A major application of micropost technology is the measurement cellular traction forces to quickly diagnose potential medical issues. Currently, polydimethylsiloxane (PDMS) is the material used to fabricate micropost arrays using an intensive three-day double casting procedure. The process begins with casting a negative PDMS mold from a very delicate silicon master array. Negative molds cast from this master require surface treatment overnight via silanization in a vacuum, and then final casting requires an additional overnight cure. The silanization step cannot be forgone in the current process; doing so would result in a micropost array that is inseparable from its negative mold. Additionally, the double casting process must be repeated for every new array in each experiment since PDMS negative molds are not reusable due to degradation by repeated surface treatments. In order to improve upon the micropost fabrication process and widen the scope of applications beyond academia, my colleagues and I are developing a technique to cast PDMS micropost arrays directly from a reusable polyurethane negative mold. Our primary goal is to determine the reusability of polyurethane negative molds in casting PDMS micropost arrays without repeated silanization. To characterize reusability, I will use scanning electron microscopy to measure and quantify the dimensions of the microposts in the arrays in comparison to the silicon master. This step will be repeated for each array reproduced from the polyurethane negative mold as a quantitative measure of its reusability. The creation of a reusable polyurethane negative mold would completely eliminate the intermediate PDMS negative mold and subsequent silanization requirements. Therefore, this replacement would reduce the time required to create a PDMS micropost array from three days to eight hours. Moreover, a polyurethane negative mold could also allow for the simultaneous fabrication of multiple arrays, thus further reducing the time requirement for fabrication significantly.

The development of drug-resistant HIV variants is an important cause of virologic failure. In order to choose effective regimens, assays for diagnosing resistance are essential. Pol gene mutations within the HIV retrovirus, which are associated with drug failure and phenotypic resistance, can be used as the assay targets. Current diagnostic assays include various genotypic and phenotypic assays, recombinant virus assays and other commercial assays. However, these assays are time consuming. To address this issue, we are applying engineering principles to convert the oligonucleotide ligation assay (OLA), developed by Frenkel et al., for point-of-care (POC) diagnostics, to significantly reduce the timeframe for diagnosing drug resistance, leading to regimen decisions. In order to detect the HIV pol gene using OLA, nucleic acid (NA) from bodily fluids need to be extracted and purified. Current extraction methods are time consuming, and require complex procedures, instrumentation, and expensive reagents unavailable in low-resource settings. We are developing novel approaches to capture NA without instrumentation through the use of stimuli-responsive polymers. These polymers are capable of responding to specific changes in their environments, such as temperature variations, resulting in a change of their physical properties; our polymer becomes cationic and aggregating. Magnetic nanoparticles (mNPs) can be utilized in conjunction with these polymers for biomolecule separation by applying a stimulus and a magnetic field. The new NA extraction strategy starts from complexing cationic stimuli-responsive polymers with NA, and then magnetically separating NA from solution via the co-aggregation with mNPs and heating (characterized using standard agarose gel protocol). This extraction method utilizes 2 reagents and takes about 20 minutes, which will reduce cost and enable POC diagnostics in low-resource settings. The extracted NA can then be amplified, tested for the presence of the mutant pol gene, and used to better diagnose HIV drug-resistance and determine patient regimens.

Self-regulation plays a significant role in an individual’s behavior and has been linked to various psychopathologies. The Behavioral Inhibition System (BIS) and the Behavioral Activation System (BAS) are two biopsychological systems that are hypothesized to affect approach and avoidance-motivated behaviors. The BIS/BAS systems provide one framework that can explain the self-regulatory mechanism. In addition, studies have found that mindfulness-based therapies can improve one’s self-regulation by increasing consciousness and awareness; however, there has been relatively little research on the effects of mindfulness training on the BIS/BAS systems. We recruited 17 youths aged 15-19 from a juvenile correction facility who are expected to possess extreme BIS/BAS levels, and placed them in a 6-week mindfulness-based intervention. We collected self-reported data on mindfulness and BIS/BAS level before and after the intervention. Using paired t-tests, we will test whether BIS/BAS and Mindfulness changed from baseline to a 1 month followup assessment.

Intra-articular calcaneal (heel bone) fractures are common tarsal injuries that can require months for recovery. There are a few different treatment options available for these fractures including nonsurgical immobilization treatments, open reduction and internal fixation (ORIF) and percutaneous fixation. Classification of fractures can be a useful tool for medical professionals for communication and treatment planning. The existing calcaneal fracture classification systems used in clinical settings are subjective and not reliable tools for determining the best treatment for patients especially for highly fragmented (comminuted) calcanei. Calcaneal fracture treatments have high complication rates which compounds the need for a better treatment planning tool. The project goal is to develop a new calcaneal fracture classification system that utilizes CT image data to objectively calculate fracture severity metrics of the calcaneus, namely free bone surface area and the volume of surrounding soft tissue swelling. These metrics will be included on a calcaneal fracture report in addition to a model of the fractured calcaneus with fracture lines indicated. The software is developed using the Visualization ToolKit (VTK)  and the calcaneal model generated based on Simpleware ScanIP segmentation of the CT images. In the future, the software can be used to develop an index for the best treatment option for each fracture classification giving medical professionals a tool for aiding in treatment planning and a means of communicating fracture severity.

Nitrogen is important for plants because it helps makes chlorophyll and keeps the plants healthy, however use of fertilizer is expensive and also environmentally damaging. Previous studies in our lab have shown that inoculating crop plants with endophytes isolated from poplar and willow trees enhanced biomass and fruit yields of peppers and tomatoes. In this study, we screened commercial varieties of rice in low nitrogen soil in a greenhouse setting and found that one out of the six variety of rice inoculated with an endophyte consortium had increased shoot biomass in comparison to the uninoculated control plants. We are further experimenting with the endophyte inoculation on this particular rice variety to identify which individual isolate has a positive impact on the growth of rice in low nitrogen conditions. With rice being one of the most important crops around the world, the impact of these findings could hugely reduce the environmental impact of chemical fertilizers in rice farms around the world.

This project investigates how to create a website system that will store service information, strengthen usability, and improve the website’s function to improve community service needs. One important aspect of creating a helpful website is reinforcing the database management, web development, and information analysis techniques. The organization, Chinese Baptist Church (CBC), hopes to improve their user-friendly website. So the CBC has decided to redesign their website. I investigated community members’ user experiences to improve the old version of the website. I reviewed the written code and analyzed the user feedback for future improvements, utilizing previous skills involving HTML, JavaScript (Ajax, jQuery, etc.), PHP, and CSS programming languages. This includes driving down the bug count before deploying the final product to CBC’s operations and teaching their team members how to use the website and its database efficiently. Currently, I have conducted research and constructed a suitable platform for the new CBC website and database. Our group is investigating the design of the website’s structure. Our goal is to improve the user experience and to make an interesting and significant website serve the local community.

Balance control, or stability, is the ability to maintain the body’s center of mass over a base of support. Measurements of stability have a wide range of applications in both clinical and research settings. Many methods have been developed for measuring stability, including functional balance tests, force plate analysis, and video analysis. The preferred method to evaluate standing stability employs the use of a triaxial accelerometer (TA) due to ease of use, portability, cost, and reliability. The aim of this study was to develop an accelerometry-based system for measuring inverted stance (handstand) stability. Using a TA, we collected accelerometry data from a group of acrobats and gymnasts who performed handstands over a period of 30 seconds in four different test conditions: eyes open/firm surface, eyes closed/firm surface, eyes open/compliant surface, and eyes closed/compliant surface. We calculated a set of performance parameters, including medial/lateral displacement, anterior/posterior displacement, and mean speed for each condition from the TA data. Our statistical analysis showed that the TA reliably distinguishes between test conditions for handstands. This suggests that accelerometry can be expanded from regular stance analysis to the evaluation of inverted stance stability. Future applications of accelerometry may include evaluating an acrobat’s handstand stability, comparing the relative stability of differing handstand techniques, or as an additional tool for those investigating the balance control mechanisms of the inverted stance.

Tree growth rate is a major driver of tree population dynamics, forest structure, and forest function. As temperature and competition (e.g., for light) can both influence growth, a clear understanding of the relationship between climate, competitive environment, and tree growth will be critical for predicting how forest ecyosystems will respond to climate change. We hypothesized that growth rates would decrease toward higher elevations as mean annual temperatures and growing season length decrease. Additionally, we expected growth to be faster in canopy gaps (low competition) than in closed canopy areas (high competition). To examine this issue, we assessed the relationship between tree height and age (based on tree ring counts) for Abies amabilis, a dominant conifer species of the Pacific Northwest, across a large environmental gradient encompassing this species elevational range. Specifically, we collected 231 tree saplings from three different elevations (600m, 1000m, 1400m) in gaps and closed canopy areas in Mount Rainier National Park, USA. We found that growth rate declined as elevation increased, suggesting that growth might be constrained by temperature and growing season length. For example, the average age of a 1 meter tree increased by 30 years for every 400 meter increase in elevation (e.g., 28, 58, 88 years from low to high elevation). However, competition for light had little effect on growth rate, possibly because gaps in the forest canopy are short lived (10 - 25 years) relative to the lifespan of these saplings. Our results suggest that global climate change may accelerate growth rates, which could shift this species geographic range, accelerate forest regeneration, and alter forest structure. To further understand this issue we plan to 1) examine the period of time these trees experienced gap and closed canopy conditions and 2) contrast growth with estimates of survival across elevation.

On September 1st, 1991 a large nation state of Soviets died, giving birth to the Republic of Uzbekistan. As a result, thousands of non-Uzbek people became citizens by default in the newly independent republic. They had no chance but to stay in Uzbekistan – now their only homeland. Uzbekistan is a highly understudied region, especially when it comes to the national identities of Russian-speaking population that used to be labeled as the Soviets. The main research question is how linguistically russified non-Russian youth identify their national/ethnic identity in Tashkent, Uzbekistan today? I hypothesize that the majority of this population in Tashkent connect themselves to the Uzbek culture while integrating Russian cultural culture in their everyday lives. Russian speaking population today, though varying in gender, ethnicity and class, has similar experiences thus forming an identity that we can name as “Central Asians.” Methods I have employed are my positionality (I am a Russian-speaking citizen of Uzbekistan), interviews (informal interviews conducted in Uzbekistan with 5 adults), and participant observation (done in the summer of 2013 in Tashkent, Uzbekistan). The research sample are young adults between the ages of 20 to 22, raised after the collapse of the Soviet Union with Russian as their native tongue, but mixed ethnic backgrounds (linguistically russified non-Russians) who live in the capitol of Uzbekistan, Tashkent. The findings provided by this research are a significant step in understanding national formations in understudied Uzbekistan. Theoretically, studying independent formation of national identity of ethnic minorities in the post-Soviet nation-states can help understand identity formation of ostracized groups at the time of the mobilized nation building promoted by the government in the region. Urgent need for such research in post-Soviet territories is highlighted by the recent events in Ukraine and Crimean territory.

Alberta’s Athabasca oil sands are one of the world’s largest petroleum reserves. Oil sands comprise sand, clay, and bitumen – a viscous form of petroleum which is more difficult to extract than conventional crude. Current bitumen extraction practices such as surface mining and steam-assisted gravity drainage are energy-intensive. This independent project investigated the potential of employing microbial biosurfactants to reduce the energy required for bitumen recovery in oil sands. Biosurfactants are employed in the industrial practice of microbial enhanced oil recovery (MEOR), which is viable in the conventional oil industry, but has not been applied to the Athabasca oil sands. In this study, a biosurfactant-producing bacterial strain was isolated from an environmental sample and its biosurfactant compound was characterized. Biofilm formation under simulated oil sands reservoir conditions was analyzed by scanning electron microscopy, and bitumen extraction was tested in a small-scale model. Potential microbial applications in oil sands extraction, upgrading, and bioremediation were explored in a literature review.

During the last glaciation (18,000–13,000 years ago) a lobe of the Cordilleran Ice Sheet advanced into the Puget Lowland, scouring and sculpting the previous landscape into the present-day topography. Prior to the Puget Lobe’s advance, Cascadian alpine glacial systems had reached their maximum extent and were depositing large volumes of sediment into the Puget Lowland basin. While the extensive glacio-fluvial (glacial meltwater) deposits that are preserved in the Puget Lowland indicate a thick sequence of sediment had filled the basin prior to the Puget Lobe’s advance, there has been no previous research on the source (or provenance) of these sediments. Much of this outwash sediment is currently attributed to the Puget Lobe, although a significant volume was likely deposited by Cascadian glacier sources prior to the Puget Lobe’s advance. I have collected samples from glacio-fluvial sediment and Puget Lobe till at bluff exposures on Whidbey Island, as well as alluvium from present-day Cascadian river drainages (a proxy for paleo-Cascadian glacio-fluvial outwash) that can be used to obtain a modal analysis of the basin fill. I analyze these samples using a microprobe to evaluate the proportional occurrence of source-indicative minerals in order to determine the provenance of Puget Lowland sediments. My expected results are for minerals present in the Cascadian drainage sediments, e.g., garnets, to occur in larger proportions than in the Puget Lobe sediments due to source-bedrock composition. For example, the result of high garnet (or other indicator mineral) content in Lowland sediments would indicate that the sediment was deposited by Cascadian glacio-fluvial source rather than by the Puget Lobe, redefining which depositional units are attributed to each source. Knowing the relative volumes of sediment deposited by these two sources will allow me to calculate alpine versus continental glacier sedimentations rates in the region during the last glacial maximum.

Trapped ion quantum computing uses trapped ions to represent information not only as 0s and 1s as in a classical computer, but also as a superposition of 0s and 1s. This provides many benefits over traditional computing, such as enhanced encryption and more efficient factorization of prime numbers. In our implementation, one of the first steps to trapping barium ions to use a qubits, the quantum equivalent of classical bits. We use a 791 nm laser to ionize barium-137, making further operations possible. In normal operations, the frequency of this laser can fluctuate unpredictably, making it unsuitable for ionization. To combat this problem, I designed and built an optical cavity to stabilize the laser. By arranging two mirrors so that the light interferes with itself, only a limited range of frequencies are allowed to pass through. First, a spacer with a low coefficient of thermal expansion keeps the mirrors at a fixed distance, while an analog circuit regulates the temperature of the spacer to insulate it from swings in ambient temperature. A second analog circuit then monitors the light coming out of the cavity and feeds back to the laser, allowing the user to lock onto a specific frequency. This locking system provides a low cost, yet effective method of stabilizing a laser  and will be easy to implement again, with other lasers. More generally, this shows how an ion trap can be improved with the relatively simple laser locking method used here.

As twilight falls on the University of Washington Bothell campus, the skies turn black with the arrival of 10-15,000 crows (Corvus brachyrhynchos) that use the restored North Creek Wetlands as their winter nocturnal roost.  The crows leave large amounts of nitrogenous waste under their roost, and the impact this has on the biodiversity of flora in the wetlands is unknown.  Using simple random sampling, we surveyed flora composition and collected soil samples at sites on- and off-roost across seasons (spring 2013, fall 2013, and winter 2014).  We analyzed soil from each site for moisture, pH, and levels of nitrates.  Across all seasons, significantly higher levels of nitrates and lower pH levels were found on the roost compared to non-roost locations; pH levels varied both with season and by location, but there was no significant interaction effect. Plant community composition was not significantly different on and off the roost, nor did it vary significantly across seasons.  Although the crows are fundamentally changing the chemical composition of the soil under their roost, they are not yet affecting plant biodiversity or abundance.  Birds, however, are vectors for invasive plant species, and these variables are predicted to change over the long term, which will have important management implications.  The North Creek Wetlands are an exemplar of ecological restoration, surrounded by rapidly-developing urban centers, so any future impacts the crows have on these wetlands could cause larger ripple effects over the remaining fragmented forests, streams, and wetlands of the Lake Washington watershed.

Myocardial infarction and resulting heart failure continues to be the leading cause of death worldwide. The heart does not respond effectively to injury through tissue regeneration because cardiomyocyte proliferation after differentiation occurs at a greatly diminished rate. The goal of this research is to study stem cell-derived cardiomyocytes to better understand the mechanisms and possible controls of cardiomyocyte proliferation. Achieving this goal would allow for the intitiation of cardiomyocyte proliferation, which would better direct new technologies aimed at regenerating cardiac tissue after injury. RUES2 human embryonic stem cells are differentiated into cardiomyocytes using a directed differentiation protocol that manipulates the TGFβ and Wnt signaling pathways, by the controlled addition of Activin A, BMP4, chiron, and Xav signaling factors. After twelve to fifteen days, cells are harvested and stained for cardiac troponin T (cTnT), which is a protein present in the sarcomere of cardiomyocytes. Flow cytometery is used to identify cardiomyocytes expressing cTnT and indicate the purity of the differentiated culture. Cardiomyocyte proliferation is measured by exposing cells to Bromodeoxyuridine (BrdU), which is a thymidine analog that is incorporated into synthesized DNA. A double immunohistochemistry stain for Beta Myosin Heavy Chain (β-MHC) and BrdU is used to indicate proliferating cardiomyocytes. RNA is isolated from cardiomyocyte cultures and analyzed using quantitative real time PCR. Specific sequences of RNA present in cells can indicate relative gene expression, and allow for genes of interest to be monitored through stages of differentiation to determine peak expression. Ongoing experiments are investigating the activity of the Notch signaling pathway during directed differentiation of cardiomyocytes and its relation to cardiomyocyte proliferation. Results from these experiments will better indicate time points for enhancing cardiomyocyte proliferation using Notch signaling and help further characterize methods for induced proliferation as a potential therapeutic after cardiac injury.

The toll-like receptor (TLR) proteins are expressed on cells of the innate immune system and recognize structural motifs present on microbial pathogens. Recognition of pathogens by TLRs activates an intracellular signaling cascade and activation of NF-kB, a transcription factor that is critical for expression of inflammatory mediators. Previous studies from our laboratory have shown that single nucleotide variations present in human TLR1 lead to altered expression of TLR1 and are associated with increased risk of death in patients with sepsis. We tested the hypothesis that variations of mouse Tlr1 which correspond to common human variants would result in comparable changes in TLR1 activity. We used site-directed mutagenesis to introduce coding variations in the Tlr1 cDNA. After transfecting the variants into cultured human cells, we stimulated them with Pam3CSK4, a triacylated lipoprotein known to be recognized by the TLR1/TLR2 complex. We then used a bioluminescent luciferase reporter system to measure the levels of NF-kB transcriptional activity. Tlr1 with an Ile allele at position 605 showed increased activity relative to a Leu allele, and the Tlr1 with a Ser allele at position 605 showed increased activity compared to Leu, but to a lesser degree than the Ile allele. Tlr1 with a Ser allele at position 251 showed increased activity compared to Asn, but elucidating its specific effect will require further investigation. These results indicate that variations in the transmembrane domain of mouse TLR1 result in alterations in function comparable to that of human variants in the same region. These findings suggest that mouse models may be used to study TLR1 variants’ effect on the immune response in humans. This could lead to a better understanding of the mechanisms leading to poor clinical outcomes for patients with sepsis and other inflammatory diseases.

As college education becomes more accessible, it has become important to examine particular difficulties faced by the increasing numbers of first-generation college students. Students who are the first in their family to go to college often enter into the university from a working-class background, where interdependent norms of family assistance and cooperation are emphasized. In contrast, university norms emphasize the independent, middle-class value of individual development and personal freedom. Do first-generation college students identify with both sets of cultural values after entering college? If so, do they feel conflict when having to choose between two courses of action, one embodying interdependent family values and the other embodying independent individualist values? Participants evaluated several vignettes in which the decision to uphold family obligations was directly pitted against the decision to pursue personal autonomy. They imagined both going with family preferences at the expense of the self, as well as going against family preferences in favor of the self. Participants then rated their emotional response for both decisions. We hypothesized that holding the dual considerations of their working-class home culture and the middle-class university culture would lead first-generation college students to experience more distress during this decision-making process. In contrast, continuing-generation college students, who are more likely to come from middle-class backgrounds valuing independence, would not be expected to experience a mismatch in cultural values and thus would feel less distressed when having to choose between family and the self. This work will shed light on the cultural challenges that first-generation college students face, and may help identify yet another barrier to first-generation college student success.

Medical implants ranging from artificial hips to pacemakers profoundly impact millions of Americans by replacing malfunctioning body parts and restoring normalcy to patients’ everyday lives. However, microbial infection is a cause of frequent implant failure, making preventative measures vitally important to patient health as well as the efficiency of the healthcare system nationwide. The Tamerler Lab research focuses on engineering the tissue-implant interface using solid binding peptides conjugated to bioactive molecules such as antimicrobial peptides (AMPs). The focus of this project has been on engineering peptides that may reduce zirconia implant failure due to bacterial infection. Our group has been generating sets of peptides that have strong self-attachment activity and high specificity for zirconia. In conjunction with our experiments, AMPs were computationally designed, synthesized, and tested against S. mutans bacteria, one of the most prevalent infectious organisms causing failure in craniofacial implants. To prevent bacterial growth, we tested various concentrations of AMPs to find their minimum inhibition concentration (MIC). Specifically, we focused on developing an engineered peptide that has both solid binding and antimicrobial activities, and testing these peptides’ effectiveness against bacterial growth. With this aim, we first test for the MIC values and then the zirconia-binding affinities of the engineered peptides. In future work, these engineered peptides will be tested on the implant surfaces as self-organized coatings to determine their effectiveness on actual craniofacial implants.

This study aimed to analyze benthic foraminiferal assemblages to determine if environmental changes from pollution and other factors have affected the health of the Bremerton area marine ecosystems. Foraminifera are shelled, single-celled microorganisms that can act as good indicators of environmental change. Sinclair and Dyes Inlets border Bremerton and are connected by the Port Washington Waterway in Kitsap County, Washington. Bremerton is the home of Puget Sound Naval Shipyard and is listed as a federal Superfund site due to accumulations of industrial pollution. Additionally, agricultural and residential pollutants have been released into Dyes Inlet from local streams. Foraminifera were collected from 40 sediment samples provided by the Washington State Department of Ecology from 1998-2012. Samples were analyzed for foraminiferal species composition, diversity, and density (number of inidividuals/gram sediment). Results show 22 species were found, however most samples had low diversities and densities. Ten samples contained no foraminifera. In addition, numerous euhedral gypsum crystals were found in 13 samples in both inlets. Gypsum is a calcium sulfate mineral, and occurrences of subaquatic gypsum crystals in non-evaporitic settings are unusual. Samples with gypsum showed either lower foraminiferal densities or no foraminifera present. Sulfur isotope analysis of the crystals indicates they formed in a sulfate-reducing environment. The comparitively high Total Organic Carbon (TOC) values in gypsym-containing samples suggests that decomposition of organic material may lead to this sulfate-reduction. Additionally, significant dissolution observed in the calcium carbonate shells of many foraminifera indicates pH values of the sediment were low. This can also result from the degradation of organic matter. Thus, foramiferal assemblages within Sinclair and Dyes Inlets are responding markedly to conditions in sediments, establishing the efficacy of using foraminifera as a tool for monitoring benthic ecosystems in Puget Sound.

Sensory Processing Disorder (SPD) is a complicated disorder of the brain that affects children and adults. Simply put, SPD is when the sensory stimuli received are confusing, upsetting, or not meaningful to a child, which results in poor adaptive responses. To achieve an adapted or successful response means that the central nervous system has been able to successfully organize incoming sensory information from the environment for use. The central nervous system is responsible for allowing the neurons to work together and communicate effectively. When this does not happen the result is sensory processing disorder. Within the field of occupational therapy, SPD is a widely accepted disorder; however some pediatricians, psychologist, and teachers question its legitimacy. As a future OT I was intrigued by the concept of SPD, however I wanted to move beyond the initial curiosity and gain a more concrete understanding. I started my research with a literature review asking the question, "What is the foundation of SPD?" and "What are common treatments?" This developed into looking at current research methods, SPD in combination with other disorders, diagnostic and assessment precessess, and treatment through occupational therapy. Through this process, I quickly realized the complexity of SPD and wanted a more tangible and personable perspective of SPD. This led me to conduct a series of thirteen interviews with occupational therapists. They were all pediatric OT who worked either in a school or clinical setting and had extensive experience working with children with SPD. The interview questions addressed the same themes in my literature review with more elaborate questions on treatment. Through my research, I hope to expose others outside the field of occupational therapy to SPD in hopes of advocating for awareness and promoting research. Because of the complexity and individualistic manner of SPD, it is important to push for more research. As more children demonstrate symptms of SPD, continuous research is vital to convey validity, support clarity and accuracy during assessment, and provide more effective treatment. 

In radiation therapy (RT) normal tissue damage and especially late-developing normal tissue damage limits the total dose that can be safely delivered to a tumor. Coupling RT with agents that selectively protect normal tissues from adverse effects or sensitize malignant tissues to radiation would improve therapeutic outcomes. Vitamin E derivatives like tocotrienol have been proposed as potential anti-tumor agents. Some studies suggest an interaction between these agents and radiation. Based on these studies, we investigated the combined effect of radiation and delta-tocotrienol (dT3) on B16 mouse melanoma cells. The toxicity of dT3 alone and in combination with radiation was measured using clonogenic survival assay. Preliminary studies show that minimally toxic levels of dT3 sensitize B16 cells to radiation-induced toxicity. Ongoing follow-up studies are investigating the effect of combined dT3-radiation exposure on normal human fibroblasts, in order to evaluate whether the radiosensitizing effect of dT3 is selective for tumors. Identifying an agent that is selectively toxic and radiosensitizing to tumors would quickly be translated into clinical studies.

X-linked Myotubular myopathy (XLMTM) is a congenital myopathy caused by mutations in the myotubularin (MTM1) gene. Clinical symptoms of the disease including muscle weakness, respiratory failure, and hypotonia. XLMTM is caused by deficiency of the protein myotubularin necessary for normal muscle excitation-contraction. Dogs harboring a canine MTM1 gene model the disease and provide valuable insight due to phenotype similarities with patients. Affected dogs exhibit muscular atrophy, weakness, and stilted gait. A neuromuscular assessment score was developed to measure neurologic impairment in affected dogs. My project involved validation of this previously tested neurological assay by an independent observer. I was the independent observer who reviewed and scored neurological examination forms. I also reviewed and scored videotaped examinations and conducted statistical analyses looking at the correlation of the results within groups of dogs and between groups of dogs. Intra- and inter-reliability were performed by another observer and myself and wild type (n=6) and affected dogs (n=4) were scored. Prior to beginning the study, I received training on the assessment scoring system, including how to perform and analyze clinical tests. The intra-reliability test involved repeated examinations of wild type dogs and generated a consistent score in 88% (46/52) of exams based on interpretation of written neurological examinations and 100% (9/9) of exams based on scoring videotaped examinations at two time-points. The inter-reliability tests generated a consistent score in 54/71 examinations (76%) based on scoring of pre-recorded neurological exams and generated a consistent score in 12/15 cases (80%) based on scoring video-taped exams. The two observers scored dogs within 1 point in 68/71 (96%) of cases based on written notes of the neurological examination and within 1 point in 15/15 (100%) of videotaped examinations. Together, these data support the reliability of the neurological assessment score in XLMTM dogs.

 Community-based organizations face numerous challenges as they develop initiatives for delivering various types of services. What are the varying challenges that small, non-profit, community-based organizations face as they aspire to become highly functioning service centers? Exploring this question exposes the true depth and variety of internal and external challenges that many community-based organizations meet. To develop an understanding of these challenges, I utilize participant observation while volunteering at a small, community-operated organization called the Somali Community Services Coalition in Tukwila, WA. Since September 2013 I have been a tutor for students enrolled in their after-school program, and recently I have also been involved in planning of the future after-school program as well as the summer enrichment program. During this time I have developed an understanding of the organization’s dependence on volunteers and the implications of that dependence; the roles of the staff, both those that belong to the ethnic community as well as the temporary staff from organizations like the Peace Corps; the difficulties regarding organizational funding through grants; issues with food provision within the organization and the community, as well as many other topics. Through furthering my time with the organization I hope to learn more about potential funding competition with other local organizations. I hope the implications of this study will develop my own as well as other’s understanding of the challenges small community-based organizations face and how understanding these challenges can assist those working with them to properly engage the multifaceted functions of these organizations to best suit the organization’s interests and development.

Impervious surfaces have increased the amount of rainwater runoff and pollution entering nearby waterways causing water quality concerns in cities throughout the United States. In order to address water quality problems, policies have been mandated at the national, state, and city scales which require water quality issues related to pollution from stormwater to be addressed. These policies have created an array of stormwater management practices and projects. My research investigates stormwater policy and how it has been applied through different practices implemented at the city-scale. While incentive programs have been implemented in order to help solve communities’ stormwater issues, this research shows that citizen participant equity has not been addressed in the various stormwater programs. First, I explore the lack of equity measures through a policy analysis of stormwater programs that are commonly used at the city-scale. Then, I make the case for why equity measurements are needed in stormwater management by identifying local initiatives that require the governments responsible for stormwater management to address issues of equitability in all programs. Finally, I suggest best management practices in order to help achieve equitability goals using my experience with King County and the RainWise program as a case study. This research exemplifies how equity can be addressed in an incentive program, and can be used with the purpose of illustrating methods in order to address equity in the future.

This analysis discusses changes over time at the Gua Talimbue site, excavated in 2013 by Sue O’Conner, Ben Marwick, Tim Maloney, Emma St. Pierre and Rose Whitau in South-East Sulawesi, Indonesia. Sulawesi, formerly known as Celebes, is one of the four Greater Sunda Islands and is considered the largest island in Wallacea. This area was a major stepping stone for the migration of peoples into Island Southeast Asia and what is now modern Australia, and understandng shifts in occupation here can shed light on human movement patterns throughout the globe approximately 18,000 years ago and onward. The research has examine the changes through time with consideration to geoarchaeological and lithic samples taken from the Talimbue site, with a specific focus on shifts in human behavior, site formation, and the environment of the site over time. Geoarchaeological testing of the site includes pH and electrical conductivity testing, loss on ignition, magnetic susceptibility, color and particle size distributions. Lithic analysis is focused on the change in distribution, size, mass and type throughout the occupation of Talimbue, first occupied by humans 18,000 years ago and continuously inhabited until recent times. Preliminary results have identified an environmental shift at the Pleistocene-Holocene transition, approximately 12,000 to 8,000 years ago, which affected occupation and has implications for the decreased complexities seen in lithics analysis. The purpose of this research is to understand human impact on environmental processes, as well as identify occupation and technological trends within the context of Sulawesi and beyond. 

Although our pre-hominin ancestors almost certainly were climbers, most species of hominins infrequently used this ability. Recently, however people have begun to climb for recreation. With the growing popularity of rock climbing, we were interested in learning how the human body changes under these new forces. This study was designed to understand the relationship between hand proportion and hand strength in rock climbers and non-climbers. We hypothesized that rock climbers have stronger hands for their size than non-climbers. A hand dynamometer was used to test hand strength. The lengths of the carpals and metacarpals from the radial and ulnar styloid processes to the distal ends of each ray and the lengths of each digit and each phalanx were measured. In addition, the width and circumference of each hand was taken from the proximal phalanges, with the fingers resting against each other, and the circumference of each digit was measured around the proximal interphalangeal joing. We also measured the basic anthropometric variables of stature, body mass, and age and asked climbers how long they had climbed, the level of difficulty they usually climbed, and how often they climbed. Unsurprisingly, Student's t-tests revealed that the hands of men are bigger and stronger than those of women (all p's </= 0.004) and climbers are stronger than non-climbers (all p's </= 0.01). The circumference of the digits is ia significant predictor of hand strength when sex, side, and climbing status are controlled (all p's </= 0.03), with 67% of the variation in hand strength explained (r²=0.67).

Brain-computer interfaces (BCIs) are an emerging technology that uses neural signals to communicate with external electronic devices. Leveraging this technology for a variety of rehabilitative purposes has become a popular focus of research. One application is in guiding spike-timing dependent synaptic plasticity, a physiological mechanism where the strength of connection between two neurons is shaped by the precise temporal relationship of firing between them. Using a BCI to exploit this principle may offer a means of strengthening spared pathways after neurological trauma. In studies using rat models of spinal cord injury (SCI), a recurrent BCI called the Neurochip has been used to translate electromyographic signals (EMG) from impaired forelimb muscles into precisely timed intraspinal electrical stimulation at associated sites in the cord. Data recorded from forelimb reaching tasks have shown significant recovery of motor function following stimulation therapy. This is thought to be due to spike-timing dependent synaptic plasticity, but the fundamental mechanisms of behavioral improvement have yet to be elucidated. This investigation explores the role of gene expression in the stimulation treatment. Rats were given the same unilateral spinal contusion at vertebral level C4 and stimulated with the Neurochip device for 2 weeks of treatment. RNA from the rats was then isolated and analyzed using either Affymetrix GeneChip® or RNA-seq technology. Certain genes that have been implicated in neural plasticity were evaluated for expression levels. Of particular interest is the regulation of the mTOR and Wnt signaling pathways. Genes for mTOR and upstream mediators, such as AKT and PTEN, are important regulators of process outgrowth, regeneration and synaptic plasticity. The Wnt family of glycoproteins play critical roles in neuronal targeting and may guide synapse formation. Results of this study will contribute to understanding the cellular changes undergone during treatment.

Cardiac cell death has been implicated in the pathogenesis of various forms of cardiomyopathy and end-stage heart failure. Here, a novel signaling molecule, TAK1 (TGFβ-activated kinase 1, gene name Map3k7), is identified as a nodal regulator of apoptotic and necroptotic myocyte death, myocardial remodeling, and heart failure propensity. I found that genetic or pharmacological inactivation of TAK1 in cardiac myocytes markedly induced programmed necrosis and apoptosis in response to TNFα (tumor necrosis factor alpha), an inflammatory cytokine. Conversely, overexpression of the constitutively active mutant TAK1ΔN (N-terminal truncation), or TAK1 plus its activator TAB1 protected cardiac myocytes from TNFα-induced cell death. Mechanistically, inactivation of TAK1 promoted formation of the necroptotic cell death complex consisting of RIP1 (Receptor-interacting protein 1), RIP3 (Receptor-interacting protein 3), caspase 8 and FADD (Fas-Associated protein with Death Domain). In addition, my data showed that genetic ablation of cytoplasmic cell death signaling molecules such as RIP1, RIP3, caspase 8, or FADD largely blocked TNFα-induced cell death in TAK1-deficient cells, whereas deletion of mitochondrial cell death proteins, Bax/Bak or cyclophilin D, showed no effects. Further, the transcription factor NFκB-mediated cell survival signaling was greatly impaired in TAK-1 deficient cardiac myocytes. Taken together, the data indicate that TAK1 functions as a critical “molecular switch” in TNFα-induced programmed necrosis in cardiomyocytes by interacting with the RIP1/3-caspase 8-FADD cell death pathway as well as the NFκB cell survival pathway. These results also reveal a new paradigm where TAK1 functions as a necessary survival factor in the heart by directly antagonizing necroptosis.

 DNA sequencing is essential to life sciences and personalized medicine. Nanopore sequencing is a single molecule sequencing technique that promises both an increase in sequencing speed and a reduction in cost compared to current sequencing technologies. In nanopore sequencing, DNA is drawn through a nanometer-wide pore while we record any changes in ionic current. These current changes depending on the nucleotide sequence. Our lab uses the nanopore Mycobacterium smegmatis porin A (MspA), which was engineered to examine DNA with single nucleotide resolution. MspA is sensitive to the four standard nucleotides (A, T, C, G) as well as small chemical changes to those nucleotides. Uracil is a nucleotide that differs from thymine by a single methyl group which we now show we can detect with MspA. Using a process called bisulfite treatment, which converts cytosine to uracil, we obtain information that helps us to determine the input DNA sequence.

One goal of aging research is to identify genes that modulate aging, in order to eventually develop therapies that delay the onset of age-associated diseases. In particular, genes that control mitochondrial function appear to play an important role in aging. Although severe mitochondrial dysfunction causes childhood diseases such as Leigh syndrome, there is growing evidence that moderate mitochondrial stress can extend lifespan in yeast, worms, fruit flies, and even mice. In the nematode Caenorhabditis elegans, a long-lived mitochondrial mutant called isp-1(qm150) has a defect in the Rieske iron-sulfur protein. The Rieske iron–sulfur protein is a primary electron acceptor located in complex III of the mitochondrial electron transportation chain. Complete RNAi knockdown of isp-1 results in larval arrest, while partial knockdown extends lifespan, further supporting the idea that milder forms of mitochondrial stress can increase lifespan while severe mitochondrial stress is detrimental. We are conducting a mutagenesis screen to identify genetic suppressors of the larval developmental arrest induced by isp-1(RNAi). Through exposure to EMS/ENU mutagen, wild-type worms will have random mutations generated throughout their genomes. These mutants will then be screened for the ability to develop on isp-1(RNAi) . We speculate the novel suppressor mutations from the screen may affect proteins that interact with ISP-1 or cause mutations that bypass the need for ISP-1 in the mitochondria. Identification of such suppressors will help to clarify the mechanisms linking mutation of isp-1 to development and aging. The isp-1 gene and its function in the electron transport chain are conserved in humans. Therefore these studies may be relevant for human aging and disease.

Prior research shows that well-functioning interprofessional (IP) healthcare teams lead to greater patient satisfaction, shared clinician understanding, and stronger teamwork, yet healthcare providers are educated in separate “silos” – inhibiting the development of critical teamwork skills. IP service learning (IPSL) is one way to integrate student learning across the health professions. There is little research on how best to use IPSL in current curricular models for the health profession. Furthermore, though student engagement is central to both service learning and IP education, there are few models for how to develop IPSL. This project will explore this important area by asking: how do students who are knowledgeable in both IP and service learning envision effective IPSL? In addition, how could IPSL support current health professional education? One proposal is the creation of a common curriculum among health professional schools, as focused, year-long IPSL experiences. To explore this idea and generate other options, student experts from four of the six health science schools (Nursing, Pharmacy, Public Health, and Social Work) participated in a focus group on the UW Seattle campus. Participants were recruited based on their availability and interest, and questions were formulated using Gitlin, Lyons, and Kolodner’s extended social exchange theory. The results of this study were used to help inform current curriculum changes in the health sciences schools, and to reinforce current understandings of IP education and ways to use IPSL in educational programs. Areas for future research include pilot testing 1-3 proposals, as well as sampling health sciences faculty to uncover systems-level barriers to IPSL.

Products of silicate mineral dissolution play important roles in geochemical cycles and biologic processes including secondary mineral precipitation and microbial and plant growth, respectively. Understanding the mechanisms and stoichiometry of silicate weathering are therefore important in relating weathering rates to global ion budget. In abiotic systems, silicate dissolution often begins as a non-stoichiometric reaction dependent on solution pH. In acidic versus alkaline solutions, forsterite leaches cations and silica, respectively, resulting in a surface enriched with the counter ion. Microbes are, however, ubiquitous in water-rock systems where active silicate weathering occurs. Recent evidence shows a dramatic increase in forsterite (Mg₂SiO₄) dissolution rate that correlates with the presence of dormant bacterial endospores. The mechanisms driving enhanced dissolution in the presence of a complex organic surface and its effect on mineral surface chemistry is unknown. We investigate the effects of Bacillus subtilis endospore enhanced dissolution on the surface chemistry of forsterite grains using Attenuated Total Reflectance- Fourier Transform Infra-red (ATR-FTIR) spectroscopy. ATR-FTIR is a technique that measures mid (MIR) to far (FIR) infrared absorption of a sample over a shallow penetration depth. Forsterite (Fo₉₀, Mg_1.8Fe_0.2SiO₄) powder was incubated with B. subtilis endospores for 40-80 days at pH ~ 7.5 under oxidizing conditions. We compare MIR and FIR ATR-FTIR spectra of un-reacted forsterite, abiotically reacted controls, and biotically weathered forsterite to determine how the surface has changed from the weathering processes. MIR and FIR ATR-FTIR spectral scans of forsterite grains incubated with endospores display peak shifts relative to the controls. Small amounts of Fe(II) present in the forsterite likely precipitated as iron oxide coatings on the mineral and endospore surfaces. Variations in FIR peaks may indicate the presence of iron oxides. Results from this research will advance our understanding of how bacterial surfaces affect chemical weathering in the environment.

Graphene, a material consisting of only one layer of carbon atoms, was isolated and electrically measured for the first time in 2004. New physics is rising that can be studied with graphene, such as half integer quantum hall and Klein tunneling and etc. Its excellent physical properties and chemically inactive nature make graphene and other 2D materials promising for both device applications and fundamental research. My project is making 2D-material based devises I learned to make tape with a variety of 2D materials (including insulators, conductors, ferromagnets), to exfoliate those materials onto silicon oxide chips, and to search for monolayers. Since the charge carriers in 2D materials are exposed to environment, they are very sensitive to the impurities from the surroundings including substrates. To fabricate high quality nano-devices, putting these materials in an ultraclean environment is very important. One strategy is to sandwich a material in two boron nitride flakes to build heterostructures. I will present the process I use for making graphene structures by van der Waal’s transfer technique. Van der walls forces are utilized to pick up a boron nitride flake on silicon oxide with a PPC (Polypropylene carbonate a thin clear copolymer) spin coating on PDMS (a clear rubber). A monolayer can then be picked up by the boron nitride and then one more layer of boron nitride completes the sandwich. The PPC is melted off the PDMS, and its residue is washed away. The devices I have made were characterized by atomic force microscopy, scanning electron microscopy, and electronic transport measurements. The measurements made on my devices pave the way to making state-of-the-art low dimensional electron systems.

Reward presentation reinforces behaviors and increases the likelihood of a particular behavior occurring in the future. This increase in behavior is one of the bases underlying reward-based learning. Reward-based learning involves several brain areas, of particular interest to this study is the periaqueductal grey (PAG). The PAG is anatomically linked to other brain structures that are involved in reward processing, such as the ventral tegmental area (VTA). When presented with rewards and novel stimuli, neurons in the VTA exhibit burst firing (brief periods of high activity), and the intensity of this firing is modulated by reward size and expectation. It is possible that incoming glutamatergic signals (excitatory signals) from the PAG contribute to this burst firing as it is the third heaviest subcortical supplier of glutamate to the VTA; however this has not been previously investigated. Previous studies have recorded burst firing in the VTA during a working memory task on a radial 8-arm maze. These past studies also showed velocity correlated firing by VTA neuron of rats as they approached or retreated from rewards. The current study had rats perform the same working memory task, but recorded neural activity from the PAG to examine whether the PAG showed similar burst firing patterns as the VTA. Recordings of single unit (one cell) responses were collected from the PAG as rats performed the task under different conditions, including omission of expected rewards or expected reward magnitude reversal. The rat’s velocity as it was approaching and retreating from the reward was also recorded. It was found that a subset of PAG neurons did show reward-related firing, similar to that of the VTA, and we anticipate also finding velocity correlates. Thus, this study will better characterize the function of the PAG and the neural processes through which reward-based learning occur.

Pharmacogenetics is the field of biomedical science that studies how variation in a person’s genetic composition affects his or her response to drugs. Glucocorticoids are a group of signaling molecules that include endogenous hormones such as cortisol, which the body produces naturally in response to various forms of stress, as well as drugs with strong anti-inflammatory properties that are used to treat diseases ranging from asthma to cancer. The goal of this study was to identify correlations between interpersonal genetic variability and alterations in glucocorticoid receptor (GR)-mediated signaling. Salivary cortisol concentrations were measured in healthy volunteers (n=66) in order to determine differences in circulating cortisol concentrations and cortisol response following simulated physiological stress. Statistical analysis indicated inter-participant cortisol (CORT) variability was not correlated to participant demographics, health history, or single nucleotide polymorphisms (SNPs) in genes encoding the GR (NR3C1) or the essential GR chaperone protein hsp90 (HSP90AA1). Two participants with highly similar demographic profiles and opposing CORT responses (a matched pair) were selected for exome sequencing—the sequencing of all coding regions in the genome. A filtering algorithm identified candidate SNPs in approximately 30 genes involved in mediating the cortisol response and GR signaling. Interestingly, corticotropin releasing hormone receptor 1 (CRHR1) showed a difference of over 20 SNPs between the matched exome pair. CRHR1 regions were then sequenced from the larger participant group. Participants with average cortisol concentrations that slightly decreased following CORT stimulation most commonly exhibited a unique SNP profile—they shared many of the same SNPs in CRHR1. We speculate participants with SNP profiles associated with altered cortisol response may share a corresponding sensitivity to corticosteroid-based drug therapies.

Positive outward remodeling of arteries and formation of arteries from arterioles in response to arterial injury is highly dependent on the proliferation and migration of smooth muscle cells (SMC). To form new arteries, SMCs also have to degrade the extracellular matrix (ECM) around the existing vessel to serve as a starting point for arteriogenesis. The p27 protein is a cyclin-dependent kinase inhibitor that stalls the cell cycle. We have previously demonstrated that mice lacking the p27 protein have improved arterial enlargement in response to arterial injury. We hypothesize that decreased expression of p27 improves SMC proliferation and migration, as well as ECM degradation. To test this hypothesis, we extracted aortic SMC isolated from p27 knockout and wild type mice and performed several in vitro assays to assess proliferation and migration. We used a growth assay to measure proliferation. We used a wound assay and a modified Boyden Chamber assay to measure migration in the absence (wound assay) and presence of a chemoattractant, platelet-derived growth factor-BB (Boyden Chamber assay). The wound assay is done under growth arrest to study migration independently of proliferation. The Boyden Chamber assay is performed with and without matrix metalloproteinase inhibitors (MMPIs) to assess migration in the presence and absence of ECM degradation. We performed these assays under varying conditions such as nutrient starvation and hypoxia to better model recovery from ischemia or under wound conditions. Preliminary wound assay data has shown that certain isolates of p27 KO SMCs exhibit a higher degree of migration. A better understanding of p27’s role in arterial remodeling may lead to improved therapies to stimulate arterial growth to treat ischemia resulting from arterial injury or to inhibit arterial growth to treat cancer.

With ‘baby boomers’ reaching late adulthood, it is growing increasingly important for the scientific community to work toward treatments and potential cures for the diseases that affect this age group most often. Included in this category are the myelodysplastic syndromes (MDS), a group of related disorders involving the production of abnormal or insufficient quantities of myeloid blood cells. MDS is relatively prevalent in the older American population, yet much remains unknown about it. One treatment option is hematopoietic cell transplantation (HCT), in an effort to “reprogram” the body to produce healthy blood cells. The focus of this research study relates to the subgroup of MDS patients who undergo an HCT as part of their treatment but relapse 18 or more months later. The hypothesis was that there exist particular chromosomal abnormalities that manifest after transplantation which contribute to the propensity of MDS patients to late-relapse.
To test this, I worked with the Fang Laboratory at the Fred Hutchinson Cancer Research Center to optimize the process of extracting DNA from frozen patient cell pellets by manual and automated methods. We then analyzed paired patient DNA samples from before and after transplantation to look for signs of genomic evolution that could help explain why these patients exhibited such a long delay before relapsing. Chromosome genomic array testing (CGAT) was conducted on two patients’ paired samples. This method of analysis uses molecular probes to identify chromosomal abnormalities that may be causing disease. Both patients’ relapse samples showed clonal evolution, suggestive of disease progression and poor prognosis. These findings are promising evidence that the continuation of this research could help contribute to the discovery of MDS late-relapse markers and a more comprehensive understanding of these conditions.

Traumatic Brain Injury (TBI) is a leading cause of mortality and permanent neurological disability. Microglial activation and inflammation contribute to the onset and progression of TBI-induced pathology. In this study we aimed to identify the factors that activate brain inflammatory pathways in the early responses to TBI, and the mechanisms by which inflammation is sustained during the late stages of pathology. We have previously demonstrated that the microglial expressed receptor, CX3CR1, regulates microglia activity. We have also shown that lack of CX3CR1 leads to deficits in cognitive function and synaptic plasticity. Here we hypothesized that the loss of the CX3CR1 receptor - and consequently, the loss of normal microglia function - would increase pathologic conditions in the brain after TBI and cause measurable defects in behavioral and cognitive abilities. To this end, three month old male CX3CR1 wild type and CX3CR1 knockout mice were subjected to a mild TBI (0.5mm depth) using a controlled cortical impact (CCI) animal model. Control mice (sham operations) received the same anesthesia and craniotomy, but were not subjected to CCI. All animals were humanely euthanized and transcardially perfused at fifteen and thirty days following TBI. I specifically participated in assisting TBI surgeries, behavioral testing, transcardial perfusions, brain sectioning, and immunohistochemistry. Our results indicate that lack of CX3CR1 at early time points protects neuromotor function, reduces cell death, and is associated with expression of microglia that have an anti-inflammatory phenotype. However, the long term effect of CX3CR1 receptor loss following TBI induces neuronal cell death, as well as neurobehavioral and cognitive deficits, which are associated with an increase in expression of microglia that have a proinflammatory phenotype. These results point to the presence of CX3CR1 in the early stages following a TBI as essential in slowing the progression of the neuropathology. In conclusion, CX3CR1, the fractalkine signaling receptor, plays a central role in regulating neuroinflammation after a TBI.

Giardia lamblia is a protozoan parasite common in mammalian hosts and belongs to one of the earliest diverging eukaryotic groups. Subsequently, Giardia has a simplified and highly divergent genome that lacks classical actin associated proteins, yet has retained a conserved microtubule (MT) cytoskeleton. This unique cell biology could provide a window into ancient cellular processes, particularly cytoskeletal function and regulation. Giardia has evolved novel and elaborate MT-arrays that mediate important stages in a complex life cycle and assemble into structures critical to its virulence, motility, and cellular organization. Like most eukaryotes, Giardia possesses a superfamily of five tubulin paralogs (gene duplicates) that constitute the MT cytoskeleton (alpha, beta, delta, epsilon, and gamma-tubulin). Interestingly, the Giardia genome contains multiple paralogs of the alpha- (TUBA1, TUBA2) and beta- (TUBB1, TUBB2, and TUBB3) types. It remains unresolved why multiple, highly conserved TUB paralogs are maintained in the Giardia genome. To address this question, we localized tubulin proteins by expressing GFP-tagged tubulin under its native promoter in a pGFPa.pac expression vector. These constructs were transfected into cells and proteins were localized using 3D fluorescence microscopy. Microscopy of fixed cells revealed that TUBA and TUBB proteins localize similarly in interphase cells and are largely interchangeable (ventral disk, median body, flagella), yet display differential levels of expression. Namely, GFP::TUBB3 and GFP::TUBBA1 signal is much brighter than that of GFP::TUBB1, GFP::TUBB2, or GFP::TUBA2. Microscopy also demonstrated delta-tubulin is diffuse throughout the cytoplasm, epsilon-tubulin localizes faintly to the ventral disk, and gamma-tubulin appears to track the internal axonemes of the flagella and concentrate at the flagellar tips. This data could suggest Giardia utilizes alpha- and beta-tubulin paralogs in different stages of the life or cell cycle, which has not yet been demonstrated. This data will also inform the development of a robust live cell marker for Giardia’s MT-arrays.

The purpose of this project is to develop and refine a technique for taking photographs to be included in an atlas of the foraminifera of Puget Sound. Foraminifera are shelled single-celled organisms that live at the sediment/water interface in marine environments. Their sensitivity to environmental conditions makes them useful in monitoring ecosystems on the sea floor. The atlas will contain photographs and detailed descriptions of each foraminifera species. Despite their importance as an environmental monitoring tool, foraminifera have never been comprehensively studied in Puget Sound. This catalogue will become a useful tool for future workers in research and environmental monitoring. Key to this atlas is the inclusion of clear, accurate photographs of each species of foraminifera. Scanning Electron Micrographs yield excellent images, but these often do not portray the organism as it is viewed through a light microscope. Thus, the inclusion of photomicrographs is essential. Photography becomes quite technical when dealing with objects of a microscopic scale while still trying to produce a clear image with depth of field. With the Puget Sound Foram Project we work with specimens that can be as small as a tenth of a millimeter in size, so producing a high quality publishable picture takes time and a specialized technique. Using a light microscope, each specimen has to be photographed several times, focusing at a different depth each time. Once all of the layers have been captured, the pictures are then synchronized and edited in such a way that we get one final image that has both clarity and depth of field. In addition, each specimen must be photographed from two or more angles to illustrate critical features. Photography for research purposes can be a complicated and lengthy process but is essential for not only supplementing data but conveying vital information.

There is debate regarding whether the Earth has the same magnesium (Mg) isotopic composition as chondritic meteorites (chondrites). Chondrites are thought to be composed of material from the original solar nebula from which the Earth accreted. Several studies have analyzed the compositions of chondrites, basalts, and peridotite xenoliths, but there is discrepancy in the δ²⁶Mg values reported for the Earth’s mantle as well as for certain geostandards. One such geostandard is San Carlos olivine, a mineral found in San Carlos peridotite xenoliths from the San Carlos Apache Reservation in east-central Arizona. The role of a geostandard is to assess the accuracy of one lab’s data compared to those of other labs, so conflicting δ²⁶Mg values raises the question of whether the difference is due to analytical artifacts or to sample heterogeneity. This study examines olivine, orthopyroxene, and clinopyroxene minerals in San Carlos peridotite to determine if the peridotite is homogeneous with respect to Mg. Mg isotopic compositions will be measured by multi-collector inductively coupled plasma mass-spectrometry (MC-ICP-MS). The results will indicate if San Carlos olivine can be used as a geostandard for Mg isotopic analysis. Once there is reliable data for the Mg isotopic composition of the mantle, it can be determined whether the composition of the Earth is indeed chondritic, which has further implications in determining a model for the formation of the Earth.

The ability to optically isolate single gallium arsenide (GaAs) electron donors has many implications both in quantum information science as well as semiconductor physics. However, conventional methods of fluorescence microscopy are fundamentally limited by diffraction, which sets a minimum resolvable feature size preventing the direct optical imaging of nanoscale structures. At the University of Washington Optical Spintronics and Sensing laboratory we are building a super-resolution optical microscope, based on ground state depletion, to isolate single GaAs donors. A critical component of the microscope is the two-dimensional methodical scanning of laser beams over the sample, for which we have elected to use a piezoelectric fast-steering mirror. I have designed a scanning system capable of under 1 nm mechanical resolution, which will be sufficient for our target optical resolution of about 100 nm to isolate single donors. I will present preliminary results on the performance of the test system using a transmission electron microscopy (TEM) grid as a test sample, which will allow us to evaluate the achieved accuracy and resolution of the imaging due to its regular features. Isolating single donors in GaAs using this super-resolution microscopy technique may lead to new insights in how to localize and evaluate impurities such as donors and acceptors for quantum information as well as classical applications.

The Yi people lives in the south west of China, leading a life of the combination of growing and grazing which has lasted for hundreds of years. They have a rather distinctive culture characters with their own writing system, religion, customs, and clothes. Jieshy and Vala are two kinds of capes that play an important role in Yi people’s daily life. They are made of wool and have several similarities in outlooks but are functioned differently, especially in social lives and celebrations.However, as modern life is eventually influencing the life of the Yi people, Jieshy and Vala are less worn in daily life. In order to know the status of these two traditional clothes in Yi people’s modern life, we did several field researches in three Yi counties in Sichuan Province, China, during the summer of 2013. Our research focused on the productions and the application of these two kinds of clothes in modern life, and the Yi people’s attitudes toward them. Through our observations and interviews with local people and scholars, we know that nowadays, Jieshy and Vala are still used in funerals and traditional weddings and have almost been considered as a symbol of the group. But there is indeed a trend of reduction in the use of these two kinds of traditional clothes. The goal of our project is to present how modernization shapes the tradition and how people react to the changes.

 In the last ten years, social media use and user base have exponentially increased in prevalence and interface with everyday social activities. In part this increase can be attributed to the advent of mobile web technology. Currently, the popular assumption that social media use causes higher levels of anxiety in users is trending throughout media platforms. A 2013 study by Ethan Kross, of University of Michigan, predicts Facebook usage may issue a decline in subjective well-being in young adults. Our study was designed to examine any association of anxiety levels between users and their most frequented social media platforms. Participants were recruited via various social media outlets, Facebook, Twitter, E-mail, Snapchat, Instagram, and SMS messaging and directed to an online self-administered anxiety survey. The study also requested participants to field several questions surveying their own social media usage and activity. For validity purposes, the order of the anxiety survey and social media usage questionnaire were alternated between participants whose biographical data remained anonymous. Nominal age and gender data were reported in planned descriptives. We used the Zung Anxiety Scale (S.A.S.) which quickly quantifies levels of anxiety making it ideal for collecting data for our study. We anticipate that our results will reflect no relative difference between anxiety levels of participants and which social media platform they indicate as their primary channel(s). Results from this study may direct methods or areas for future research on the topics of social media and technology use and anxiety or mood disorders.

The world’s largest dam removal project is now occurring on 283 hectares of sparsely vegetated sediments once inundated by the dams in Olympic National Park. In 2011 two dams on the Elwha River were removed to open the waterway to salmon and restore the riparian ecosystem. The objective of this research is to quantify the amount of nitrogen in the soil at former Lake Mills reservoir of the lower Elwha River Basin and determine if this amount is sufficient for native vegetation. In the summer of 2013, 174 soil samples were obtained from 77 plots in Lake Mills using hand tools and a bulk density corer. Samples were collected at depths of 0-20cm, 20-50cm, 50-100cm, 2-3m, and 4-5m. The samples were then run through a carbon, hydrogen and nitrogen analyzer, and the concentrations of nitrogen measured. The mean soil nitrogen concentration was 0.092% with a standard deviation of 0.026. The mean soil bulk density was 1.39 g/cm3 with a standard deviation of 0.31. To a depth of 0-50cm, the mean soil nitrogen percentage was 0.08 with a standard deviation of 0.03. Soil nitrogen concentration increased with soil depth, ranging from 0.077% at 0-20cm to 0.129% at 4-5m. The mean fine sediment nitrogen concentration was 0.100. The mean coarse sediment nitrogen concentration was 0.057. Total content of nitrogen was 1610 kg ha-1. The data suggests that the amount soil nitrogen in Lake Mills is insufficient for native vegetation. Nitrogen is an essential nutrient necessary for plant growth, and the most frequently limiting nutrient in most Pacific Northwest soils. Determining the concentration and spatial distribution of soil nitrogen will aid the Park Service in the restoration of Lake Mills; an effort being undertaken to reduce erosion, prevent the growth of invasive plant species, and enhance the natural beauty of Olympic National Park.

 Organic thin-film solar cells have received considerable attention on account of their flexibility, ease of production, and light weight. Recently, modifications of these solar cells’ nanoscale structures has yielded efficiencies higher than 10%. To further improve device efficiency and durability, detailed understanding of the pathways of charge generation and loss (through recombination of opposing charges) is essential. Electron paramagnetic resonance (EPR) spectroscopy, which directly observes radicals and unpaired electrons, including the charge carriers in these solar cells, is a useful tool for the characterization of these pathways. EPR analysis of relevant materials such as P3HT:PCBM blends indicate aspects of charge carrier lifetime and recombination rate that are temperature-dependent. Also, analysis of the variation of EPR spectra with modulation frequency allows us to directly determine the lifetime of excited charge carriers. Combination of this data alongside theoretical modeling of charge generation and recombination allows us to determine parameters such as charge recombination rates. For this purpose, a new method for simulation of an EPR spectrum that allows for computationally-efficient incorporation of radical life times, relaxation times and saturation has been created.

 Embryonic stem cells can be used to study development and diseases. Mouse embryonic stem cells (mESC) are used as a model in developmental research and display several pluripotent properties not seen in the current human embryonic stem cell (hESC) lines. A newly derived line of naïve hESCs is thought to recapitulate an earlier stage of development, and resemble mESCs. To understand how the naïve hESCs differ from typical hESCs we are investigating DNA methylation, a form of epigenetic regulation, on a global scale at nucleotide resolution. The underlying genomes of cells are highly similar, yet their epigenomes change with cellular differentiation and development. Epigenetic modifications are important factors controlling gene regulatory elements. To analyze whole genome DNA methylation of the naïve hESCs, I am using MethPipe, a computational tool, to determine how the naïve hESC line differs from a later stage hESC line, and if the naïve line is more similar to mESCs at the epigenetic level. I will compare global methylation levels, hypo/hyper-methylated regions, and partially methylated domains. I will determine which regulatory elements, such as promoters, enhancers and CpG islands, show similarities and differences. We also expect to determine differential non-CG methylation (a characteristic of ESC). This characterization of naïve hESCs will provide a foundation for studying the earliest stages of human development.

The incidence of esophageal adenocarcinoma (EAC) is increasing world-wide. Early and specific detection of EAC and its precursor non-cancerous stages, particularly Barrett’s esophagus (BE), is crucial to treatment and long-term survival. Currently, endoscopy coupled with serial biopsy is the gold standard for diagnosis. Standard endoscopy lacks specificity. BE presents as a salmon-reddish discoloration against the pale-pink color of healthy esophageal tissue. Cardia intestinal metaplasia (CIM) arising from gastric tissue is visually similar; both develop in response to prolonged acid reflux and are often confused during histological analysis. However, CIM is unrelated or even negatively correlated to EAC development. Distinguishing between BE and CIM at the gastroesophageal junction is crucial to minimizing the use of invasive biopsy. Targeting molecular markers specific to disease state can enhance specificity. Fluorescent dyes attached to targeting molecules are used to label these biomarkers. Instrumentation designed for in-vivo fluorescent imaging, like the ultrathin, flexible, multimodal scanning fiber endoscope (SFE) can then excite and image the fluorescent dyes, effectively red-flagging abnormal regions for both enhanced biopsy capture and point-of-care diagnosis. Reliance upon one biomarker alone is ineffective due to patient heterogeneity, as expression varies between patients due to factors including genetic background and lifestyle. Multiple biomarkers are necessary for specificity. The purpose of our work is to first demonstrate that the three cell surface biomarkers, EGFR, C-MET, and ERBB2, well established in other cancers, are appropriate for early EAC or BE diagnosis. We plan to demonstrate the simultaneous application of all three biomarkers to provide added specificity and sensitivity for diagnosis and ultimately prognosis. These goals will be achieved through immunofluorescent staining of cell lines and paraffinized tissue samples excised from patients. The efficacy of using multiple markers will be assessed through image processing and statistical analysis.

It’s important that the public heeds warnings issued by the local government during a natural disaster, despite costs associated with taking preventative action which are not guaranteed to have been necessary if the threat fails to occur. This can lead to a low compliance rate for warnings issued for natural disasters. To begin to understand the characteristics of warning-compliant citizens, we aim to see if previously experiencing a catastrophe influences future behavior. We believe the disaster would be more salient and easily recalled the next time a warning is issued, potentially making people more cautious. This is called the availability heuristic (Tversky & Kahneman, 1974). In a controlled computer simulation, we ask participants to act as a road maintenance company president and judge whether to take precautionary action in preparation for freezing roads. Each of 90 trials will represent a single night, and one or three of these trials will be manipulated in the middle of the experimental sequence so participants experience a large and sudden temperature drop from what was forecasted. For those trials, we expect participants not to take precautionary action, leading to a large penalty. We then anticipate that after experiencing these “Big Miss” trials that the participants will take precautionary measures more frequently, because of the activation of the availability heuristic. If this happens, we may propose that victims of previous disasters are more cautious with similar scenarios in the future, and we may then move forward in finding ways to create the same mentality in residents of disaster warning zones.

Purkinje cells are large neurons with intricately elaborate dendritic spines inside the cerebellum of the brain, where they play an important role in controlling motor movement. The migration of Purkinje cells during morphogenesis of the cerebellum requires multiple signaling pathways. Prior research has shown that the extracellular matrix protein Reelin promotes Purkinje cell migration by triggering tyrosine phosphorylation of the cytoplasmic adaptor protein Dab1. Reelin signaling is terminated by the proteasomal degradation of phosphorylated Dab1. Cullin5/RING ligase (CRL5) is the E3 ubiquitin ligase that targets active Dab1 for degradation and its absence causes Dab1 accumulation and promotes Purkinje cell mispositioning. SOCS (suppressors of cytokine signaling) proteins are substrate adaptors for CRL5 that bring phosphorylated substrates to the ligase. SOCS7 has been previously described to be the substrate adaptor that recruits phosphorylated Dab1 to CRL5 during neocortical layering. However SOCS7 is not required for ubiquitylation and turnover of Dab1 in the cerebellum. Thus, the identity of the SOCS protein that regulates Dab1 in Purkinje cells during migration remains to be explored. A novel in utero electroporation method in the cerebellum enables us to manipulate expression level of different genes without the need to generate mutant animals. We will inject specific shRNAs to silence SOCS expression in Purkinje cells to address which SOCS recruits phospho-Dab1 to CRL5 in the cerebellum. Our expectation for this experiment is that knockdown of one or more SOCS proteins will mimic the same misposition phenotype of the Purkinje cells as described for CRL5 mutant animals. Thus we will identify the SOCS-CRL5 complex that regulates the migration of Purkinje cells during cerebellum development. Purkinje cell mispositioning may contribute to diseases like spinocerebellar ataxias and autism, pointing to the exigency of conducting experiments on the mechanism of Purkinje cell migration.

Previous research done in our lab found that poplar and willow trees grow in nutrient-poor soil, and they obtain their nutrients through partnership with microbes that live within them (referred to as Endophytes). We then have successfully isolated endophytes and are now exploiting them for plant growth promotion, stress tolerance. My research question is to test if inoculating poplar clone OP-367 with endophytes can help the plant survive drought condition. The experiment was set up by controlling watering schedule that simulate drought condition and compared the results of the uninoculated control group and the inoculated group. We quantified the results using measurement of wet and dry biomass, chlorophyll (SPAD readings) which quantifies the health of the plant by measuring the greenness, and maximal photochemical efficiency (Fv/Fm), which quantifies the photosynthesis happening in the plant. The implication of this project will permit better growth of plants under water stress condition. The potential of endophytes to increase drought tolerance in OP-367 poplar hybrid and or other plant species can become a potential remedy for rescuing agriculture problems in relation to drought weather condition.

Stroke is the fourth leading cause of death and the leading cause of long-term disability in the United States. Microglia are the resident immune cells in the central nervous system and play critical roles in stroke pathophysiology. During ischemic stroke, cytokines known as interferons (IFNs) are released from CNS cells and bind to microglial cell surface receptors. This binding initiates a signaling cascade that leads to increased expression of Interferon-Stimulated Genes (ISGs). We hypothesize that increased expression of ISGs contributes to post-stroke neuroprotective pathways. There are three types of IFNs, and Type 1 includes IFNs α and β, which are well-known immunomodulatory factors. Type 2 IFNs include IFNγ, a prototypical pro-inflammatory activator. Type 3 IFNs (IFNλs) are the most recently discovered and least well characterized. IFNλs are effective in treating several viral illnesses and they are better tolerated (fewer side effects) than Type 1 IFNs. We have recently shown that Type 1 IFNs can induce increases in ISG expression in microglia. My project goal was to characterize the ability of IFNλs to induce ISG expression in microglia and compare their pharmacological potency to that of Type 1 IFNs. To do this, I stimulated primary microglia with varying dosages of IFNλ3 and for varying amounts of time. I extracted RNA and prepared protein lysates from the stimulated cells. ISG expression was quantified by qRT-PCR and immune (Western) blotting. I also collected conditioned medium and assayed for the release of specific ISG chemokines, such as CCL5 and CXCL10 using ELISA. I obtained dose response curves and time course data. Our results showed robust up-regulation of multiple ISG mRNA transcripts and increases in release of specific chemokines following stimulation of microglia with IFNλ3. These studies will help us continue to characterize important cellular and molecular targets for pharmacological intervention in stroke.

According to the World Health Organization, cardiovascular diseases are the leading cause of mortality worldwide. The low regenerative capacity of the heart renders it ineffective in healing itself after heart failure. Our lab has developed efficient methods to derive cardiomyocytes (cardiac muscle cells) from human embryonic stem cells (hESCs). However, it has become apparent that the cardiomyocytes derived using current differentiation methods exhibit immature morphological and functional properties. An important characteristic of mature cardiomyocytes is the presence of T-tubules, which are invaginations of the plasma membrane into the cell interior. These subcellular structures are critical for synchronized calcium release and contraction of mature cardiomyocytes. The protein caveolin-3 is a critical component of T-tubules, and thus its visualization can be used to assess the development of these important subcellular structures. I am currently evaluating the expression of caveolin-3 in hESC-derived cardiomyocytes using immunostaining techniques. For these studies, I expose the sample to anti-caveolin-3 as the primary antibody followed by a complementary secondary antibody conjugated to a fluorescent marker for visualizing the protein. Titration of several parameters, such as time of incubation and dilution of the antibodies allows for staining optimization. I have obtained detailed images of T-tubules aligning with striations in control heart tissue, but the staining is diffuse for the hESC-derived cardiomyocytes grown in standard 2D cultures. I am currently exploring whether this is a technical issue or the result of developmental immaturity. To assess the latter, I will stain hESC-derived cardiomyocytes that are more mature due to mechanical conditioning in 3D constructs. Optimization of this technique will facilitate our efforts to find novel agents that can accelerate the maturation of hESC cardiomyocytes. Finding ways to mature these cells will then provide a better platform for therapeutic applications such as drug screening, disease modeling and cardiac repair.

 The spins of electrons bound to donors are promising candidates as qubits for quantum information processing. The quantum properties of these donor-shy bound electrons are studied in high magnetic fields while immersed in liquid helium at 4K. For stability reasons, the level of liquid helium in the experiment must be monitored with a sensor. An effective way of implementing a sensor is with a superconducting wire. Super conductors have the property of having zero electrical resistance when they are cooled below a critical temperature. Above this critical temperature, the superconductor has significant electrical resistance. Using these characteristics, a superconducting wire can be used to construct a liquid helium level sensor. When current is run through a thin wire submerged in liquid helium, a relationship can be established between the voltage drop across the sensor and the amount of wire submerged in the liquid helium, allowing the liquid helium level to be measured. Results will be presented on the sensor design and performance.

“Fetal alcohol spectrum disorders” (FASD) are considered to be the leading preventable cause of mental retardation. Among this spectrum Fetal Alcohol Syndrome (FAS) represents the most extreme consequence of maternal alcohol consumption and as such associated facial dysmorphologies and neurological defects have been heavily investigated. In this project, our lab is interested in assessing the extent craniofacial development is affected due to chronic paternal and pre-conception maternal alcohol consumption using a mouse model. For the paternal exposure project, male mice were first bred with normal females as control and then given a 10% ethanol water solution for six weeks, which covers a full cycle of spermatogenesis in mice, and mated with normal females again as experimental compare. For the pre-conception exposure project, female mice are divided into three groups given different solutions respectively, four days a week for ten weeks: 10% ethanol water exposure, calorie-matched sucrose solution, and water. Upon completion of exposure, they were bred with normal males. When litters are at 60 days of age, the craniofacial phenotype of the specimens will be assessed using high-resolution microCT and their shape differences will be statistically assessed using a set of landmark and geometric morphometric methods (GMM). By the time of the Symposium, we expect to complete the image acquisition of the control group, as well as some of the initial GMM analysis. We will also be able to report differences in litter size, birth weight, and growth rate of litters between groups in both projects. As a whole, we expect to see subtle but significant differences in craniofacial development between mice that are born to fathers or mothers consuming alcohol and their controls. From there, the leading mechanism and degrees of alteration of craniofacial development can be related to the effects of chronic alcohol exposure in humans.

 In eukaryotic organisms, regulation of gene transcription depends on the presence or absence of transcription factors, which are proteins that bind to promoter sequences on DNA. One common promoter element is the TATA box, and transcription is initiated when a protein complex called TFIID (which stands for transcription factor for RNA Polymerase II, D subunit) binds to it. TFIID is a multi-subunit machine, made of a TATA-binding protein (TBP) and TBP-associated factors (TAFs). Our lab has previously shown that an interaction between TAF1 and TAF7 inhibits TFIID's ability to initiate transcription of select genes. If TFIID cannot function properly, TATA-box controlled genes will be transcribed when they should be repressed. Some of these genes are cyclins, which are important for regulating the cell cycle. If they are misexpressed, a cell could proliferate with damaged DNA. This type of uncontrollable proliferation is a key component of cancers. Therefore, I am studying the interaction between TAF1 and TAF7 to determine the nature of TFIID inhibition. TAF7 is the proposed inhibitor in this model. I have cloned wild-type and mutant TAF7 genes, expressed and purified protein, and plan to use co-immunoprecipitation to determine if TAF1 and TAF7 associate. If the wild-type and mutant molecules have different affinities for TAF1, then I can conclude that the residue I've mutated is essential for TAF1 association. Later, I would like to study the difference in TAF1 activity in the presence of wild-type or mutant TAF7. This research asserts its relevance through the ubiquity of TFIID-mediated transcription regulation in every one of our cells, as well as the implication that TAF7 misexpression could lead to aberrant cell growth and possibly cancer. 

Polydimethylsiloxane (PDMS) micro and nanoposts have been widely used to measure traction forces in cells, however, the mechanical properties of PDMS micro and nanoposts remain unclear. The current measurements of our lab’s PDMS posts are calculated based on the bulk or global properties of PDMS versus the local modulus, which matters at the cellular scale. It is important to denote this difference because cells actively sense their local physical microenvironments, not the global environment or bulk modulus. In this project, I fabricated specified polydimethylsiloxane micro and nanoposts, which are used in lab to measure cellular forces, and examined the mechanical properties (spring constant and Young’s modulus) of these micro and nanoposts, taking into account the fact that cells only sense their local microenvironments which means experimentally finding the properties of PDMS at the micro and nanoscale. To fully characterize this hydrogel, I utilized atomic force microscopy (AFM) to provide a local constant force (0-60nN for calibration), scanning electron microscopy (SEM) to calculate the dimensions of specified posts of the micro and nanoposts at 110⁰ C at different curing times. Future research utilizing PDMS micro and nanoposts to measure cell forces will take into account these correction values for exact force measurements. These findings provide an update and more thorough way to calculate the mechanical properties of the PDMS micro and nanoposts used in lab.

Behavior in mammals follows a 24 hour clock, wherein some activities like locomotion and feeding are prominent in certain periods and inconspicuous in others. These behaviors are said to follow a circadian rhythm and are controlled primarily by a region in the brain called the Suprachiasmatic Nucleus (SCN), known as the “Master Clock” of the body, that uses light as a zeitgeber, or entrainer, for its circadian output. Previous work has shown that non-photic entrainability of feeding is possible, suggesting that light to the SCN is not the only zeitgeber affecting circadian activity. Additionally, other studies have shown stress to alter certain circadian activities, implicating it as a possible zeitgeber. Although much research has been done on how the SCN affects bodily functions, not as much work has been done identifying other zeitgebers that may directly influence or compete with the SCN. Our experiment sought to find the effects fear has on circadian rhythms and if it could act as a zeitgeber in the absence of the SCN. We studied rats living in a closed environment with a safe bedding area and an aversive foraging area consisting of an electrical shock grate where food and water could be obtained. The room was kept on a strict 12 hour light/dark cycle, with random shocks occurring during either the light or dark cycle. We found that compared to control animals, SCN lesion rats had significant changes in their circadian feeding behavior but not locomotion. Although our results show that fear cannot act as a zeitgeber in the absence of the SCN, they do indicate fear as a modulator for circadian behaviors. Given the close link between fear and stress, our findings are especially relevant for students, showing the negative effects of stressful environments.

Testosterone plays a major role in aggressive and territorial behavior in many male vertebrates during the breeding season. Song sparrows, Melospiza melodia morphna, are not only aggressive in the breeding season but also in the non-breeding period, when their testes are regressed and plasma testosterone levels are low. Previous studies suggest that this territorial aggression is mediated in part by high levels of an androgen and estrogen precursor, dehydroepiandrosterone (DHEA), produced by the adrenals. In this study, we aim to determine whether DHEA exerts these effects via an androgen or estrogen receptor mediated mechanism, or both. Male song sparrows were captured, kept on a short day cycle (8L: 16D) typical of the non-breeding season, and implanted subcutaneously with either empty or DHEA-filled silastic implants for fourteen days. DHEA implanted birds had significantly higher circulating DHEA and testosterone levels and showed increased aggression in a laboratory-based aggression test. Brains were extracted and in situ hybridization was used to label androgen receptor (AR) and aromatase mRNA. Aromatase is the enzyme that catalyzes the transformation of androgens to estrogens. Using dark field microscopy, AR and aromatase mRNA expression in brain regions previously associated with aggressive and reproductive behaviors were quantified. In the ventromedial nucleus of hypothalamus (VMH), AR expression was significantly greater in DHEA birds compared to controls. However, in the preoptic area (POA), aromatase mRNA was significantly higher in DHEA birds over controls. In the medial preoptic area (mPOA), neither AR nor aromatase expression differed between DHEA and control birds. We conclude that DHEA may cause non-breeding aggression via both androgen and estrogen receptor mediated mechanisms. Deeper understanding of how exogenous DHEA affects the brain and behavior of song sparrows may provide useful insights for biomedical researchers interested in investigating the health benefits and side effects of DHEA supplements.

Childhood epileptic encephalopathies (CEE) are a subset of rare epilepsies, largely characterized by seizures and mental disabilities. Recent studies, including those by the Mefford lab, have shown that about 30% of CEE is caused by mutations in specific genes.These mutations are ‘de novo’ – they arise during fertilization or the development of the embryo. While many types of epilepsy have a hereditary basis, de novo mutations are increasingly recognized as significant risk factors in some epilepsies. The mutations also change the amino acids that make up proteins and are mostly in exons – parts of the DNA that code for protein. Few studies focus on synonymous mutations that do not change the amino acids of a protein or mutations in introns –pieces of DNA that have regulatory function but do not code for protein. We have previously sequenced 29 known epilepsy genes in a cohort of 800 CEE patients, but focused primarily on exonic mutations that change the protein sequence. My goal is to understand whether intronic or synonymous mutations that we have already found in these patients are inherited or de novo. We hypothesize that these classes of de novo mutations disrupt the typical formation of RNA transcripts, which in turn results in an absent or mutant protein. I will use molecular inversion probe (MIP) capture to determine the inheritance of intronic variants in patients with CEE. A MIP is a piece of DNA that is designed to target specific regions of DNA which are then amplified and analyzed; the technique enables us to capture a large number of variants at once. Discovering de novo variants in intronic regions of genes will deepen our understanding of the cause of CEE. These variants will inform us of affected biological pathways and contribute information for future CEE treatments.

There has been concern about the effect of anthropogenic deposition on high-elevation alpine ecosystems such as in the North Cascades and Mount Rainier National Parks, as plant uptake of this nitrogen could alter species composition. The ability to identify roots of alpine plants will enhance the ability to study nitrogen deposition effects on these important ecosystems. The objective of this project was to determine methods to identify alpine plants by their roots in soil samples. Plant species of roots were recorded and soil samples were collected beneath that plant. Lupine and heather soil samples were collected as the dominant alpine species at the park sites. Soil samples were brought to the lab to be examined for root morphologies and mycorrhizal relationships. The lupine roots had constant morphological features including a tap root system, straightened woody roots, less branching, some striations and deep roots. White heather and pink heather were observed to have a fibrous root system, branching of roots, lower nodes of root development, few striations on the roots, and were variable in root length. The morphological features of each root varied depending on the soil the plant species grew in, with deeper root systems in more fertile alpine soils. The roots were taken from shallow soil cores therefore only short sections of roots were available. Mychorrihizal associations were examined from ink-stained alpine plant roots and assessed for the presence of ectomychorrizal or arbuscular mychorrhizal. With the development of these reliable methods to identify alpine roots, data such as root biomass per unit of soil will document specific plant impacts of nitrogen deposition on alpine ecosystems. With these specific plant impacts from nitrogen deposition known, the National parks can acess ecological problems caused by nitrogen deposition and discuss laws to lower nitrogen pollution.

Understanding what people know about the topic of climate change is important to develop better strategies for informing and teaching the public about important environmental concerns. To understand people’s knowledge and personal perspectives on climate change, I used a series of surveys to answer four important research questions regarding this subject matter. 1. Do people with the most negative opinions about climate change tend to be less informed about the science surrounding this subject? 2. Is the college/university experience by itself enough for people to have an understanding about this subject matter? 3. Do people who think that climate change is a serious problem have a strong background in environmental sciences or studies? 4. What are the media or literary sources predominantly used by the people with the most misinformation about climate change? The surveys were given to several classes across the UW campus as well as to an outside adult population and to two local high schools. The survey data provided important information regarding the diversity of the sample group, including age, major, educational level, where they grew up and previous classes. This study should also provide important information regarding potential relationships between educational level and academic background to their respective knowledge about climate change. This study may also provide information about the potential information sources that people use as the basis of their understanding of climate science.

Dravet Syndrome (DS) is a rare form of epilepsy characterized by severe seizure phenotype beginning in early childhood. It is primarily caused by loss-of-function mutation in SCN1A gene which encodes a voltage-gated sodium channel Nav1.1. DS is resistant to most commercially available anti-epileptic drugs. However, ketogenic diet (a high fat and low carb diet), has been shown to provide considerable seizure protection to patients with this epilepsy. Its mechanisms of action are not completely known. One of the hypotheses explaining the efficacy of ketogenic diet is that the elevated level of ketone bodies leads to the modification of Tricarboxcylic Acid Cycle (Krebs Cycle), increase GABA (Gamma Amino Butyric Acid) synthesis, and promote energy production in brain tissues. This research study focused on understanding the effects of ketogenic diet on epiletiform activity in DS mice. We recorded field potentials from hippocampi of wildtype and DS mice in brain slice preparations. We added ketogenic diet metabolites to the Artificial Cerebrospinal Fluid (ACSF) and investigate their effects on brain activity.

H5N1 Avian Influenza is a global threat. Given the globalized nature of our world as well as the universal human susceptibility to disease, an influenza outbreak in Southeast Asia poses a concern for the rest of the world. Although there are no major transmission cases reported among human populations, the virus might evolve to become lethal over time. Despite it being a global threat, only the West was well situated for preparedness during the 2005 outbreak in Asian poultry farms. Vaccines created were targeted towards developed countries, and few drug companies were willing to work with Indonesia and Thailand because of lower profit potential. This policy report is designed to accommodate the needs of high-risk countries. Due to the potential destruction of what the avian influenza carries, this report outlines the fundamental causes of avian influenza. We question how the avian influenza virus is evolving to become infectious to humans, and investigate necessary changes to avoid future outbreaks. From our research through news articles, public health journals, microbiology textbooks, and a telephone conference with Dr. Suraya Afiff of the University of Indonesia, it is apparent that food sovereignty is the most crucial issue in preventing future outbreaks. We address our policy-recommendation report to La Via Campensina (LVC) an Indonesian Non-governmental Organization attempting to regulate the poultry industry and working to minimize the negative consequences of large agriculture. Through this report to LVC, we hope to convey a specific set of strategies that will mitigate future pandemics. Above all, we show LVC the importance of considering future outbreaks in the fight for food sovereignty.

This study is using Hamilton Search (HS) memory task and Set breaking (SB) task to assess working memory, cognitive flexibility and inhibition control of a variety of domestic dog breeds reflecting differences in evolutionary lineage. HS and SB have been tested on young pigtail macaque monkeys as well as other species. The HS task consisted of each dog searching 4 containers side by side to find a randomly placed food reward in 5 sessions of 16 trials each. SB followed HS and the location of the food was fixed based on performance in the HS trials. Trials continued until a dog reached criterion, establishing that they had learned the position of the food. The latency and search pattern was recorded to determine performance of the task. Results compared to pigtail macaque monkeys as well as across dog breeds. The primary variables analyzed were number of containers examined, probability of selecting an outer container and right/left side bias. Preliminary results show that dogs in general are approaching this task in a remarkably systematic manner, searching from one end to the other, which is considerably different than the performance of macaques. This approach leads to few recheck errors that, rather than indicating perfect memory, actually reflects an efficient alternative to memory. Results of this study can be used to establish the validity of these tasks, and provide interesting new perspectives on both optimal foraging strategy models and the refinement of training techniques.  

 Widespread use of tetracyclines in marine and fresh water aquaculture has resulted in tetracycline resistant (Tcr) bacteria in these environments. Previously, 15 of 25 Tcr Chilean bacteria were shown to carry seven different tet genes. The tet(A) was the most commonly identified gene [n=6]. In the present study, we reexamined nine bacteria that were previously negative for the 22 tet genes tested using PCR assays for the identification of tet(W), tet(X), and tet(39). Three of the isolates’ genera designation differed between phenotypic and 16S sequencing identification. The 16S identifications were used in the current study. Six (67%) of the isolates including 3 Acinetobacter spp., 1 Pseudomonas spp., 1 Psychrobacter spp., and 1 Corynebacterium spp. carried the tet(39) gene. None of the isolates carried either tet(W) or tet(X) genes. In the present study, 6 of the isolates carried the tet(39) gene making it as common as the tet(A) in the original study. Of the 4 genera identified 3 [Pseudomonas, Psychrobacter, and Corynebacterium] represented new genera not previously identified as carrying the tet(39) gene. We are currently testing the strains for their ability to transfer tet(39) by conjugation and/or transformation. The tet(39) gene was first identified in Acinetobacter spp. from freshwater trout farms, sewage and human clinical samples from Europe between 1986-1997. Since then the tet(39) gene has been identified in fish farms in Thailand, a polluted river in Nigeria and clinical samples in the US. This study shows that the tet(39) gene has been present in bacteria from Chilean fish farms since 1999. The tet(39) gene has wider host range in water associated bacteria than previously recognized and should be considered when examining Tcr water bacteria in the future. The current study illustrates that aquaculture contaminates the local environment into surrounding soils and water sources with antibiotic resistant bacteria and genes.

Immunohistochemistry (IHC) is a valuable technique used to validate biomarkers, not only because it provides direct visualization of biomarker expression in histologically relevant regions of examined tissues, but also because validated IHC assays may be implemented readily into clinical practice. IHC assays have traditionally been semi-quantified by pathologists, but this method is significantly costly, inherently subjective, and produces ordinal rather than continuous variable data. Computer-aided analysis of digitized whole slide images may address these serious limitations. Using tissue microarrays (TMAs) representing 578 prostate cancer (PCa) specimens stained for estrogen receptor β2, I evaluated the degree to which data obtained using computer-aided methods correlated with data obtained by pathologist visual scoring. Digital images were obtained using a whole slide scanner. Under pathologist supervision I annotated TMA spots using annotation software. Images were preprocessed using the Genie Histology Pattern Recognition software suite to segment tissues into three user-defined Image Classes (tumor, stroma, glass). Malignant epithelial DAB staining was separately quantified within cytoplasmic and nuclear regions using the Cytoplasmic algorithm using a continuous variable metric that incorporates both the staining strength and the percentage of positive pixels in cytoplasmic/nuclear malignant epithelial areas. I compared the IHC intensity data derived from the Cytoplasmic algorithm with pathologist visual scores and identified high Spearman correlations. This study demonstrated that computer-aided methods to quantify IHC staining within carcinomatous areas of PCa tissue specimens can produce highly similar data to visual evaluation by a pathologist. This will allow further research studies to be performed using less time consuming and more technician based image anaylsis methods instead of costly pathologist consultations.

 In neurodegenerative diseases, such as Alzheimer’s disease, tau protein aggregation is observed. In Alzheimer’s disease and related conditions, accumulation of insoluble tau impairs physiological function that ultimately leads to neuron loss. Using a transgenic Caenorhabditis elegans model that expresses human tau protein in neurons, we are able to see accumulated, insoluble, phosphorylated tau, and uncoordinated movement (Unc) phenotypes in these organisms. To identify genes participating in aggregated tau and Unc phenotypes, we conducted a mutagenesis; an estimated 20,000 worm genomes were screened and we isolated seven promising strains of C. elegans that carried loss of function genes suppressing the tau-induced phenotypes. These strains are currently being sequenced to determine the identity of the loss of function genes, which may represent novel suppressors of tau pathology. Currently, the Kraemer lab has identified two genes suppressing tau-induced Unc phenotype and called them suppressor of tau 1 (sut-1) and suppressor of tau 2 (sut-2). Animals with sut-1 and/or sut-2 mutations are resistant to the negative effects of tau, meaning that the proteins made by these genes are necessary for tau phenotypes. With this knowledge and the additional information to be gained from the sequencing of seven novel sut alleles, we hope that potential neuroprotective approaches for treatment can be developed.

Learning and memory are influenced both by adult neurogenesis and circadian rhythmicity; however, it is unclear if and how the biological clock influences neurogenesis. The BMAL1 protein is an essential component in the molecular clock. Transgenic Bmal1 -/- (knock out) mice, which lack the gene to produce this protein, are completely arrhythmic. Furthermore, the Bmal1 knockout mouse demonstrates poor learning and memory compared to its wild type littermate control which retains both Bmal1 alleles. As an animal that is both arrhythmic and exhibits deficits in learning and memory, the Bmal1 knockout mouse provides the opportunity to investigate the potential interaction of the molecular clock and neurogenesis. I hypothesized that the rate of adult neurogenesis in the hippocampus of the Bmal1 knockout mouse would be reduced as compared with wild type littermate controls. To quantify this, I used immunohistochemistry to fluorescently mark proliferating cells with bromodeoxyuridine (BrdU) in the subgranular zone (SGZ) of the hippocampus. BrdU is a synthetic nucleotide, intraperitonealy (IP) injected prior to brain collection, and incorporated into the DNA of newly dividing cells. I found that the Bmal1 knockout hippocampus had significantly reduced rates of cell proliferation and survival compared with wild type controls. Thus, BMAL 1 is required for adult neurogenesis in the hippocampus. This finding now invites the consideration of whether BMAL 1 protein plays a direct cellular role in adult neurogenesis, or if it is the disruption to systemic circadian rhythmicity which compromises neurogenesis.

Colorectal cancer (CRC) is the third most commonly diagnosed cancer among US men and women, and it disproportionately affects Korean-Americans (KAs). Currently, CRC related deaths are decreasing among all racial/ethnic groups, but increasing among KAs. Prevention and early detection of CRC, through cancer screening reduces disease incidence and death. However, research shows that KAs may not be adhering to the current CRC screening guidelines. The purpose of this study was to investigate whether KAs residing in Washington State are up-to-date with the current CRC screening guidelines and identify demographic predictors of CRC screening up-to-date status. We conducted a cross-sectional survey using convenience sampling among KAs ages 50-75 residing in three WA State counties (-Snohomish, King, and Pierce). Participants were recruited from Korean community organizations including churches, community health centers, and senior centers. The interviewers administered the 15 minutes survey in person. The total sample size was 170 participants. We analyzed data to investigate the number of KAs in the WA State who reported being up-to-date with CRC screening, based on current medical guidelines. We also examined demographic predictors that relate to CRC screening up-to-date status such as insurance status, having a primary care provider, annual household income, English proficiency, and education level. The findings highlighted the CRC screening up-to-date rates among KAs in WA State and demographic predictors that are associated with CRC screening up-to-date status. Identifying these predictors will allow researchers to understand the barriers and facilitators of CRC screening and develop interventions targeted to a particular high-risk group within KAs.

Public health concern in the U.S and across the world regarding the impact of environmental hazards on human health is becoming more evident as we develop tools to see disparities in exposure and health outcome. Pollutants in the air and water can have lifelong consequences for the quality of lives of many people. Therefore, our toxicology laboratory is examining the result of pesticide exposure and toxicity in children, and how it may cause harm to normal development. Our research centers on studying exposure using blood, urine, and oral buccal samples obtained as part of a Children’s Health study on Yakima Valley. We have determined blood pesticide, urinary metabolite concentrations, cholinesterase activity, paraoxonase activity, and genome wide variation in 200 families. We have analyzed buccal samples from farmworker and non-farmworker families that were collected during different pesticide spray seasons. We are studying these buccal swab samples of children and their parents to identify bacteria present within them. We are utilizing targeted 16S ribosomal polymerase chain reaction (PCR) to determine the phylogenetic composition of the microbiome in saliva samples. The microbiome refers to the vast colonization of microbes within the body. There are more bacterial cells in our bodies than human cells even though they are only about 10% of the total mass. We hope to find a difference in microbe composition and metabolism for samples from agricultural seasons (pesticide use and non-spray seasons), and for child and adult samples. We have already analyzed pilot samples and found consistent results. Recently, we have begun PCR-based analysis and sequencing the remainder of the cohort buccal samples to characterize the diversity of oral microbiomes. If a link is found between the microbiome, pesticides, metabolic traits, and host genotype then we can more fully describe risk and metabolic biomarkers for exposed populations. 

Adolescence through emerging adulthood is a peak period for risk taking and impulsivity matched with a significant increase in emotional reactivity. This increase in stress response makes those in this age group vulnerable to psychopathology and act more impulsively. One way to diminish this is using an effective mean of coping. Mindfulness meditation has been found to increase attentional control, cognitive control, and executive function, and these increases may reduce the tendency toward impulsive action when distressed. This current study will be comparing focused breathing –a component of mindfulness meditation– to rumination and unfocused attention to investigate whether focused breathing is an effective strategy for reducing impulsive tendencies. Using an expected n = 40 undergraduate students, we will test whether focused breathing produces reductions in impulsiveness and reward sensitivity following a stress induction, compared to a neutral or a rumination condition.

One of the goals of modern longevity research is to find pharmacological means to increase lifespan and/or healthspan without deleterious physiological effects. Berberine is a yellow, bitter-tasting natural alkaloid with a long history in Eastern Asian medicinal use in treatments of a variety of age-associated diseases and infections. In human cell culture models, berberine treatment increases AMPK activity, resulting in delayed senescence through inhibition of the mTOR/S6 pathway. Here, we use the budding yeast Saccharomyces cerevisiae and the nematode Caenorhabditis elegans to investigate the effects of berberine on healthspan and lifespan. For the yeast S. cerevisiae, growth rates of haploid and diploid yeast cells, as well as chronological (a measure of population viability through time) and replicative (a measure of mitotic potential) yeast aging paradigms were assayed in the presence of concentrations of berberine ranging from 0.1 - 100.0 ng/uL. For C. elegans, organismal lifespan as well as various healthspan measures are being analyzed. Currently, we have found that concentrations of berberine greater than 10 ng/uL in haploids are sufficient to inhibit cell growth and lower concentrations (0.1-10 ng/uL) result in increased yeast viability when shifted to respiratory metabolism, a means of extending lifespan in yeast. Our lifespan studies are ongoing. Our experiments will provide valuable insights into the role of this long-used medicinal substance on cell aging and health using two important longevity models.

Increased atmospheric CO2 has the potential to have fertilization effects on plant growth, as long as there are no limiting effects of insufficient Nitrogen, among other potential limiting factors. However making chemical fertilizers is energy intensive, producing more green house gases. The NIFA project seeks to use Nitrogen-fixing endophytes to minimize the need for chemical fertilizers. Endophytes are bacteria or fungus naturally found in plants, some of which have the capability to fix Nitrogen. Preliminary experiments show that some crop plants that do not normally have this association with Nitrogen-fixing endophytes can be inoculated with the endophytes to create this symbiotic relationship. For our experiments, we will be testing the possibility to inoculate Conifer species with endophytes for improved growth without the use of artificial Nitrogen fertilizers. We will compare the growth of conifers that have been inoculated with endophytes to those without. If inoculating the conifers with Nitrogen-fixing endophytes was successful, inoculated conifers should show more overall growth, as well as higher overall Nitrogen concentrations than the control varieties. If our experiments show these results, there is a potential for Nitrogen-fixing endophytes to be used in place of artificial chemical fertilizers when growing conifers, making commercial forestry more sustainable.

The North Creek Wetlands restoration on the University of Washington Bothell campus harbors a seasonally variable roost of American Crows (Corvus brachyrhynchos), of between 2,000 and 16,000 crows. I investigated the seasonal changes of crows’ movements within and around the North Creek Wetlands roost by making observations of their evening arrival times, numbers, and directions of origin. I investigated seasonal fluctuations in area of the roost by mapping the perimeter on foot; the nocturnal activities of crows in this roost by placing trail cameras; and the distances traveled daily by crows commuting to this roost by making field observations of their flight vectors at varying distances and directions from the roost. The nightly times of arrival of crows coming to their nearby pre-roost (R² = 0.955), their move from pre-roost to roost (R² = 0.992), and the time that >99% of crows are in the roost (R² = 0.9942) are strongly correlated with photoperiod. The roost area changes both seasonally and annually, averaging approximately 45,330m² in Winter of 2013, 127,880m² in Fall 2013, and 65,960m² in Winter of 2014. Population density also varies seasonally. Field cameras show little nocturnal movement of crows after arrival in the tree tops, and never show birds moving to the understory, shrub layer, or forest floor. While flight paths to and from the roost are highly variable in direction, observations suggest that some crows are commuting to this roost nightly from at least 30 kilometers, and possibly greater than 40 kilometers away. This study may help us to understand why the crows have chosen this, or other, wetland roost sites, and may be useful in crow management decisions and the planning of future restoration sites.

This project examines the ethical and social frames which facilitate environmental conditions that fuel the rampage of tuberculosis in India. Three key components of the basic biomedical outbreak narrative that are often overlooked will be brought forth to attention: the tuberculosis outbreak as more than just a bacterial spread, social constructs which shape the bacteria’s accessibility to victims, and socioeconomic determinants of treatment (and lack thereof). India accounts for approximately one-fifth of global tuberculosis occurrences, and with over 2 million cases in the country, tuberculosis is a continuously growing health concern, posing a major threat to the well-being of the population. Due to India's developing health care system, pollution, and poverty, these conditions amplify the ease of transmissibility and rate of infection. As treatment regimens are abandoned due to social stigma, medical side effects, and overwhelming price tags, the bacterium which causes TB (mycobacterium tuberculosis), tends to mutate and become drug-resistant to varying degrees; increasing the difficulty of containment and scientific development of effective medicines. Furthermore, the capitalist structure of the pharmaceutical industry contributes to the financial challenges which prevent the eradication of tuberculosis. Drawing on the theory of biopolitics while using sources of statistical information and analytical discourse,  my presentation seeks to explore various properties of the disease and discover how the complex interconnectedness of these underlying issues attribute to forms of structural violence, perpetuating the growth of tuberculosis in India.

Cortisol, the main glucocorticoid hormone in humans, and testosterone, an androgenic hormone, are secreted from the adrenal cortex and gonads respectively in response to physiological and social stimuli. Cortisol and testosterone both display diurnal patterns with peaks in the early morning and gradual declines throughout the day. These morning peaks have been suggested to play a role in preparing the body for the day. The hypothalamic-pituitary-adrenal and hypothalamic-pituitary-gonadal axes regulate stochastic, homeostatic processes, which respond to short and long-term caloric changes. Although studies have determined the effect of fasting on cortisol and testosterone, the effect of the time of feeding is currently unknown, particularly the effect of eating close to bedtime on subsequent morning hormone levels. We hypothesized that the morning awakening response of cortisol will be resistant to changes in the timing of feeding before bed and testosterone will be more responsive to these changes. To test for the effect of timing of eating the night before, twenty male and female participants, aged 20 – 35, followed a four-day regimen. This protocol included three cycles of controlled time of evening feeding and next day saliva collections upon waking up and 30 minutes after waking up. The first cycle required consumption of dinner four hours before sleeping, the second cycle required consumption of dinner one hour before sleeping, and the third cycle was a repeat of the first. Salivary cortisol and testosterone were measured with enzyme immunoassays. Morning cortisol levels following late night feeding are expected to display no difference from those following normal dinner times, while morning specimens following late night feeding specimens may show increased testosterone awakening responses. This research may provide insights into the timing of eating and homeostatic regulation in humans.

Merkel Cell Carcinoma (MCC) is a rare, yet aggressive skin cancer with a disease-associated mortality of approximately 40%. Although surgery is typically a component of standard care, it is unclear how often patients should also receive radiation to control disease at the primary site. One retrospective study of 1254 patients found that of the patients who received surgery alone, 39% recurred locally compared to only 12% of the patients who received surgery in combination with radiation therapy. In contrast, in a cohort of 243 patients from New York treated with surgery alone, only 3.8% recurred locally. In order to establish a better standard of care for low risk MCC patients and attempt to resolve this discrepancy in the literature, we identified low risk patients within our cohort based on the following inclusion criteria: 1) primary tumor ≤1cm in diameter, 2) microscopically negative margins on the surgical excision, 3) absence of lymphovascular invasion in the primary tumor, 4) absence of profound, chronic immunosuppression, and 5) no microscopic evidence of nodal metastasis in the sentinel lymph node biopsy. Of the 29 patients who met these criteria, 17 were treated with surgery alone, while 12 received adjuvant radiation to the primary site after surgery. We found that 4 patients recurred locally (24%) among the patients treated with surgery monotherapy, while there were no recurrences among patients treated with surgery and adjuvant radiation. Even patients with low-risk MCC still have an appreciable chance of local recurrence if treated with surgery alone. While this risk may be acceptable to some patients, adding radiation therapy can lower the chance of local recurrence.

Sepsis is a medical condition characterized by potentially fatal generalized inflammation, typically resulting from infection. Despite previous attempts to identify mechanistic therapies, no targeted intervention has impacted survival rates. Prior hypotheses focused on immunologic mechanisms, but this project investigates metabolic responses in sepsis using a mouse model. The current target of investigation is a nuclear hormone transcription factor called PPARalpha. PPARalpha regulates the expression of proteins and enzymes associated with inflammation, stress, lipid metabolism and is related to both human and mouse survival in sepsis. We hypothesize that PPARalpha is necessary to maintain energy production sufficient to support organ function in sepsis and that deficient PPARalpha activity results in organ system failure. Because cardiac tissues preferentially use fatty acids as an energy source, we hypothesize that it is the organ that will be most affected in sepsis. Sepsis was surgically induced in mice, tissues were harvested at specific time points and gene expression was quantified using real time qPCR. Seven genes involved in fatty acid metabolism were analyzed at time points 0, 24, 48 and 72 hours. PPARalpha expression was higher in wild-type tissues, but it was down-regulated in both wild type and knockout mice. In addition, wild type mice showed greater expression of fatty acid oxidation genes than PPARalpha-knockout mice at every time point though this group of genes was down-regulated in both cohorts. Knock out mice also showed increased cardiac PPARgamma expression at 24 and 48 hours, which is a marker of reduced survival in other models. Our findings indicate that wild type mice are better able to maintain energy production necessary to preserve tissue function. These results support the hypothesis that the major cause of death in septic mice is cardiac failure due to impaired PPARalpha expression and therefore inability to sufficiently metabolize fatty acids.

 Little is known about the conditions that permitted photosynthetic life to survive the global glaciation events known as Snowball Earth events. During this time, sea glaciers covered the Earth’s oceans, prohibiting the transmission of light. Without liquid water and sunlight, where did photosynthetic life survive? Research has shown that under certain climatic conditions, areas of sufficiently thin ice known as refugia could have existed at the landward ends of narrow channels where sea glaciers would be unable to penetrate. The idealized channels used in sea-glacier flow models allow for refugia under a restrictive range of climate conditions. However, experiments varying the channel entrance width have shown that a less restrictive range of climate conditions are needed for these refugia than has been previously thought. I hypothesize that moderate variations in channel entrance and sidewall geometries could both 1) expand the range of climate conditions that allow for a refugium at the landward end of the sea glacier and 2) generate thin ice zones in additional locations within the channel, likely near the entrance and on leeward sides of promontories. Utilizing finite element modeling techniques to simulate rectangular, trapezoidal, and round-sided channel entrances and promontories,  I will determine the locations of regions of zero or near-zero ice thickness in idealized channels. Combined with reconstructions of real-world Neoproterozoic geomorphology, these results could help us understand the conditions under which photosynthetic life survived Snowball Earth and even help to locate evidence of Snowball Earth-era photosynthetic life on the modern Earth.

In this project, I will compare river channel patterns, both in profile and in planform, of drainages that flow across mapped faults in Peninsular Malaysia. The purpose of this study is to evaluate the degree of fault-slip activity throughout the region. Peninsular Malaysia is a tropical country located ~300km to 600km from the seismically active Sumatra subduction zone. Despite this close proximity, Peninsular Malaysia is considered a tectonically stable terrain because of low seismic activity. Based on previous seismotectonic maps, most of the earthquakes are less than magnitude 4 and concentrated in the southwest of Peninsular Malaysia, with exception to the magnitude 3.8 Baling earthquake, which occurred in the northwest. Recent fault reactivation that is triggered by the active Sumatra subduction zone may have caused these earthquakes, but this hypothesis has been difficult to test due to overall rapid erosion rates and the small magnitude of the earthquakes. I will use open source digital elevation models (ASTER GDEM V2), along with previously published geologic and seismotectonic maps of Peninsular Malaysia to complete a regional GIS assessment. Fault activity will be assessed using fluvial and topographic metrics useful in identifying active structures. In particular, I expect to find knickpoints and overall steeper rivers as well as changes in hillslope and local relief near to active faults. This reconnaissance is a part of a larger project in Malaysia to incorporate geomorphic data with Southeast Asia active fault mapping in Earth Observatory of Singapore (EOS). The results of this work will be incorporated further with the Global Earthquake Model (GEM) for earthquake hazards.

Due to small position uncertainties within internal systems, it is difficult to accurately predict the movement of cable-driven robots. By studying what happens within the pulley-cable system, we will be able to propose a model that accounts for the specific properties of the system, such as friction, tension, and viscosity. This model can be used for more accurate control of robot movement.

A pulley board was built with similar mechanical parameters (same pulleys, cables, tensions, and dynamics) as the RAVEN II surgical robot of the UW BioRobotics Lab. The board was fitted with velocity sensors which measured the movement of the cables when predetermined forces were applied to the ends. Position, velocity, and acceleration functions were obtained from these data. A proposed model was formed from physical predictions of the cables, and by fitting the data from the pulley board into the proposed model, functions for a viscous damping coefficient and friction force were found. From the findings, the model was refined and applied to the RAVEN II, transforming sensor inputs into a more accurate estimation of the robot pose.

Millions of Americans experience inhibited motor function due to neural damage caused by stroke and spinal cord injuries. The Fetz Laboratory has developed methods to stimulate neurons based on current neural activity at other sites. They use recurrent brain computer interfaces to record neural activity at one location and use the information to logically dictate stimulation at another location. This has been shown to be useful in strengthening neural connections and has been hypothesized to be helpful for patients who have experienced neural damage that results in a decrease in motor function. However a limiting factor for application to humans is the longevity of electrodes and the fidelity of the signal recorded after prolonged use. My project focuses on monitoring cortical electrodes, post implant, and characterizing the electrochemical changes over time. I am testing the effects different amounts of stimulation have on the electrochemical properties of the implanted PtIr electrodes, as well as how different surface coatings affect the degradation timeline. I am adjunctly working on creating a spinal cord electrode array for use with the Neurochip series, one that allows for higher current densities and voltages without the rapid degradation that has been previously seen.

This is an ethnographical account about a topic of little exploration, indigenous martial arts of the Philippines. Filipino cultures are a plurality of multi-stratum genealogies that merge and occasionally, disrupt one another. Filipino martial arts (or Eskrima) are a collection of Indigenous martial arts that has existed in the Philippines since pre-contact to today. Over the centuries, these arts have distilled into various systems, each with their own styles and codes. Through the methods of ethnographic data collection such as direct participant observation, shadowing, interviewing, literature review, journaling and self-analysis, I will document the shared phenomenological experiences and present a grounded theory based on the sorted material. My qualitative focus will be on one particular Filipino martial art system, Balintawak Cuentada. My presentation will convey the significance of heritage practices along with analytical insight into embodied anti-colonial perspectives while asking; how do practitioners, while living our everyday lives as Americans of Filipino decent maintain this embodied perspective? My study population is a group of practitioners dedicated to the Balintawak Cuentada system. Nearly all of the participants of this research, including myself, began our training because we wanted to connect to our ancestry in a way that could pragmatically fit into our modern lives. In our quest to reconnect with our heritage we have connected with each other. The preliminary results of this ethnography will have implications of significance to other social sciences concerned with place-making, identity, postcolonialism, embodiment, cultural memory, health and politics of the body including gender norms and performances as well as asking the philosophical questions of what are movements of the body without meaning, and what is a community without traditions. My expectation for this research is to document an example of deepening an understanding of the self through the physical engagement of ancestral habitus and ceremony.
 

Although patients diagnosed with pulmonary hypertension (PHTN) are at a higher risk for postoperative complications, studies to date have not distinguished whether this applies equally to all PHTN patients, nor have they determined the severity of postoperative complications should they occur. Pre-operative screenings take place in the form of echocardiograms (ECHO), which are accurate but rather expensive. This study, one of the largest pulmonary hypertension studies to date, is headed by Dr. Gail van Norman, director of the Pre-Anesthesia Clinic at the University of Washington Medical Center (UWMC). We look to explore the correlation between the severity of pre-existing pulmonary hypertension and post-operative complications. This study additionally explores the correlation between metabolic equivalences (METS) and PHTN, which could yield a new, more cost-effective method of screening for the disease. As research interns on this project, we abstract data from the medical records of eligible patients at UWMC. We focus on patients who have undergone surgery with moderate to severe PHTN, and collect pre-op and post-op data using hospital affiliated databases. Patients with these specific conditions are believed to have <4 METS and are at an elevated risk for post-operative complications. Should these hypotheses be proven correct, the results of this study will benefit both patients and providers, reducing the cost of screening, as well as improving patient care by allowing doctors to better manage the disease in the peri- and postoperative settings, reducing both complication and mortality rates.

Phylogeography is the study of the geographic and spatiotemporal distributions of populations, and it is an important tool for investigating the evolutionary history of species. Determining when and where species originated, merged, or moved is critical in understanding the factors that generate biodiversity. Novel Bayesian phylogeographic methods provide a useful tool for obtaining accurate estimates of these important population dynamic parameters. West Africa is a global biodiversity hotspot with a high number of endemic species, and Agama lizards are a diverse and prominent member of this distinctive ecoregion. To investigate the phylogeography of Agama lizards, I constructed phylogenetic trees from over 300 unique samples of Agama lizards from 20 countries using Bayesian phylogeographic inference. Molecular genetic data from a quickly evolving gene (16S rRNA) provided sufficient variation for estimating the divergences between distinct populations. Visualization of the phylogeographic history for the populations was conducted with Google Earth. This group of species is predicted to have originated in the central part of West Africa (i.e., Cameroon, Nigeria, and Equatorial Guinea). At least five distinctive species formed across the region from that one ancestral population, and one paraphyletic group requires further investigation. Outlining the history of species formation in subtropical West Africa, with an emphasis on when and where populations divided, moved, and reconnected, serves to show the benefits of phylogeographic studies as they can reveal patterns that are otherwise too difficult to detect using other approaches. These studies also serve as tools in providing information for the detection of distinct species.

Roux-en-Y gastric bypass (RYGB) surgery, which promotes major, sustained weight loss, is the most commonly performed bariatric operation in the United States. A ramification of RYGB is the remission of type 2 diabetes mellitus (T2DM) in patients. With 25.8 million people in the U.S. suffering from diabetes and 90-95% of adult diabetes being T2DM, we hope to understand the underlying mechanisms of RYGB on glycemic improvement in T2DM patients. The two leading hypotheses for the impact of RYGB on T2DM resolution are the lower intestinal hypothesis and the upper intestinal hypothesis. The lower intestinal hypothesis postulates that increase in nutrient delivery to the distal intestine stimulates glucagon-like peptide-1 and increased insulin secretion. The upper intestinal hypothesis postulates that elimination of contact between the ingested nutrients and the proximal small intestine exerts direct anti-diabetic effects. To test the two hypotheses, we use a porcine model and perform various bariatric gastrointestinal (GI) operations to assess the effects of each operation on glucose homeostasis. Ossabaw pigs are used for their similarities in gastrointestinal anatomy to humans. We feed them a high-fat diet to induce obesity and insulin-resistance resembling those seen in T2DM patients. The four GI operations - Gastrojejunostomy (GJ), GJ with Duodenal Exclusion, Roux-en-Y Gastric Bypass (RYGB), and RYGB with Vagotomy - rearrange various parts of the stomach and small intestine. To assess the effects of surgery on glucose homeostasis, we perform two types of blood tests: intravenous glucose tolerance test and meal tolerance test. The monitoring of the post-operative changes in insulin and gut peptide hormone levels provides insight into the mechanism of T2DM remission. Although more data is needed to support either hypothesis, the results of this study will help identify new targets for surgical and medical interventions for T2DM patients.

Gradiometer is a powerful instrument for detecting the underground condition. It works by measuring the gradient of the magnetic field so no physical contact is needed. For this reason, gradiometer is usually employed by archaeologists to figure out the condition of buried sites when excavation is not necessary (sometimes not allowed). We wrote a manual about how to use the instrument to collect, analyse, and interpret data in very detail.Considering the quality of the survey result will be affected by factors like the distance between the object to the sensors, the amount of the object, the material of the object, and the operation of the instrument, we did a series of control variable test at the Denny Yard, UW campus. Our purpose is to explore how those factors can affect the survey result, and find ways to improve the result in the real archaeological survey.According to our result, the most important factor is distance, which suggests we can get a better result by shortening the distance between the sensors and the ground. And the amount of the object has a positive relation to the strength of the signal, according to that we can estimate our result before a survey. Whether a kind of material can be detected depends on its magnetic contrast to the soil as well as the factors mentioned above. With this study, we can know when and how should we apply gradiometers into archaeological study.

Recent evidence has indicated that immune activation in mucosal surfaces is a critical determinant of HIV infection risk. HVTN914 Trial aims to quantitate immune activation in foreskin, blood and rectosigmoid mucosa from HIV-negative, uncircumcised men who have sex with men and who are at high risk for HIV acquisition. RNA of high quality and quantity must be isolated to quantitate rare immune transcripts present in these tissues. I isolated RNA from human foreskin, colon, rectum and blood samples to evaluate the RNA yield and quality. Results indicated that 91% of the samples met the required quality and quantity standards for future tests and could therefore be assessed by Reverse Transcription Polymerase Chain Reaction (RT-PCR). I then amplified an endogenous control mRNA to determine if expression of an endogenous control would be different among isolations. The blood samples amplified twice as rapidly as the other tissue types, indicating normalization to the endogenous control would be required. Our data suggests one possible cause for differential amplification is the increased phenol residue in the tissue isolations compared to blood, which can reduce reverse transcription or polymerase activity.

The current standard eyes-free speech dictation mechanism available on mobile phones is Siri's Eyes-Free mode. There are only three functions available: reviewing the current message, redoing the entire message, and sending the message. This makes speech-input of a long text-message on a mobile device very tedious. I created an app that automatically detects potential errors by aligning alternative dictations of the message and allows users to swipe through different portions of the message. I did a study on 6 users to dictate and edit sentences using Siri's Eyes-Free mode and the app that I created, and I did another study on 10 users to dictate and edit paragraphs using Siri and my app. I compare the resultsof Siri and my app in the two studies. I found that Siri's Eyes-Free mode was generally acceptable for short messages, but that my app increased efficiency for sending paragraphs. My app decreased the amount of time users spent editing paragraphs and also decreased the average word error rate.

Recent developments in research on Human Papillomavirus (HPV) have allowed for an increase of strategies that prevent the onset of cervical cancer within communities of little economic privilege. This project will compare preventative measures taken in India and those currently called for within the U.S., primarily examining how cultural and economic limitations are taken into consideration. As a “space of negation and the socially dead, produced by expropriations and alienations in and outside European nation-states” necropolis or low and middle income countries have historically been perceived as a necessity in order to maintain the priorities set by biopolis. However, organizations such as PATH are beginning to look into more cooperative methods. I will analyze the onset of preventative practices that promote self sufficiency in a format that maintains economic functionality and decreases potential dependency for aid. One significant limitation however is that western corporations still hold much of the control regarding economic leeway of vaccination use and price on a global scale. In this project I explore alternative methods that advocate information as an alternative or supplemental method for prevention. This project will continue by examining how circumstances in low economic environments are taken into consideration through efforts of vaccination or treatment. This includes the incorporation of cultural awareness, provision of knowledge and communication within the community, and how this leads to a stronger awareness of what limitations remain.

Every year millions of Washingtonians depend on winter snowfall contributing to their main source of drinking water. Seattleites alone make up 22% of the states population, and 70% of those residents use water from the protected Cedar River Watershed in their homes and businesses. I am analyzing and monitoring snow depth and hydrologic sensors in western Washington maintained by the state Department of Ecology to create a graphical and quantitative model of available water resources that will predict the availability of clean water for years to come. My study will provide a model for the maintenance of sustainable usage for our area as population increases and water availability due to climate change over time decreases. The Cascades’ annual snowfall is diminishing enough to raise concerns about future availability of water and its effects on daily life. Regions of Washington that depend on ice and snowmelt are the most affected by global warming, and some models predict that western Washington may have a problem with water shortages as early as 2040. I will be providing an independent look at the shortages we can expect to face in the near future based on trends over the last 40 years.

Aging is a degenerative process characterized by progressive deterioration of cellular components resulting in mortality. The nematode Caenorhabditis elegans has been used extensively to study the biology of aging, and several determinants of C. elegans longevity are conserved in mammals. The specific mechanisms of longevity in C. elegans have not been completely identified, and many pathways are poorly understood. Dietary restriction (DR) is one promising pro-longevity intervention, though its mechanism is still poorly defined. DR, a reduction in caloric intake in absence of malnutrition, extends lifespan in C. elegans and in a wide range of other species including yeast, flies, mice, and primates. To investigate the genetic determinants of the response to DR, we are performing a genome-wide screen using the Vidal RNA-interference (RNAi) Library, which contains 11,511 unique RNAi clones. The C. elegans are fed E. coli that express dsDNA that, when ingested, form ribonucleic-protein complexes that bind to and facilitate cleavage of target mRNAs; effectively preventing expression of specific genes. By knocking down specific genes within the C. elegans genome through RNAi, we are able to observe extension or reduction in longevity in response to DR. We have screened over 8,143 RNAi clones; 71 of which reduce lifespan under DR conditions and 85 of which increase lifespan. We are currently following up on our data by verifying the effect of these RNAi clones on lifespan under both control and DR conditions. We hope to verify how significantly these genes are affected by DR and follow up on those specific pathways to further investigate the interaction between DR and other potential longevity pathways. These findings will provide insight into fundamental mechanisms of aging and may facilitate the development of therapies to delay the onset and progression of age-related diseases in people. 

Autism spectrum disorder (ASD) is a developmental disorder characterized by deficits in communication, social skills, and repetitive behaviors. Statistics show that one in every 52 boys are diagnosed with Autism while only one in 252 girls are diagnosed with the disorder. Research shows that girls display lower communication skills independent of their age and cognitive functioning compared to their male counterparts. This study aims to identify if these gender differences in language ability can be identified prior to diagnosis. In this study, infants at high or low risk for autism were assessed via the Communication and Symbolic Behavior Scales (CSBS) at 12 and 18 months of age to identify various communication modes used by each infant. The videotaped interactions between the infant, clinician, and parent were coded offline by a group of trained undergraduates. The following communicative acts were defined and coded: (1) behavior regulation: the infant’s attempt to control some behavior of an individual around him/her; (2): social interaction: the infant’s ability to bring attention to him/herself; and (3) joint attention: the infant’s ability to bring attention to an object. Infants were evaluated for elevated autism symptoms at 24 months of age and differences in rates of communicative acts in males with elevated ASD at 24 months were compared to female infants with elevated ASD. We hypothesize that girls who go on to develop ASD will display significantly lower rates of communication acts at 12 and 18 months of age across all three communication domains compared to boys who go on to develop ASD. This study will contribute to a better understanding of how these symptoms might be different based on the sex of the infant. Understanding of how symptoms differ in males and females may have implications for the low rate of recognition of the disorder in females.

The HOPE Study is a randomized control trial conducted by investigators from the Kenya Research Program at the University of Washington Department of Global Health, Kenyatta National Hospital, and the Kenyan Ministry of Health. The clinical trial investigates the impact of home-based HIV testing and education for partners of pregnant mothers in Kisumu, Kenya. Couples are randomized to either home-based partner education and HIV testing (HOPE) as part of routine pregnancy services or to standard antenatal care (ANC). The intervention provides education and HIV testing to men in stable relationships with pregnant HIV-infected and HIV-uninfected women in order to improve overall health of women and infants, reduce risk for vertical and horizontal transmission, and increase identification of men living with HIV and link them to care. Couples will be followed up for uptake of HIV testing, facility delivery, exclusive breastfeeding and postpartum contraceptive use, as well as linkage to HIV care. Cost-effectiveness of HOPE will be evaluated in order to inform future scale up of the intervention in Western Kenya, a region with high HIV-1 incidence during the pregnant/postpartum period and high seroprevalence among men (~10%). A custom Open Data Kit (ODK) Tables application developed at the University of Washington Computer Science and Engineering Department serves as the entry-point for data collection, allowing users to collect client health information, send data to a server, and view aggregate data on their Android device. The HOPE Study Tables application allows the nurses and community health workers to screen patients for eligibility, follow up with the study participants and collect geo point data for home visits. The HOPE Study demonstrates the capabilities of ODK Tables as a data browser and manager, highlighting the importance of developing customizable data-driven applications for disconnected environments.

Superconducting Josephson Junctions may be an enabling technology for the next generation of supercomputers. Breakthroughs in nanotechnology have allowed for transistors to close in on their theoretical limit of several nm in size, allowing for tens of billions of transistors per square inch to be fabricated on integrated circuits. As the transistor count increases processor power consumption will become a limiting factor. Josephson Junctions are a promising candidate to replace silicon transistor based switching as they consume several orders of magnitude less power than current silicon transistors while promising even faster switching speeds. Furthermore Josephson Junctions have been shown to be a possible basis for a scalable quantum computing architecture, increasing their usefulness beyond advanced classical computing applications. We characterize Josephson Junctions fabricated at the University of Washington using both room temperature and cryogenic measurements as part of a preliminary effort to further develop this technology.

Fire damage is a stressor that plants have evolved around (to varying degrees) for thousands of years, and is known to induce a signaling/repair pathway that is not fully understood. Illuminating the transcriptional regulatory mechanisms of this pathway could not only give us insight into a basic biological system that is applicable to all forms of life that experience aging, cancer, and disease, it could also provide us with information about plant-specific fire resistance and healing that could be utilized to fortify our crops in a warming world. This study explores possible interactions between histone acetyltransferase (HAT) proteins produced by HAC1 and RNA synthesis enzyme RNA Polymerase II (RNAPII) in Arabidopsis thaliana plants damaged by fire. HAT proteins upregulate gene expression by chemically loosening histones (tightly-wound "spools" of DNA) and exposing coding regions to increased transcription, and RNAPII regulates overall transcription by recruiting and manipulating numerous factors of RNA transcription and synthesis, but current understanding of complementarity or antagonism between HAC1 and RNAPII remains tenuous at best, especially in plants. In this experiment, I either exposed both Arabidopsis thaliana mutants (deficient in functional HAC1) and their non-mutated counterparts (containing normal HAC1) to flame wounds, or left them alone, and then collected non-burned leaf samples twenty minutes later. Because gene expression can be measured by quantifying proteins, I used Western Blotting to detect levels of RNAPII in an attempt to elucidate any interaction between these two transcriptional regulators. My preliminary results suggest that less RNAPII is produced when plants are burned, but that the HAC1 gene is not necessary for the regulation of RNAPII after burning, though replications are necessary to confirm this.

An estimated 500-2500 gigatons of methane carbon is sequestered in the seafloor along continental margins and these deposits are associated with overlying methane seeps. The growing interest in these dynamic systems stems from 1) their importance to future energy reserves, 2) their novel microbial communities and 3) the importance of methane as a greenhouse gas. In order to constrain the impact that seeps have on methane concentrations in overlying ocean waters and to characterize the bubble plumes that transport methane vertically into the water column, water samples were collected above Southern Hydrate Ridge, a well-studied methane hydrate seep site ~90km west of Newport, Oregon. These samples were co-registered with acoustic imaging of the plumes and with robotic vehicle observations, informing more accurately on the origin of the seeps, on the rise height of the plumes above the seafloor and on the temporal variability that characterizes plume propagation. Dissolved methane concentrations exceeding background levels were observed in the upper water column, indicating that methane is transported much further vertically than previously recorded. These data provide the first detailed investigation of the short-term variance in plume dynamics and methane release into the overlying ocean, and will improve understanding of the role that sub-seafloor methane deposits play in oceanic carbon cycles. 

Graphite Oxide combined with selective laser scribing were used to create laser scribed graphene based electrochemical electrodes. Combining multiple electrodes with varying types of gelatinous and aqueous electrolyte a highly conductive, flexible, and robust capacitor was produced. The primary goal of this research was to discover an optimal combination between variables including graphite oxide density, substrate type, laser scribing patterns, and electrolyte porperties. A range of possible combinations were selected to be tested based on prior resaerch and their probability to produce an increase in energy density. To insure consistency, isolation of a single variable insured each capacitor was constructed with the same methods and regiment. In order to isolate a single variable and maintain uniformity within the test capacitor, a single sheet of laser scribed graphene electrodes were used to construct capacitors of a similar size and shape while simultaneousy adjusting a single parameter at any one time. Initial research was used to select starting values for variables, which were then individually adjusted, with a new capacitor constructed for each test run. Further research with seek to discover the feasibility of maintaining the combinations discovered through current work.

By detecting interesting subatomic particles, physicists can understand fundamental constituents of the natural world. In detection processes, however, unwanted particles can mingle with those which are targets of investigations, and thus in any experiment it becomes necessary to extract the data corresponding to the wanted particles from the rest of the measurement results. In my research project, I analyzed data that were known to come from neutrons and those from alphas, both of which were obtained in a subterranean detector located in eastern Canada. Specifically, I tested the efficacy of a proposed method to discriminate the two types of particles. When detected, each particle gave rise to an electrical pulse. Based on the hypothesis that the shapes of the pulses are qualitatively different for different particles, I have mathematically analyzed the pulses by performing integrals on the pulse shapes and finding the numerical difference between each pair of the pulses. As a result, I found that about only 25% of neutrons could be clearly distinguished from alphas, but the others were indistinguishable, solely on the basis of this pulse shape analysis. Since this result is not satisfying, my goal of the next phase of this project is to devise an improved method that can distinguish the particles more efficiently.

The Navajo Nation Department of Forestry (NDF) manages 596,728 acres of commercial forests and 4,818,815 acres of woodlands located along the Defiance Plateau and Chuska Mountains in northeastern Arizona. In previous years, the NDF managed the forest under an uneven-age forest plan, but today the NDF combines even- and uneven-age silvicultural treatments. Since 1991, there have been no commercial harvests and the amount of timber removed has been reduced from 15.9 million board feet to 4 million board feet. Concurrently, wildfires across the west have been increasing in frequency, severity and size over the last several decades. My research asks how changes in forest management have modified the composition of the forest as well as fire severity and occurrence. To answer this question, I am evaluating data from multiple sources including primary literature, forest inventory, maps and data sets from Monitoring Trends in Burn Severity project and the Navajo Forestry Department. I am comparing the growth/density of the forest over the past twenty years to that during previous management regime, and analyzing the potential impact on species composition, regeneration, mortality and the frequency and severity of fire (adding fuelwood, medicinal plants, food, wildlife, etc). I anticipate the reduced timber harvesting has resulted in increased tree density in the forest stand, changing the forest type from Ponderosa pine to mixed conifer forest and resulting in increased fuels and fire hazard. This project will deepen our understanding of the impacts of forest management decisions in this region, thus aiding the Navajo Nation and setting the groundwork for developing better strategies to sustain its natural resources and livelihood.

Disruption of the nervous system has been widely associated with aberrant bone function. The capacity for zebrafish to undergo rapid bone regeneration following fin amputation confers unique opportunities to examine neuroskeletal signaling within a highly tractable experimental system. In this study, we developed a model of cholinergic disruption in the regenerating tail fin using intramuscular injections of Botuliunum toxin B (BTxB). The tail fin is muscularized only at the base. By administering BTxB into the base, we were able to examine remote effects of BTxB at the distal tip while minimizing the influence of impaired muscle near the regenerate. BTxB resulted in multi-faceted effects on bone regeneration in adult zebrafish. For example, BTxB fish exhibited a significant decrease in the % length of bone regrowth compared to saline controls (sal: 74.8±8.1%, btx: 35.2±4.6%, p<0.001 for treatment, n=5). The degree of inhibition was significantly more pronounced in the middle of the regenerate compared to the dorsal/ventral regions (p<0.05 for region: treatment), suggesting that the trophic influence of fin base nerves can be spatially focused to distinct fin regions. Finally, when fish were calcein stained, we found that BTxB dramatically decreased osteoblast activity in regenerating bone. Interestingly, the effects of BTxB appeared to be spatially restricted within individual rays, as evidenced by marked differences in osteoblast activity between adjacent rays. This suggests a potential role for intra- and/or inter-ray nerves in mediating the effects of nerve disruption in the fin base on osteoblast activity in the distal tip. Collectively, these studies demonstrate the capacity for focal cholinergic disruption to remotely alter bone regeneration in a multi-faceted manner. Given the genetic and optical tools for dissecting neural pathways in zebrafish, this model provides a powerful platform for identifying novel mechanisms underlying neural control of bone growth.

Duchenne muscular dystrophy (DMD), is a degenerative disease caused by a recessive X-chromosome mutation, resulting in lack of the protein dystrophin. Affected males experience profound muscular weakness in childhood and by adolescence the heart and respiratory muscles are affected. The Golden Retriever muscular dystrophy (GRMD) dog model displays phenotypes analogous to patients and is considered the premier preclinical model. We evaluated changes in limb function by assessing gait in GRMD (n=3), wildtype (n=2), and in GRMD dogs injected with an adeno-associated virus carrying a dystrophin replacement gene in a single limb (n=3). Dogs were walked at a self-selected pace along an instrumented carpet to measure gait parameters. The average velocity over time of non-injected GRMD dogs was 60.39% of wildtype. Treated dogs displayed slightly increased velocities (67.17% of normal). By the final timepoint, velocity in all groups was comparable. The average paw pressure of the non-injected GRMD group increased 8.05%, while the average paw pressure for the wildtype decreased by 12.75%. The ratio of mean paw pressure for untreated GRMD: wildtype increases from 0.819 to 1.014 over time. We therefore determined that paw pressure might be a sensitive indicator of a treatment effect. To estimate the minimum sample size required for future studies, a power of analysis was performed. Results indicate that n=14 is required to detect differences between groups with a confidence level of 5% and power of 80%. Given that human patients suffer multiple symptoms of muscular weakness analogous to the dogs, including reduced limb function, changes in treated GRMD dogs as measured through gait analysis may evaluate the efficacy of the gene therapy as treatment for DMD. Human patients suffer multiple symptoms from muscular weakness (non-ambulatory, cardiac impairment, etc). Future studies hope to transition treatment to human patients to improve ambulation and increase quality of life.

We present preliminary results of an analysis of sedimentary deposits from Malakunanja II, a rock shelter and archaeological site located in Arnhem Land, Australia. Malakunanja II is the oldest known site of human occupation in Australia. Previous dating of the site has shown the first human occupation to have occurred at approximately 50,000 years BP (Roberts RG, Jones R and Smith MA 1990). Presented here are the results of laser particle size, color and magnetic susceptibility analyses on sediments taken from deposits at the site during the 2012 excavation. These analyses will serve to further the understanding of site formation processes, environmental changes and human behavior at the site.

Some symptoms of Parkinson’s disease, a movement disorder characterized by neuronal death in the basal ganglia, may result from or be exacerbated by low-level long-term exposure to manganese. Manganese-mediated impairment of normal glutamate uptake and metabolism may also contribute to abnormalities in the neuroprotective functioning of astrocytes, including manganese and other toxicant storage. Misfolding and aggregation of the protein α-synuclein, a product of the SNCA gene, has been implicated in neuronal degeneration in Parkinson’s disease, and various studies have proposed links between manganese exposure or defects in manganese metabolic systems and synuclein toxicity. Preliminary data from our laboratory suggest that knockout (KO) of SNCA in astrocytes attenuates some physiological effects of manganese exposure. The present study seeks to compare toxicant attenuation in SNCA KO and wild type (WT) astrocytes after increasing low-concentration 24-hour manganese pre-treatments. Imaging with fluo-4 dye will be used to observe glutamate-induced calcium waves in astrocyte cultures exposed to 1, 5 and 10 μM Mn2+. We predict significant deviations from a control waveform profile in both SNCA KO and WT cultures treated with higher concentrations of manganese. We further predict WT cultures to demonstrate significantly altered waveform profiles compared to SNCA KO cultures treated with equal concentrations of manganese.

College student drinking continues to be a concern due to high consumption rates and negative consequences associated with excess use. Both male and female students are at risk for heavy alcohol use and related consequences. Symptoms of depression are also reported frequently amongst college students, particularly female students who tend to be at higher risk than males for depression. Evidence suggests that depressive symptoms are often correlated with increased alcohol use. Alcohol consumption has also been related to perceived social norms of peer drinking, i.e. the perceived amount of drinking that other students and peers typically engage in. Injunctive norms, which refer to perceived approval or disapproval of a behavior by other students, may be highly regarded by students as important for decisions regarding their own behavior. Furthermore, it is hypothesized that this relationship may shift with an increase in depressive symptomology in females. Injunctive norms-indicating approval from others-have been linked to increased alcohol use in this population. Prior research also suggests that female drinking tends to be more strongly correlated with typical same-sex student alcohol injunctive norms more than their male counterparts. Thus the proposed study examines the moderating role of depressive symptoms and gender on the relationship between injunctive norms and alcohol use in college-age students. The goal of this study is to further understand the nature of the relationship between injunctive norms and alcohol use, and whether depressive symptoms enhance risk from injunctive norms in females more than males.

The precision of a measurement can be enhanced by reducing the experimental cycle time, thus allowing a greater number of measurements for a given amount of time. In our apparatus, we utilize a coherent atomic source, Bose-Einstein condensates (BEC), for a precision interferometric measurement of a fundamental physical constant - the fine structure constant, which lies at the heart of atom-light interactons. The cycle time and therefore precision of these experiments is limited by the production time of BECs. A critical number of atoms must be collected in our atom trap for BEC production. This project explores the effectiveness of doppler cooling and collimating the transverse motion of a ytterbium atomic beam in reducing experiment cycle time. Transverse cooling is done using four lasers oriented orthogonally to the path of the beam and detuned from the resonant frequency of the 1S0 to 1P1 transition in Ytterbium. The optical pressure alters a significant fraction of trajectories to lead into the trap, as opposed to being lost due to large velocity components orthogonal (transverse) to the beam path. We have created a computer simulation of the doppler cooling technique to motivate design parameters such as the detuning frequency of the lasers. We will report on the experimental implementation of transverse cooling and compare it to our theoretical simulation

Burkholderia thailandensis uses acyl homoserine lactones (AHLs) as quorum sensing (QS) signals. Broadly, QS is a communication system that, at high cell densities, allows bacteria to coordinate group activities. Recent work from our lab has shown that in B. thailandensis, QS controls contact dependent growth inhibition (CDI). CDI is an intraspecies bacterial competition system  where CDI-proficient bacteria (CDI+) contain a three-protein system of CdiA, CdiB and CdiI. CdiA is a toxic protein displayed by CdiB, and CdiI neutralizes the toxic activity of CdiA. We predict that QS-control of CDI is a means of eliminating QS mutants from the population. B. thailandensis has three QS-systems; QS-1, QS-2 and QS-3, which each comprise of a specific transcription factor and a cognate AHL signal synthase that produces a specific AHL. A B. thailandensis mutant in all three synthase genes (btaI1-3) is defective at growth, inhibiting a cdiAIB mutant. However, we do not know the specific contribution of each AHL signal synthase. Thus, we sought to test the CDI activity of synthase mutants against cdiAIB mutants within a competition. We compared the starting and ending ratios of the different strains tested in each competition. Preliminary results have demonstrated that the btaI1-3 mutant is CDI defective, relative to wild type. However, signal synthase mutants are not defective at CDI. QS is required for WT levels of CDI. Additionally, we are investigating the dynamics of how CDI controls the growth advantage of QS mutants. A btaR1 mutant has a growth advantage when co-cultured with wild type in liquid but not during growth on a solid surface, which allows for CDI to occur. Thus, we are evaluating if CDI is responsible for restraining the growth advantage of the btaR1 mutant during growth on a solid surface.

Bats are a critical component of ecosystem function. For this reason it is important to acquire information about bat abundance, species composition, and activity of bats around Seattle City Light property. The study sites are two sloughs located near the Skagit River: Harrison Slough and Barnaby Slough. This research is in preparation for a habitat restoration project on Barnaby Slough geared towards salmon. Seattle City Light is interested in acquiring baseline habitat information to determine the impact of the restoration project on bats. My work in analyzes bat echolocation calls using sonograms to estimate species presence, and understand their daily and seasonal activity patterns. Preliminary analysis shows that I detected nine species of bats. Myotis lucifigus was most commonly detected (>3000), followed by Myotis yumanensis (>2000). Lasionycteris noctivagans and Myotis californicus were also detected at high (>500) rates. Other species detected were Eptesicus fuscus, Lasiurus cinereus, Myotis evotis, Myotis thysanodes, and Myotis volans. Bat activity was highest between mid April and mid May. However, unlike other species, Myotis lucifugus activity continued at a high rate into June. These data will provide a baseline for the effects of future salmon habitat restoration efforts on bat activity.

The following project examined the ways in which HIV/AIDS treatment in St. Petersburg, Russia has been shaped by geographical narratives of blame centered on stereotypes of the United States. Currently the majority of HIV/AIDS in St. Petersburg is spread by intravenous drug use (IDU). When compared to countries with a similar GDP per capita, Russian HIV/AIDS mortality rates rank significantly higher. With Foucault’s concept of biopower as theoretical framework, my thesis focused on the ways in which the United States is negatively associated with heroin and the disease of HIV/AIDS itself. Heroin use greatly increased after the fall of the Soviet Union when heroin was considered to be fashionably new and ‘American’ by youth in St. Petersburg, and the United States invasion of Afghanistan in 2001 was perceived as exacerbating levels of heroin abuse. HIV/AIDS itself seemingly originated in the United States, and only later traveled to Russia. These associations have been used instrumentally by Russians to avoid having to address the epidemic and to minimize treatment for affected Russians. Through an exploration of biopolitics, media, and personal ethnographic research of Russian culture and political policy, it is apparent that the United States is perceived as the origin of blame for the spread of HIV/AIDS.Drawing on the concept of biopolitics, explorations of media representations, and research (including ethnographic interviews) about Russian politics and health policies, I argued that the United States is perceived by the Russian government and people as the origin of blame for the spread of HIV/AIDS.

Following the discovery of graphene, other two-dimensional (2D) materials have been identified including a variety of truly 2D semiconductors. Because of their unique optical properties including a direct band-gap in the visible spectrum, these new semiconductors are exciting candidates for next-generation devices that can translate an electric signal into light or vice versa, known as optoelectronic devices. One such important device is the light emitting diode (LED), since they are essential components in displays and optical interconnects, logic, and circuits. Further, despite being atomically thin, techniques have recently been developed to transfer and stack these monolayer materials and produce arbitrarily complex devices and heterostructures, a structure composed of layers of different 2D materials. Here, we present our adaptation of these transfer techniques in conjunction with electron beam lithography to produce the first monolayer LED using 2D crystals of tungsten diselendide. Future work will focus on cleaner “dry” transfer techniques and the fabrication of LEDs based on heterostructures of two different 2D semiconductors. This new transfer process will allow for more efficient LED’s free from debris and contaminants. This research also enables the development of brand new nanoscale optoelectronic devices and the investigation of new 2D materials.

Does magic practiced on Pacific Islands migrate to the US, and if so, is it different than the forms practices on the islands? How are ideas about magic transmitted in Pacific Island cultures? We will be engaging in interviews, discourse analysis and legends analysis and also film analysis, the Pacific Island Legends book is a body of text we’ll be looking into also. We’re expecting to find that magic is a misunderstood entity and somewhat secret and sensationalized. We’re comparing Pacific Island culture to American culture because it’s important to compare cultures that often cross-over, especially in the US, being culturally respectful and learning more about it is an important tool.

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 hypothesized that 1) commuting (in general) would have a noticeable impact on salivary testosterone and cortisol levels, 2) that testosterone levels would be highest shortly after commuting for both busing and driving, and 3) that salivary cortisol and testosterone levels would be higher when study participants drove than when they took the bus. Ten male and female participants selected two days to commute home via bus and two days to commute home via driving their own car or vehicle. Shortly before and after all four of these commutes, each participant completed mood surveys and stress scales and collected 2 mL of saliva specimen. Cortisol and testosterone were assayed in the specimens using enzyme immunoassays. Bus commute day hormone levels were compared to car commute day hormone levels. Findings from this research project could have implications for future research on stress and transportation which could 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.

Chronic heart failure as a result of damaged myocardium affects millions of individuals around the world. Our laboratory has shown that human embryonic stem cell derived cardiomyocytes (hESC-CMs) are capable of improving contractile function and decreasing arrythmias in a guinea pig model of myocardial infarction (Shiba et al. Nature, 2012). We have also shown that hESC-CMs migrate in response to gradients of the soluble extracellular matrix protein Wnt5a in vitro (Moyes et al. Stem Cells Dev, 2013). We hypothesize that a gradient of Wnt5a, delivered by sustained release from gelatin microspheres, will drive the migration graft hESC-CMs towards healthy host tissue and increase the physical contact between graft and host cardiomyocytes. We fabricated gelatin microspheres by an oil/aqueous phase emulsion and controlled their diameter to range from 4-6 um in order to prevent capillary obstruction. We loaded Wnt5a into the microspheres overnight and characterized the release kinetics over 14 days through an ELISA assay of samples taken throughout that time. We verified the bioactivity of the eluted protein with a cell-based fluorescent response to a sample at a late time point. The migration-inducing effects of the sustained Wnt5a gradient on hESC-CMs can be modeled in future via in vitro, three dimensional collagen gel seeding experiments. The resultant in vitro data will inform future in vivo transplantation studies in which we will test the ability of Wnt-5a loaded microspheres to direct the migration of grafted hESC-CMs to locations where they can couple with host myocardium and more effectively contribute force generating units to a failing heart.

The aim of this study was to further the understanding of mechanisms that underlie the pruritic and antipruritic effects of kappa opioid receptor drugs. The kappa opioid receptor (KOR) has been studied for its potential therapeutic benefits. It has been previously shown that the kappa opioid receptor antagonist, 5’-guanidinonaltrindole (GNTI), causes compulsive, hind-leg scratching in mice, while the kappa opioid receptor agonist, nalfurafine, inhibits this pruritic effect. There are applications for this antipruritic effect in medicine, to counteract symptoms in kidney-failure patients. By learning more about the mechanisms of this anti-pruritic effect, better drugs can be developed in the future with increased specificity and decreased side effects. We pre-treated male C57/BL6 mice with either saline or nalfurafine (0.05mg/kg, s.c. flank), and challenged them 20 minutes later with 5’-GNTI (0.03mg/kg, s.c. behind the neck) or saline. The number of hind-leg scratches in the 30 minutes following GNTI injection were counted as the measure of itch. We compared the results between kappa opioid receptor (KOR) knock-out mice, mu opioid receptor (MOR) knock-out mice, and wild-type mice, to determine the role of KOR and MOR in GNTI-induced itch and nalfurafine inhibition of itch. We concluded that while the pruritic effects of 5’-GNTI are not mediated by KOR or MOR, the antipruritic effects of nalfurafine was KOR dependent.

Are Pacific Islander Diets changing in the US because of public health education about health related concerns. For this Research we will be using prior research of Barbara McGrath, and from Neil A Palafox. The methods we will be using are; interviews, online research and surveys. I expect to see improved diets, decrease in numbers of people with health issues, increased education on the topic. This research is important because of the growing health concerns within the Pacific Islander Communities. 

The Western honey bee (Apis mellifera) is the principal pollinator for commercial agriculture and is estimated to provide over $200 billion in pollination services worldwide per year. In 2006, observations of widespread honey bee colony collapses initiated research on the cause or causes of their decline. Since then, pesticides, such as neonicotinoids (a class of newly popular agricultural pesticides), have received attention as a possible leading component in colony collapse disorder (CCD). However, regulations and research on neonicotinoids have been inconsistent in the United States (US) and the European Union (EU). The aim of this study was to compare registration and labeling standards for pesticides, and to review recent regulations to prevent bee declines in the US and the EU. I researched US pesticide regulations in chapter 40 of the Code of Federal Regulations and EU pesticide regulations published in the Official Journal of the European Union. I also compared published CCD research in the US and the EU in order to find correlations between pesticide regulations and instances of CCD. From my research I found that CCD is a complex issue with multiple compounding causes; neonicotinoids appear to be only one of the factors involved in colony collapse. I also found that some of the major differences in pesticide regulations are caused by the speed and extent of invoking the Precautionary Principle, which would support limiting the use of a pesticide with unknown effects; the EU tends to use the Precautionary Principle more often and implements it earlier. This research is important because the success of honey bee colonies has a large effect on worldwide agriculture and economics. By evaluating different approaches to CCD in the US and the EU, I have found certain regulations and farming methods that appear to have an effect on the number of cases of CCD.

Spinal cord injury (SCI) is a devastating condition that impacts the lives of people around the world. In order to develop cures for SCI, it is necessary that researchers are able to produce sustained and repeatable injuries in experimental animals. We induced a cervical contusion injury with a 4th generation Ohio State injury device that utilizes tissue displacement as the controlled variable during impact. We then quantified the resulting in behavioral and morphological changes in adult rats from 2 weeks to 12 weeks after injury. Quantitative behavioral assessment employed Irvine, Beatties and Bresnahan’s (IBB) rating scale, a forelimb-reaching task (FRT), and a cylinder exploration task. Morphological assessment involved observation of bruise evolution and quantitative evaluation of histology from the spinal cord. All animals showed significant loss of forelimb function after impact to the cervical spinal cord, followed by prolonged and relatively stable functional deficits. Histological evidence revealed significant loss of both white matter and gray matter near the injury epicenter. This device therefore provides a tool for SCI research by delivering a displacement-defined impact that results in a relatively repeatable and sustained injury.

Ongoing research has recently identified three glacial kettles on the campus of North Seattle Community College in North Seattle. These kettles were established as the Puget Lobe retreated from this area some 13,000 years ago. They were subsequently filled with lacustrine deposits, in a post-glacial lake which occupied this area up until the late 1800’s. The kettles are unusual in that they are not formed by burial in recessional outwash. Instead, they appear to be "ice presses" depressions in the landscape. This study has mapped the lateral and vertical extent of the northernmost of these kettles, informally identified as the 103rd Street Kettle. This mapping was done by borehole analysis, revealing a kettle up to 50 m wide, 4 m deep and over 80 m in length.

Vitamin D is an essential hormone in the body that helps regulate calcium and phosphate homeostasis. The effects of vitamin D in the body depend on dietary intake, sun exposure, and the ability of the body tissues to convert vitamin D into biologically inactive and active forms. One of the metabolic processes involves the sulfation of 25-hydroxyvitamin D₃ [25(OH)D₃], a form of the hormone that is a major circulating vitamin D metabolite in humans and can be excreted into the bile to affect the function of intestinal epithelial cells. In this research experience, I studied the relative efficiency of the 25(OH)D₃ sulfation reaction by liver sulfotransferase enzyme (SULT2A1) and the effects of the sulfate metabolite on intestinal epithelial cell gene expression. Using purified SULT2A1 and pooled human liver cytosol, I helped to determine the rate of the sulfation reaction over various 25(OH)D₃ concentrations. The Michaelis-Menten equation was used to determine kinetic parameters, which established the efficiency of 25(OH)D₃ sulfation. In order to determine which enzymes are regulated by 25(OH)D₃-sulfate, I tested the effects of 25(OH)D₃-sulfate on the expression of different gene targets in human intestinal cell systems. LS180 cells were used as a human cell model in order to examine how changes in concentration of phosphate solution and 25(OH)D₃-sulfate affect the enzyme gene expression in the cells. The experiments consisted of LS180 in various concentrations of phosphate and 25(OH)D₃-sulfate solutions. The cells were incubated, and lysed to isolate their RNA. Then, the RNA was converted into DNA to be retrieved via a PCR machine, which amplified the difference in relative mRNA expression among different enzymes. This study is expected to provide in vitro evidence that 25(OH)D₃–sulfate plays an important role in regulation of intestinal gene expression, which contributes to optimization of individualized drug therapies using vitamin D.

Microfluidic devices containing microposts, which are used to measure platelet contractile force, are fabricated using a flexible polymer called polydimethylsiloxane (PDMS). PDMS possesses many advantageous characteristics including being non-toxic, optically clear, and typically desirable mechanical properties. However, PDMS is limited as a microfluidic device given its hydrophobicity, or strong ability to repel water. This characteristic is especially problematic when PDMS is used in microfluidic devices intended to analyze blood samples. Blood is protein rich, so the hydrophobicity in PDMS leads to significant biofouling, or biological accumulation, on the surface. Moreover, the range of mechanical properties available with PDMS is limited. This limits the utility of microposts as a platelet force sensing tool since the microposts may be either too stiff or too soft to adequately measure contractile force. The work presented herein sought to circumvent these challenges by replacing PDMS with polyurethane (PU). PU was chosen because its mechanical properties can be modified to adapt to a range of platelet contractile forces from, for example, hyper- and hyporeactive platelets. First, a range of PU types was explored. From these, a commercial grade PU entitled ClearFlex was found to be the best. As a result, it was used in subsequent characterizations, and an optimized protocol for casting microfluidic devices from ClearFlex was developed. The next steps are to test the mechanical, optical, and biological properties of ClearFlex compared to PDMS. The final PU microfluidic device will be developed into a diagnostic tool specifically for clotting disorders.

BTX/A is often used in human masticatory muscles to treat TMJ disorders or reduce muscular mass. Although its multiple applications are common practice, the consequences of its neuromuscular blockage in masticatory muscles are unknown. The goal of this study is to investigate these consequences by analyzing the compound muscle action potential or evoked electromyography (eEMG) of bilateral masseter muscles (MAs) in rabbits. Eighteen female rabbits were injected with either 10 units of BTX/A (n=13) or the equivalent 0.9% saline (n=5) into three locations on one masseter at weeks 0, 12 and 24. At week 36, bilateral MA nerve trunks were exposed through surgery, and eEMGs from 11 sites of injected and non-injected MAs were examined through inserted fine-wire electrodes when the nerve was electrically stimulated at the threshold intensity (5-6V) with 2Hz frequency. Ten consecutive evoked pulses from each recording site were identified and filtered. Time domains were analyzed to calculate onset, duration, peak and integral values of these pulses. Paired t tests and non-parametric statistics were used to test differences between injected and non-injected MAs, and between BTX/A and saline injected MAs, respectively. The preliminary results show that although the overall values of stimulated pulses were smaller in the BTX/A injected than non-injected MAs, significant differences were only found in a few sites, and these sites were mainly located at the middle portion of MAs. However, there was no difference between saline injected and non-injected MAs. The comparison between BTX/A and saline injected MAs are ongoing. These results suggest that multiple applications of BTX/A in MAs may not lead to severe neuromuscular blockages over a long term as seen in the single application of BTX/A over a short term. The new information provided by these findings can lead to a better understanding and use of BTX/A in clinical dentistry.

Patients with Alzheimer’s disease experience progressive loss of memory and thinking skills, gradual declines in the ability to carry out simple tasks and, in severe cases, full blown dementia. The brains of patients with Alzheimer’s accumulate extracellular plaques of a family of beta amyloid peptides (strings of amino acids cleaved from a larger protein called Amyloid Precursor Protein) in multiple regions of the cerebral cortex resulting in losses of synapses and neurons. Given the fact that beta amyloid peptides have been evolutionarily conserved, our group became interested in the potential normative roles of these peptides. Observations by a member of the Martin lab led to the hypothesis that small concentrations of beta amyloid may have a proliferative effect on cells. My project explores this idea with experiments that compare the proliferative effects of various members of the beta amyloid family and dose-response experiments that determine optimal concentrations for proliferative effects. In my experiments, various concentrations of beta amyloid 1-40 (and, as controls, comparable levels of a scrambled beta amyloid peptide 40-1) are added to a standard culture medium along with 10% embryonic serum (serum without detectible levels of beta amyloid peptides) and established required growth factors. The cells’ division rates within those two week periods are determined for individual clones though crystal violet staining. From these initial experiments, we have concluded that cells are indeed able to divide more rapidly in media that has beta amyloid 1-40 present and that further attempts to determine optimal concentrations should focus within the range of 1-10 nM and above 10 nM to the point at which we anticipate evidence of toxicity. Future experiments will probe changes in gene expression resulting from treatment with optimal concentrations of beta amyloid for the enhancement of cell proliferation.

Amongst the Pacific Island community tattooing is a vital tool of cultural preservation. Each tattoo portrays specific values that carry cultural significance for Pacific Islanders. This study aims to depict the importance of the tattoo master sharing acquired knowledge about their community to their subject through the act of tattooing. Through qualitative research from articles such as Tattoo: An Anthropology by Makiko Kuwahara, interviews with Pacific Islander tattoo artist, and photo analysis of the styles of Pacific Island tattoos, this study will show how the tattoo master continues to connect ancient traditions to contemporary culture through the art of tattooing.

Multiple sclerosis (MS) is an immune-mediated disease whereby myelin surrounding nerves degenerates, disrupts nerve signaling, and forms scar tissue plaques, referred to as sclerosis. People living with multiple sclerosis can experience a wide range of symptoms including balance and gait issues, fatigue, vertigo and dizziness, cognitive sensory problems, tremors, and visual impairment. These symptoms can affect how individuals living with MS engage in society. There are currently an estimated 2.3 million people living with multiple sclerosis worldwide. Regionally, the National MS Society, Greater Northwest Chapter, serves 15,000 people living with MS and 89,000 friends, family members and caregivers in Alaska, Northern Idaho, Montana, and Washington. The current research study focuses on the Financial Assistance Program, offered by The National MS Society is an intervention that provides guidance, support, and resources to help manage the financial impact of MS. The intervention is aimed at supporting independence, safety, health and quality of life for clients. People living with MS can receive financial assistance for rent and utilities, medical equipment, transportation, and gym memberships, among other wellness needs. The study was conducted by using a survey assessing the perceived impact of the program. Outcome measures included self-efficacy in advocating for needs, awareness of community resources, independent living, and perceptions of social support. Quantitative and qualitative analysis offer a holistic understanding of the perceived impact of the Financial Assistance Program. The findings will inform program improvements and expansion of other services offered by the National MS Society.

Aging is associated with increasing degrees of drifts in gene expression, thought to be epigenetic in origin. The resulting variegated gene expression would have the potential to lead to a loss of proliferative homeostasis, resulting in multifocal tissue atrophies and hyperplasias. Because of enhanced rates of DNA replication, the latter could lead to neoplasms via mutations in oncogenes and tumor suppressor genes. As an initial test of this hypothesis, I immunostained leiomyomas (a benign uterine tumor)of six patients for nuclear proteins, including p53, sirt 1, Cdk4 (Cyclin-dependent kinase 4) and p16/Ink4a (CDKN2A), and examined the degrees of variegated gene expression at the three sites: cells within benign leiomyomas of the uterus and of smooth muscle immediately contiguous to and distal to these lesions. The preliminary results showed that the majority of specimens gave results consistent with the hypothesis of clonal expansions derived from precursor populations of cells with more extensive variegation. There were several exceptional results, however, perhaps related to technical issues. These pilot studies encourage additional studies, including the use of alternative methodologies for the detection of neutral mutations associated with the predicted clonal expansions. In addition, these studies may provide new prospectives for diagnostic approaches to tumor formation in uterine.

Arm swing length has been observed to vary with walking velocity. The mechanical function of arm swing as a counterbalance to leg movement has been suggested as an explanation for these observations. If this explanation is valid, then step length should be a better predictor of arm swing length than velocity, even though velocity and step length are correlated. We hypothesize that the variation in arm swing length can be explained more by step length than by walking velocity in shod and unshod walking conditions. Thirteen women completed 30 shod and 30 unshod trials at self-selected slow, medium, and fast velocities. An eight-camera system collected kinematic data by tracking infrared reflective markers placed on participants. For each trial, we calculated average walking velocity and step length from the calcaneal tuberosity and measured two distances that the distal arm travelled in a step: arm swing length measured from the travel of the radial styloid process (RSAL) and the ulnar styloid process (USAL). Arm swing length does not differ between shod and unshod conditions (RSAL: p<0.69; USAL: p<0.74). Using linear regression, arm swing length is significantly associated with both step length (RSAL: p<0.001, r2=0.46; USAL: p<0.001, r2=0.53) and walking velocity (RSAL: p<0.001, r2=0.37; USAL: p<0.001, r2=0.43). In stepwise regression, walking velocity and the interaction of walking velocity with step length were removed in favor of step length. Our hypothesis is not rejected: arm swing length is more closely associated with step length than walking velocity in both shod and unshod walking conditions.

The role of the core of the nucleus accumbens (NAcc) in foraging behavior will be explored. After a period of habituation to a semi-naturalistic foraging environment, fifteen male Long-Evans rats will undergo electric lesion of the NAcc core and fifteen will undergo a sham surgical procedure. Animals will then be measured on attempts to retrieve a food reward in the presence of a simulated predator three, six and nine days after surgery. It is predicted individuals in the lesion group will make fewer attempts at retrieving the food reward. Implications that illustrate the role of the NAcc core in goal-oriented behavior, predator and risk assessment will be discussed.

M dwarfs are small stars with masses between 10 and 50% that of the Sun. They are also the most common, making up about 70% of the stars in our galaxy, and are noted for their high levels of magnetic activity. Stellar flares are a dramatic manifestation of this activity, and occur frequently on M dwarfs. Flares are explosive events resulting from rearranging magnetic fields on the stellar surface. We used data from the Kepler satellite to search for flares from a binary M dwarf system, named GJ 1245. With more than 8 months of continuous imaging, taken every 1-minute, we detected over 3,000 flare events from these twin stars. We conducted a by-eye search of this data to validate our automatic flare-finding code. We present the energy distribution and morphology for flares on these stars, and compare them to other well studied flare stars from Kepler. The long term goal is to train the automatic flare-finding code to run without the need for human validation, and thus to study tens of thousands of stars. By studying the statistical properties of flares on many M dwarfs, we will further our understanding of stellar magnetic activity and how flares are affected by the internal magnetic dynamo.

In this research project we will examine the use of the coconut in popular culture in comparison to its place across many Pacific Island cultures in order to look at the question: What misconceptions are created about the Pacific Islands as a result of the use of the coconut in popular culture? Through a combination of interviews, photo analysis, legends and object analysis we hope to show that Western ideas of the Pacific Islands are romanticized and inaccurate due to their misrepresentation in popular culture. With these finding, we will show the importance of proper education on the many nations, people and cultures that make up the Pacific Islands as well as show the significance of the coconut in the Pacific Islands.

 This research project focuses on the ecosystem at the sediment/water interface of Possession Sound, Central Puget Sound. This area, off Everett, has been subjected to major anthropogenic pollution from manufacturing, paper and pulp mills, and agricultural run-off channeled down the Snohomish River and Steamboat Slough. The investigation reported here utilized sediment samples from Possession Sound, collected by the Washington Department of Ecology (WADOE) in the summers of 1997, 2009, 2011 and 2012. The sediment samples were processed and benthic foraminifera were separated and analyzed. Foraminifera (forams) are shelled single-celled organisms ranging in size from 0.1 to 0.5 mm. They live in a wide range of marine environments with adaptations to their tests (shells) to best suit their needs. Forams can be powerful bio-indicators of marine ecosystem health, and different species assemblage compositions can reflect changes in nutrient levels, organic and metal pollutants, and decreased oxygen levels. In addition, deformation and degradation of their tests may be indicators of water quality. From the Possession Sound samples, 15 species of foraminifera from 12 genera have been identified. Diversity and density are low in all samples, but results are mixed. One pair of samples demonstrates improved species richness from 1997 to 2012, which may indicate improving water health. Another pair of samples, however, demonstrates a severe decrease in density and species richness from 1997 to 2012, which may suggest a decline in the condition of the water/sediment interface. Comparison of these results with sediment quality parameters from the WADOE may clarify the reasons for these results.

5-fluorodeoxyuridine (FUDR), hydroxyurea (HU) and thymidine are all mutagenic agents that have been shown to alter cytosolic deoxyribonucleotide triphosphate (dNTP) pool levels in human cells. FUDR and HU affect the synthesis of dNTPs, whereas thymidine directly affects deoxythymidine triphosphate levels, one of the four types of dNTPs. Starving cells of dNTPs has a particularly large effect on cancerous cells, killing them more specifically than healthy cells since they multiply at a faster rate, making FUDR, HU and thymidine popular chemotherapy agents. Inhibiting dNTP synthesis therefore plays an important role in starving tumor cells of the proper resources they need to multiply. Studies have suggested that imbalanced cytosolic dNTP pool levels additionally play a role in the generation and accumulation of mutations in aging and large numbers of mutations found in human cancers. However, there have not been any concrete links between imbalanced mitochondrial dNTP pools and increased mutation frequency in mitochondrial DNA (mtDNA). The objective of this study is to determine whether FUDR, HU and thymidine alter mitochondrial dNTP pools, and whether the resulting imbalance affects mutation frequency in mtDNA. The method used is an extension assay with a single nucleotide overhang: a DNA duplex containing one 18-nucleotide strand tagged with Phosphorous-32, a radioisotope, and one 19-nucleotide strand. The samples are run through gel electrophoresis, which separates DNA molecules by size and allows us to determine the concentration of each dNTP in the treated cells. So far, I have found that FUDR and HU do in fact perturb mitochondrial dNTP pools and cause mtDNA mutations. My findings will help improve our understanding of the mechanisms of DNA mutagenesis and how mitochondrial dNTP pools contribute to the mutator phenotype in cancer and aging.

After a myocardial infarction (heart attack), affected areas of the heart have limited function and capacity for regeneration. Injectable therapies with human embryonic stem cell-derived cardiomyocytes (immature heart cells derived from stem cells) can improve the function of damaged tissues, but graft-cell integration with the host is limited. Studying the migratory behavior patterns of these cells may provide insight to understand the basic biology of how the heart develops and may suggest new approaches to improve graft-cell integration. Toward this end, I have tested the hypothesis that a low-cost, high-throughput gap closure assay can be used to identify novel small-molecules that will promote the migratory behavior of human embryonic stem cell-derived cardiomyocytes. In particular, I developed a modified gap-closure assay that can be employed in the University of Washington's small-molecule screening core. Using a bottomless 384-well cell culture plate as the main platform, I used surface-modifying plasma lithography techniques to create a bottom surface that has alternating high- and low-affinity regions. Cells seeded into the multi-well plates with this patterned bottom initially adhered in the high-affinity regions and I used microscopy to monitor their migration into the low-affinity areas during exposure to various small-molecules introduced by robotics. To ensure these results were valid, I tested the assay using previously identified pro-migratory factors as a positive control. This assay can be a customizable and low-cost alternative to commercially available products, which is beneficial for studies requiring mass screening. Although this high-throughput assay was used to study the migratory behavior of stem cell-derived cardiomyocytes, it can be used for a variety of cell types.

 Most kinematic studies using modern human gait to provide insights into the movement of extinct hominins collect data from shod participants. Given that our hominin ancestors were habitually unshod, this methodology is potentially problematic if the kinematics of shod and unshod gait differ. We investigate whether or not differences are present by comparing the movement profiles of the lower limbs in shod and unshod walking. Kinematic data was collected on 20 women using an eight-camera motion capture system. Participants walked at self-selected slow, medium, and fast velocities, performing 30 shod and 30 unshod trials. Average velocities were calculated for each trial, as well as the maximum and minimum angles made by the foot, calf, and thigh during the stride cycle relative to the horizontal (foot) and vertical (calf, thigh) axes in the sagittal plane. Paired t-tests indicated that for all individuals, velocities differed between shod and unshod trials (p<0.01). Linear regression comparing shod and unshod lower limb angles was thus performed including velocity as a variable. Foot (p<0.001) angles differed between shod and unshod trials at all velocities. Additionally, the magnitude of the differences is positively correlated with velocity in all individuals (foot: p<0.001). These observed differences warrant a second look at previous kinematic studies. Future studies should consider using unshod participants to obtain data that is applicable to extinct bipeds.

 Eptesicus fuscus, an insectivorous bat, vocalizes extensively in social communities and during nightly foraging. In their natural environment, bats are frequently exposed to calls from other members of their species. These calls are processed in the bats’ central auditory system, which is highly specialized and adept at processing sounds within the frequency range of its own species’ vocalizations. To investigate the effects of altering the characteristics of the calls that bats were exposed to during postnatal development, we reversibly deafened infant bats with earplugs, presented calls that were higher or lower in frequency range than normal calls, and looked at calls emitted by the bats and physiological changes in the inferior colliculus (IC), the auditory midbrain center. We hypothesized that bats exposed to calls that differed in frequency range from normal calls would learn to vocalize in a frequency range consistent with that to which they were exposed and that there might be a corresponding alteration of the frequency map in the IC. Experimental pups’ earplugs were removed only during the presentation of pre-recorded bat calls that were manipulated in frequency. Following call presentation sessions, vocalizations were recorded. After eight weeks, electrophysiology was used to study the activity of neurons in the IC. The IC is organized tonotopically, meaning that neurons that respond to a particular frequency are grouped together, forming a systematically organized frequency map. While recording neuronal activity, auditory stimuli were presented to determine the preferred frequency of neurons. Using data from recording sites throughout the IC, we found that the tonotopic organization of the experimental pups differed from that of control pups. This implies that after birth, the auditory centers of the brain remain plastic for a period of time and that communication in Eptesicus fuscus is not a purely innate behavior.

The Fu lab, in collaboration with the Yager and Lutz labs, develops low cost disposable diagnostic assays, much like the common pregnancy strip, but capable of performing more complex sample processing. Our paper-based assays are capable of automatically sending multiple reagents over a detection zone in sequence. These assays have been developed to perform the detection of markers for diseases such as dengue fever and malaria. My project is to further expand the capability of these assays by providing methods to passively control the flow rate through the detection zone. By optimizing the flow rate for each reagent delivery step, the time to result of the assay can be reduced without sacrificing signal strength. Devices are designed in AUTOCAD and fabricated from nitrocellulose and cellulose using a CO2 laser system. Flow speed within a device is quantified using a fluid marking system, in conjunction with custom video tracking software composed in MATLAB. Our experiments demonstrate several preliminary designs, in which flow is modulated by modifying the geometry of the assay substrate. In one design, a steady flow rate eventually transitions to a secondary, slower steady flow. A second design performs the opposite, where a steady flow rate eventually transitions to a secondary faster steady flow. The third preliminary design incorporates both these results to have a slow steady flow, which eventually transitions to a faster steady flow, and then finally falls back to a slower steady flow again. These flow profiles are well tailored for common protocols currently in use, where fast flowing rinse steps alternate in between slow flowing reagent binding steps.

Social media in this day and age has become a network of information both relied upon and consumed by the general public. Of these, studies have shown that Youtube.com, a video sharing site, has more than a billion unique visits each month, and that this network is proliferated by decades worth of videos watched and shared every day. Thus, Youtube.com becomes a valuable platform for the inquiry of contemporary public issues such as the legalization of marijuana. The controversial social, economic, political and health consequences of the legalization of marijuana has recently moved into the spotlight due to Washington and Colorado State’s new marijuana legalization laws. By analyzing content of public Youtube videos and their user comments on the topic of “legalization of marijuana” – categorized by key subjects (i.e., political, health, crime, etc.) and attitude towards legalization – information and opinion both relayed and consumed are shown. A maximum number of comments are chosen in order of date posted, and videos are selected in terms of relevance pertaining to the search results of key word: “legalization of marijuana.” Content analysis included are subject and attitude towards marijuana displayed in comments, likes on user generated comments, and video details such as views, orientation towards legalization of marijuana, published date, and length. I anticipate my results will include a comprehensive evaluation of approximately 900 comments generated on at least forty different videos relating to legalization of marijuana to illustrate contemporary issues, concerns, beliefs and attitudes towards marijuana and legalization. This data could prove useful in determining public attitude, knowledge and responses towards critical issues.

The discipline of Pathology has an enormous scope and students are constantly challenged to determine which topics are essential for clinical patient care and for the National Medical Board exams. There is a significant gap between the material presented in pathology textbooks and the material outlined in medical board review books. In addition, much of the practical knowledge that is most useful for medical students and physicians falls in between this gap. To address this need, we created PocketPath, a web-based modular teaching system, to help students and physicians consume Pathology material more efficiently. PocketPath is designed in an easy to read format that delivers an optimal amount of high yield pathology content, which includes USMLE Step 1 Board Review-style questions and important research papers. Using student surveys, interviews and web usage data, we determined how students consumed PocketPath’s content and improved their pathology test scores. Initial data suggests that PocketPath may replace lecture time, promote active learning, and potentially change the way students prepare for Board exams.

Measurement of tumor proliferation following cancer therapy can provide information on the effectiveness of that therapeutic treatment. One method for measuring tumor proliferation in vivo is Positron Emission Tomography (PET). In PET, a patient is injected with a radiolabeled compound that is selectively taken up by tumors. The radioisotope used decays with the ejection of a positron, which subsequently interacts with an electron to produce two 0.51 MeV gamma rays traveling 180 degrees from each other. The two gamma rays then strike a detector surrounding the patient. The in vivo location of the positron-electron interaction can be determined by comparing the energies of the two coincident photons. The PET tracers used to measure proliferation are either thymidine or thymidine analogs. Recent studies on one of these tracers, 4’-[methyl]-thiothymidine (4DST), found that the kinetics of 4DST uptake in tumor cells was much slower than thymidine (TdR) or 3’-deoxy-3’-fluorothymidine (FLT). Differences in the kinetics of uptake could be due to variations in transport kinetics, different rates of phosphorylation, or different rates of nucleotide incorporation into DNA. Understanding the factors that underlie different tracer uptake kinetics improves our ability to interpret PET information and thereby develop more effective treatment options. In the present study we examined whether the different rates of uptake observed for 4DST were due to differences in the first step in nucleoside metabolism, phosphorylation by thymidine kinase-1 (TK1). The kinetics of phosphorylation for 4DST was determined using purified TK1 in an in vitro assay. Reactions were stopped at various time points and phosphorylated products were collected by ion-exchange filtration. Results were compared to assays of TdR and FLT phosphorylation. Preliminary results suggested that 4DST phosphorylation was slower than either TdR or FLT phosphorylation. Slower phosphorylation by TK1 could explain the slow kinetics of 4DST uptake in cells.

Bilingualism has become increasingly prevalent, but few studies have been done to specifically examine how language diversity might affect children with Autism Spectrum Disorder (ASD). The purpose of this study is to analyze whether home language affects how children with ASD respond to an interactive book reading intervention. The Preschool Autism Literacy research project at the College of Education has developed an early intervention program that would prepare young children with ASD to improve their language skills and be more successful in later reading instruction. Forty-nine students with ASD in their last year of preschool (ages 4-5) met individually with a tutor 4 times a week, for 15 minutes per each session for an interactive book reading session designed to improve the language development of young children. To examine the effects of home language differences, we compare the data of four groups of participants: (a) a control of monolingual English students, (b) a control group of students with a home language other than English, (c) a treatment group of monolingual English students, and (d) a treatment group of students with a home language other than English, on receptive and expressive language outcomes controlling for pre-intervention scores. We hypothesize that bilingualism would have no negative impact on the response of the bilingual students. Thus, we expect to see no difference or a slight increase in improvement of reading skills of bilingual children compared to monolingual English children, because of the intervention program. This study has implications for emergent literacy instruction for young children with ASD from a variety of language backgrounds.

Neural plasticity in adult brains has much potential for therapeutic treatment of neurological diseases such as Alzheimer's and Parkinson’s diseases, age-related dementia, and neural insult from stroke or traumatic brain injury. We investigated mechanisms underlying plasticity in adult White-crowned Sparrows. Seasonal changes in photoperiod and hormones lead to plasticity in areas of the brain that control learned song behavior. We focused on seasonal changes in dendritic branching and spine density of neurons in the song nucleus HVC. For this approach we visualized neurons transduced with lentiviral constructs encoding fluorescent markers and evaluated their branching pattern and spine density. Preliminary data suggest that the branching pattern of dendrites may be identical in breeding and non-breeding condition birds, while the spine density increases in the HVC neurons of breeding birds.

Autism Spectrum Disorder (ASD), including previously diagnosed Autistic Disorder, Asperger’s Syndrome, and Pervasive Developmental Disorder-Not Otherwise Specified, are all biologically based neurodevelopmental disorders that are characterized by an increased lack of social communication, social interaction skills, and restricted, repetitive behaviors or interests. College students who are either diagnosed with ASD or have similar behavioral traits might have a tougher time coping with the demands of a college lifestyle. In the past, little empirical data have shown if there is a link between Autistic traits found in the general population of college students and academic factors. For my project, I will briefly survey approximately 200 college students using similar questions taken from the Autism Quotient and Social Responsiveness Scale to test and see if the presence of Autistic traits affects college students academic performance and adjustment to college life. Additionally, I will interview students who are registered with Center for Disability Services with an ASD diagnosis at Everett Community College to better understand their challenges. Because of the difficulties Autistic students have to face in a college campus, such as social communication, navigating the campus, and a change in their once regular routines, it is hypothesized students who display Autistic like characteristics will decline in their academic performance compared to those who show an absence of Autistic traits. Suggestions for transitioning and coping to college life are discussed

The Kaeberlein laboratory uses the budding yeast Saccharomyces cerevisiae as a model organism in the study of aging. S. cerevisiae is a serviceable model organism for this task because its lifespan is relatively short and simple to assess. Furthermore, many genetic tools are available to study the effect of individual genes on lifespan. Two experimental paradigms have been extensively used to quantify lifespan in yeast: chronological and replicative. In the chronological lifespan model, cells are maintained in a non-dividing state and viability is determined over time by assessing the ability of cells to re-enter the cell cycle upon exposure to growth promoting conditions. In the replicative lifespan (RLS) model, mitotically active cells are incubated until they divide and produce newly budded daughter cells, which are then micro-dissected away from their mother cells and counted. This process is repeated until daughter cells are no longer produced by the mother cell, and the total number of daughters produced prior to irreversible cell cycle arrest is quantified. After analyzing numerous strains of S. cerevisiae, many different genes and environmental conditions that modulate replicative lifespan have been identified. Importantly, many longevity modifiers identified in yeast have been found to be conserved in higher eukaryotes.