Fluorodeoxyuridine, FUDR, is a widely used treatment for many different types of cancer. It starves cells of deoxythymidine triphosphate, dTTP, which causes an imbalance in the deoxyribonucleotide, dNTP, pool sizes in the cell during DNA replication. We are predicting that with a lack of dTTP, the DNA polymerase sometimes will substitute an incorrect nucleotide in place of dTTP causing mutations, and we believe that this will affect the number of mutations in the mitochondrial genome. To study this, we are using combinations of FUDR and O4-ethyldeoxythymidine (O4-EtdT). This second mutagen, a modification of Thymidine, is used during the salvage pathway for the synthesis of dTTP. It can compete with dTTP and causes a loss in specificity for the DNA polymerase during further rounds of replication. We are treating colon cancer cells with combinations of these mutagens and using the random mutation capture (RMC) assay to quantify the mutations in the mitochondrial genome. The mutations are enriched with a restriction enzyme that cleaves the wild type DNA leaving only mutated DNA to be amplified using qPCR. We are then using TOPO cloning to sequence the mutations. Many believe that uracil will be used as substitute when the cell is starved of thymine because of their similar structures. However, incorporation of uracil will not result in mutagenesis, while incorporation of the other three nucleotides will. With the results from TOPO cloning, we will be able to take a closer look at this hypothesis. Further on, results will be analyzed using the Student T-Test and One Way ANOVA. So far, the data shows a change in the quantity of mutations in mitochondrial DNA when either O4-EtdT or FUDR are added. Results can help us to understand and respond to post-chemotherapy health problems that may be due to mutations in the mitochondrial genome.

Arsenic-contaminated groundwater poses a major environmental hazard and adversely affects the health of over 100 million people worldwide. Prolonged consumption and exposure leads to arsenicosis, which can result in skin lesions, impaired cognitive development, cancer and death. One current groundwater remediation technique involves injecting microbial nutrients (i.e. sulfate and carbon) into the subsurface of contamination sites to promote the formation of minerals (through microbial-induced reducing conditions) that either integrate arsenic into their matrix or adsorb arsenic onto their surface. However, studies have shown that post-treatment arsenic levels can increase beyond pre-treatment levels if the predominant removal mechanism is adsorption, and if sufficiently high levels of organic carbon are present to further reduce the adsorbing minerals, resulting in the re-mobilization (release) of arsenic. Therefore, it is important to determine the dominant removal process in order to develop effective long-term remediation techniques. Presently, we are working to determine the effectiveness of techniques involving induced microbial sulfate reduction through nutrient injections, which are currently being used for arsenic removal near Tacoma, WA. To simulate field conditions I pumped contaminated groundwater, extracted from the site, through sediment test columns to determine the arsenic retention capacity of the system. Once effluent and influent arsenic concentrations matched, we used the method of sequential chemicals extractions, along with μXRF and μXRS (microscale x-ray fluorescence and absorption, respectively) to determine the speciation and prevalence of the various arsenic-utilizing and -adsorbing minerals created. Next, we will pump uncontaminated groundwater through the columns to determine the post-treatment concentrations expected for this site due to re-mobilization. If our results show that the dominate removal mechanism is the incorporation arsenic into precipitates, preliminary treatment will be sufficient. However, if adsorption is the primary removal mechanism, a long-term treatment plan will need to be developed to minimize the remobilization of arsenic.

In rural Kyakitanga village of the Luweero Triangle in Central Uganda, inequalities resulting from structural violence of colonialism and the post-independence regimes of Milton Obote and Idi Amin trickle down as negative effects on the local food system, which in turn degrade the environment and peoples’ health. Locals express a desire to remedy this food insecurity through modern farming techniques. My research explores the potential role of Traditional Ecological Knowledge (TEK) in the village’s pursuit of food sovereignty and the formation of a future food system. TEK is the usually place-based intergenerational knowledge of indigenous populations about how to thrive in a mutually evolving and beneficial relationship with the environment; as an informer of subsistence methods, TEK tends to be more environmentally sustainable and socially equitable than modern farming. My methodologies included literature review of colonial and post-colonial political economic history of Uganda, current food insecurity in rural Sub-Saharan Africa, TEK in East Africa, and examples of TEK recovery in other communities. Additionally, in Summer 2013, I will conduct participant observation and a community-based participatory action research (PAR) study involving multigenerational semi-structured individual and group interviews in the village. I will collaborate with locals in the research process and share all results with Kyakitangans as a resource for pursuing food sovereignty. My research is significant because it addresses upstream issues of local autonomy, historical trauma, and ecological factors that are often neglected in public health and development discourses in the Ugandan context, which is historically dominated by verticalization and neoliberalism. As applied anthropology, my research will promote the autonomy and agency of an oppressed community, and lead to relevant broader implications for global health and development work and discourse.

Currently, various methods of creating engineered heart muscle cell, or cardiomyocyte, patches are being researched. These cardiomyocytes patches have shown therapeutic potential for implantation over injured or killed heart tissue. Current research often focuses on deriving these cardiomyocytes from stem cells, but these stem cell-derived cardiomyocytes are often not as mature as adult cardiomyocytes. Growing these cells on patterned as opposed to flat surfaces, or substrates, also affects the maturity of the cells. This project is the development of automatic video analysis software for cardiomyocytes. This provides a method of automatically quantifying contraction strength and orientation of cardiomyocytes, therefore providing an indicator of the maturity of the analyzed cells. Furthermore, the analysis program can simultaneously measure voltage-sensitive dyes in order to provide information about calcium levels as electrical signals called action potentials travel throughout the cells. This computational analysis will be performed upon human embryonic stem cell (hESC) derived cardiomyocyte tissue, as well as primary adult cardiomyocyte tissue isolated from rat hearts, grown on both flat substrate as well as nanopatterned substrate. Primary cardiomyocyte tissue is anticipated to show greater contractility, higher levels of alignment, and faster action potential propagation than the hESC-derived cardiomyocyte tissue due to its greater maturity. Furthermore, cardiomyocytes grown onto nanopatterned substrate are expected to show greater contractility, alignment, and action potential propagation than those cells plated onto flat substrates. The program in development can be used to automatically create a quantitative measurement of cardiomyocyte maturity levels across a wide array of various experiments. Being able to use the same computational analysis on different experiments both allows for easy comparison of results as well as substantiating the resultant data. Ultimately, this project provides simple automatic quantification of certain key properties of cardiomyocytes based upon videos of their contractions.

Duchenne muscular dystrophy (DMD), an incurable disease that causes severe morbidity and mortality, affects one in every 3600 male children in the United States. Stem cell therapy and cell replacement therapy have been utilized as possible solutions to the disease; however, there has only been limited success due to two major deficiencies: the lack of blood vessels in the regenerating tissues resulting in oxygen deficiency, and the lack of proper structural cues resulting in the disorganization and un-differentiation of the stem cells. Previous research has demonstrated the angiogenic and myogenic potentials of sphingosine-1-phosphate (S1P). Moreover, muscle tissues in human body have highly organized nanoscaled structures that guide the organization and differentiation of muscle cells. We designed nanopatterned muscle patches conjugated with S1P, and proposed if the patches would allow better cell survival, retention, and long-term functionality. We developed biodegradable S1P-conjugated nanopatterned muscle patches with capillary force lithography (CFL), chemical conjugation, and culturing of undifferentiated muscle cells in vitro. The patches’ biochemical, structural, and functional properties were characterized. We also evaluated the force generation, the long-term potential of regeneration, and the synchronization of the engrafted tissue of the patches within the limb muscle of mice. We anticipated proper vascularization of the patches and that the implanted cells would develop into differentiated, mature, and functional skeletal muscle cells due to the bio-mimicking mechanical cues provided by the nanopattterns. This project aimed to develop fully functional muscle cell patches which could be engrafted into patients with DMD and thereby treating the disease. In addition, the experiment would also improve our understanding in the response of muscle cells to topographical cues.

Capillary Force Lithography (CFL) is one of the novel techniques that established recently for fabricating biomimetic substratum of tissue engineering. This approach of making nano-scale scaffold allows us to fabricate complex design of polymer substrates at low-cost. One practical challenge to further application of CFL to biomedical science is biocompatibility of the materials used in CFL. Most of the materials used for CFL are only biocompatible in vitro, but not in vivo. 2-hydroxyethyl methacrylate (HEMA) is introduced as a good alternative biomaterial with exceptional biocompatibility. We recently succeeded in fabricating nano-structured pHEMA via CFL. We confirmed the structure of pHEMA nano-substrate with SEM imaging and AFM measurement. We hypothesized that biocompatibility of HEMA will enhance the cell contact with the nano-substrate, which will let cells penetrate through the grooves. Cells penetrating through the nano-grooves can benefit from increased surface area for cells to contact with polymer substrate, and allow cells to efficiently interact with glycan present on the surface to achieve more biomimetic environment.

Does gubernatorial leadership matter? Declining revenues associated with the great recession encourage state governments to use their limited resources more efficiently. However, elected officials will often disagree about priorities, which can make enacting necessary reforms difficult. The goal of this research is to investigate whether state variations in gubernatorial powers impact the ability of governors to respond to important challenges, such as the effects of the great recession. I study fifty states from 1960 to 2012, and ask whether unified government versus divided government, the strength of the governor’s veto power and the strength of the governor’s budget powers is related to a state’s economic performance. Controlling for other factors, I expect to find that states where governors have more veto and budget power, and where the government is unified under same party also tend to have lower unemployment rates. By improving our understanding of the relationship between governors and state economic performance, this study may also shed light on how national reforms strengthening the executive powers might impact presidential economic performance.

Active learning generally increases student achievement, but not all instructors experience the same magnitude of gains. This variation in student achievement is likely due to differences in how active learning is implemented in the classroom as well as differences in total classroom time devoted to active learning. Although multiple tools exist for characterizing active learning in the classroom, none have been used to explain variation in student achievement. We are developing a new type of Classroom Observation Tool (COT) that characterizes how closely an instructor’s use of active learning matches the best practices suggested in education literature and can be correlated with observed variation in student exam achievement. Currently, we are using the COT to analyze 3 randomly selected classroom sessions from 27 different introductory biology instructors using archived footage. We will create a linear model that combines instructor’s COT scores with controls for variability in student ability and exam challenge between instructors to predict student exam performance. This model will allow us to identify which elements measured by the COT are most strongly correlated with variation in student performance. With this baseline data, we will be able to recommend the use of the COT to help Faculty assess the effectiveness of their instruction, thereby enhancing teaching and student learning not only within Biology education at UW, but also across all postsecondary education.

Consolidated bioprocessing is the creation of one engineered organism that can handle a chain of chemical reactions that take a low cost feedstock, such as plant biomass, and synthesize high value products such as biofuel or medicine. This project aims to address the first stage of consolidated bioprocessing which is the digestion of the feedstock. Our design is an E. coli strain that can be in one of two states: the consumer state or the altruist state. In the consumer state, the cell carries out normal function by growing and dividing. In the altruist state, the cell produces a large payload of digestive enzymes that degrades plant biomass then lyses to release the payload to the environment where it can act. The consumer cells can then utilize the products of the digestion process to grow and divide. This system is made up of three major genetic components. The first component is a plasmid that designates whether the cell is in the consumer or the altruist state. This plasmid carries a genetic toggle switch that has two operons that produce transcription factors that co-repress each other. The second component is a plasmid that handles the production of digestive enzymes coupled with a lysis gene. Expression of this plasmid is only active if the cells are in the altruist state. The third component a genomic set of enzymes that handles the digestion of cellobiose, the product of the breakdown of cellulose. Initial results demonstrate that the engineered strain is able to grow solely on cellulose and current work focuses on the optimization of the system.

Trypanosoma cruzi is a protozoan parasite prevalent in South and Central America that causes life-long infection in humans. Approximately 30% of infected individuals develop a condition called Chagas disease which usually manifests as life-threatening cardiomyopathy or pathologies in the gastrointestinal system. Over 8 million people are believed to be infected. Unfortunately, current drugs for treating Chagas disease have low antiparasitic activity, are expensive, and are known to cause harmful side effects. As a result, research needs to focus on discovering new drug targets in Trypanosoma cruzi to lead to improved drugs. Most eukaryotic organisms synthesize sterols for essential biological functions that when manufactured incorrectly can lead to cell death. This research is investigating the effects of blocking the synthesis of ergosterol by deactivating an enzyme called sterol 14-demethylase. This enzyme catalyzes one of the intermediates of 20 steps of ergosterol synthesis. Inhibitors of sterol 14-demethylase, known as azoles, have been shown to be extremely active on T. cruzi in vitro and are now the center of research regarding anti T. cruzi drug discovery. We are developing novel azole compounds that target the sterol 14-demethylase homolog of T. cruzi to deprive the cell of its necessary sterols. However, further evidence shows that blocking sterol 14-demethylase can lead to an accumulation of sterol intermediates that are converted into toxic agents in the presence of another enzyme, ERG3 enzyme, which is normally active in a later step in the biosynthesis of ergosterol. The objective of this research project is to analyze the ERG3 homologs of T. cruzi. The putative homologs have been cloned and sequenced. No significant mutations were found. The research now is focused on engineering genetic knockouts of the ERG3 genes to test the hypothesis that the knockout parasites may be resistant to treatment with azole drugs.

Chagas Disease is a tropical disease caused by the parasite Trypanosoma cruzi. An estimated 100 million people are at risk in 21 endemic countries and ~8 million persons are currently infected. Left untreated, Chagas disease can cause fatal heart rhythm abnormalities, a dilated heart, a dilated esophagus, and/or a dilated colon. The efficacy of current treatments decreases the longer the person is infected, and 40% of treated patients experience adverse reactions, demonstrating the need for new and improved drugs. A drug target has been identified in T.cruzi: the Methionyl-tRNA synthetase (MetRS). Like tRNA synthetase enzymes in other species, the T.cruzi MetRS is crucial for protein production and survival. To exploit this target in T.cruzi we developed a robust assay (Z' factor ≥ 0.5) to identify hits against the MetRS enzyme using a luminescence ATP detection method (Promega's Kinase-Glo™). Compounds that inhibit the T.cruzi MetRS enzyme will be tested for specificity by assaying the compound with the human MetRS homolog. Specific T.cruzi MetRS inhibitors will be tested for potency against intracellular T.cruzi amastigotes and cytotoxicity against mammalian cells. Compounds that are toxic to T. cruzi, not cytotoxic to mammalian cells, and have good pharmacological properties will be selected for hit--to--lead drug development. 

Kaposi’s sarcoma-associated herpesvirus (KSHV) is a human pathogenic γ-herpes virus and is the infectious cause of Kaposi’s sarcoma (KS), primary effusion lymphoma, and plasmablastic multicentric Castleman’s disease. The KS tumor is highly vascularized and is characterized by latently infected spindle cells of endothelial origin. While in the KS tumor 1-5% of the cells are lytic and producing more virus, greater than 90% are latently infected and not producing virus. The goal of this project is to investigate whether the latent genes expressed by KSHV are sufficient to alter endothelial cells to induce angiogenesis, metabolism, and oncogenic cell signaling. We will determine if latent genes alone induce these effects or if the small percentage of lytic genes play a paracrine role. In order to do this we are cloning the entire KSHV latent locus into a gutted adenovirus expression vector that eliminates the contamination from expression of adenovirus genes. The latent locus includes the 4 genes expressed during latency and the 12 virally encoded microRNAs. The adenovirus vector expressing the latent locus will be used to infect endothelial cells and we will examine the cells expressing the KSHV latent locus for changes in factors involved in angiogenesis, metabolic pathways and oncogenic cell signaling. If the latent genes recapitulate the pathways of a KS infection, this will indicate that latent gene expression alone, without the low percentage of lytic genes in the culture, are sufficient to induce many cellular pathways induced by KSHV and are relevant for the pathogenesis of KSHV in KS tumors.

Public transit plays an important role in the daily lives of blind individuals. Unable to drive, blind commuters must rely on buses, subways, and other methods of public transportation to travel from one destination to another. Although transit schedules and even real-time arrival information are now readily available and accessible, there are still barriers to overcome. For example, a blind bus rider may find it difficult to locate a designated bus stop. Prior research has shown that, given a set of information about a bus stop, blind commuters will find their bus stop with significantly greater ease and lesser time spent. Our goal is to create a database of useful and reliable information regarding every bus stop in Seattle and make that information accessible to blind individuals. In order to collect such a vast amount of data, we have turned to crowdsourcing, a process that involves dividing work amongst a large group of workers over the Internet. By providing workers with Google StreetView images (ground-level visuals on the appearance of a location) and simple, multiple-choice questions, we hope to collect accurate information to answer questions such as “How many benches are at the bus stop?” or “In what direction is the bus stop relative to the intersection?” After acquiring a sufficient set of bus stop data, we will test the usefulness of this information to blind commuters by providing them with access to our database and analyzing their feedback. We believe that our work in this field will improve the public transit experience for blind individuals by providing them with a feeling of confidence and independence during their commute. Our work in this field may lead to further research in utilizing crowdsourcing to solve accessibility issues.

The evolution of multicellular animals required the development of epithelial tissues that function in controlling the transport of molecules from environment to organism. Collagen proteins are crucial to the formation of epithelial tissues, and are therefore critical in understanding the origins of multicellularity and Metazoan evolution. We characterized the collagen complement from the sequenced genome of the ctenophore Pleurobrachia bachei. 1) We discovered that P.bachei has 7 distinct type IV collagen genes, an expansion unseen in any organism sequenced to date. These genes show unique distribution across the genome: four were in an in-line pattern (αAand αB, αD and αE); two were found independent of other genes (αF, αG), while another two were aligned in a head-to-head fashion (αB,αC). This exceptional arrangement suggests both traditional and inverted gene duplication events in the ctenophore lineage. Ctenophore collagen intron/exon arrangements were also uniquely diverse, with a range of 14-40 exons, depending on the gene. 2) Phylogenetic analysis revealed two distinct collagen groups, the α1-like and α2-like sub-families that are common in bilaterians. Yet, gene to gene comparisons reveal a more diverse type IV collagen complement than found in chordates, including humans. 3) We found widespread expression of type IV collagen. Our findings imply that the common ancestor to all Metazoa might have contained a much more developed collagen complement than was previously appreciated. At the same time, there is extensive parallel evolution of ancestral collagens with remarkable functional specification and diversity of body plans in ctenophores.

One role of the state is to help foster economic growth. Financial incentives and regulations are thought to play important roles in industry location and investment decisions. Industries related to bioscience address important problems in society, from how to increase food production to how to defend against viruses. These industries are projected to see large levels of growth over the next 20 years. To explain differences in bioscience industry growth across states, I created a data series that analyzes the incentives available in each state against the impact of each of their bioscience industries. I then defined the quantitative and qualitative effects of these policies in order to show which provide the best areas for growth of bioscience firms and in-turn State economies. I expect to find that high levels of state incentives and reduced regulation directly lead to increased levels of employment, increased number of bioscience establishments, and an overall more impactful bioscience industry.

Since gaining the right to vote, women’s influence in the political sphere has been on the rise. While voters have started to embrace women as political leaders, research done by Huddy and Terkildsen on gender stereotypes and their effects on candidates finds that female leaders are not given the same respect as male leaders when it comes to military and police issues. Their research also finds that the inverse is true; male leaders are not perceived to have the same competence as female leaders on social welfare issues such as health care and family support programs. I will investigate this question of gender issue ownership by examining the impact of gender on gubernatorial power in 50 states over ten years. I will compare the changes in state spending across policy areas. I hypothesize that, other things equal, a change in the gender of the governor will predict changes in program spending across policy areas. I expect that governors who are perceived to be more competent in a specific policy area will have a greater effect on the spending in that area and therefore that a change in gender of the governor will predict a change in spending.

Jane is blind and in need of a new wall oven for her apartment. As she shops for the new oven she discovers that almost all ovens have digital displays for showing the state of the oven, its temperature, and the time left on its timer. They also have buttons flush with the display panel for providing input. She knows from past experience that she can put Braille labels on the buttons and will eventually memorize which button does what, so input will not be a problem. However, how will she read the digital display? Through a friend she learns of a smartphone application called Appliance Reader, which can read digital displays for various appliances including some wall ovens. In order to implement the appliance reader, we have to utilize computer vision algorithms particularly those that match one image against another. The two images can be taken from different perspectives so geometric transformation needs to be inferred from the two images. Once the two images are matched properly, the digital data has to be interpreted to form what would be told to the user. The interpretation requires a machine learning algorithm because standard optical character recognition is not sufficient. A user interface suitable for blind users is needed to make the Appliance Reader practical. As a part of CSE Mobile Accessibility group, which develops smartphone application to aid handicapped people, the two of us have been developing Appliance Reader from the ground up. Under the guidance of Professor Ladner and our mentor, an Intel researcher, Bryan Russell, we have been researching and building Appliance Reader, which is a computer vision-based smartphone application.

 A new non-visual method of numeric entry into a smartphone is designed, implemented, and tested. Users tap the smartphone screen with one to three fingers or swipe the screen in order to enter numbers. No buttons are used—only simple, easy-to-remember gestures. A preliminary evaluation compares the method to a standard accessible numeric keyboard with a VoiceOver-like screen reader interface. Preliminary results indicate that users can enter numbers faster and with higher accuracy when using this method. The Tapulator, a complete calculator based on this non-visual numeric entry that uses simple gestures for arithmetic operations and other calculator actions is described.

Most procedures for DNA transformation of chloroplasts involve the use of circular DNA as vectors and green leaves as the target tissue. Although such procedures are successful with some plants, they are inefficient and do not work with other plants, especially monocots. In maize, plastid DNA (ptDNA) is comprised largely of linear molecules. Furthermore, ptDNA is rapidly degraded as the cells become green. In consequence, the introduction of transgenes into maize chloroplasts has been problematic. In order to create vectors for monocot transformation, the plastid chromosome is most efficiently targeted in its natural form. Previous work suggests that the plastid chromosome is found primarily in a complex, branched linear form with defined ends. We chose one of these ends, “End 1,” which is near an origin of replication, as our target for transgene integration. We constructed vectors with the “End 1” gene sequences and a mannose-metabolizing transgene, PMI, as the selectable marker. Plastid transformation was performed by particle bombardment using gold microcarriers coated with either the circular or linear vector. The successful introduction of a transgene using the “End 1” DNA sequence indicates that the transgene integrates either by end joining of the linear vector to the linear ptDNA or by homologous strand invasion. Thus, using a linear vector with an end sequence produces optimized results when engineering monocot plastid transformants.

Liver transplantation is a widely performed treatment for patients with end stage liver disease. Each day, the liver transplant waiting list grows longer due to a shortage of acceptable organs. Hepatic steatosis in donor livers is an increasing problem that contributes to discarding otherwise-transplantable livers. Numerous studies and surgeries have shown that high fat content in livers correlates with poor graft function and lower patient survival. Longer waiting lists are causing transplant centers to consider moderately steatotic livers as a part of extended donor criteria. However, a method is needed to objectively quantify fat content in livers in order to select suitable organs. Many studies over the past 30 years have characterized a correlation between fat content of livers and acoustic properties such as sound speed and attenuation. This research seeks to employ an acoustic caliper device to acquire acoustic measurements that can be used to quickly, accurately, and noninvasively evaluate the fat in transplant donor livers. Sound speed and attenuation were measured by transmitting short pulses of ultrasound at frequencies from 1-10 MHz through excised pig and cow livers. Liver thicknesses were measured with a dial caliper to permit sound speed estimates, and reference measurements were taken in water to calculate the additional attenuation created by the presence of the liver. In addition, a thermocouple was used to take precise temperature measurements in liver and water, since sound speed varies with temperature. Although fat content was not quantified, sound speed and attenuation did correlate with the presence of visually observable fat. However, significant variabilities between measurements were also found with regard to the preparation of the liver samples and tissue inhomogeneities. This work has led to ideas for an improved acoustic caliper device and for testing lipid emulsions as tissue phantoms for evaluating measurement capabilities.

In Africa, emergency humanitarian relief operations, in response to ongoing conflict and natural disasters, continue to struggle with violent, targeted attacks on humanitarian workers, aid supplies and refugees. While food aid plays an important role in addressing food shortages, previous studies suggest that food aid can contribute to the spread of civil conflicts under certain conditions. The purpose of this study is to examine the possibility that certain levels of food aid could exacerbate political violence against refugees in Africa because armed groups are likely to be motivated to prevent food aid distribution to an opponent refugee population, and to achieve material gain. To evaluate this claim, I analyze the relationship between three different types of food aid and delivery methods reported in the World Food Programme between 1988 and 1998, and data on affected refugee population by political violence against refugees in African countries. Food aid, defined as emergency food aid, programme food aid and project food aid, is delivered through direct transfer, local purchase and triangular purchase. In this study, the basic conditions of food aid consist of all commodities and all donor countries. There are fifty-three African countries that reported the amount of food aid they received, and twenty-six African countries that reported the number of refugees affected by political violence during the years of study. Based on the available data, I create several regression models that show the effects of each food aid and delivery method on rate of violence. I expect that emergency food aid and direct transfer have a strong positive correlation with violence involving refugees because they are directly delivered to targeted refugees who are at risk for insurgent attack.

Proteins perform a vast array of functions within all living organisms. These functions are heavily dependent on the protein’s 3-D structure. The loss of protein structure can lead to a wide variety of problems, one being the formation of insoluble protein aggregates, which can result from cellular stress. Aggregates can inhibit proper protein function, disrupt cellular homeostasis, and are implicated in many diseases. In order to combat aggregate formation, a family of proteins exists that interact with misfolded and aggregate prone proteins (clients). This family, known as the small heat shock proteins (sHSPs), delays the formation of insoluble aggregates in the cell. Their expression and activity as molecular chaperones have been seen to increase under stress conditions. However, how sHSPs delay aggregation is not well understood. We seek to better understand binding between sHSPs and their clients. Interactions between the sHSP αB crystallin and the model client Δ131Δ (a mutant of staphylococcal nuclease) have been characterized by Nuclear Magnetic Resonance (NMR). We hope to test the observations made from this simplified system in more functionally applicable assays. sHSPs not only interact with clients, but also amongst themselves. There are ten known human sHSPs, some of which have been shown to interact with each other. While this interaction is not well understood, we do know that sHSP monomers associate non-covalently to form dimers, which in turn form higher ordered oligomers. To begin understanding these interactions, we seek to investigate whether sHSPs can exchange their monomer subunits to form heterodimers. The ability to heterodimerize would suggest an even greater diversity of oligomer structure and function. We are interested in characterizing the properties of this proposed heterodimerization. Through site directed mutagenesis, experimental based assays, and gel electrophoresis, we seek to gain a detailed description of these proteins that play such a critical role in cellular health.

 A significant proportion of the 23 known human-specific duplicated genes are implicated in neurological processes. The Rho GTPase activating protein 11 (ARHGAP11) is one such gene that is also found in an area associated with schizophrenia, intellectual disability, and autism. We determined that the human-specific duplicated copy (ARHGAP11B) duplicated 5.3 million years ago (mya) shortly after the human-chimpanzee divergence (~ 6.5 mya). The human-specific duplicated copy is not found in all humans, and represents an incomplete version of ARHGAP11A, suggesting a potentially alternative function. The goal of my project was to identify sequence differences between A (ancestral) and B (duplicated) paralogs (i.e., genes related by duplication within the same species) and use these differences to assess gene variation and expression. By sequencing 64 different transcripts (i.e. protein coding regions) we found nine “paralog-specific variants” (or PSVs) allowing us to distinguish between A and B. We assessed the level of gene expression (how much a gene is “used”) for each paralog in human fetal brain samples and found that A was expressed approximately two times more than B. Interestingly, we found similar results from other tissue types including blood. Further, we have used these PSVs to identify potentially pathogenic mutations of ARHGAP11A from targeted large-scale sequencing data of a collection of children with intellectual disability. We are using these data to understand the function of different members of the ARHGAP11 gene family both on neurological development and potentially on the evolution of traits that make us uniquely human.

America has become a world leader in medical research because of the achievements in research and technology made by pioneering physicians in various specialist medical fields. In America, specialist physicians have come to dominate the physician population, more than in other developed nations. For government health insurance programs such as Medicare, specialist physicians already receive substantially higher reimbursements than primary care physicians, namely because of the higher costs of running their practice, in addition to complex and risky nature of their field of medicine. Medicare calculates reimbursements based on the costs, skills and liability of a physician's practice at the same rate throughout the country. However, Medicare provides for regional adjustments of these costs through the Geographic Practice Cost Index. I hypothesize that the geographic index amplifies the difference in how much specialist physicians make compared with their primary care counterparts. As the index increases in value, the compensation gap between the two groups widens, providing additional incentives for specialists to practice in areas where the geographic index is high. Using physician population statistics and the Geographic Practice Cost Index, I ask whether the income advantage of specialty physicians where the index is high leads to an undersupply of primary care physicians compared to other areas.

In conjunction with Sustainable Seattle, my project seeks to identify barriers to recycling and composting in Seattle in order to ultimately increase the amount of recycling and composting that takes place. My research questions attempt to address this in two parts. First, if Seattle residents participate in recycling and composting, what is preventing them from recycling and composting more? Second, what changes can be made by the government or policy makers in order to encourage more recycling and composting? To identify barriers, I have read past reports studying difficulties in implementing recycling, and also behavioral studies identifying which methods can lead to behavioral change. From this, I discerned that the ideal methodology would consist of conducting interviews of consumers and testing multiple waste infrastructure systems at Seattle Tower using different plans on each floor to determine obstructions to commercial recycling in Seattle. Next, to identify possible behavioral modifications for more renewable action, I will conduct consumer interviews as well as the Seattle Tower project, both of which are still in progress. Initial research suggests that common recycling problems for consumers are a lack of standardized labeling on packaging and products. The results of this study and recommendations about labeling will be posted on the Sustainable Seattle website, and the Seattle Tower will be given infrastructure recommendations for improved efficiency based on the results of what was most effective. Ideally the implications of this project will lead to increased recycling and composting rates in Seattle. This can lower the costs of waste disposal for the public, businesses, as well as the city. Recycling and composting is much more cost effective and environmentally responsible than hauling trash each day to a landfill, and therefore should be utilized as much as possible. 

“It is useless to consider the house except as a part of a community owing to the interaction of these on each other.” This was the opening sentence to the Doorn Manifesto written by Team 10, a collective of architects in the 1950’s who discussed the future of modern architecture and the modern city. My research begins with delving into Team 10’s theories on urbanism and Aldo van Eyck’s writings on the architect’s role on modernization and the relationship between a house and a city. With those theories as the driving force of my research, I delve into the development of housing projects as architectural responses to political and social impacts and modernization. I begin with the rise of housing projects in Algeria and Morocco as colonialism has made Northern Africa the experimentation grounds of new architectural types that differ from the traditional casbah and impoverished bidonvilles. From there, I move to France and investigate the banlieue as a reaction to post-war migration into the city and how the architecture has fallen to the banlieue’s socio-economic image. Using Team 10’s utopian ideals, I address the successes and failures of the housing projects up to this point and examine how architecture can act as a solution instead of a reaction to urbanization and modernization.

Abnormal perinatal environment, commonly manifested as low birth weight, has been associated with birth defects as well as chronic disease. In particular, studies have shown low birth weight to have a deleterious association with type II diabetes, hyperlipidemia, hypertension, cardiovascular disease, renal disease, and renal failure. Of specific interest are the mechanistic details concerning low birth weight subjects’ altered excretion. Maternal low protein diet (LPD) during gestation and lactation results in offspring with significantly lower birth weight. Using the maternal LPD rat model, we determined renal excretion of select endogenous compounds by collecting blood and urine at various times of age. Clearance of creatinine and uric acid are indicators of glomerular filtration rate and reabsorption, respectively. High Performance Liquid Chromatography assays were used to simultaneously quantify creatinine and uric acid in the urine and in the blood of the rat. It was found that creatinine clearance and uric acid clearance decreased in LPD rats in an age-dependent and sex-specific fashion. Specifically, there were observed significant decreases in both creatinine clearance and uric acid clearance in LPD male offspring at both age’s day 90 and day 120 old. No differences were observed in both creatinine clearance and uric acid clearance in LPD female offspring until age day 120 old indicating an age-dependent programming of females as opposed to males. The results strongly correlate with decreased and altered excretion observed in low birth weight subjects and suggest that perinatal LPD has a long-term programming of renal excretion. These findings could partially explicate the underlying mechanisms that lead to higher incidence of renal disease/dysfunction in perinatally growth restricted subjects. Furthermore, altered renal function could affect the renal handling of therapeutic agents and thus affecting the overall treatment outcomes in low birth weight subjects.

Pathologic enlargement of the esophagus, termed megaesophagus, results in a failure to complete peristalsis leading to vomiting, severe weight loss and potential death. Megaesophagus is characterized in a number of disease states including parasitic (Chaga’s disease), autoimmune (myasthenia gravis), and neuromuscular (muscular dystrophies), however, a detailed cellular and mechanistic understanding is lacking. To address this deficit, we performed a comprehensive examination of the onset, pathology and cellular composition of megaesophagus in the mouse model of Duchenne muscular dystrophy (DMD). DMD affects ~1/3500 male births and is a catastrophic and ultimately fatal muscle wasting disease resulting from a mutation in the gene encoding the integral skeletal muscle protein dystrophin. Although poorly defined in human DMD patients, megaesophagus is documented in canine and mouse models of DMD. Advancements in DMD therapies has increased patient lifespan and underscored a need to assess the impact of the disease in a range of organ systems, including the esophagus. We find that older dystrophic mice can develop severe megaesophagus, but that the timing of onset depends on the nature of dystrophin expression in muscle and non-muscle tissues and the genetic background of the mice. Further studies are in progress to delineate the precise dystrophin expression pattern in different cell types of the esophagus that contribute to pathology. Work presented here will be essential for future work aimed at identifying new therapeutic targets for DMD.

Among writers both ancient and contemporary there is great interest in magic and wonder --- thaumata, thelxis, and cognates in the Greek. From the shining gods of the Homeric epics, through the second-century collection of spells known as the Papyri Magicae Graecae, down past the the occultism of Yeats, to the popularity of the Harry Potter series today, the western literary tradition has long been held fascinated by the imaginative power of such factors. Beyond popular culture, today's wealth of scholarly articles, anthologies, and monographs on the study of magic attests to a lively academic discussion around magic's cultural import. This paper aims to participate in that perennial discussion. Although it can be traced back to Platonic philosophy, it has become something of a commonplace that stories themselves work magical effects. This paper seeks to unpack the claims about narrative magic through a literary analysis of its representative deployment in selected works of Classical Antiquity. The Greek Magical Papyri are my main source for the internal terminology of magic, as it was employed in the Imperial period. For language scrutinizing magic, external to the practice itself, and for the fictional representation of both registers, I primarily examine two particularly important oeuvres: those of Lucian of Samosata, in Greek, and Apuleius, in Latin. Lucian's Philopseudes and Alethe Diegemata are key examples of fantastic stories, containing as they do the first versions of the ``Sorcerer's Apprentice'' and the ``Voyage to the Moon'' tales, respectively. Equally significant, Apuleius' Metamorphoses tells a series of magical accounts in a well-wrought masterpiece of early novelistic prose. Furthermore, both these authors are careful and clever, expressing within their texts a self-awareness of the narratological process itself. Outside these decisive thaumata, further comparative examples will be brought to bear as tangental explorations of storytelling's magical properties.

Fatty acids (FA) are commonly used as biomarkers to infer contributions of different basal energy sources to consumers. However, modification, catabolism, and storage of dietary FA are poorly understood for most animals. Controlled feeding experiments are critically needed to determine the appropriate use of FA as dietary tracers. We conducted two such experiments with juvenile isopods to investigate their use as a model organism for FA signature analysis in a direct herbivore. Newly hatched Idotea wosnesenskii were raised for 10 weeks on five different macroalgal diets at two different temperatures. Broods were raised from hatching on a single food source. Highest growth rates were on Ulva spp., fresh Nereocystis luetkeana, and aged N. luetkeana. Animals grew significantly slower on diets with chemical (Fucus distichus) and structural (Mazzaella splendens) anti-herbivore defenses. Temperature did not substantially affect growth rates. Preliminary analyses indicate that FA in Idotea tissues reflect those of their diets, and that FA driving the patterns include common biomarkers for the algae consumed. Juvenile Idotea are a promising model organism for FA trophic ecology because they generally thrive in a laboratory setting and can be hatched and quickly grown on a variety of diets.

Drift algal material is expected to play a significant trophic role in ecosystems adjacent to kelp beds. The contribution of drift algae to the overall nutritional state of various invertebrates can be assessed using phenotypic traits such as gonad index (larger gonad mass with better nutrition) and for sea urchins, jaw diameter (larger jaws with inconsistent food supplies). In this study, we collected green urchins (Strongylocentrotus droebachiensis) within kelp beds (5m depth) in the shallow subtidal photic environment (SSPE) and outside of kelp beds in nearby deep subtidal environments (DSE; 100m depth) during spring and winter seasons. SSPE and DSE sampling stations were less than 1 km apart. In the initial spring analyses, urchins in the DSE had significantly larger gonads but also larger jaws than SSPE animals. Winter gonad indices showed a reverse pattern, in which SSPE urchins had larger gonad indices, but still had smaller jaw indices than the DSE urchins; gonads in all urchins on this sampling date were much larger than those in the previous spring. These results suggest that food supplies may vary among seasons, especially in deep habitats, where the subsidy of algal material exported from shallow water may be inconsistent; the jaws from DSE urchins concur with this hypothesis. Gut content analyses from winter samples will be used to evaluate the types of foods available to deep vs. shallow-water animals.

 In the desert Southwest, predation and monsoon thunderstorms are determinants of aquatic invertebrate communities. Recent research has focused on the potential of non-native species, like bullfrogs (Rana catesbeiana), to change communities into which they are introduced. Bullfrogs were introduced into Arizona in the mid-1900s and have established widespread populations that predate on and compete with native fauna. In an effort to control bullfrog populations, state and federal agencies have focused on the eradication of bullfrogs in Southeast Arizona – a biodiversity hotspot with many endemic species. Although the effects of bullfrogs on vertebrate species are documented, how and whether bullfrogs alter aquatic invertebrate communities remains uncertain. We examined how the effects of bullfrogs compare to the seasonal effects of the monsoon in influencing invertebrate communities. The objectives of this study were to 1) examine and quantify the effects of bullfrogs on aquatic invertebrate communities by comparing ponds with and without bullfrogs; 2) determine the effect of monsoonal seasonality by sampling invertebrate communities during the early- and late-monsoon, and 3) observe the interaction of bullfrogs and monsoonal seasonality to explore the relative importance of these two factors in shaping aquatic invertebrate communities. We hypothesized a lower invertebrate abundance and richness in ponds with bullfrogs. We expected to see taxonomic differences in community composition based on body size, dispersal ability, habitat use, feeding group, and defense. We anticipated smaller invertebrates and a higher number of defended invertebrates in ponds with bullfrogs. Finally, we expected that the relative abundance of high dispersing insects would increase in late monsoon samples. Preliminary analyses suggest that invertebrate abundance and richness is negatively related to the number of bullfrogs present at a pond. By studying the effects of bullfrogs on aquatic invertebrate communities, we can begin to understand and quantify the impact of this heavily managed non-native predator.

Ongoing neurogenesis in the adult brain is a fundamental process of neural plasticity. The songbird is an established model for studying neuroprotection, neurogenesis, and neuronal turnover due to dramatic plastic changes in morphology and function in the nuclei that control avian song production. HVC, a region of the brain that times the production of song, doubles in size at the beginning of the breeding season, largely as a result of an increase in new neuron incorporation. In the nonbreeding season the song control circuit rapidly regresses in size. Within 7 days HVC regresses to non-breeding condition volume, and neuron number decreases by around 25% (> 68,000 neurons) via neuronal apoptosis. We tested the hypothesis that regression of HVC via neuronal apoptosis upon transition into nonbreeding conditions is tightly linked to proliferation of neural stem cells (NSC) in the nearby ventricular zone and changes in singing behavior. We also asked whether new neurons recently incorporated into the song control system were retained or lost through transition into non-breeding conditions. We rapidly transitioned birds from breeding to nonbreeding conditions and measured neuronal death and survival, NSC proliferation, and song behavior over a time-course of 28 days. We found that some but not all neurons incorporated into HVC during the previous breeding season persist at least 28 days into the nonbreeding season, suggesting that both new and mature neurons must undergo apoptosis during transition into nonbreeding conditions. Interestingly, we found that proliferation in the VZ was tightly linked to the amount of cell death occurring within HVC. We also quantified song degradation following rapid transition into nonbreeding conditions and correlated the observed changes in behavior to the cellular changes occurring within HVC. These findings demonstrate a relationship between cell death and neural stem cell proliferation.

Evaluation of the current Israeli-Palestinian conflict by analysis through a historical framework, centered on the events and ideologies of the anti-apartheid movement in South Africa in the early 1990’s, provides a template for enhancing human rights efforts in the present-day Middle Eastern crisis. Comparison of this conflict against a successful internal political shift to promote peace presents the foundation for research in which the synergistic combination of knowledge gained through human rights coursework applies to a current humanitarian dilemma. The cooperative relationship between theories of international studies, historical analysis, and political structural components will help contribute to the ongoing dialogue about the paradigm shift in Israeli-Palestinian relations that is so urgently needed. Methodology for this research relies heavily on the historical framework of the grassroots movement to overthrow the apartheid system in South Africa. Theories of international and national conflicts from coursework in the Jackson School of International Studies, as well as firsthand accounts of the state of affairs in the Middle East gathered through field research in sociology, contextualize the problem within a larger historical and analytical picture. The implications of this research therefore centers on human rights promotion and enhancement with regards to a modern-day conflict. This critical investigation allows for active understanding rather than passive scholarly observance of a deadly and drawn-out battle for land, identity, freedom, and dignity.

Once subjected to an extractive history of British colonial and Indian national imperatives and ignored by development institutions, the mountainous region of Kumaon, Uttarakhand now thrives in its current development trends. Given that developers and donors disregarded the region while much of the rest of India prospered, it seems incongruous that the region should now be succeeding in its own development. This project asks why the isolated and “underdeveloped” region of Kumaon that was “left behind” by mainstream development and has only recently emerged is now able to succeed in its own grassroots development. My thesis posits an answer grounded in the notions of agency and identity politics in regional social movements and development vision. I examine the interdependency between the environment and social and historical processes, which informs my analysis of local development strategy regarding regional preferences for grass use in livelihood practices. The analysis employs data collected through qualitative, ethnographic interview processes and participant observation in the field. Supplementing the primary data with literature on identity assertion as a response to colonial and neocolonial processes, I argue that Kumaon has thrived and created significant agricultural development, notwithstanding the lack of outside aid, because development there is inextricably linked to regional movements for social change. These social movements have distinguished Kumaon from national trends of development and have led to the emergence of a conservationist and rights-oriented regional identity, which has helped form a particular vision for Kumaoni development focused on the environmental rights of the people.

This project strives to address the surprisingly high rates of eating disorders (ED) in southern Italy. Determinants for ED in southern Italy are characterized by perfectionist tendencies and parental criticism, rather than the low self-esteem and body image problems most often associated with ED. The high prevalence of ED and unique composition of symptoms and psychological determinants among southern Italian women point to a compelling paradox: Given the widely supported theory that the development of eating disorders in a population may be ascribed not only to media-stimulated body image concerns but also to processes of modernization and industrialization, why do women in southern Italy, which has not conformed to or successfully undergone these processes, exhibit relatively high rates of eating disorders? This thesis argues that ED in southern Italy is a symptom of transgenerational transmission of trauma related to collective regional fear of the end of WWII and loss of the precarious economic certainty of the Allied occupation. This fear and eventual loss could not be openly mourned because of its conflict with the Resistance and liberation myths that sustain WWII memory in the popular national narrative. To substantiate this claim, I will further propose that both culture-specific ED pathologies and acclaimed films represent ciphered articulations of the psychically “indigestible” nature of this conflict. I will analyze selected films of acclaimed southern Italian directors: Ettore Scola’s La famiglia, Giuseppe Tornatore’s Cinema Paradiso, Francesco Rosi’s Tre fratelli, and Mario Martone’s L’amore molesto. Each film is analyzed anasemically—that is, completely wthin the bounds of its auditory, visual, and cinematographic language—before connecting it to historical and social contexts.

Tribal communities often lack access to high-quality healthcare, prevention programs, cancer education, cancer screening tests, and cancer clinical trials. As a result, American Indians and Alaska Natives experience the worst cancer-related disparities and the poorest survival rates of all racial and ethnic groups in the U.S. Comic books have long been used as an educational tool to improve public health. In 2008, the Native Comic Book Project was launched as a youth-focused community education project of Native People for Cancer Control, a Community Network Program Center funded by the National Cancer Institute. The purpose of this ongoing project is to use comic book creation as a way to educate Native youth about cancer, especially methods of cancer prevention. Along with basic art skills, participants learn about traditional foods and wellness, non-ceremonial tobacco use, human papillomavirus, and obesity prevention. Modeled after Dr. Michael Bitz’s Comic Book Project, the Native Comic Book Project has been adapted for both urban and reservation-based youth by incorporating Native storytelling and traditional values. Its ultimate goal is to promote healthy decision-making for Native youth and their communities. We are currently conducting a formal evaluation of this project by using pre- and post-intervention assessments to measure knowledge, habits, and decision-making among youth participants regarding tobacco use, healthy eating, exercise, and human papillomavirus. The Native Comic Book Project has been implemented at 10 sites and has enrolled 55 participants. Data will be fully analyzed in Summer 2013.  Future planned activities for research and education include developing more youth-oriented health interventions.

Immigrant farmworkers have been an integral part of Eastern Washington communities for decades. Their health and well-being are essential for the health of our communities. But for many farmworkers, health care access is unavailable or limited by a lack of insurance, health providers, discrimination, and social marginalization, making it difficult to assess their health status and burden of disease. A lack of basic information about farmworker health complicates the decision-making of state and local officials, physicians, and other advocates dedicated to improving farmworker health. In partnership with the Migrant Clinicians Network and the Pacific Northwest University of Health Sciences, I used community-based research techniques to design a farmworker public health study which could provide the necessary data to inform policies to improve farmworker health. Our survey research was designed to assess the serious injuries, diseases and causes of death among farmworkers and their families.

In The Left Hand of Darkness by Ursula LeGuin, the humans inhabiting the planet “Winter” are genderless, and through the monthly phase called “kemmer” choose to develop either male or female sexual reproductive organs. With one equal gender either member of a relationship can choose pregnancy, and “no one is quite so thoroughly ‘tied down’ here as women, elsewhere, are likely to be- psychologically or physically.” Since there is lack of gender subjugation on Winter, nature is not regarded as being connected to the womanly tasks of nurture and reproduction as it is here on earth. Nature is not viewed as an “other” and is instead an active part of a human ecology rather than something to abuse or command. In similar regard: “Winter is an inimical world; its punishment for doing things wrong is sure and prompt: death from cold or death from hunger.” On Winter humans better survive by working with Nature and not against it. Reading this novel as an ecofeminist text involves uncovering details regarding the relationship between feminism and ecology. Viewing the planet Winter as a utopian example of the absence of dualistic genders provides a fascinating scope into what the impact of a non-gendered society has on its environment: a planet where humans advance slowly and cautiously with respect for their natural world. This project examines the comparison between this model and that of ours on Earth. Winter’s slow technological development is accented by its low level of significant cultural change, whereas on Earth technology develops rapidly as culture blends, infuses, and diversifies. From this example I find that dualistic gender, or the lack thereof, greatly impacts not only the behaviors of a society, but determines its rate and quality of technology, egalitarianism, and ecology.

Ecofeminism is a movement which shows how the marginalization and subjugation of women by men parallels the domination and exploitation of nature by human beings. Interdisciplinary at its inception, ecofeminism draws from a wide range of fields including philosophy, ecology, english, and gender studies. Sometimes though, the values of these different fields conflict with one another in ways that question whether some theories can (or should) be put into practice. Queer ecocriticism, pioneered by Timothy Morton, is one such theory. Queer ecocriticism attempts to fuse ecological criticism with queer theory to challenge essentialist views of women and nature in existing environmentalisms. In Queer Ecology, Morton suggests that subjects only appear to fit neatly into stable categories like “women,” “humans,” and “nature” because we fail to perceive them on larger timescales. The best approach to environmentalism, he argues, is one which contemplates “ecology's unfathomable” and “almost unbearable” intimacies—that is, one which recognizes our world as a cacophony of interrelated, messy, and untotalizable “strange strangers” rather than a mere collection of discrete beings who can be categorized. What this “contemplation” means in practice, however, is unclear. Moreover, queer ecology appears difficult to reconcile with environmental activism, which often organizes strategically under essentialist labels like "women" or “human beings” to protect "nature." So what is the interdisciplinary-conscious ecofeminist to do? Drawing from the work of ecofeminists Sherry Ortner and Val Plumwood, queer theorists, and environmental activists, my project investigates ways in which ecofeminist activism might incorporate queer ecocritical theory into practice. If it’s true that, as Morton says, ecofeminism “is grounded on binary difference and [is] thus unhelpful for the kinds of difference multiplication that is queer theory’s brilliance,” activism informed by queer ecocriticism might be able to achieve its aims without relying on problematic, essentialist views of women and nature.

Force-feedback (haptic) displays are becoming increasingly common in consumer technology. Perceptual thresholds for haptic interaction are important for determining how much force output is required from a haptic display to provide detectable feedback to the user. In this presentation, I will talk about an experiment measuring force perceptual thresholds for single and multi-finger interaction with haptic targets. Using the UW Multi-Finger Haptic Display, subjects feel two targets on a virtual surface. One target produces center-attractive force when in contact with the subject’s finger while the other produces no force. A thresholding algorithm presents the subject a new set of haptic targets with a smaller maximum force after correctly selecting the force-producing target twice. Eventually the subject can no longer detect a target and the max force is adjusted upward. This type of forced-choice algorithm causes the threshold force to converge to where the user successfully detects the stimulus 71% of the time. In this experiment, three new methods of presenting time-correlated haptic feedback to the subject were tested. Statistical analysis shows significantly lower minimum force thresholds for multi-finger methods providing time-correlated feedback (mean thresholds of 22.94, 21.75, 22.13 mN) when compared to the single finger and multi-finger spatially correlated methods (29.24 and 33.37 mN respectively). These results suggest a neural summation effect when haptic targets are arranged to provide time-correlated stimulation to multiple fingers at once.

While timely and specific diagnosis can enable optimal treatment of patients, modern-day diagnostics are not optimized for the needs of low-resource settings due to: 1) low sensitivity, 2) slowness in providing results, and 3) dependence on electricity, chemical reagents, and trained clinicians, at a high cost. The result is frequent poor diagnosis, which can increase the rate of negative health outcomes such as patient mortality and evolution of drug resistant pathogens. There is therefore a pressing need for diagnostics that are sensitive, accurate, rapid, and inexpensive. The field of synthetic biology holds much promise in this area due to the ease and affordability of genomic re-engineering and replicating of single-cell organism like Escherichia coli and Saccharomyces cerevisiae. To address this need and opportunity, we have engineered a prototype detector in E. coli that utilizes two DNA-based modules to sense a low concentration of target analyte and produce a large output response. Module 1 detects small concentrations of target analyte by switching cells from an ON to OFF state and Module 2 significantly amplifies an output signal by enabling ON cells to grow rapidly. By seeding these cells in an environment with a non-detector strain that grows faster than OFF cells but slower than ON cells, we hope to demonstrate that this genetic logic enables a detector strain that conditionally and significantly takes over the population after detection event. This approach holds much promise for a multiplexed diagnostic, as different strains of detector cells sensing a specific pathogenic marker could be co-cultured, with the result that only the cells that detect a target analyte grow. As a cell that is a diagnostic could be easily replicated and simple to use, this approach could be extremely feasible for a low-resource setting.

Miller syndrome is a rare autosomal recessive disorder resulting from mutations in the DHODH gene. DHODH encodes the enzyme dihydroorotate dehydrogenase, which is involved in de novo pyrimidine biosynthesis. Clinical characteristics of Miller syndrome include craniofacial abnormalities, coloboma of the eyelids, cleft palate, and missing postaxial digits on the hands and feet. To broaden the spectrum of causal mutations, DNA samples obtained from two unrelated affected probands were Sanger sequenced to screen for mutations in DHOHD. In family 1, the affected individual was a compound heterozygote for a mutation in one of the canonical splice bases of intron 6-7 (c.820+1G>A), and a missense mutation in exon 8 (c.C1036T, p.R346W). DNA samples obtained from both parents of the affected proband were screened using Sanger sequencing and each parent was found to be heterozygous for one of the DHODH mutations. The c.C1036T mutation has been reported now in three families with Miller syndrome and functional studies have demonstrated a reduction of DHODH activity by 40%-70% in yeast with this mutation. No pathogenic mutations in DHODH were found in family 2. The next steps we will take include whole genome sequencing of the proband and parents in family 2 to determine if there are mutations in other genes that cause Miller syndrome. Our results mark the first report of a splice site mutation in a patient with Miller syndrome and further expand the spectrum of causative mutations in DHODH.

Speciation is the process by which one species becomes two species that can no longer interbreed. A common hypothesis is that infertility between closely related species is due to the rapid evolution of DNA double-strand-break (DSB) hotspots that are active in meiosis. DNA DSBs play an important role in meiosis by initiating recombination after chromosome replication and can be repaired as crossovers. During DNA DSB repair, the broken strand of a homologue invades and repairs its break with the strand of the aligned homologue by matching complementary nucleotides. The repair can result in a heteroduplex which refers to a mismatch, such as adenine paired with cytosine. Heteroduplex rejection occurs when the cell’s recombination machinery rejects recombination at sites where heteroduplex forms, and it is known to require the msh2 gene. We hypothesize that heteroduplex rejection inhibits DSB repair in interspecies hybrids and can result in infertility and thereby the isolation of species. Schizosaccharomyces kambucha and Schizosaccharomyces pombe are two very closely related fission yeast species that are 99.5% identical at the DNA sequence level. Despite their similar genetic makeup, we found that their hybrids show decreased fertility and decreased recombination frequencies. We are testing whether inhibiting heteroduplex rejection will increase the fertility and recombination rates of the fission yeast hybrids. For this experiment, we have created msh2Δ Schizosaccharomyces pombe and Schizosaccharomyces kambucha mutants that cannot do heteroduplex rejection so the cells should go through DNA DSB repair even if there are mismatches. We expect to see an increase in fertility and recombination in the msh2Δ hybrids. This research may be extended further to explain speciation in other organisms or unexplained infertility between human couples.

Recent reports have shown that metabolic signatures are highly characteristic for a cell and may act as a leading cause for cell fate changes. In particular, we have shown that during development from naïve to primed embryonic stem cells, the cells undergo a dramatic transition from metabolically bivalent to highly glycolytic. However, this state of inert mitochondria rapidly changes to highly potent mitochondria during further differentiation. It is not yet understood how and why the pluripotent cells enter the highly glycolytic, metabolically cancer-like (Warburg effect) stage and how a differentiating cell leaves this stage. We have now performed metabolomics profiling of naïve and primed pluripotent cells using mass spectrometry. Interestingly, primed cells accumulate the tryptophan degradation product kynurenine. Kynurenine (KY) can act as a ligand for the transcription factor AHR. In cancer cells AHR activation by KY incudes growth while in surrounding T-cells KY based AHR activation will inhibit T-regulatory cells. My hypothesis is that kynurenine is a key metabolite in determining stem cell stage and in helping protect the primed stage embryo against the mother's immune cells. I will now test this hypothesis by analyzing the levels of the key KY metabolism enzymes by qPCR. My prediction is that the tryptophan degrading enzyme IDO1 will be unregulated in the primed stem cells. To test whether kynurenine can change the state of naïve stem cells, I will add different concentrations of kynurenine to the stem cell media and test potential cell fate changes. Since primed ESC-like metabolism is also observed in cancer cells, it is of great interest to understand how the cellular transitions in and out of this metabolic state are controlled during normal development. These studies will shed light on the function and regulation of metabolism during normal and disease developments.

Fandango is a centuries old community based musical tradition from Veracruz, Mexico dating from the first half of the XVIII century. With Indigenous, African, and European influences, this tradition takes place around a tarima, a wooden platform where mostly women improvise foot percussion as a group, involving each other and engaging the community in a musical dialogue. The intent of this research is to explore the tarima as a space of female empowerment and communal decolonial praxis. This research uses as its primary method the collection of oral histories from women participants in the fandango tradition in the U.S and Mexico. Considering the community building nature of Fandango and the tarima as an important space for women, it is anticipated that the results of this research will reflect the empowerment women and communities have found through this practice. Moreover, this research focuses on the collection of personal narratives that support this argument to further explore their implications for resistance and healing in response to gender, institutional and social opression. My interest in theorizing and writing about these oral histories is to elicit examples and create community for other women and individuals who might want to explore community-based arts as a mode of healing and resistance. This research is an important part of a longer project that explores and proposes community-based arts as tools for recovery from trauma and violence prevention for individuals and communities at large. This research will also open the door for other scholars in the field to consider alternative ways of responding to trauma.

With any particle physics experiment, precision is paramount. In experiments with electromagnetic shields, the inherent magnetic field noise from the shield itself can impede the accuracy of high precision experiments. For this research project I designed a model of an electromagnetic shield to find the magnetic field noise of the shield; the results of which can be used to increase the precision of a particle physics experiment being conducted at the University of Washington, searching for a potential source of time reversal symmetry by looking for a permanent electric dipole moment of a specific isotope of mercury (the existence of which would suggest a source of CP-violation). The cylindrical electromagnetic shield was modeled in the finite element analysis application, COMSOL Mulitphysics. The model is initially configured with a single magnetic dipole arranged on the axis of symmetry of the magnetic shield. The accuracy of the model is then compared to an analytic solution found by S.-K. Lee and M. V. Romalis at Princeton University. After refining the model to reflect the analytic solution, the number and orientation of magnetic dipoles is changed, producing the first, second and third derivatives of the magnetic field noise of the shield being used. Accounting for the magnetic field noise will increase the sensitivity of the experiment being conducted.

The neurotransmitter dopamine is a key component of the limbic sysem, which plays a critical role in emotional processing, learning and memory, and reward processing.  Disruptions in dopamine synthesis, release, and reuptake underlie many neurological disorders, learning deficits, and diseases.  Previous work from our lab established the importance of the glutamatergic NMDA (N-methyl-D-aspartate) receptor in maintaining normal phasic activity in dopamine neurons of the ventral tegmental area (VTA) and showed that mutations of this receptor cause deficits in fear-related learning, highlighting the importance of dopamine in fear-related learning.  In this project we use in-vivo electrophysiology to record from dopamine neurons of the VTA during Pavlovian fear conditioning.  The paradigm consists of two types of trials where one cue (CS+) is paired with a mild footshock (0.3mA, 0.5 sec), while the other trials utilize a cue (CS-) which predicts the absence of a footshock.  Our preliminary data show plastic differences in neuronal activity patterns across training days associated with both conditioned and unconditioned stimuli, where initially there is increased activity in response to the fearful stimulus, but across days of conditioning, this response shifts to being associated with the conditioned stimulus.

Palladium is a part of the Platinum Group Metals (PGM), and is highly used as a catalyst in many applications such as in an automobile’s catalytic converter. It has also been successfully used for hydrogen gas sensing devices. Employing low-cost, efficient means to synthesize palladium nanowhiskers in the lab has been the focus of my research thus far. Hydrothermal synthesis and dislocation mechanisms have been used for the growth of palladium nanowhiskers using a palladium (II) chloride charge. The hydrothermal synthesis involved chemical reactions as well as thermal reactions in the growth of the nanowhiskers. In the dislocation mechanism, the growth of the whiskers is the result of the thermal decomposition of palladium chloride to palladium plus chlorine gas. Scanning electron micrographs from the first four runs of the hydrothermal synthesis showed little to no nanowhiskers. The first run of the dislocation mechanism grown samples is currently awaiting characterization. Future work for my research involves many more dislocation mechanism growths of the palladium nanowhiskers, as well as an attempt to use electrochemical deposition as a third technique in producing these whiskers. The application of these nanowhiskers in hydrogen gas sensing is based on the change in the conductance of the whiskers upon absorption of hydrogen. This allows for highly sensitive sensors to be made.

Quantitative mass spectrometry (MS)-based proteomics is a powerful approach to measure relative expression changes across thousands of proteins in a complex mixture. Commonly, stable isotopes like ¹³C and ¹⁵N are incorporated into proteins and peptides by metabolic labeling (ML) or chemical labeling (CL) to produce “heavy” labeled forms. Mixtures of light and heavy peptides can be identified and quantified by MS. In ML, proteins are labeled in vivo by growing cells with isotope labeled metabolic precursors, while peptides are chemically labeled in vitro in CL. These two labeling approaches have distinct workflows and their own strengths and limitations. In a typical proteomics experiment, complex mixtures of peptides are separated on reversed phase liquid chromatography (LC) and sampled in real-time by the mass spectrometer for peptide identification and quantification. We propose to analyze mixtures of chemically and metabolically labeled peptides in single LC-MS runs, comparing each method’s quantitative accuracy and precision, and other factors, such as analytical losses from sample processing without errors introduced by stochastic sampling. We compare stable isotope labeling by amino acids in cell culture (SILAC) and reductive dimethylation CL by mixing equal amounts of peptides from each approach.Preliminary results indicate minimal differences between ML and CL in quantifying proteins (median of >2 peptide ratios). With individual peptides, however, CL showed 13% larger variance than the ML. We observed signal losses in CL samples compared to ML samples, and fewer quantified peptides and proteins in CL (6005 peptides, 897 proteins) than in ML (7448 peptides, 1059 proteins). We attribute the differences to the extra labeling step in CL, and will perform further analyses with ratio mixtures of heavy and light labeled peptides to clarify the source of variance. Our comparison of CL and ML will guide future optimization and applications of both quantitative proteomics labeling methods.

In aquariums and zoos across the world marine mammals are among the most popular exhibits. Caretakers strive to provide the least stressful and natural environments for the animals. Analysis of (steroid) hormones in excrement has proven to be an effective, non-invasive method. It allows animal keepers to monitor physiological health associated with reproduction and the stress response in captive animals. Prolactin (PRL) is a non-steroidal, non-tropic peptide hormone most commonly associated with milk production in nursing females. However, it has a multitude of normal physiological functions such as: regulation of the immune system, osmotic balance, angiogenesis, an indicator of parental care (in both males and females), sexual maturity in males, and near-term pregnancy in females. Chronically high or low levels of PRL have been associated with a number of ailments. For instance, impotence reduces the secretion of estrogens and testosterone. To date, no studies have been documented on prolactin PRL levels in captive northern fur seals, (Callorhinus ursinus), harbor seals (Phoca vitulina), or sea otters (Enhydra lutris). Using an enzyme-immunoassay (EIA) kit, previously tested only in human and elephant serum analysis, we investigated PRL levels of several samples collected from captive marine mammals (Northern fur seal blood serum, harbor seal saliva, and sea otter urine/fecal samples). Due to the helical structure of PRL, fecal samples were not expected to survive intact after transit through the digestive tract; therefore, were not expected to produce positive results. Here we report preliminary results of the EIA PRL assay on several different types of samples, as well as several longitudinal samples from the same individual, to show significant changes in this hormone over time. If detectable in urine, fecal matter, and/or saliva, this assay could prove to be a useful non-invasive method for determining health in captive marine mammals.

We  present an extension of the Astronomy Department’s curriculum for visually impaired junior high  school students. Our lesson plans are a complementary addition to  Noreen Grice's Braille and large print  readings. These lessons satisfy the Washington State Essential Academic Learning Requirements (EALR's) for junior high school students, the level at which students are introduced to astronomy. We  present here a sample of new material that covers the composition, formation and evolution of the Sun.   Our lessons include activities, audio tracks, 3D models, and online links to advanced topics. We outline additional curricula that we are actively  developing, such as biographies of visually impaired astronomers and topics including the formation and evolution of the Solar System's terrestrial planets.  We will test our curricula during this coming school year, as we did in  2007, at the Washington State  School for the Blind,  with the  expectation  of deeply  strengthening the  students' understanding of the  evolution of the Sun beyond what is available in their textbooks.  Ultimately, this curricula will be distributed and utilized throughout the United States as part of the on-going national and international efforts  to promote astronomy for those who are visually impaired.  We conclude with a description of our  role in building and testing additional educational tools.

Current research in Himalayan geodynamics focuses on the complex relationship between surface erosion and active rock uplift. This study focuses on the Eastern Himalayas, where the Yarlung-Tsangpo River abruptly turns southward, incising over a 2 km deep gorge through the easternmost Himalayan mountains to join the Brahmaputra River in Assam. Because geologists have primarily concentrated on just the surface geology, they have struggled to explain this massive gorge despite over a decade of research. Our research takes a new approach to this aging problem by examining the sedimentary record immediately downstream of this gorge. In fact, sedimentary rocks of the eastern Siwalik foothills preserve the full record of Himalayan uplift as well as the evolution of the river systems draining the Tibetan Plateau. Here we focus our research, interpreting the sedimentary record through both detrital zircon U-Pb dating, which is a radiometric dating technique, and modal analysis, which is an evaluation of the minerals present within the samples from these Siwalik units. Past research results suggested a stable sedimentary provenance sourcing from both Himalayan and Tibetan source rocks, but new higher resolution modal analysis will add valuable insight into the competitive influences of progressive roofing of the high Himalayas and sediment recycling in the Himalayan foreland. Furthermore, this research provides valuable new data for a region of the Himalayas where there is currently a deficiency of information from the sedimentary record. This information is necessary for a full comparison of landscape evolution westward across the great Himalayan arc, and also shows a regional relationship between climate and tectonics.

Two strains of Botryococcus braunii algae were grown in three different sets of conditions to compare growth characteristics and lipid production. One species, UTEX LB 572, was a Race A Botryococcus braunii from the culture collection of the University of Texas in Austin. The second species, AC 762, was a Race B Botryococcus braunii from the culture collection of the University of Caen, France. UTEX 572 was found to produce two hydrocarbon chain lengths of C:16 and C:18 in a 2:3 ratio respectively. The AC 762 was found to produce hydrocarbons called botryococcenes of chain lengths between C:30 and C:36. The lipid chain lengths of both strains were evaluated by mass spectrometry. The UTEX 572 lipid content was quantified by analytical spectrometry after direct transesterification of the lipid content to biodiesel. The AC 762 lipid content was quantified by a photospectrometry technique developed by UC Berkeley researchers Eroglu and Melis (1985). Cells of both species were harvested during exponential growth phase from 400 mL of growth media and then freeze dried and analyzed gravimetrically. Nine flasks of each species were grown with 3000 lux average light intensity on a 12h:12h light:dark cycle at room temperature averaging 22° C. Three flasks of each species were grown with 1.5% CO2 in BG-11 media, three flasks of each species were grown in BG-11 media with air only and three control flasks were grown in respective carrier medias as those used in the algae collection of origin. Knowledge of the rates at which AC 762 and UTEX 572 produce hydrocarbons as well as the specific lengths and proportions of those hydrocarbons contributes to data needed for comparison of economic considerations in industrial applications.

The Salish Sea is critical habitat for the endangered population of Southern Resident Killer Whales (SRKW). One of the primary threats to SRKW recovery is underwater noise pollution from boat and ship traffic. Killer whales rely on acoustics to communicate, navigate, and to forage in their underwater environment. The increasing levels of underwater noise have the potential to mask the SRKW calls and echolocation clicks, resulting in compensation (i.e. increasing the loudness of calls and using echolocation more or less frequently). This study investigates the potential effects that boat and ship traffic may have on SRKW echolocation. If SRKWs are experiencing masking while actively using echolocation, it is expected that there will be a significant difference between echolocation click rates in a quiet underwater environment and click rates produced in an underwater environment filled with noise pollution. Passive acoustic monitoring was used to record SRKW acoustics and measure the received levels (RL) of underwater noise. Visual boat and ship counts were conducted during acoustic recordings to quantify the amount of traffic in the study area (Whale Watch State Park, San Juan Island, WA). These data sets are being tested to determine if there is a significant correlation between the RL of underwater sound and boat and ship traffic, and if echolocation click rates change in respect to the RL of underwater sound. The anticipated results are that boat and ship traffic will be directly connected to the increases in RL of underwater sound and that masking will be impacting SRKW use of echolocation. Southern Resident Killer Whale population recovery is faced with a number of threats, however underwater noise pollution is one that we, as a society, can directly impact. Consequently, it's extremely important for us to understand how underwater noise pollution can affect this endangered population of killer whales. 

Colloidal zinc oxide nanocrystals (NCs) become charged with electrons upon reduction with decamethylcobaltocene. Understanding this redox chemistry can further develop research on quantum dot sensitized solar cells and other various applications of NCs. The goal of my project is to determine whether this electron transfer can be characterized by an equilibrium constant K_eq, and if so, to see which variables in the reaction affect the K_eq. Two models were tested: (1) in which the concentration of NCs would affect the K_eq, and another (2) in which it would not. Using data from optical titrations, I found that the K_eq is affected by NC concentration and is therefore better described by model (1). Similarly, other variables are currently being tested in order to refine a working model that can accurately describe the reaction of NCs with reductants. The contribution of a simple model for these charge transfer processes can be applied to various areas.

Cardiovascular disease is the leading cause of death worldwide, and current treatments for heart failure are limited to slowing the disease progression or transplanting a donor heart. One potential approach to restore heart function is to transplant human embryonic stem cell-derived cardiomyocytes (hESC-CMs). An independent and similarly promising approach is to overexpress the enzyme ribonucleotide reductase (RR), the enzyme responsible for the production of the small molecule deoxy-ATP, which has been previously shown to improve contractility and diffuse between cardiomyocytes via gap junctions. We hypothesize that by overexpressing RR in hESC-CMs and then transplanting the latter into infarcted hearts, we will improve global cardiac function by increasing dATP content throughout the myocardium. To begin testing this hypothesis, I will test the efficacy of an RR adenovirus in hESC-CMs in vitro using quantitative PCR and immunostaining, and I will then correlate the subsequent improvement in fractional shortening following in vivo transplantation to the amount of successfully engrafted cells. To date, I have successfully developed an immunofluorescence staining protocol for RR. I am currently working on designing an immunohistochemistry protocol to identify graft tissue and stain for RR and GFP and localize RR in relation to the infarct. After confirming the protocol, I plan to apply it to the fixed rat heart slices to assist in the proof of concept of the transplantation of hESC-CMs overexpressing RR as a novel therapeutic method.

The mission of the Cell Biomechanics Lab led by Professor Nathan Sniadecki is to “understand how mechanics affects human biology and disease at the fundamental level of biological cells.” To do this, microstructures are created in this lab to interact with cells on their native micrometer length scales. These microstructures, made out of polydimethylsiloxane (PDMS), take the shape of micropost arrays on which cells are placed. When the cells attach to the posts they exert forces which bend the microposts. This bending is measured and used to calculate the forces generated by individual cells. It is important to measure these forces to fundamentally understand cell interactions within living tissue. This research increases the sensitivity and function of micropost arrays by decreasing the diameter (thus reducing the stiffness) of the posts to the nanometer scale using PDMS etchant, tetrabutylammonium fluoride (TBAF). With nanometer diameters, researchers gain the ability to measure forces produced by individual filopodia. Difficulties with this approach include ensuring homogenous etching of the arrays, and controlling the etch rate. This research overcomes both of these difficulties. To ensure a homogenous etch, the etchant is circulated evenly among the posts using a flow chamber. Etching rate is controlled by adjusting the etchant flow rate and time. Experiments are currently in progress. So far, etch rates have been determined for different flow rates using single block and post structures. This knowledge has been used to design the micropost array experiments. These experiments will be conducted in the coming weeks.

When cells migrate, they undergo a complex process of signaling pathways coordination and interaction with their surrounding environment. Actin filaments polymerize at the leading edge of the cell membrane, pushing it forward and allowing for the cell to bind to new sites in the extracellular matrix. In order to sophisticatedly study the process of cell migration, one must pay close attention to the initial binding points, or nascent adhesions, that can either fall apart early and cause the front of the cell to collapse, or grow into the larger focal adhesions that provide the mechanical support for the cell in the extracellular matrix. Though tools have been created to study the mechanics of cell forces, there is a distinct lack of ability to measure the extremely small forces created by cells, such as the contraction of a single filopodia. This work serves to develop such a tool, in the form of polydimethylsiloxane (PDMS) nanopost arrays, via soft lithography manufacturing techniques. These nanoposts can be treated as tiny cantilever beams by which deflection can be immediately translated to force, using known dimensional and material properties of the posts. Fabrication of the posts consists of a single-cast of PDMS onto silicon negative molds and curing the posts at 110° C at low vacuum pressure. It has been determined that vacuum pressure is necessary to achieve the maximum post aspect ratio, due to surface tension issues between the liquid PDMS and the silicon chip. The gas permeability of PDMS allows for the air contained in the holes of the master chip to escape and be replaced by liquid PDMS while curing. The success rate of nanopost fabrication is characterized by relating post aspect ratios to vacuum pressure of the curing environment, which are found to correlate.

Organisms can develop normally and thrive in dynamic environments. The interplay between the environment and the biomechanics of morphogenesis can help us understand how development operates in a changing world. Effects of salinity on blastula expansion in the sand dollar Dendraster excentricus were used to investigate interactions among the environment, development, and mechanics. Salinity fluctuations, which are common in Dendraster's habitat, affect embryonic cell size. The hyaline layer, an extracellular matrix layer closely associated with the outside of the blastula, is hypothesized to resist blastula expansion. Four models describing how the hyaline layer responds to blastula expansion were compared. In three models the hyaline resists expansion as either an elastic, plastic, or viscous layer. In these models, salinity-driven changes in cell size were predicted to cause changes in the ratio of blastocoel volume to cell volume. In a fourth model, the hyaline is a perfectly accommodating layer that allows the embryo to expand freely as cells divide in a single layer. In this model, salinity changes were predicted not to affect the blastocoel-to-cell-volume ratio. To test these hypotheses, Dendraster embryos were placed in either 25‰ or 32‰ seawater wells and switched to wells of either the same or the other salinity. In a second experiment, Dendraster embryos were raised in 32‰ seawater until the 16-cell stage, at which point a subset of embryos were moved to 25‰ seawater for one cleavage stage and then moved back to 32‰ for the duration of the experiment. In both experiments blastocoel-to-cell-volume ratios were not affected by the salinity treatment. These results suggest that the hyaline layer does not resist blastula expansion in Dendraster, thereby maintaining a constant blastocoel-to-cell-volume ratio.

For thermoelectric (TE) applications, oxides possess certain advantages over conventional intermetallics, including excellent high-temperature stability, low toxicity, and a complex structure, which aids in lowering the thermal conductivity. Recently, it has been found that SrxBa1-xNb2O6 (SBN) holds potential for use in high-temperature TE applications (e.g., waste heat recovery). In our work, a solution combustion synthesis (SCS) process was devised to fabricate SBN with a range of Sr:Ba ratios in order to ascertain an ideal SBN composition for optimum TE behavior. The SCS method, which is highly amenable to mass production, allowed for good control of the SBN stoichiometry, and the as-processed powders were found to possess good single phase purity. Powders processed with different SBN compositions were subject to conventional sintering followed by reduction. The TE properties of each SBN composition were then tested and compared. Preliminary results of our acquired data suggest a decrease in the thermal conductivity as the Ba content is increased, likely due to the larger mass of Ba when compared to Sr. From the obtained results, an ideal SBN composition was determined having the optimal TE performance. Field assisted sintering (FAST) was then employed to processed SBN powder having the optimum composition, and the TE properties of the SBN from both conventional sintering and FAST were compared.

CO₂ separation from flue gases constitutes to nearly 40% of the CO₂ emission in the United States. Developments resulting from this work address factors key to increasing the transport properties in polymer membrane systems. In particular, this work explores method to engineer porous structure to enhance the selectivity of polymeric membrane. The polymer system of interest in this study is poly(l-trimethylsilyl-1-propyne) (PTMSP), a glassy polymer of extraordinary high free volume compared to conventional polymers. It is “reverse-selective” for CO₂ separation from flue gases such as nitrogen or hydrogen, i.e., transports CO₂ gas faster in respect to lighter flue gases. Based on prior studies that showed that enhanced transport properties for PTMSP if constrained to thin films, here we present our data for PTMSP if constrained in nanoporous functionalized anopore alumina oxides (AAO) “sieve” membranes. Impregnation was achieved via vacuum solution impregnation. The overall CO₂ permeability of impregnated membranes was found to be ~80,000 Barrer, which is almost two times higher than that of bulk PTMSP membranes. Selectivities of CO₂/Helium and CO₂/Nitrogen in impregnated membranes were found to be ~7.0 and 9.0, respectively; these are also noticeably higher than the corresponding selectivities of 6.0 and 6.6 found in bulk membranes. Furthermore, we investigated the temporal stability of these membranes that are notorious for their fast aging in the form of ultrathin films. The origin for physical aging is the collapse of the free volume. A two week aging study of PTMSP revealed a significantly slowed down physical aging process if confined in nanoporous AAO by about a factor of 20 over thin films. The exposure to the radial constraint within the pores helped stabilize the free volume and reduce the occurrence of free volume collapse.

One of the most vital roles of mitochondria is to provide the cell with energy through oxidative phosphorylation (OXPHOS). Mutations within the mitochondrial DNA (mtDNA) can affect gene products that function in OXPHOS. While maternally inherited mtDNA mutations often affect the nervous system and musculature, and somatic mtDNA mutations are associated with aging and age related diseases, the cellular mechanisms that influence the frequency of mtDNA mutations are poorly understood. We are using the fruit fly Drosophila to address this matter because our recent work indicates that the mtDNA mutational patterns in flies share many features with vertebrates, including a similar somatic mtDNA mutation frequency, a prevalence of transition mutations, and an accumulation of mutations with age. A high prevalence of mtDNA mutations can lead to dysfunctional mitochondria, and previous work in Drosophila has shown that a mitochondrial quality control mechanism exists to degrade dysfunctional mitochondria. Although it is clear that this mitochondrial quality control mechanism can detect and degrade damaged mitochondria, whether it can influence the mtDNA mutation frequency by detecting and degrading mitochondria with a mtDNA mutation is unknown. To address this matter we will use a high-fidelity Duplex Sequencing approach to detect rare mtDNA variants. Genetic manipulations in mitochondrial quality control factors will be used to probe their effect on mtDNA mutation frequency. We predict that loss of these quality control factors will cause an increase in mtDNA mutations and that overexpression of these factors will cause a decrease in mtDNA mutations. The results from these studies will lead to improved understanding about factors that influence mtDNA mutation frequency and also diseases caused by mtDNA mutations.

The amygdala, specifically the basolateral amygdala (BLA), has been implicated in fear-related processing and learning and is known to receive domaminergic input from the midbrain, although the specific nature of these connections is unclear. Mutations in functional glutamatergic NMDA receptors have previously been shown to impair phasic dopamine activity, leading to hindered fear-related learning and an onset of anxiety-like phenotypes following fear conditioning. To investigate how phasic dopamine activity modulates neuronal signaling in the BLA , I implanted mice with (controls) and without (KOs) a functional NMDA receptors in dopamine neurons with electrode microdrives bilaterally targeting the BLA and used in vivo electrophysiology to record neuronal activity during fear conditioning. The fear conditioning paradigm consists of 10 CS+ trials with a 10 second cue associated with a 0.3mA, 0.5 second foot shock and 10 CS- trials with a pulsed cue predicting the absence of a foot shock for a total of 20 trials in each session. Preliminary results show that the magnitude of the neuronal response to the unconditioned stimulus changes across days in the controls, while remaining constant in the KOs and that the KOs seem to show differences in tonic levels of activation during the trials compared to the controls. These results suggest that dopamine plays a role in modulating tonic activity in the BLA which may be implicated in influencing fear and anxiety related states. 

We estimate a Gaussian graphical model corresponding to a covariance matrix using machine learning techniques. A motivating application is to identify genes associated with cancer in the human genome. Suppose we have genetic data from two people, one of whom is healthy, while the other has cancer. Our goal is to identify gene expressions that differ between datasets and contribute to cancer. We model each dataset as a Gaussian graphical model, which is a network of edges and nodes. Each node represents a random variable, or gene. Each edge represents the dependence between the two variables it connects. An edge with zero value indicates the two variables are independent when conditioned on the rest of the variables. Such networks are analogously represented as inverse covariance matrices. Our goal is to use empirical genetic information from cancer patients to recover the true inverse covariance matrices. We suppose that the inverse covariance matrices have a certain structure and we set up a problem to encourage that structure. Structure refers to the relative magnitudes of matrix elements. A structure of interest is a hub node, which is highly connected to other nodes. These nodes play a regulatory role in gene expression and may be related to cancer. Our goal is to identify hub nodes in the difference between two inverse covariance matrices. We formulate this question as a convex optimization problem. We encourage the solution to have a certain structure by adding regularizer terms to the objective function. We recover the overall structure of the inverse matrices, while simultaneously encouraging the solution to have specific structures of interest. An alternating directions method of multipliers (ADMM) algorithm is used to solve the optimization problem. We run the algorithm on synthetic genetic data, and on real gene expression data to identify cancer related genes.

The field of tissue engineering is becoming increasingly aware of the role that the extracellular matrix (ECM) plays in driving cellular structure and function, especially in highly anisotropic skeletal muscle. Skeletal muscle is a cell dense tissue composed of bundles of aligned myotubes responsible for lateral contraction. Replication of this organization is critical for engineering physiologically-relevant tissue constructs. The fibrous nature of the ECM has led us to the use of bio-inspired topographical line patterns on the nanoscale. We employ UV-assisted capillary force lithography to generate polyurethane-based substrates which stimulate cell elongation and alignment. By incorporating glycidyl methacrylate into our substrates, we present a novel method for engrafting a brush polymer coating of the temperature responsive polymer p(N-isopropylacrylamide) (NIPAAm) onto nano-topographically defined substrates. Seeded with C2C12 mouse skeletal myoblasts, these substrates allow for the harvest of intact cell sheets without the use of enzymes. Using a gelatin coated plunger, these cell sheets are stacked to form tri-layer thick tissue constructs. This study explores the effects of a 3D microenvironment on mature myotube formation and the ability of our NIPAAm-based system to control myotube dimension in 3D by altering nanotopography dimension as well as layer number and the timing of differentiation, contributing to the understanding of the 3D microenvironment in regulating the skeletal muscle cell niche. In addition to paving the way for clinically-significant cell-based therapies, this work may prove vital to the development of advanced in vitro drug screening platforms for patients suffering from muscular dystrophy.

One of the most universal problems faced by persons with transtibial amputations is volume fluctuation of their residual limb. Since the shape and size of a prosthetic socket stays the same, fluctuations in limb volume affect the fit of a socket. For example, if a prosthesis is too tight; pressure spots, skin irritation, and muscle atrophy can occur. Conversely, a loose prosthesis creates gait instability and compromises the comfort of a patient’s prosthetic. Prosthetic socks of different materials and thicknesses are worn to compensate for the changes in volume and manage the discomfort created by daily and monthly volume changes. Properly fitting a residual limb to a socket is further confounded since the industry standard measurement of sock thickness is Ply, a non-linear unit of measurement. Our research examines how absolute thickness and material properties of prosthetic socks change with regular use. We measured the thickness and compression response of new out-of-box socks at three distinct points while in a donned condition. Measurement locations corresponded to area traditionally related to high interface stresses. Thickness and compression response were measured by a custom instrument. Participants walked for two hours while wearing a new sock, after which thickness was measured again in all three locations. Afterwards, volunteers took the socks home, along with a activity monitor, for regular use and returned every two days for one week, and then weekly for a month to re-assess sock thickness and response. The compression response after two hours of walking was used to validate cyclic testing conducting on an Instron material testing machine. Thickness response over long periods of normal use was used to characterize sock fatigue. We hope our study will help practitioners create more accurate volume management recommendations for their patients with a new understanding of the sock stiffness and durability.

Polymers represent a versatile class of chemical compounds that have a wide range of impact in the field of medicine as implantable devices, tissue engineering scaffolds, and drug delivery vehicles. Synthetic polymers such as polyurethane acrylate (PUA) have been used to provide mechanistic cues in the form of altered nanotopography to help guide migration and differentiation in stem cells. To promote cell-biomaterial interactions to elicit desired cellular responses, these surfaces still must be modified. Chemical crosslinking reagents have been applied to engineer surfaces to enhance biological performance; however, they have limitations due to non-ideal reaction environments as well as their low coupling efficiencies with bioactive molecules and their potential converse effects under physiological conditions. Therefore, controlling the surface properties remains a challenge to induce enhanced interaction at the biomaterial interface. Material binding peptides were shown to possess specificity and high affinity to several inorganic surfaces such as gold or silica while offering easy conjugation with biomolecules. This peptide-based surface functionalization, if applied to polymers, can provide a more efficient and robust method of providing biological cues to synthetic polymers. Herein, we demonstrate a simple single-step incubation process that promotes noncovalent binding of bifunctional peptides on polymer surfaces. One end of the peptide possesses high binding affinity for the polymer substrate while the other end contains a bio-functional motif such as the RGD domain, which allows integrin-mediated cell adhesion. When combined with the topographical cues of an anisotropically nanopatterned polymer substrate, a flexible platform can be established to pursue many different research thrusts, such as enhanced stem cell differentiation. Specifically, we plan to use the established platform to uncover the underlying mechanisms of adhesion, morphology, and differentiation in C2C12 mouse myoblasts.

Millions of people in developing countries die from infectious diseases(e.g. malaria and tuberculosis), yet many of these deaths could be prevented if tools for accurate diagnosis were available. The current method for diagnosis of infectious diseases in low-resource settings is the lateral flow test (LFT), which has the appropriate usability (i.e. equipment-free, rapid, and easy to use), but lacks the required sensitivity to have clinical utility for some disease targets. Thus there is a need for diagnostic tools with improved clinical sensitivity and with the appropriate characteristics for use in low-resource settings. I have designed a novel two-dimensional paper network (2DPN), which enables more sophisticated chemical processes for improved sensitivity and usability in low-resource settings. The novel device utilizes commercial enhancement solutions for signal amplification via a metal catalytic reaction to increase sensitivity. The set of reagents and reagent volumes have been optimized for sensitivity and incorporated into a 2DPN assay device, where the programmed delivery of reagents is achieved through the design of the paper geometry and a fluidic on-switch. 

How domestic tasks are shared among household members is an important issue because the time women spent on housework translates to time spent not acquiring valuable skills and experience that enhance aspects of women’s lives which are external to the family such as their career and political involvement. This paper studies the effects of the distribution of intra-household bargaining power on families’ time allocation using cooperative bargaining model. I hypothesize that the wife’s time spent doing domestic chores is directly proportional to her relative bargaining power in the family. I test this hypothesis using instrumental variable approach —I exploit the enactment of equitable distribution of marital property in Florida in 1988 as a natural experiment which redistributes bargaining power while holding household’s preference and price constant. This method overcomes the problem of simultaneous causal relationship between family’s time allocation and household’s characteristics such as wife’s wage. Prior to the law enactment, marital property was divided according to the name on the property title which is disadvantageous to women who generally earn less than their husbands. Thus, the enactment of equitable distribution of property in divorce increases women’s bargaining powers by improving their prospects outside marriage. They can use the ‘threat’ of leaving the marriage as leverage when negotiating with their partners to do less housework. Other literature which studies the intra-household bargaining process focuses on the effects of a shift in bargaining power on married women’s labor supply and family’s expenditure on physical goods such as clothing. Instead of replicating previous research, I instead examine the effects of a change in the bargaining position of each spouse on their allocation of non-market labor time. I anticipate that after the enactment of equitable distribution of marital property statute, the allocation of household chores between the partners becomes more equal.

The fear of being unimportant, being erased, and of being finite, grips humans and motivates us to prove that we matter, that we exist. In an attempt to create truth and authentic understandings, our society seeks evidence, proof, and facts, even while acknowledging (and often ignoring) that these “facts” may be hard to establish. In my paper I explore how post-modern novels, Everything is Illuminated and Extremely Loud and Incredibly Close by Jonathan Safran Foer and Austerlitz by W.G. Sebald, utilize multi-modal textual souvenirs to function as lodestones and guide readers through fictionalized versions of real historical traumas. These novels can be read as scrapbooks of memory which investigate the social importance and definition of “truth” in the face of traumatic events. My project questions whether or not truth can be found in fiction, non-fiction or even photographs as it looks at the way modern technologies have impacted how authors choose to create historical fiction. The multi-modal nature of these novels forces readers to reflect upon authorial intent and authority in the construction of memory and post memory. Furthermore, by looking at the narratives the authors create about traumatic events such as the Holocaust, the bombing of Dresden, Hiroshima, and the attacks of 9-11, my paper reveals the ways in which collective memories and national narratives can arise from trauma and how these events in turn create social identity for individuals within and across groups. An analysis of the postmodern methods the authors use in an attempt to create this authenticity, along with the expectations society holds concerning their construction, and the impact of individual and cultural desires on the creation of these narratives, uncovers the many fallacies and complexities that come with testimony and the social construction of authentic truth.

Like their Victorian forebears, Modernist authors have always placed keen emphasis on women’s hair, often presenting their luscious locks in vivid and scrupulous detail. Aldous Huxley's Crome Yellow (1920) uses hair as a tool of social commentary and trope of the fairytale epic to challenge and even reverse traditional Victorian gender roles and sexuality norms. The tension between the private and public self is also explored; what Modernist women could not say aloud, they expressed through the style, color and length of their hair. Crome Yellow constantly alludes to the hair of the female protagonists Jenny Mullion and Mary Bracegirdle, yet the intentions behind the loaded descriptions of their physical appearance are left up in the air. The characterization of an infamous Fallen Woman and modern flapper girl allows distinctions to be made between the Victorian and Modernist eras that determine the emergence of new gender ideals. This paper will use a close reading of Crome Yellow and Freudian theories on sexuality to explore the multifaceted significance of hair in the Victorian era so we may fully appreciate the past’s influence on modernist eras. A woman’s hair became a phallic image used to engender fear in men, which posits a move towards gender equality and the emergence of the suffragette. Hair was no longer just a physical attribute but a body politic that reflected changes taking place in modernist British society of the entre deux guerres period.

Spinocerebellar ataxia type 7 (SCA7) belongs to a family of neurodegenerative diseases. It is characterized by degeneration of the cerebellum and brainstem. The disease is caused by an expanded polyglutamine tract in ataxin 7. This type of polyglutamine mutation is a common feature in many neurodegenerative diseases. Other neurodegenerative disease including those involving polyglutamine expansions demonstrate altered microRNA (miRNA) expression. Given the supporting evidence that miRNA are differentially expressed in other polyglutamine diseases, I seek to determine the role of miRNAs in SCA7. My proposed research will analyze altered miRNA expression in Purkinje cell neurons, a selectively vulnerable neuronal population that degenerate in SCA7. I will first identify miRNAs that are upregulated or downregulated in Purkinje neurons from SCA7 mice. Next using predictive databases I will identify their potential mRNA targets. I will then test identified miRNAs in cell culture to see if they will inhibit expression of their predicted target. If expression is inhibited, I will then measure the level of mRNA and protein of the predicted target in transgenic mouse tissue. By identifying miRNAs that are differentially expressed in SCA7 mice and confirming their activity, we can build our understanding of the mechanisms of disease progression in SCA7.

Schizophrenia is a debilitating neuropsychiatric disorder with a worldwide prevalence of about 1%. Although the heritability of schizophrenia is high, there are many sporadic cases; that is, affected individuals with no family history of mental illness. We are sequencing persons with schizophrenia in the absence of family history of mental illness (probands), their unaffected parents, and whenever possible, an unaffected older sibling, in order to identify de novo mutations that may be responsible for the disorder. Our hypothesis is that de novo events will be observed in many different genes, and that the functions of the impacted genes will differ between probands and their unaffected siblings. From exome sequence, we filter variants to include only those predicted to be both de novo and damaging to the protein product. Next, we validate all candidate variants passing these filters by Sanger sequencing, using diagnostic PCR primers to amplify genomic DNA of probands, siblings, and parents. Results to date suggest that approximately 50% of probands and siblings carry at least one damaging de novo mutation. The analysis of pathways in which the genes converge is in progress. Next, we will target these candidate genes and determine their complete variant profiles in 5000 cases and 5000 controls. These results can help us identify critical pathways that are affected in schizophrenia.

The use of a fluorometer as a proxy for cell counting is a common method to track phytoplankton culture growth. A fluorometer works by emitting a beam of light, which excites the electrons in chlorophyll a, causing it to emit residual light. The fluorometer then measures the intensity and wavelength of that light. This information helps identify the relative abundance of cells present without giving a direct cell count. When conducting experiments involving the growth rate of photosynthetic cells it would be more beneficial to know the direct cell count as opposed to relative chlorophyll abundance because it is possible they may not correlate exactly as cells have the ability to adjust the ratio of chlorophyll in response to differences in light used in culturing. The marine picocyanobacteria, comprised of the genera Prochlorococcus and Synechococcus, are the most abundant photosynthetic organisms on Earth. I will culture five strains of picocyanobacteria in batch cultures using artificial seawater media in varying light and temperature conditions. I will measure the relative fluorescence of the chlorophyll a in the cultures at regular intervals throughout the growth cycle of the batch cultures. At the same time I will also measure the cell abundance using the flow cytometer. The flow cytometer will individually count and characterize the cells by chlorophyll fluorescence, accessory pigments, and cell size. My hypothesis is that the fluorescence based proxy of cell abundance measured by the fluorometer will highly correlate with the cell count from the flow cytometer, but that exact cell counts will vary under different culturing conditions. My results will be able to provide scientists who use the fluorometer as a proxy for growth a better understanding of the actual number of photosynthetic cells growing in each tube.

With the rise of atmospheric carbon dioxide, understanding the various facets of the carbon cycle is critical. In temperate coastal systems, the phytoplankton group of diatoms play a key role in the carbon cycle by sequestering carbon from the atmosphere into deep ocean sediments. Many scientists turn to laboratory culturing experiments to make predictions about the response of phytoplankton to changes in climate. But do these laboratory studies serve as realistic proxies for environmental responses? This study will compare growth rates of phytoplankton communities from Barkley Sound, British Columbia using field and laboratory experiments. Field experiments will comprise an in situ community growth rate measured using diffusion chambers in addition to on-ship incubations measuring the community growth rate in response to added nutrients. Field samples will be used to isolate the three dominant diatom species into a mixed-species culture for laboratory experiments. An analogous culture using laboratory isolates of the same species will be created, and the two cultures will be kept in a semi-continuous batch culture experiment tracking the growth rates in response to added nutrients. Comparisons of the growth rates in situ, shipboard, and in the laboratory will indicate how growth rates differ between natural phytoplankton assemblages and laboratory settings. Growth rates will be calculated using changes in chlorophyll a levels, a proxy for cell densities. If growth rates are much higher in the laboratory cultures than in the wild cultures, the implication is that there are limitations to growth in the natural environment that should be factored in when correlating laboratory experiments to the responses of natural phytoplankton assemblages. However, if there is no significant difference in growth rates between the experiments, then the laboratory conditions can provide similar conditions as in the field and laboratory culture experiments and can be easily used to predict environmental responses.

Viruses require host cell metabolism to provide the necessary energy and biosynthetic precursors for viral replication. Recently, our lab conducted a global metabolic study to determine how vaccinia virus (VACV), a large double-stranded DNA virus, alters host cell metabolism. VACV is a model system to study poxvirus infections and was used as the live vaccine against smallpox. In our study, dual mass spectrometry platforms were used to analyze a broad range of metabolites from mock and VACV-infected fibroblast cells harvested at multiple time points within 24 hours of infection. One of the most notable metabolic alterations observed during VACV infection was increased glutamine utilization, or glutaminolysis. We have since shown that under conditions of glutamine starvation there is an approximate 90% decrease in VACV production in fibroblasts, thus revealing an essential role for glutamine during infection. Furthermore, this decrease in virus production could be rescued by the addition of tri-carboxylic acid (TCA) cycle intermediates, suggesting glutamine’s use as an anaplerotic, or replacement, substrate for the TCA cycle during infection. To determine at what stage(s) in the VACV life cycle glutamine is required, quantitative real-time RT-PCR and western blot analyses were performed to examine early and late viral transcript and protein levels. We have found that glutamine deprivation does not inhibit viral gene expression, but does significantly impact viral protein synthesis. Importantly, the addition of TCA cycle intermediates recovered viral protein levels, highlighting the need for glutamine carbon to maintain the TCA cycle at this stage in the VACV life cycle. This study reveals the robust metabolic program implemented by vaccinia virus during the course of infection and provides insight into the role metabolic reprogramming plays during viral replication. Ultimately, studies such as these may help to identify novel therapeutic methods to specifically target infected cells and inhibit viral replication.

Angiogenesis is the development of new blood vessels from pre-existing vasculature and is a hallmark of Kaposi’s sarcoma (KS), a highly vascularized neoplasm prevalent among immunosuppressed individuals. The etiological agent of KS, Kaposi’s sarcoma-associated herpesvirus (KSHV), induces angiogenic phenotypes in endothelial cells during latent infection. Interestingly, KSHV infection leads to down-regulation of transforming growth factor-beta 2 (TGF-β2), an anti-angiogenic cytokine. In addition to six viral genes, KSHV also encodes 17 recently discovered microRNAs (miRNAs) during latent infection. These endogenously encoded RNAs of 19-23 nucleotides post-transcriptionally regulate gene expression through basepairing interactions with the 3’ untranslated region (UTR) of target messenger RNAs (mRNAs) and either mark them for degradation or prevent ribosomal binding. These viral miRNAs appear to directly contribute to pathogenesis and angiogenesis by targeting host gene expression. We screened the miRNAs of KSHV to determine if any are involved in the down-regulation of TGF-β2 during KSHV infection. We found that two KSHV miRNAs, miR-K3 and miR-K8, are independently sufficient to downregulate TGF-β2. To determine if these miRNAs directly target the 3’UTR of TGF-β2 as opposed to altering genes that affect TGF-β2 expression, we used a TGF-β2 3’UTR luciferase reporter construct. We also used competitive inhibitors of miR-K3 and miR-K8 to determine if these miRNAs are necessary for the down-regulation of TGF-β2. By identifying specific targets of KSHV miRNAs such as TGF-β2, we can improve our understanding of how KSHV is able to maintain latent infection and target host gene expression to alter endothelial cells.

Earmark spending has proven to be a popular political tool for members of Congress because it allows them to claim credit for a concrete economic benefit for their districts. I ask whether expectations of political benefits similarly influence large-scale stimulus spending allocations and, more importantly, whether these allocations have political benefits for the incumbent president. This research project examines the link between federal spending and shifts in election outcomes. It asks whether spending in the stimulus package of 2009 substantively influenced President Obama’s share of the popular vote in 2012. Other things equal, did competitive regions of the nation receive disproportionately more spending, and was that spending associated with increased vote shares for the incumbent president between 2008 and 2012? To answer these questions I examine government spending and popular vote shares in over 3,000 counties from 2008 to 2012. I also control for underlying social and economic issues by county, such as foreclosure and unemployment rates. Finally, I control for other political influences on stimulus spending, including whether certain counties had representatives in powerful congressional roles. Overall, this study will promote understanding of the political implications of large-scale federal spending. It will also shed light on the political dynamics of “swing counties” and whether stimulus spending is a useful political tool for winning these counties.

Epilepsy is a neurological disease characterized by recurring seizures and it affects millions. We hypothesize that modulation of p38 mitogen-activated protein kinase (p38 MAPK) will influence hyperpolarization-activated cyclic nucleotide-gated (HCN) channel activity, affecting the frequency of seizures in an animal model of epilepsy. p38 MAPK is a kinase activated by conditions of cellular stress. It strongly stimulates the HCN channels, which are voltage gated ion channels that are highly expressed in the cortex and hippocampus, brain regions where seizure onset occurs. A decrease in HCN channel expression or function has previously been observed in epileptic animals associated with a loss of p38 MAPK activity, which produced neuronal hyperexcitability. We hypothesized that increasing p38 MAPK activity might decrease neuronal activity and decrease seizure frequency. In this experiment, Epilepsy was induced in Sprague Dawley rats through treatment with pilocarpine, a convulsant drug. Recurrent, unprovoked seizures were seen after three weeks of treatment. An osmotic pump was placed in the rats, delivering drugs that modulate p38 MAPK. The drug SB203580 (SB), a specific inhibitor of p38 MAPK, was administered, as was anisomycin, a non-specific activator of p38 MAPK. Animal brain activity was analyzed with video electroencephalography through electrode implants in the skulls of the rats. Inhibition of p38 MAPK by SB increased seizure frequency by 179 % as predicted, however, anisomycin also increased seizures by 236%. We hypothesize that this may be due to non-specific actions on other signaling pathways like the Jun N-terminal kinase pathway (JNK). We’re investigating the role of JNK in modulating seizure frequency, and this may point us to some new therapies. The implications of this study may indicate a new approach to the control of epilepsy via anti-epileptic drugs targeting p38 MAPK.

The circadian system controls daily rhythms of behavior and physiology, including locomotor activity and motor-task performance. The master regulation of these rhythms is achieved by a circadian clock located in the suprachiasmatic nucleus of the hypothalamus; however, it is not clear how motor tasks are programmed by the motor cortex at different times of the day. Our hypothesis is that similar motor tasks executed at different times of the day may require different motor programs to account for the daily variance introduced by the circadian system. How and where in the brain this variance in motor control manifests has not been established. We propose to identify specific primary motor cortex activity patterns associated with specific motor outputs across the 24-hour day. We implant electrocorticographic (ECoG) electrodes onto the motor cortex of mice and record brain wave activity during wheel running and rest. We first test whether the motor cortex may show a brain-wave signature for wheel running, and second whether this signature may change predictably across the 24-hour day. Initial results indicate a broad-spectrum power increase in brain-wave output from the motor cortex associated with wheel running. Furthermore, there appears to be a circadian modulation of the power of specific ECoG frequencies. The decoding of motor cortex signals is at the core of the design of brain-machine interfaces (BMIs). These devices decode signals from the conscious brain to drive the execution of specific tasks by a machine, such as an artificial limb. Our work will directly contribute to our understanding of how the motor cortex decodes circadian time. This knowledge is essential to create BMIs that can operate effectively throughout the 24-hour day to execute tasks by brain-operated artificial devices.

Previous research has shown that adults demonstrate better memory for novel music from their own music culture as opposed to music from a foreign culture. This passive learning imprinted upon kids as they grow up in a certain musical culture its known as “enculturation”. Visual information has also been shown to impact musical memory in adults. Pertinent visual images may augment the encoding process of music and lead to greater recall abilities. In this study, we will include a visual component to the audio information presented. This addition will test the facilitation of visual information to the musical memory of culturally unfamiliar music in adults. Using four video/audio excerpts of musicians performing music in a classical Chinese style and four video/audio excerpts of musicians performing western (classical) style music, we tested the audio and visual memory of adult human subjects in culturally unfamiliar and familiar music. To test them we used twenty-four test items (twelve true tests and twelve false tests) in audio or video format. These different test items can be prepared in three different combinations: audio/audio, AV/audio, AV/AV. Preliminary data demonstrated significantly better overall memory for culturally familiar music. However, music memory scores were higher in the audio condition than in the AV condition in culturally unfamiliar music tests. These results indicate that adding visual information does not aid in musical memory of culturally unfamiliar music. Dichotomous from our initial hypothesis, it is possible that this occurred due to the dominance of visual memory in encoding, or that auditory and visual stimuli provided too much novel information for effective recall.

In typical ascidian development, a tailed larva hatches, swims, becomes competent, and then responds to settlement cues, that initiates metamorphosis. Metamorphosis triggers apoptosis, which is required for tail loss. At this time, the notochord is lost as the tail reabsorbs and the juvenile settles to a substrate. In the ascidian family Molgulidae, species have evolved tailless larval development at least four times independently from a tailed ancestor. We hypothesized that a heterochrony in metamorphosis is responsible for the evolution of tail loss in the tailless ascidian larvae. We show that there is early expression of metamorphosis genes in Molgulidae, including genes involved in the Mannose Binding Lectin (MBL) Complement pathway of innate immunity. We hypothesize that the maturation of the immune system early in tailless larvae could be a progression to competency and trigger to metamorphosis. Jeffery (2002) showed programmed cell death happens in cells that form a tail early in development. We further hypothesize that a heterochrony in metamorphosis promotes the early activation of programmed cell death in the tailless ascidian larvae. Using next generation sequencing techniques, we sequenced transcriptomes of a tailed and tailless species and their hybrid through development. We examined genes that have been shown to be expressed during metamorphosis in other ascidians and saw earlier expression of genes critical for metamorphosis in both species. Our ultimate goal is to elucidate the ascidian molecular trigger for metamorphosis and uncover whether heterochrony of metamorphosis leads to tailless ascidians.

In evolutionary history, distantly related taxa have often independently evolved the same or similar complex structure as a solution to a common problem presented by their environment. One such example is the plagiaulacoid tooth—a laterally compressed shearing blade that convergently evolved in four clades of mammals, some extant and others extinct. To better understand the selective forces that led to the convergent evolution of this specialized trait in multiple lineages, I investigated how this tooth form correlates with diet in a sample of extant taxa. Specifically, I measured complexity of lower cheek tooth rows of 15 species from five families of extant marsupials with plagiaulacoid teeth. To quantify dental complexity I created digital 3-D models from microCT scans of these specimens and used GIS software to generate orientation maps of the tooth surfaces. Contiguous pixels with the same orientation are considered a patch on the tooth surface, and the number of patches approximates the number of shearing surfaces available to mechanically process food. Previous studies of placental mammals have shown that increases in Orientation Patch Count(OPC) (i.e., increases in tooth surface complexity) correlate with increased consumption of plant material. However, I found no correlation between OPC value and diet as recorded in the literature for these taxa. I then compared complexity of the blade to complexity of the molars to investigate the functional contribution of the plagiaulacoid tooth and again found no correlation with diet. Broader sampling will help determine the pattern of tooth complexity and diet in marsupials, but the range of OPC values generated in this study suggests that the convergent evolution of the plagiaulacoid phenotype is not a simple case of one morphological adaptation to a particular environmental problem.

Wnt signaling is known to be critical for proper embryonic development in most animals studied to date. But key evolutionary questions on the origin and evolution of this pathway in the metazoan common ancestor are still unresolved. Recently, the genome of Pleurobrachia bachei, a member of the early branching metazoan linage ctenophora, has been sequenced. Insights into the function of the Wnt pathway in P. bachei will provide information on early evolution of this key pathway. Three Wnt ligand genes were identified in P. bachei and cloned for in situ hybridization. These genes showed expression in the combs, tentacles, mouth, ciliated grooves and polar fields of the adult P. bachei. In a genomic search for other members of the canonical Wnt pathway, components of the destruction complex and antagonists were incomplete or missing from the genome. High levels of Wnt expression in the adult P. bachei indicates that Wnt could also be playing a role in signaling in the adult.

Extrasolar planets are being discovered at an incredible rate today. We have yet to discover another planet that can support life as we know it, but that will very likely change soon. Currently, most exoplanets are discovered indirectly by the influence they have on their host stars. Therefore, we only know the temperature and size of an exoplanet as well as we know the temperature and size of the star it orbits. For low-mass stars in the spectral range between K5 and M2, models do not provide the desired accuracy for these parameters. I improve this accuracy for a sample of planet candidates discovered by the Kepler Space Telescope by using a technique that assigns the parameters of nearby and well studied "stellar twins" to the planet hosting stars. I find these stellar twins by comparing the spectra of planet hosting stars to the spectra of similar nearby stars. By assigning the parameters of the nearby stellar twins to the planet hosting stars, I reduce the uncertainties in size and temperature for the planet hosting stars. This will also reduce the uncertainties in the size and temperature of the exoplanets, and thus allow for a better determination of their habitability.

Since the development of the Drug Recognition Expert program in the 1970s, police officers are trained to recognize impairments and have the expertise to identify these influential drugs opposed to, or along with alcohol. This research is a part of an ongoing partnership between the Washington State University Division of Governmental Studies and Services and Washington State’s DRE program. The goal of the work entails determining whether the current evaluation standards set forth by the Washington State Patrol are sufficiently strict and assessing the accuracy of the DRE officers’ assessments of impaired drivers. The analysis will be based on the coding and digitization of all 5000+ DRE assessments done over the period 2006 to 2010 (5 years) and will include the identification of Type I errors (failure to detect) and Type II errors (false attention) as determined by toxicology reports on blood samples. An examination will be made to determine whether the Type I and Type II errors detected are related to driver racial and ethnic background, gender, and age. With this research an understanding of how law enforcement training on “drugged driving” can be improved in Washington State. Resolving DRE training issues will ensure that individuals’ constitutional rights are fully protected while law enforcement safeguards their ability to enforce the states driver safety laws in Washington.

 With an international focus on limiting greenhouse gas production, advancements in biomass conversion are needed. Conversion of biomass into more convenient fuels can lead to carbon neutrality. Biomass has the potential to be converted to transportation fuels after undergoing oxygen-free thermochemical decomposition, known as fast pyrolysis, and upgrading in a Fluid Catalytic Cracking reactor. Wood, one form of biomass, is largely made up of cellulose, hemicellulose and lignin. Lignin is comprised of polymers constructed of p-coumaryl alcohol, coniferyl alcohol and sinapyl alcohol monomers. In the present project, pyrolysis of these monomers will be modeled using phenol, guaiacol and syringol, respectively. Individual monomers and mixtures of monomers will be pyrolyzed using a Pyroprobe and the bio-oil contents will be quantified by gas chromatography-mass spectroscopy. It is expected that when combinations are pyrolyzed, the bio-oil production will be suppressed due to secondary reactions of each monomers’ product. As combinations of these monomers are characterized, pyrolyzed lignin can be characterized. Eventually, all aspects of pyrolyzed wood will be characterized and optimum conditions for pyrolysis of woody biomass can be assessed, thus yielding the highest quality bio-oil.

Mussels attach themselves to hard substrates by extruding thin, strong, and flexible hair-like attachments known as byssal threads, which are anchored in place with small adhesive plaques. While many studies have examined how multiple abiotic variables affect mussel attachment strength, few have performed extensive single variable tests within a controlled lab setting. In this study, we investigated the effects of temperature on byssus production in the native Northern Pacific intertidal blue mussel, Mytilus trossulus, and expected to see a decline in production with increasing temperatures (specifically, between 18°C-25°C). We also hypothesized that threads produced in warmer temperatures would show a visual decline in quality, either through thinning, degradation, or tearing. Mussels were placed in a temperature controlled flume for 24 hours. Six different trials were run at 10°C, 14°C, 18°C, 20°C, 22°C, and 25°C. Byssal threads were counted for the duration of each trial to determine production and rate of attachment. Threads produced under these different temperature regimes were analyzed using scanning electron microscopy to visually compare differences in quality. Overall, there was a significantly negative correlation between thread production and increases in temperature, with the highest amount occurring at 18°C (11.25 threads +2.0 s.e.m.) and the lowest at 25°C (0 threads). Acclimatization and rate of thread production was also negatively affected by temperature increases. In addition, we saw a significant difference in percentage of mussel attachment between temperatures, with up to 93% attached in the 10°C and 18°C treatments and 50-0% in the 20°C-25°C treatments. However, visual analyses of thread quality differences were inconclusive. Our findings indicate that climatic temperature increases may negatively affect both the range of M. trossulus within Northern Pacific rocky intertidal zones as well as potentially setback the aquaculture of the species.

Ceftaroline fosamil, otherwise known by the name ceftaroline (trade name Teflaro) is a cephalosporin antibiotic indicated for the use against MRSA skin and soft tissue infection and bacterial pneumonia. Though approved by the FDA, the drug is thought to be safe and thought to be well tolerated, however, our team has found it to cause neutropenia in patients. Neutropenia is defined as the abnormally low number of neutrophils, which seems to be present in some patients during and after taking ceftaroline. It is our objective in this study to determine the incidence of neutropenia among patients using ceftaroline. The drug has been approved for more than a year but the post marketing experience has been limited. We have seen two cases of agranulocytosis, which has led us to conduct this study. The study will be conducted in the Tri Cities, which is comprised of Kennewick, Pasco, and Richland. The Tri Cities has a diverse population of 193,000 individuals and there are three major medical facilities: the Kadlec Regional Medical Center, Lourdes Medical Center and the Kennewick General Hospital. The study will determine the incidence of neutropenia among patients receiving ceftaroline. We also hope to describe the side effects of the drug, and track patients’ health over the course of a 7 month period by reviewing patients' charts and through phone interviews. All patients that are included in the study received ceftaroline from August 2011 to March 2012. The patients’ data will be secured from hospital records from the three local hospitals. Our hope is to find a link between usage of ceftaroline and incidence in neutropenia in patients.

Acute myeloid leukemia (AML) is a cancer characterized by the accumulation of abnormal immature white blood cells in the bone marrow. This proliferation impairs normal blood production and leads to suppression of normal blood counts, increasing susceptibility to infection and bleeding. Adhesion of leukemia cells within the bone marrow confers resistance to chemotherapy drugs. E-selectin is a molecule on the surface of endothelial cells lining blood vessels; it mediates the adhesion of white blood cells to vessels within bone, and may play a role in the residence of leukemia in the bone marrow “vascular niche.” Interaction of cancer cells with E selectin has been shown to activate survival pathways such as PI3K-NFκB, making cells less susceptible to apoptosis (programmed cell death). Using an assay for E-selectin and analysis via RT-PCR, we have preliminary data suggesting that binding of cells from a specific patient sample to E-selectin up-regulated genes in the TGF-beta, Wnt and Hedgehog signaling pathways, the latter of which may play a role in drug resistance in CD34+ leukemia cells. Further work aims to demonstrate that the activation of these survival pathways is consistent across a range of cell samples, and to investigate whether up-regulation extends to other pathways known to inhibit apoptosis in cancer cells. Knowledge of the mechanisms by which leukemia cells increase their survival upon binding ligands in their environment such as E-selectin could aid the design of therapies aimed at treating AML, and potentially help overcome the persistent problem of resistance to chemotherapeutic agents.

Atherosclerotic cardiovascular disease is the leading cause of death in North America.  Hepatic lipase (HL) a liver enzyme that hydrolyzes lipoprotein triglyceride, plays a critical role in lipid metabolism and may influence the development of atherosclerosis.  Hepatic lipase may serve dual roles in the development of atherosclerosis: 1) HL may be pro-atherogenic by converting low-density lipoprotein (LDL) to small-dense LDL, causing build up of cholesterol in arteries. 2) HL may be anti-atherogenic by increasing the cholesterol-poor form of high-density lipoprotein (HDL) that removes cholesterol from the arteries, decreasing build up of cholesterol in arteries.  Previous studies have reported that elevated levels of human hepatic lipase (hHL) reduce the development of atherosclerosis.  However, those studies were done in transgenic mouse models with supra-physiologic levels of hHL and may not accurately reflect the role of physiologic levels of HL on atherosclerosis.  Therefore, we created a physiologic model of hHL expression to analyze the role of hHL in the development of atherosclerosis.  This “humanized” mouse model expresses hHL at physiologic levels in the liver.  We hypothesize that physiologic levels of HL will reduce the development of atherosclerosis compared to mice lacking HL.  To test this hypothesis we bred the transgenic hHL mice onto a low-density lipoprotein receptor knock out background (which is an established model for atherosclerosis).  Littermates with and without the hHL transgene were fed a high-fat, cholesterol enriched (“Western”) diet to accelerate atherosclerosis development, after which the aortas were harvested, dissected longitudinally, and pinned to expose the inner aortic walls for visualization.  Cholesterol buildup was stained and the ratios of atherosclerotic accumulation to total aortic surface area were quantitatively measured via Adobe Photoshop.  Results from this study will clarify the contribution of hHL to atherosclerosis development.  Ultimately, our results will guide the development of new treatments to prevent and treat atherosclerosis.

In most eukaryotic organisms, sexual reproduction is a critical component of the life cycle. Through meiosis, diploid cells containing two copies of each chromosome divide to produce genetically diverse haploid gametes (e.g. sperm) that contain one copy of each chromosome. This process is fundamental in sexual reproduction. Generally, meiosis results in a 50:50 transmission of heterozygous alleles into gametes. For example, human males make approximately 50% sperm with an X chromosome and 50% with a Y chromosome. Because only one copy of each chromosome can contribute to a given offspring, there is an evolutionary pressure on alleles to compete for preferential inclusion in gametes. Sometimes alleles can “cheat” to gain a greater than 50% transmission through meiosis. This is known as meiotic drive. Meiotic drive could have significant implications: altering fertility, speciation, and the evolution of chromosomes in eukaryotic organisms. For example, competition between alleles may be driving the rapid evolution of chromosome segregation factors acting during meiosis and mitosis, potentially driving chromosome segregation away from a theoretical ideal. Though meiotic drive has not been well understood due to limited model systems, we have developed a novel system utilizing genetically tractable fission yeast S. pombe and S. kambucha, to investigate meiotic conflicts between competing alleles. In S. pombe, heterozygous alleles are widely thought to segregate fairly in meiosis, suggesting that active meiotic drive loci are not present in this organism. We have found that silenced meiotic drivers can be de-repressed in aged germ cells, leading to greater than 90% transmission of certain alleles through meiosis. This provides experimental evidence in support of the evolution of cheating in meiotic processes that could ultimately result in decreased fertility amongst eukaryotic organisms, including humans. 

The goals of this project are to produce transgenic maize and wheat tissue expressing a gfp marker in chloroplasts and to regenerate homoplasmic plants from the transformed tissue. There are several advantages of transgenic chloroplasts, including multiple copies and high expression of the transgene without gene silencing. In addition, plastid DNA (ptDNA) is maternally inherited so that the transgene will not be present in pollen, thus providing containment for the transgene. We used circular and linear vectors that contain ptDNA sequences corresponding to the ends of the plastid chromosome and the gfp transgene. We used particle bombardment to deliver the vector to plastids of maize and wheat. Transgene expression was assessed by fluorescence microscopy and imaging of GFP in plastids. In addition, both standard PCR and quantitative PCR were performed using DNA extracted from the tissue and primers for the gfp transgene. Although we achieved successful plastid transformation for both maize and wheat, the tissues were heteroplasmic, containing both wild type and transgenic ptDNA. We found that a linear vector gave higher transformation efficiency than a circular vector. The next step is to develop procedures for generation of homoplasmic maize and wheat plants.

The project I worked on was part of a larger project in which we examined the activity of O6-methylguanine-DNA methyltransferase activity, CpG promoter methylation, and progression-free survival following alkylating agent therapy in glioblastoma and anaplastic glioma brain tumors. For my part in the research I performed standard biochemical assays to quantify O6-methyalguanine-DNA methyltransferase (MGMT) in extracts of glioblastoma (GBM) and anaplastic glioma (AG) tumor samples from adult patients operated on with consent at the University of Washington Medical Center between 1991 and 2008. MGMT is a DNA repair protein that excises methyl adducts from the O6 position of guanine residues. Chemotherapy and other alkylating agents such as methylating triazene derivative temozolomide (TMZ) are responsible for methylating DNA, or damaging the DNA and thus destory the tumor cell. There has been poor prognosis for GBM due to the intrinsic resistance mechanisms in the MGMT that limit the efficacy of TMZ and other alkylators. GBM and other gliomas display a wide range of MGMT activity and thus the question for research was whether the amount of MGMT in tumors correlated with progression-free survival following alkylator-based chemotherapy. Overall the data strongly supported that MGMT activity in GBM and AG tissue mediates resistance to alkylator therapy.

The pupil is an important component of the visual system, changing in size to regulate the amount of light entering the eye and affecting vision quality. Light is the primary determinant of pupil size (the pupil constricts with light increments, a response known as the Pupillary light reflex or PLR), but there are also well known effects of internal states such as cognitive effort and arousal. Recently, our lab discovered an interaction between light and cognitive factors at the level of the pupil: selectively paying attention to brighter areas in the visual field, without looking at them, enhances the PLR. This finding prompts a revision to the neural substrates of the PLR, suggesting that cortical attentional influences modulate the subcortical PLR circuit. To better characterize this Attention-PLR interaction, we tested whether the effect is limited to spatially-directed attention to bright regions, or if it generalizes to cases where attention is directed to bright features from a single localized stimulus. Preliminary data using a psychophysics setup of overlapping dark and bright surfaces shows an effect of feature based attention. Specifically, two overlapping surfaces defined by white and black moving dots were presented at fixation and subjects were cued to attend to either surface. Attending to the bright surface induced a relative pupillary constriction compared to attending to the dark surface. Moreover, ongoing work attempts to quantify the magnitude of the attention effect at different levels of brightness and contrast of the stimuli, and for different spatial extents of the attended regions. In this way we investigate the characteristics, dynamics, and anatomical substrates of the Attentional-PLR effect, while establishing the pupil as a convenient marker of the direction of attention.

The lift-to-drag ratio (L/D) of a wing or propeller is the most important measure of its performance, as a higher ratio defines more efficient designs. The goal of my group’s project is to show that sufficiently strong and persistent vortices along the top of a wing will positively benefit its aerodynamic efficiency. Previous results in the UW Fluid Dynamics Lab show that a wavy wall modeled after the Kelvin-Stuart Separatrix shape -  which intends to more accurately outline the vortice's flow - further minimize boundary layer turbulence compared to previous wavy wall shapes. We believe that modeling the upper half of our wing along the Kevlin-Stuart Separatrix will keep the boundary layer attached even at very high angles of attack, with relatively low drag coefficients. Two model wings are being tested in the wind tunnel, a control wing with a familiar smooth top half and our experimental model outfitted with vortex generators and the top half shaped to match the Kelvin-Stuart separatrix. Further research may include attaching surface tufts to both models, which qualitatively aid in visualizing flow. The models are mounted to a force balance system, which returns the lift and drag. Our experiments test various angles of attack and vortex generator configurations to compare the wings in different situations, such as take-off and cruise. Achieving the desired L/D will have broad implications for both wing and wind turbine propeller design. More efficient wings could drastically lower fuel costs for airliners, and more effective wind turbines would return more energy at lower cost.

The emphasis of my research is the treatment of the Holy Family created by the artists Francisco de Zurbarán and Bartolomé Esteban Murillo. The subject of the Holy Family in a genre setting is a response to the efforts of the Church during the Counter-Reformation to appeal to common people, thereby encouraging their loyalty to the Papacy. However, different areas portrayed the Virgin Mary, Christ, and Joseph in regional styles. I observed how Spanish Baroque artists created a “fusion” of both genre and religious artworks by comparing Zurbarán’s “Christ and Mary in the House of Nazareth” and Murillo’s “Holy Family with the Little Bird” with other artworks from Netherlandish and Italian artists. To help prove the fusion style, I answered three questions. First, how would a viewer recognize the figures as the Iconic Family? We know this through symbolic objects found in both paintings. In Zurbarán’s work, the white cloth lying on Mary is referring to Michelangelo’s “Pieta.” The boy is pricking his figure on the crown of thorns is symbolic for Christ. Murillo uses a carpenter’s bench right behind Joseph and the basket of garments as symbol for the Virgin Mary. Secondly, how does each work portray everyday moments like genre paintings? We see this in Zurbarán’s work by Mary glancing over towards her son with a single tear running down her face. Murillo has caught a moment were Mary is glancing over her shoulders with a “semi-smile.” My last question is how these paintings showed religious and family values? The morals are seen in both paintings by the body language of the figures and their interaction with one another. These Spanish Baroque artists created a hybrid style that is visually proven through the mixed elements of both religious and genre artwork.

Burma is now undergoing an incredible transition toward peace, stability and democratic governance – a transition that we found requires a fundamental re-thinking of U.S. policy relations. While we discovered Burma’s commitment to reform to be genuine, its challenges are numerous. Instability and underdevelopment continue to dominate the Burmese landscape, defined by a weak constitutional government, repressed civil society, unreformed military, ethnic violence and an underdeveloped economy. This past year marked President Obama as the first American president to visit Burma, extending a hand of friendship, committed to strengthening political and economic ties between the United States and Burma. As well as a real commitment by the United States to reduce economic sanctions, aid in the democratization process, and lift the Burmese economy out of poverty -  elements we proved to be essential in ensuring a successful transition in Burma. Our task force recommends greater engagement, aid, and targeted policy consultation to help the country emerge from decades of military-junta rule and rejoin the international community. Ultimately, Burma offers a new roadmap in the United States in re-thinking the reform process of developing countries, one that now requires trading the tools of isolation and international pressure for those of integration and international cooperation. The world is now watching Burma as it prepares to chair the ASEAN council in 2014 and hold democratic elections in 2015. No one knows how long this moment for reform will last, but the time for action is now.

Double stranded DNA (dsDNA) viruses such as herpesviruses, adenoviruses, and many bacteriophages, have common development pathways. In these cases, a packaging motor (the terminase enzyme) inserts the viral genome into a capsid shell. We study the DNA packaging motor from phage lambda as a model system. Lambda terminase plays four critical roles in virus assembly: (1) It has a site specific DNA binding activity where it binds to the cos sequence in the lambda genome, (2) it has an endonuclease activity where it symmetrically nicks the duplex at the cosN site, (3) it has a strand-separation activity that separates the cleaved DNA strands, and (4) it has “packaging” activity where it translocates the “matured” viral genome into a capsid shell through a portal ring structure. The lambda gpFI protein is required for virus assembly in vivo but the function remains unknown. In this study, we use a defined in vitro system to study the effects of gpFI on the endonuclease, strand-separation, and DNA packaging activities of lambda terminase as our first steps towards an understanding the function of gpFI in virus assembly. Our data demonstrate that gpFI affects the cos-cleavage and strand-separation activities of terminase. In contrast, the protein significantly increases the efficiency of DNA packaging in vitro. In addition, we demonstrate a direct interaction between gpFI and the procapsid shell. The significance of these results with respect to the biological role of gpFI in virus assembly is discussed. The information obtained from these experiments will be useful in understanding similar processes in the eukaryotic herpesviruses and adenoviruses.

Pseudomonas aeruginosa can cause chronic and latent infections within many human diseases especially those related to immunocompromised individuals such as cystic fibrosis. This species also engages in a density dependent communication known as quorum sensing, which can often increase the species' virulence through the production of extracellular products such as proteases and components for the production of biofilms. Quorum sensing is a highly conserved process in nature yet quite interestingly we find that cystic fibrosis lungs provide a unique environment for the profileration of quorum sensing mutants (LasR mutants) as opposed to free living forms of the species elsewhere in the general environment. I am working to characterize 100 environmental isolates of Pseudomonas aeruginosa to test the hypothesis that quorum sensing mutants are infrequent within free living populations of the bacteria and to this point, I have not conclusively found an environmental mutant. Any information will in turn help us understand more about the selective pressures leading to the emergence of LasR mutants in the cystic fibrosis lung and consequently the therapeutic significance or insignificance of targeting quorum sensing in cystic fibrosis chronic infections.

French films about World War II tend to fall into either the category of heritage film, which promotes a narrative of reconciliation in relation to the collaboration, or another group of films that serves to implicitly or explicitly question that narrative of reconciliation. Films of the latter category have probed the memory of the Vichy collaboration since the end of the war despite Charles de Gaulle’s efforts to repress narratives that challenged the idea of a country united against Germany. In the 1980’s, however, heritage film became the prevalent genre and offered a Gaullist view of World War II in France. This thesis asks, why, given the plethora of films dealing with the realities of the French role in World War II dating from the late 1940’s, a rubric of heritage film emerged as one way of classifying filmic discourse on the Vichy collaboration. I hypothesize that the heritage cinema rubric, specifically the Vichy cinema rubric, asserts a legibility of French identity, which was compromised by France’s collaboration with Nazi Germany, and attempts to thwart filmic narratives of identity loss. Through tropes of the uncanny and characters that experience repressed trauma and blocked mourning, films such as Le Silence de la Mer (1949), Lacombe, Lucien (1974), Mr. Klein (1976), and Le Dernier Metro (1980) reflect the French collective experience of loss during and in the aftermath of the collaboration. WWII films produced in the decades leading up to the emergence of this genre in essence dramatize a troubling disappearance of French cultural identity, which the heritage rubric sought to control.

Circadian rhythms like the sleep-wake cycle are governed by a circadian clock located in the suprachiasmatic nucleus of the hypothalamus (SCN). This clock is synchronized by the light-dark (LD) cycle and the cyclic-guanosine monophoshate (cGMP) pathway is known to mediate its phase advances. Phase-advancing light pulses increase the levels of cGMP in the SCN and inhibition of phosphodiesterase-5 (PDE5), the enzyme that degrades cGMP, by artificial drugs such as sildenafil, enhance light-induced phase advances. These inhibitors act by inducing PDE5 phosphorylation and converting it to phosphorylated-PDE5 (phPDE5). This evidence suggests that the inactivation of PDE5 through its phosphorylation could be a mechanism through which light induces phase advances, and that sildenafil utilizes the same mechanism to enhance phase advances upon light stimulation. We are testing this hypothesis by measuring PDE5 and phPDE5 in the SCN of mice treated with a phase-advancing light pulse. Mice are administered either a light or a dark pulse at the time that light causes phase advances. Animals are sacrificed at different times after the presentation of the pulses, their brains are dissected and alternate sections are processed for immunohistochemistry against either PDE5 or phPDE5. Our hypothesis predicts an increase in phPDE5-expressing cells in light-treated animals. Future experiments will determine if sildenafil- induced enhancement of phase advances are mediated by further increasing phPDE5. Our findings may lead to the development of pharmacological treatments to treat circadian rhythm disorders such as insomnia, fatigue, and jet lag, as well as to manipulate circadian rhythms to achieve periods of extended wakefulness and alertness.

Mobility is a sensitive indicator of overall health in older adults. Objective mobility data can provide insight into patterns of function including sleep, behavior, and activity, which is valuable information for both individuals and their healthcare providers. By visualizing objective mobility data, providers can assess for trends that could indicate a shift in overall health in order to intervene earlier, whereas older individuals with mobility goals may benefit from feedback in order to better meet goals or sustain behavior change. The purpose of this project is to perform a secondary data analysis and determine methods for visualizing objective mobility data for both individual users and healthcare providers. Participants of the original study were older adults with a recent history of mild traumatic brain injury. Data was collected with an active sensor system requiring participants to wear a sensor device during waking hours. Working through an interface designed by the University of Washington Ubicomp Lab I aim to create a composite profile of a single participants mobility and movement within their life-space. I am using Excel to sort and create visual representations of data. Key factors to include in visualizing these data are total amounts of active versus passive time spent per participant and the participant’s mobility within his or her life-space. Data analysis is in process and prototypes for visualization are under development. With any visualization effort, it is important to address the different stakeholders’ needs and expectations. Visualizing mobility data for individuals to better understand their own activity levels requires an approach that may be different from visualization targeting clinicians to support clinical decision making. Differences include preferences for granularity of data, level of abstraction, and annotation of context specific details that complement the visualization data. This study demonstrates that visualization can maximize the utility of health monitoring data.

The 1960s was an era of political and social transformation in America. Seattle, as most people do not know was a segregated city for most of its history. Racial discrimination applied not to just African Americans but also Asians, Chicanos, etc. People of color could not attend certain schools, restaurants, and could not live outside certain city boundaries. The Northwest African American Museum preserves local artifacts from the Civil rights movement in Seattle. Maid Adams, a Seattle C.O.R.E member during the 1960s donated two protest signs used during the movement. These posters were extremely crucial in the movement. They represent how protesting instigated social change and paved the way for future activism. 

Next generation DNA sequencing, with high throughput and low costs, has already greatly benefitted life sciences and personalized medicine. Nanopore sequencing is a new single molecule technique with potential to further revolutionize DNA sequencing. In this technique DNA is electrophoretically drawn through the nanopore, an integrated membrane channel protein, while an ionic current is recorded. We use the nanopore Mycobacterium smegmatis porin A, MspA, a porin engineered to provide a robust nanopore with the necessary features for nanopore sequencing. To control DNA translocation, we use a phi29 DNA polymerase to pull single stranded DNA through MspA in single nucleotide steps. For each step, the recorded current can be mapped the four standard bases, A, C, G, and T, present in the pore’s narrow constriction. Additionally, the modified nucleobases associated with suppressed gene expression and a number of diseases, 5-methylcytosine and 5-hydroxymethylcytosine, may be directly detected with MspA. We compare two mutants of MspA and examine how different amino groups within MspA’s constriction affect the nucleotide-specific current. We find that additional modifications to MspA may be used to tailor the pore for specific sequencing applications, such as increased accuracy in detecting and, other modified DNA detection.

This research examines the cultural climate that existed in Seattle’s Central District and on Jackson St. in the 1940’s-1960’s and the music produced at the time, jazz music. I examine Seattle’s changing laws (like racial restrictive housing covenants and prohibition), newer freedoms in the Black community (like desegregation), the influence of the Great Migration, and changing national attitudes toward racism and equality. I parallel those changes with evolutions in the styles of jazz produced throughout this period, and the social, political and economic landscape becomes evident in the music, to the extent that the way jazz sounded at a certain time reflected the social, political and economic atmosphere. I look at these issues through the lens of Floyd Standifer, a well-known Seattle trumpet player, who began his career in the 1940’s and was active in the Seattle jazz community until his death in 2007. Standifer witnessed some of the most radical changes in the socio-political landscape of Seattle throughout the ‘40’s through ‘60’s that I examine. The malleability he demonstrated in his music is testament to the changing social climate, population increases in the Black community, new freedoms, and the changing desires of community members and music venue patrons. In the shift from “rags” to jazz to rock ‘n’ roll, I demonstrate how jazz can be seen as political and reflects how music lies at the heart of the African American resistance to oppression during this period.

A stepper system is a device that uses multiple small-scale steps to achieve large ranges of motion. There are several fields where stepper systems can be used as positioners such as micro/nano-fabrication, scanning probe microscopy, and alignment of optical components. This stepper design enables a motion stage, whose motion is generated by friction induced with piezoelectric actuators underneath, to move horizontally without vertical displacement. The objective of this research is to investigate a way to support the load of a motion stage while limiting any restriction of motion in the horizontal direction. This restriction of vertical load bearing and horizontal freedom has led to investigating bearings as a component of the support structure. There are many types of bearing designs (i.e. ball, magnetic, and air) that may satisfy the design constraints, so I have performed analysis on various types of bearings and investigated their impact on the design of a new stepper system. For instance, ball bearings support radial loads or thrust loads with forces normal to the ring pathways, but continuous compression, along with expansion, leads to fatigue of the rings, which shortens their lifetime; while magnetic bearings provide support with a distance force that is controlled by the current flow in the circuit, maintaining a more stable and friction-free motion. However, applications may be sensitive to magnetic interference and therefore magnetic bearings may not be suitable for use in some cases. Taking into consideration the benefits and limitations of the different bearing systems, my research has proposed and introduced the design of a stepper support system.

Quantum dots (QDs) are the precursor for numerous burgeoning technologies, including more efficient solar panels and high definition LED displays. QDs are small, soluble crystallites of a semiconductor material that have unique qualities since their electrons are confined in all three dimensions in a region of space smaller than the de Broglie wavelength of their electrons. This allows the electrons to occupy spectrally sharp energy levels like an element or a molecule. Their electronic properties can be tuned as a function of their size. In our lab, indium phosphide QDs are prepared under an inert atmosphere of dinitrogen by hot injection of a phosphine precursor into an indium salt and fatty acid solution. Taking aliquots of the reaction mixture at different time intervals during QD growth allows us to chart the rate of particle evolution and provides information about particle size and uniformity. Indium phosphide is a member of the III-V semiconductor class, meaning they are made from elements from column three and column five of the periodic table, and have desirable properties like direct band gaps which allow these materials to efficiently absorb and emit light as well as faster carrier mobility than II-VI quantum dots. The synthesis of III-V QDs has not allowed for size- uniformity or tunability to be achieved because of the uncontrolled reactivity of the precursors, in particular the group V precursors. Our group has recently uncovered an important acid-catalyzed pathway leading to phosphine speciation during indium phosphide QD growth. My project focuses on addressing this issue by incorporating new, more stable precursors and chelating capping agents to better stabilize the reaction products and intermediates, which would be crucial in controlling the formation and therefore the properties of the resulting quantum dots.

Alcohol use consistently correlates with negative physical and psychological consequences like dependence, memory loss, impaired judgment, etc. Studies also link the use of sleep medications with similar negative consequences. Yet, no study examines the interaction of sleep medication and alcohol use onto these negative consequences. Therefore the present study investigated the relationship between peak blood alcohol content (BAC) in the past 6 months, negative consequences of alcohol use in the last 12 months as measured by the Rutgers Alcohol Problem Index, and lifetime, 3- and 12-month non-prescribed sleep medication use in Washington state and Swedish high school seniors participating in a larger study (N = 3,352, 56.6% Female). Regression analyses revealed a significant main effect of BAC and lifetime use of sleep medication onto negative consequences (β = 18.69, F (3, 2875) = 115.86, p < .05) such that individuals with a higher BAC and who had a lifetime incidence of sleep medication usage experienced significantly greater negative consequences than those who only drank or used sleep medications. There was no significant moderating effect of frequency of sleep medication use in the last 3 or 12 months onto BAC and negative consequences (p >.05). While these results suggest that use of sleep medication and alcohol interact to contribute to increased risk for negative consequences, there are limitations. Lifetime use of sleep medication may reflect a tendency to act impulsively or self-medicate (e.g. for sleep/depression/anxiety), not a direct effect of the medication per se. Additionally, analyses did not control for other drug usage or mood disturbances, and all sleep medication drugs were grouped under one survey item. Given our observations, additional research testing combined effect of sleep mediation and alcohol use as related to negative consequences is warranted.

There are many challenges to creating high performance diagnostic tests for use in low resource settings. One approach is to use a paper network format. These paper networks operate by capillary flow and do not require dedicated instrumentation for transporting fluids in them. However, in order to carry out more advanced, multi-step, processes in these networks there is a need for well-characterized paper fluidics tools to precisely control fluid transport within the networks. To address this need, we have designed and characterized a cellulose valving mechanism to produce precise fluid delays, over a wide range of delay times, in both a bench-top system and a disposable card format with less than 10% variability in reproducibility. We achieved this by systematically investigating the effects of valve parameters such as valve dimensions and materials, on the delay time and reproducibility. Finally, the cellulose valves are demonstrated in a paper network card that can automatically sequentially deliver a series of fluids.

Impulsivity is often found to be related to alcohol use (Pearson and Henson, 2012; Simons, 2003). Additionally, a correlate of alcohol expectancies is increased alcohol use (Hull and Bond, 1986). However, no research to date has looked directly into the association of impulsivity, alcohol expectancies, and alcohol use. Therefore, the present study looks into impulsivity, alcohol use, and alcohol expectancies in high school seniors (N = 3,352, 56.6% Female) from Washington and Sweden. Participants completed a baseline online survey as part of a larger trial that assessed peak blood alcohol content (BAC) in the past 12 months, impulsivity as measured by the sensation seeking scale (Zuckerman, 1964), and alcohol expectancies as measured by the brief Comprehensive Effects of Alcohol questionnaire which consists of positive (sociability, tension-reduction, liquid courage, and sexuality) and negative (cognitive/behavioral impairment, risk/aggression, and self-perception) classifications (CEOA; Fromme, 1993). Regression analyses revealed a significant main effect of impulsivity, cognitive/behavioral impairment, sociability, risk/aggression, and self-perception onto peak BAC (p < .05). However, significant interactions were only found for impulsivity onto risk/aggression and self-perception. A significant moderating effect of risk/aggression onto the relationship between impulsivity and peak BAC (β = .002, F (3, 2913) = 114.71, p < .05) was found such that those with lower impulsivity and lower risk aggression scores (alcohol seen positively) have higher peak BAC. A significant moderating effect of self-perception onto the relationship between impulsivity and peak BAC was also found (β = .001, F (3, 2908) = 132.04, p < .05) suggesting that individuals with lower self-perception scores (alcohol seen positively) and higher impulsivity have higher peak BAC. These results suggest that individuals who are both highly impulsive and have higher positive expectancies for alcohol drink more. Future research should continue to investigate the relationship between alcohol expectancies and impulsivity in vivo.

Select proteins from the HSP70 gene family were studied in Anthopleura artemisia and Anthopleura elegantissima, two intertidal sea anemones found in Puget Sound. A. elegantissima are clonal aggregates that are found on rocks while A. artemisia live buried, solo within sand. HSP70 are a family of heat shock proteins that are present in prokaryotes and eukaryotes that regulate various protein functions such as heat stress or protein folding. PCR techniques were utilized to determine the presence of HSP70 genes. It is predicted that both species have similar or identical genes that produce HSP70 heat shock proteins, but differ in how the HSP70 genes are expressed. In prior studies, A. elegantissima are found to express HSP70 proteins depending on position in a colony (edge versus central positions). A. artemisia, allows for an interesting comparison as to how living conditions would affect the expressions of HSP70 proteins. RT-PCR techniques were utilized to determine HSP70 gene expression in both species. Variations of HSP70 expression can be used to generate models to predict the effect of climate change in the Puget Sound area.

MicroRNAs are a class of post-transcriptional regulators that silence messenger RNAs. MicroRNAs are interesting molecules to study because they are stable at room temperature and are long lived unlike messenger RNA. These characteristics make them useful molecular biomarkers to monitor disease and exposure status. For example, urine microRNAs have been used clinically as potential diagnostic markers of bladder cancer. However, their utility in non-clinical settings has yet to be elucidated. The goal of this study is to compare total RNA isolated from fresh and archived urine field samples to investigate their quality and quantity over time. RNA will be isolated from both whole urine and urine sediments and will be analyzed using multiple techniques. To date we have optimized the methodology required to isolate total RNA from the whole urine sample and the sediment. In a preliminary study, we isolated total RNA from samples under various conditions (fresh vs. field; whole urine vs. sediment) and compared them using the Nanodrop and Bioanalyzer. Our results demonstrate that RNA can be isolated from both fractions under both conditions. The molecular profiling of the RNA suggests that the urine samples contain only small RNAs because they lack ribosomal RNA peaks. Interestingly, the profiles look similar for all urine samples examined. In light of this, we performed an additional microarray experiment to characterize and identify the small RNAs present in the whole urine and sediment samples. These results are currently in progress. Using the information obtained from the preliminary microarray study, we will select the 3 most abundant microRNAs present in the urine for further analysis using RT PCR techniques to determine whether archival duration has an effect on their stability. The results of this experiment will provide valuable insight on the utility of archived field samples for the future development of urine biomarkers.

Exposure to environmental stress can impact health and affect well-being. Specific populations, such as low-income and minority women, may experience increased exposure to environmental stress and be at a higher risk of related health impacts. In order to better characterize susceptibility to stress and the related health effects, it is necessary to quantify the exposure. Exposure to stress triggers the release of the hormone cortisol, allowing it to be used as a biomarker. The objective of this study was to evaluate acute and chronic salivary cortisol levels within- and between-individuals and communities specifically characterizing the stress responses within a Hispanic agricultural community and a low-income non-Hispanic urban community. Saliva samples were taken from 27 Hispanic women living in an agricultural setting and 28 non-Hispanic women living in an urban environment. To measure short-term variability within- and between-individuals, four saliva samples were collected five times per day for two consecutive days. To measure long-term variability, a second set of samples was collected ten weeks later. Samples were collected by placing an oral swab in the mouth for one minute. Cortisol concentrations were quantified using a cortisol enzyme-linked immunosorbent assay (ELISA). We evaluated daily average salivary cortisol concentrations and diurnal indexes from samples taken consecutively and ten weeks apart to characterize acute and chronic within-individual variability. Similar evaluations were made within the agricultural and urban communities to better understand between-individual variability. On average, we found evidence of diurnal variation in both the agricultural and urban communities. Within individual variability was substantial in both communities as well, both on the short and long-term. These analyses support evidence that in order to evaluate differences in environmental stress exposure between as well as within individuals, robust biomonitoring methods are needed to fully characterize different levels of cortisol.

Nicotinamide adenine dinucleotide (NAD) is used in many biological processes such as metabolism and cellular signaling. Recently NAD is found to have an important role in many signaling pathways. As NADs role in cellular pathways is being discovered, it is important to be able to detect and quantify the amount of NAD in vivo and in vitro by nuclear magnetic resonance (NMR) analysis. NAD is found in cells at low concentrations and its peak intensity in a phosphorus spectrum is much smaller than that of other phosphorus containing molecules like ATP. By decoupling and Insensitive Nuclei Enhanced by Polarization Transfer (INEPT) experiments we hope to isolate and increase the signal of the characteristic phosphorus nuclei found in NAD molecules. Isolating and increasing the signal to noise of the NAD peak will prove that we have the right peak and allow us to quantify the amount of NAD in vivo. This will provide information about health risks of a subject.

Throughout their lives, children in the United States public education system are bombarded with messages of equality and the promise of the American dream (i.e., no matter who you are, you can succeed through merit and hard work). Yet, the field of science is still dominated by White men in both number and power. Young girls of color are thus faced with two conflicting narratives: "You can achieve anything you set your mind to," and, "There are few people like you in the field of science." My research asks how middle school girls of color process this dichotomy. Based on my past experience as a female minority pursuing a career in science, it is my hypothesis that these girls will consciously believe and express the first narrative, while internalizing and acting according to the latter. One of my research methods is the Draw-a-Scientist Test, a tool used to gauge a student's perceptions of scientists by asking her to make a drawing of one. I expect the girls to express their belief that anyone can become a scientist, while drawing stereotypical images of scientists (i.e., people unlike themselves). I will also analyze and look for themes in observations, recorded interviews, and interactions of girls participating in an after-school program called Chemical Oceanography After School Time (COAST). I believe that as the girls gain firsthand experience with science in the COAST program, they will begin to interact in ways that show growing confidence in their own potential to become scientists. These findings indicate a need for more science intervention programs, like COAST, for girls of color. My research aims to open up dialogue about how the prevailing ideologies and realities of race and gender in the United States impact the potential in science for girls of color.

The ZaP Flow Z-Pinch Project researches the stabilizing effects of sheared axial flow on Z-pinch plasmas. Stable plasmas are necessary to harness nuclear fusion. Nuclear fusion is an essentially limitless source of power, but the difficulty of creating long-lived plasma stability has hindered the development of controlled nuclear fusion. The ZaP experiment forms a Z-pinch plasma by ionizing hydrogen gas into plasma and dynamically forming a 100 cm long column approximately 1 cm in radius. Z-pinches are extremely susceptible to instabilities, which tend to disrupt the plasma on the order of 10-8 s. With a sheared flow, the ZaP experiment has observed a Z-pinch plasma lifetime of 10-5 s. These successful research results have led to a new experiment, ZaP-HD, the goal of which is to produce and observe high energy-density plasma pinches. This new experiment will be modeled with MACH2, a two-dimensional magnetohydrodynamic equation-based simulation code. The design that is modeled will use a similar design to the current ZaP Flow Z-Pinch experiment, however it will have significant modifications to its geometry and operating conditions. These will be reflected in the simulation by changes to the code’s input file, which consists of a matrix of points to form the geometry and parameters that determine the running conditions. Two major design changes include the addition of a third electrode and a second power supply. These modifications allow for independent control of the motion of the plasma through the two phases of the experiment. The simulation data collected will provide the team with a complex and detailed view of how this new experiment, ZaP-HD, will operate as well as any changes that need to be made to the planned design.

UVC is ultraviolet light in the wavelengths 280 – 100nm. UVC’s germicidal ability has been employed to prevent forward contamination, or the contamination of a pure planetary body with terrestrial organisms. Scientists have used UVC to sterilize equipment on land and water, but the effective dose of UV needed to prevent contamination when probing a pure ice environment is still unknown. In order to safely explore pure icy environments, data of bacterial response to UVC in ice are needed. To create these data, we first irradiated Escherichia coli in water as there is a known decimation curve for this experiment, which serves to standardize our irradiation set up and techniques. After matching the decimation curve of Escherichia coli of previous UV sterilization experiments, we will then proceed to irradiate bacteria in ice. In addition to irradiating commonly tested bacteria such as Escherichia coli and Bacillus subtilus, we will also irradiate samples of glacial isolates that would be more likely contaminants in an icy environment. Each sample of bacteria will be assessed before and after radiation using the Most Probable Number technique, or MPN. The MPN method uses serial dilutions of a sample bacterial culture until no colony forming bacteria are present to determine the concentration of bacteria present in the original sample. From our irradiations, we will create curves of UV dose versus survival of bacteria that will allow future equipment used in the exploration of pure icy environments to employ the correct dosage of UVC light to minimize forward contamination.

Heart disease and stroke are the first and third leading causes of death for men and women in the U.S. and together account for more than a third of all U.S. deaths. It has been shown there is a correlation between decreased intercellular forces in the endothelial cells in the artery and higher permeability of the arterial lining. This can lead to atherosclerosis, a disease defined by hardening of the arteries, a major underlying cause of heart attacks and strokes. The focus of this research was to determine how different blood flow rates affect the intercellular forces of the endothelial cells that line the arterial wall. The first step was to fabricate microposts. Negative molds of the posts were cast off of a master array of microposts for which the spacing, height and diameter of the microposts were known. A positive mold was then cast off of the negative mold to produce a substrate on which the cells could be cultured. Next a flow chamber was designed for these experiments. Human Pulmonary Artery Endothelial Cells (HPAECs) were then cultured and transferred onto the microposts which were stamped with fibronectin, a protein that would help the cells bind to the tips of the microposts. The stamping portion of the experiment required much trouble shooting and ultimately the experiments were halted here. If the research had continued the experiments would have run for 14 hours, after which the cells would be fixed in place, stained with a fluorescent dye and imaged using microscopy. These images would then be transferred to a custom written Matlab code that would calculate the deflection of the microposts and the forces induced by cells. This data could then be used to determine the effect of varying blood flow rates on the permeability of the arterial wall.

Mogadishu, the capital of Somalia, is overcoming over twenty years of violence, political chaos, and lack of economic growth. Now, for the first time in twenty years, with a recognized government and renewed international investment, Mogadishu has a chance to escape the cyclical patterns of poverty and hunger. However, this cannot be simply done through a top to bottom format of political changes; the political inequalities ingrained in the population need to be alleviated. The people who are most vulnerable, women, particularly young women, and minorities have to be given economic opportunities. I went to Mogadishu to find out more about the economic situation of the city, particularly, the economic situation of young women in Mogadishu through observation and conversations with this population. I visited busy market places and very small kiosks. I spoke with women who graduated from Universities and women who were illiterate about their situation and perspective of Mogadishu's economy. I found out that the Mogadishu economy is alive, changing rapidly, and there are many people who are living decent lives. However, there are many more women who are poor, illiterate or with little education, and/or come from tribes that are politically marginalized. If these women, and others in similar economic situation, are not systematically given beneficial economic opportunities, the cycle of poverty, violence, and disease will continue.

Different groups of smokers are commonly discussed in the research literature on tobacco use and nicotine dependence. Examples include heavy smokers, light smokers, intermittent smokers or social smokers. While forming such groupings may be scientifically useful, there are not standardized definitions for group membership, leading to a lack of clarity and comparability across studies. In tobacco research, groups are commonly formed via cutoff scores. That is, if one is below a certain score, one is in group A, otherwise one is in group B. However the lack of clear group definitions has lead to different researchers using different cutoff scores. For example, one researcher may use smoking five or fewer cigarettes a day as a cutoff to define the “light smoker” group, where another researcher may use smoking ten or fewer cigarettes a day to define the same group. A further potential weakness of the use of cutoff scores is that they enforce absolute distinctions and thereby ignore within group variability as well as measurement error. Rather than using cutoff scores to define groups, we can use statistical models, such as latent class analysis (LCA), to empirically group observations. LCA is a multivariate statistical model often used for exploratory analysis, It identifies homogeneous subgroups (or classes) in a population on the basis of item associations. The purpose of this study is to compare group differences between groups formed via cutoff scores with groups from LCA. The question of interest is if results in these analyses differ or parallel each other. Specifically, we examined the differences in the associations between group membership and relevant demographic covariates as well as measures of nicotine dependence.

Porous membrane-based lateral flow assays have become extremely popular for their inexpensive and user-friendly diagnosis of many conditions. However, commercial lateral flow assays typically only run single-step processes that may lead to inaccurate results for low concentration analytes. Our goal is to create more sophisticated assays that can perform automated multi-step processes to achieve higher sensitivity detection. In order to achieve this goal, porous fluidic valves for the precise regulation of fluid flow in these devices are needed. We have developed a simple shut-off valve that passes a well-defined volume of fluid before permanent shut-off. These dissolvable bridges offer a way to automate the metering of reagent volumes that would otherwise require the user to pipet exact volumes into the device. We have characterized the operation of the valves and demonstrated their tunability using parameters such as geometry and composition of the dissolvable bridge. In addition, we have demonstrated the utility of dissolvable bridges in the important context of automated delivery of multiple volumes from a common source to different locations in an assay for simple device loading and activation. Dissolvable bridges have the potential to help bring advanced testing using porous membrane networks to limited-resource settings.

Multi-Legged terrestrial biological systems demonstrate complex forms of locomotion by utilizing different gait patterns and leg motions for different environmental conditions. Typically, the footpath of biological systems consists of a smooth flat region when the foot is in contact with the ground where the forward motion of the body occurs. The following research focuses on designing a mechanical linkage-based leg mechanism to effectively mimic the step of a biological system. A simulation of the linkage was performed using MATLAB in order to analyze the dynamics of the foot path and duration of stepping phase that the foot was in contact with the ground. After analyzing the foot motion, the linkage-based leg designs were then constructed using the LEGO Mindstorms NXT robotics kit and tested with various walking patterns. Further analysis of the linkage-based leg motion was done by comparing the resulting motion to biological leg motion; successful linkage-based leg designs should display similar stepping characteristics to the biological motion.

Chitin is the second most abundant polysaccharide extracted from the cuticles of insects and crustaceans. Chitin is known for its biocompatibility and superior mechanical strength. Silk fibroin is produced by the Bombyx mori silkworm and is known for its biocompatibility and superior toughness. Composites of chitin and silk fibroin would possess each biomaterial's unmatched mechanical strength and toughness, making them ideal candidates for biomedical applications requiring such mechanical properties. Here, squid pen β-chitin and Bombyx mori silkworm cocoon silk were blended in Hexafluoroisopropanol (HFIP) solution in specified ratios to synthesize the nanocomposites. As a result, chitin/silk hybrid nanocomposite thin films were produced to test the mechanical properties. Studies of both X-Ray Diffraction (XRD) and Fourier Transform-Infrared (FTIR) Spectroscopy revealed a successful effective interaction between the chitin nanofibers and the silk proteins matrix, which was most likely caused by hydrogen bonding. Results of the self-assembled chitin nanofibers embedded in the silk matrix showed a noticeable enhancement in mechanical properties. The tensile strength, flexibility, and toughness of the biomaterial nanocomposites were enhanced compared to just pure chitin or pure silk fibroin. Due to the improvement in mechanical properties, these chitin/silk engineered nanocomposites can be utilized in biomedical applications such as hybrid scaffolds for bone tissue engineering or microneedles to serve as a new methodology for drug delivery systems.

The objective of this research is to produce bio-oils from biomass at high temperature in water or ethanol at different pressures. The high pressures and temperatures takes the solvents near or into supercritical conditions where small changes in pressure have great effects on the solvents' physiochemical properties. Conditions are tailored to increase the yield of bio-oil that can be hydrogenated and then refined in existing petroleum refineries to produce high-quality level jet fuel. Char, coke, and tar - unwanted byproducts - must be decreased to a minimum as they deactivate catalysts and clog reactors. These unwanted byproducts are a ubiquitous problem in biomass themro-chemical conversion; removing them would help the industry develop. It is possible to get no char, coke and tar produced, as I have been able to attain, though a better understanding of how to reproduce it consistently and at larger scales is necessary. Temperature are adjusted within the sand-bath - apparatus for heat transfer- and to increase pressure more reactants are placed into the chamber to which the pressure can be calculated.  

Individual bees often show flower constancy, specializing on a floral resource while avoiding switches to newly available resources. The familiar plant species benefits by receiving high rates of visitation even when competing floral resources become available. However, the phenology of some plants is shifting more rapidly than others in response to climate change, resulting in a changed sequence of floral resources bees encounter. To determine whether this affects plant fitness, we asked how pollinator visitation to competing plant species depends on the order in which those species bloom. We used two plant species with overlapping phenology and a common pollinator, Hypochaeris radicata and Campanula rotundifolia. We allowed Bombus impatiens to forage in floral arrays representing three phenological progressions: (1) CR Early, with C. rotundifolia decreasing from 100% to 0% while H. radicata gradually increased from 0% to 100%; (2) HR Early, with H. radicata decreasing from 100% to 0% while C. rotundifolia gradually increased from 0% to 100%; and (3) a Control with both species at 50% throughout. We quantified visitation rate to both species, and seed production by H. radicata. Overall, the number of bee visits received by both species was significantly higher when that species was introduced first than when it was introduced after its competitor. In treatments where the proportion of the early species decreased over time, the visitation rate per flower to that species increased, for both species and across all stages of experimental treatments. Seed set for H. radicata showed a similar pattern to visitation. Our results corroborate other work suggesting that individual bees learn and become specialized on a single floral resource and are slow to switch to a newly emerging resource. As a result, the flower species that blooms first can benefit from higher bee visitation and thus, potentially higher reproductive success.

Within the last 20 years, global climate change has become well accepted. Research on plant dynamics has documented species shifting their ranges to higher elevations in order to remain within physiologically optimal conditions. This general observation has motivated my investigation of elevation as a driver of life history traits within alpine and subalpine plants. Specifically I’m interested in sex-ratios among the subalpine flower Erythronium montanum. This species of flower has been shown to exhibit a unique trait known as androdioecy, wherein some individuals produce male-only flowers instead of hermaphroditic flowers. In other species this trait has been attributed to sexual dimorphism, the variability between the sexes in terms of morphology and/or phenology. The discovery of this trait in E. montanum has inspired my research project with two distinct goals: To determine whether 1) abiotic factors such as elevation, soil moisture and time of flowering and 2) biotic factors such as canopy cover, and flowering density, influence sex-ratios within E. montanum. Surveys done at Mt. Rainier in 2011 and 2012 by the Hille Ris Lambers lab provide data on multiple factors which should allow me to test a number of possible hypotheses. Preliminary data analysis spanning both elevation and flowering time, show biased sex-ratios where the number of males to hermaphrodites decreases at higher elevations, late in the season. A future look at the extent of sexual dimorphism within the population can help to shed light on the mechanism behind sex-ratio bias and increase our understanding of plant reproductive-ecology over an altitudinal gradient. As many species begin a slow but steady march up the mountain, it’s paramount that we have a better understanding of how and why life-history traits fundamental to reproduction (e.g. sexual dimorphism) vary with elevation. Ultimately, future generations and plant communities rely on what is happening today.

Climate change is likely to affect the altitudinal range limits of conifers in the Pacific Northwest as population performance declines at lower elevations and populations expand into upper elevations. However, it is possible that populations are locally adapted, thus influencing range shift dynamics. To assess the potential for local adaptation for three conifer species, Abies amabilis, Tsuga heterophylla, and Tsuga mertensiana, we monitored bud set and bud dormancy in seedlings collected from two to three different source elevations at Mt. Rainier National Park. Phenotypic differences in bud set and bud dormancy influence the length of the growing season, and thus, have impacts on fitness. However, source elevations from which we collected seeds differ strongly in climate, meaning that the optimal climatic triggers for bud set and dormancy are likely to differ. Observations on apical (terminal) bud formation were taken for one year in a common garden at the UW Greenhouse, to correct for environmental differences. Our study thus provides a glimpse at whether or not populations of one species in different locations on Mount Rainier are producing seeds with similar or different phenological traits when observed in the same climate. If seeds of the same species from different locations have inherent differences in the timing of key ecological events, a species may experience difficulty shifting its range when between-population interactions from different elevations become necessary because of climate. Dormancy and bud set differences at the seedling level could influence how conifer population dynamics on Mount Rainier and in other places around the Pacific Northwest will change. In preliminary analyses, we found that seedlings differ in budset and dormancy by species, and there is a trend for differences by elevation within species.

The nematode Caenorhabditis elegans is a powerful model for the analysis of the conserved mechanisms that modulate healthy aging, from organismal to molecular levels. As animals age, tissues deteriorate at different rates. It has been shown previously in the Kaeberlein lab that deletion of the hypoxia-inducible factor, HIF-1, extended lifespan of worms at high temperature (25°C), but did not do so at low temperature (15°C). Accordingly, it was speculated that at low temperature, worms were not as healthy. At low temperature, we noticed an abnormal vulva phenotype and called it age-related vulva integrity defect (Avid). As worms age, their vulvae protrude and rupture, causing internal organs such as the intestine to erupt out of their bodies. Through investigation, we realized that animals lacking oocytes tend to show more Avid phenotype, compared with the animals that produce enough oocyte. It has been shown before that at 15°C worms produce around 300 fertilized and unfertilized oocytes, while at 25°C this number increases to 400. My preliminary results suggest that germline is involved in the progress of Avid. I studied animals with germline defects and observed 100% Avid phenotype in germless mutants. Avid is a sign of healthspan as animals age, and decline in germline fecundity due to aging can induce Avid. Healthspan is defined as the period of life spent in relatively good health, and maintenance of healthspan is an important component of aging. Our data indicate that Avid can be used as a measurement tool of the age-related healthspan of worms.

Over the past year, I have conducted ethnographic and oral history research here in the Puget Sound-area to understand the role of urban parks and organic farming in the development of a bioregional concept of sustainability. Throughout the research process I have maintained a focus on the study of sense of place, as this is one important aspect of bioregional ethics and design principles. This study was completed using collaborative ethnographic methods including participant observation, talking circles, surveys, and interviews with forest stewards at Seattle’s Carkeek Park, and farmers at Viva Farms in Skagit Valley. Through this I discovered the role of autotopography, the expression of self through place shaping practices, a process that contributes to a developing sense of place and is often overlooked. Some members of the Carkeek Park forest stewards, or WEWO as they call themselves, and some of the farmers used the act of shaping landscapes to express themselves in ways unique to the individual, and my data supports that this act was, and is, important to their developing a sense of place.

Stress can have a major impact on the cognitive functions of the brain in both humans and animals. One of the possible areas affected is the hippocampus, a structure known to be important in consolidation of short and long-term declarative memory and spatial memory. Previous studies have shown that stress can reduce long-term potentiation, impair spatial memory, and affect the firing rate of location-specific place cells in rats. This study aims to investigate the effects of temporary hippocampus impairment during stress on risky foraging behaviors in rats using APV, an NMDA receptor antagonist that inhibits the synaptic plasticity of local neurons at the site of injection. Rats were trained to retrieve food pellets from a simulated foraging environment and return to a safe nest, and APV infusions were then made via implanted cannulas into the rats’ hippocampus 30 min prior to stress. Foraging behavior in the presence of a robot predator was assessed in comparison to control rats infused with artificial cerebrospinal fluid. Hippocampal dysfunction is hypothesized to contribute to faulty contextual fear conditioning and memory formation, thus rats with an impaired hippocampus may exhibit excessive, contextually-inappropriate fear behavior in the presence of an artificial predator. The results of this study may help us understand the affects of stress and its role in the behaviors of individuals with post-traumatic stress disorder.

The late Oligocene-early Miocene fossil record (30-18.8 million years ago) of North America includes an abundant fauna of entoptychine gophers (Rodentia: Geomyidae: Entoptychinae). These burrowers make up as much as 20% of the mammalian individuals of the faunas they are part of, and can be involved in habitat partitioning or competition with other burrowing herbivores. The wide geographic range of these animals and their use in constraining the age of the fossil assemblages of the Cabbage Patch beds (western Montana) and John Day Formation (eastern Oregon) makes them critical to understanding the biogeographic relationships among these fossil faunas. To understand these spatiotemporal patterns, I am evaluating the taxonomic richness of entoptychine gophers. Previous studies have discriminated among these gopher species by using morphological features of skulls and dentaries. However, anatomical elements are rarely preserved in fossil assemblages, and isolated teeth are commonly the only material available for identification. Since the occlusal surfaces of the cheek teeth are usually worn down as a consequence of feeding, they lack discrete features for species identification. Instead, some researchers have hypothesized that the more continuous variation in the enamel outline of select cheek teeth could be used for discrimination. Here, I use semi landmark-based geometric morphometrics (an arithmetical comparison of geometric coordinates) on the upper last premolar and molar. Applying this method to 98 modern geomyine gopher specimens (Geomyidae: Geomyinae), representing 10 taxa, helps quantify intra- and interspecific tooth shape variation and offers guidelines for recognizing fossil gopher species. Preliminary analyses indicate that tooth shape is species-specific in modern gophers, supporting the use of this method to differentiate fossil gopher taxa. Applied to 96 specimens of fossil gophers, I found a strong overlap of fossil taxa in the morphospace, suggesting that previous researchers may have overestimated the species diversity of some fossil gophers.

Motivated by current interest in one-dimensional materials for their ideality and potential in the next generation of electronics, a device consisting of a microscopic four-point probe was developed with electron beam lithography (EBL) to investigate the conductance and resistivity of manganese-doped copper oxide (Mn:CuO) nanowires. Mn:CuO nanowires were chosen because they are known to be ferromagnetic and conductive below a critical temperature of 80K, while maintaining semiconductor properties and low resistance at room temperature. Nanowire samples were obtained from the Yang Group in Nanjing, China, and were deposited with ethanol onto silicon dioxide wafers to obtain a rough monolayer for easy isolation of individual nanowires. A pattern consisting of four contact lines was created with DesignCAD and conditions were optimized to obtain an adequate dose (charge/unit area). A subsequent process of precise alignment was performed to ensure complete overlap between the nanowire and pattern on a nanometer scale. With accurate alignment, the pattern was then etched onto a single nanowire, developed, and coated in a 50 nm thin film of gold to enable conduction between the nanowire and the four-point pattern. From this point, the current and voltage can be measured through the nanowire macroscopically. Consequently, the anisotropic resistivity of the material as well as gate voltage effects can be studied to verify the material’s applicability to future spintronics devices such as field-effect transistors.

In adults, acute myeloid leukemia (AML) generally remains difficult-to-treat, and most patients are currently expected to die either of their disease or treatment-related toxicities. The need for novel therapies is thus unquestioned. With increasing availability of specific small molecule inhibitors, one line of research has focused on targeting signaling pathways that are pivotal for the in vivo growth, survival, and propagation of AML cells as a means to improve therapeutic outcomes in this disease. Emerging evidence suggests that the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway, a key physiologic regulator of transcription, translation, cell cycle progression, differentiation, metabolism, and suppression of apoptosis, could provide a rational drug target in AML. Constitutive activation of AKT signaling is indeed observed in many patients with AML, and several groups have reported inferior relapse-free and overall survival in AML cases with AKT activation. Given these data, we recently conducted an early phase clinical trial testing MK-2206, a highly specific allosteric AKT inhibitor, in patients with relapsed AML. However, MK-2206 has insufficient anti-AML activity at tolerated doses. Therefore, the goal of my project in the preclinical studies that I am currently carrying out is to identify signaling pathways that are critically important for AML cell survival in the presence of MK-2206 with the goal of developing combination therapies that optimize chemosensitization that can be achieved with AKT inhibitors. This is being done by mixing the MK-2206 inhibitor with various conventional chemotherapic drugs and measuring their toxicity on different AML cell lines by use of flow cytometry.

Diatoms are major players in Earth’s biogeochemical cycling, accounting for approximately 20% of global primary production. Their dominance in several aquatic ecosystems is a testament to adaptations that provide an advantage in nutrient acquisition and growth. Ocean fertilization experiments have demonstrated that iron is the primary limiting nutrient in many open ocean ecosystems. This metal is critical to the functioning of numerous important proteins, particularly in photoautotrophs such as diatoms. An excess of intracellular iron, however, can cause oxidative damage to the cell. This project investigates genes related to iron metabolism in the recently sequenced transcriptomes of 21 diatoms and select outgroup species. Of particular interest are genes coding for ferritin subunits. Ferritin is a multimeric protein that serves as an iron storage unit, safely sequestering iron atoms in an oxidized state and releasing them back into the cytoplasm when iron supply is low. Ferritin in diatoms was previously thought to exist only in pennate diatoms. Here we show that ferritin-coding genes are expressed in members of every major diatom group. 13 out of 21 diatom transcriptomes contain full-length sequences, forming a clear monophyletic clade most closely related to cyanobacterial ferritins. Additionally, 7 of the 21 transcriptomes surveyed reveal two distinct ferritin genes, possibly due to an ancient duplication event. The presence of ferritin helps to explain the success of some diatom species in iron-poor waters, and our expanded understanding of which diatoms possess ferritin genes will help to predict the responses of this important group to a dynamic ocean environment.

Observations suggest that the intracluster medium in galaxy clusters is strongly influenced by supermassive black holes residing in cluster centers. We investigate the consequences a black hole can have on the thermodynamics of the gas in galaxy clusters by comparing the properties of a simulated galaxy cluster without black holes to X-ray observations of real clusters, which have black holes. For our data we used Python to manipulate a cosmological simulation made with ChaNGa, a code capable of simulating gravity, thermodynamics, and other things. Our cluster simulation includes gravity, hydrodynamics, gas cooling, star formation, and stellar feedback in a fully cosmological setting. However, this simulation lacks black hole formation, growth, and feedback, allowing us to pinpoint the effects of these processes on the cluster by comparing to observations. In our investigation, we found only minor differences in the density, temperature, and pressures of the gas between the observed and simulated galaxy clusters in the cluster outskirts. However, we found significant deviations between the simulation and observations near the cluster centers, where a large spike in star formation is present in the simulated cluster, not seen in the observations. Our results suggest that black hole formation and feedback in galaxy clusters most strongly affect the cluster in the central regions, where it can significantly decrease star formation by heating the gas.

The field of synthetic biology aims to modularize gene-networks within microorganisms for the purpose of using them as well characterized parts in building novel, useful biological systems. Realizing this goal is vital because biology’s inherent practicality holds the key to needed energy efficient technologies. As the human population increases, energy efficient technologies will become more and more a necessity. The first step in realizing this vision of adapting biology into an engineering discipline is through properly characterizing gene expression and enzyme function within engineered and natural biological systems. To do this, I have developed a turbidostat with mixture control and fluorescence sensing capabilities. A turbidostat is a continuous culture bioreactor that maintains constant culture volume and microbe density through a feedback loop between turbidity measurements and dilution with fresh media. The constant culture environment and microbe density allows the cultured microbes to reach a “steady state” where growth and all environmental conditions are kept constant. By maintaining microbes at steady state, variables that effect microbial processes are minimized, which facilitates accurate examining of gene-network dynamics. The mixture control ability of the turbidostat allows for the concentration of a given molecule within the culture environment to be modulated through time without altering any other environment variables. Through mixture control, a molecule that acts as an input to a gene-network (i.e. an inducer molecule that initializes a gene-network) can be varied in concentration through time within the culture environment. Since most gene-networks utilize fluorescent protein production as their output, the turbidostat’s fluorometer (a fluorescence detector) can monitor the output of a gene-network while the input to the network is modulated. We believe that the ability of this turbidostat to look at the input-output dynamics of gene-networks under defined environmental conditions will yield enhanced behavioral understandings, aiding to the modularity of these systems.

Over the past decade, measurements of hydrogen isotopes of lipid biomarkers, in particular leaf waxes, have been used to reconstruct past climate change. This method is appealing because lipids are well preserved in sediment and their hydrogen isotope ratio is primarily controlled by the hydrogen isotope ratio of their source water, which itself is controlled by factors such as precipitation amount and temperature. However, it has been shown that the isotopic difference between source water and lipids is distinctive among different plant species. In particular, it has been shown that salinity strongly affects the hydrogen isotope ratio of lipids produced by mangroves, but the mechanism that controls this relationship is unknown. Plants, including mangroves, take in water mainly from surrounding source water, but the deuterium to hydrogen ratio within the plant may be impacted by differences in transpiration rate, photosynthetic rate and respiratory water production. The photosynthetic rate (mol CO2 m-2s-1), internal carbon, (μmol CO2/ mol air) and conductance (mol H2O m-2s-1) of three species of mangrove, Rhizophora apiculata, Avicennia germinans, and Xylocarpus granatum, were measured at six different salinities from 5 to 30 ppt. The date, time, and leaf number were also recorded as additional measurements. Determining whether salinity, time of day and/or differences among species affects photosynthetic rate, internal carbon, and conductance is important in comparing the biosynthetic pathways of the three species of mangroves. It will allow us to investigate the cause of increased deuterium discrimination in mangroves at high salinity.

Fertility rates amongst women in Romania have been declining since the collapse of Communism in 1989. Data provided by the World Health Organization has shown the rapid growth of impairing diseases that have rendered some women infertile. With only 4% of Romania’s GDP allocated to the public health system, Romanian healthcare has struggled in providing women with quality treatment. The current study performed in Romania, investigates the effects of Romania’s Communist past on the ideologies of Romanian women. It considers how social networks, entrenched by these ideologies, have come to affect how women view the healthcare system and seek treatment. Using qualitative methods such as participant observation, in-depth interviews, and discourse analysis, this research will explore how Communist and post-Communist health policies have influenced attitudes towards fecundity and reproductive health for women and medical practitioners. By considering the ways in which ideologies construct the reproductive health network of Romanian women, this research shows that residual expectations of State responsibility in its role as a healthcare provider have prompted women to discuss and construct public healthcare as an illegitimate and inadequate government institution. Ultimately,  the emergence of this type of discourse in the female sphere is beginning to influence cultural notions of reproductive care and treatment. 

Using the lens of contemporary Egyptian archaeology, this research examines how the roles of Egyptian archaeology and archaeologists have changed since George Reisner first excavated the Menkaure Valley Temple over a hundred years ago in 1910. By examining the changes in archaeological practices, and the profession of archaeology in Egypt, this study seeks to explore a new perspective on how the application of archaeology has changed within the last century. There are many different ways of excavation, and methodologies have advanced dramatically over the past century. By comparing and contrasting old archaeological practices used by George Reisner in 1910 while excavating the Menkarure Valley Temple, Ancient Egypt Research Associates (AERA) is re-excavating the same structure and using Reisner’s old work combined with new archaeological practices to develop a new understanding of the temple and its many modifications, and changes of use throughout the history of Ancient Egypt. By using myself as an archaeologist and Reisner’s work along with the research conducted by the AERA team members as the investigative perspective, I will present a story that is relatively unknown, as well as introduce a new perspective on how the methods and practice of archaeology in Egypt have evolved. The project has the following steps:1. Compare Reisner’s methods of excavation strategy, artifact analysis, dating (etc.—focus on methods) to 21st century AERA methods. 2. Discuss how contemporary archaeologists in AERA use old data collected by Reisner—problems, challenges, and solutions 3. How does all of this change the way we interpret the Menkarure Valley Temple?

Huntington’s disease (HD) is a devastating hereditary neurodegenerative disease characterized by motor and cognitive dysfunction. One of the earliest molecular events in HD pathogenesis is the down-regulation of the cannabinoid 1 receptor (CB₁) in medium spiny neurons (MSNs), cells that play a key role in movement and degenerate in HD patients. Our goal is to rescue the specific loss of CB₁ in striatal MSNs to understand the role of this event in HD pathology. To do this, we inserted flox-stop CB₁ (fsCB₁) into striatal MSNs of R6/2 mice, the best characterized HD mouse model, and confirmed its localization. To validate the functionality of the knock-in fsCB₁ protein, we crossed fsCB₁ mice into a CB₁-/- background, so that the resulting mice lacked endogenous CB₁ receptors, and crossed them with globally-expressed Cre, so that fsCB₁ receptors became globally-expressed. We then demonstrated that replacing endogenous CB₁ with fsCB₁ did not alter responses in four classic cannabinoid behaviors (analgesia, hypolocomotion, catalepsy, and hypothermia) by injecting these mice with cannabinoid agonists. Once the knocked-in mice were validated, we measured changes in motor phenotype and synaptic integrity between R6/2 and fsCB₁-R6/2 mice. Motor phenotype was measured using rotarod, open field, clasping behavior, and Catwalk assay biweekly from 4 to 12 weeks of age. Synaptic loss was measured in the striatum, globus pallidus and substantia nigra reticulata by costaining with synaptophysin and either PSD95 or neuroligin 2 to resolve excitatory and inhibitory synapses, respectively. Preliminary results indicate that CB₁ rescue did not improve motor phenotype, but did reverse loss of excitatory synapses in the dorsal-medial striatum. Our future goals are to identify the specific subset of excitatory synapses being lost. By understanding which synapses are lost and how CB₁ rescue improves this loss, we hope to obtain greater insight into the pathophysiology of HD.