes in sphingolipids and phospholipids may contribute to LD homeostasis. Future studies will use genetic screens to elucidate the mechanisms that underlie the accumulation of LDs with age. Because LDs are evolutionarily conserved, these studies should provide insight into how age-dependent changes occur in lipid metabolism and potentially contribute to diseases of aging in humans.

 Nitrous oxide (N2O) is a pharmacologically active gas used clinically in dentistry and anesthesiology. It is also an abused inhalant. Inhalants are generally separated into: 1) volatile alkyl nitrites, 2) volatile solvents, fuels, anesthetics, and 3) N2O. Many drugs of abuse activate stress responses. Our first study addresses whether N2O elicits responses in two main stress systems: the hypothalamic-pituitary-adrenal (HPA) axis and/or the autonomic sympathetic system. To assess HPA axis responses, mean corticosterone (CORT) values during a 2-h 60% N2O administration (N=5 male rats) were measured: Pre-N2O onset, at baseline = 38.0 + 25.3 ng/ml; post-N2O onset, at 20-30 min = 514.9 + 242.9 ng/ml; at 60-90 min = 613.6 + 223.4 ng/ml; at 120 min = 455.1 + 103.4 ng/ml. To assess sympathetic responses, norepinephrine (NEPI) and epinephrine (EPI) plasma responses were similarly measured: NEPI, Pre-N2O onset, at baseline = 237.0 + 34.9 pg/ml; post-N2O onset, at 20-30 min = 375.0 + 53.1 pg/ml; at 60-90 min = 351.0 + 35.4 pg/ml; at 120 min = 434.0 + 76.5 pg/ml; EPI, Pre-N2O onset, at baseline = 170.0 + 51.4 pg/ml; post-N2O onset, at 20-30 min = 430.0 + 118.4 pg/ml; at 60-90 min =315.0 + 144.0 pg/ml; at 120 min = 202.0 + 71.5 pg/ml. These preliminary data indicate N2O causes an elevation of these stress hormones. N2O causes an initial hypothermia in rats that becomes an allostatic hyperthermia over repeated N2O administrations. Because stress mechanisms impact thermoregulation, we conducted a second study using telemetric and infrared (IR) measurements in rats to assess the effects of nine repeated N2O administrations on thermal measures in the interscapular brown adipose tissue and tail, two regions under neuroendocrine influence. These studies will provide a foundation for investigating whether stress responses play a role in drug-induced allostasis observed in the thermoregulatory system.