Genetic and molecular analysis is important to understand the evolutionary trends and relationships between chordates and hemichordates. Hemichordates share significant characteristics with chordates, including pharyngeal gill slits, a worm-like body plan and a post-anal tail. We have been studying the development of the central nervous system (CNS) in hemichordates to compare development to chordates. During early development in chordates, Shh and BMP play major roles in neural tube formation and dorsal ventral CNS specification. Elav has been used as a neuronal marker in chordates and is expressed within neural plate during early neurulation. We looked for expression of Shh, BMP and Elav in hemichordates to compare the development of the CNS in each group. We cloned and sequenced Shh, BMP and ELAV in S. bromophenolosus and P. flava, using primers designed from S. kowalevskii. Our purpose is to examine the expression of Shh, BMP, and Elav in the developing neural tube in S. bromophenolosus and P. flava by in-situ hybridizations. We see expression of Elav in the CNS of S. bromophenolosus, and expect to see Shh expression in the hemichordate endoderm. BMP expression has already been reported in the dorsal ectoderm and neural tube of S. kowalevskii, but those results were interpreted as a dorsal-version in chordates. We suggest instead that this hemichordate expression pattern is identical to that of chordates during neurulation. These results suggest that hemichordate and chordate central nervous system develop by similar genetic pathways.

Ascidians are invertebrate chordates that live in the ocean and have an interesting life history. Normally, a tailed chordate larva hatches from the embryo, swims, becomes competent, and then responds to settlement cues, which initiate metamorphosis. Metamorphosis triggers apoptosis and tail loss. 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. 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 the tailless species. The number of transcripts were quantitated and we have cloned many of the genes. We show that there is early expression of metamorphosis genes in Molgulidae, including genes involved in the Mannose Binding Lectin (MBL) Complement pathway, such as MBL and MASP. We hypothesize that the maturation of the immune system early in tailless larvae could be a progression to competency and trigger to metamorphosis. We further hypothesize that a heterochrony in metamorphosis is responsible for the early activation of programmed cell death in the tailless ascidian larvae. In situ hybridizations were used to examine the localization of expression of these genes and quantitative PCR was used to independently validate RNA counts from our transcriptome data. Our ultimate goal is to elucidate the ascidian molecular trigger for metamorphosis and uncover whether heterochrony of metamorphosis leads to tailless ascidians.

A continuous measurement of wall shear stress can be achieved using shear sensitive liquid crystal coatings (SSLCC). Utilizing thin films of polymer-embedded birefringent liquid crystals as sensors, they deform elastically when subjected to shear stresses. The light passing through the SSLCC is recorded with a charge-coupled device camera and is processed by the specialized birefringence detecting Milliview. Shear magnitude and direction are determined from the results. A single coating comprises of polymers, solvents, and liquid crystals in varying proportions. When applied to a surface, solvents vaporize, leaving liquid crysals embedded inside polymer structures. By adjusting the proportions of the components, the viscosity and curing rate of the coatings can be controlled to match the method of application. A series of formulations were applied to a flat, glass substrate using methods such as airbrushing and direct deposition. Each combination was tested repeatedly at free-stream speeds ranging from 5 m/s to 36 m/s. The best coatings were determined by optical clarity, surface smoothness, signal strength, and reusability both through visual observation and testing in a small wind tunnel. Few combinations yielded both a high signal strength and reusability.

The effects of temperature and salinity on growth rates and toxicities of a dinoflagellate species in the genus Alexandrium will be determined. Species of Alexandrium can produce a suite of dangerous neurotoxins, the most potent being saxitoxin, that cause paralytic shellfish poisoning in humans if contaminated shellfish are consumed. These potentially lethal toxins are dangerous to humans and marine mammals alike. Toxins accumulate in both wild and farmed shellfish during a harmful algal bloom (HAB), and can result in shellfish harvesting closures causing economic losses to commercial shellfish growers. Climate change scenarios project increased freshwater input to the Puget Sound region (causing decreased salinity) and warming of surface waters. These changes were simulated in the laboratory using a temperature gradient bar – a cell culturing apparatus designed to facilitate cell growth over a range of uniformly distributed temperatures (5-28°C) and salinities (10, 15, 20, 25, 30, and 35 ppt). Nutrient concentrations in the culture media is locally relevant and light is on a 12:12 h light:dark cycle. These experiments will be conducted for two Puget Sound isolates of Alexandrium collected in 2010 – one from Cypress Island in the north and one from Quartermaster Harbor in the south. Preliminary results for the northern Puget Sound isolate indicate zero to low growth at all temperatures for the 15 ppt salinity treatment, while greatest growth occurred between 15°C and 22°C in the 20 and 25 ppt salinity treatments. Toxin analyses are yet to be performed. The growth and toxicity response surfaces generated from this study will be used in a model to assess potential range and timing of future blooms under projected climate change scenarios.

Duchenne Muscular Dystrophy (DMD) is a degenerative disease leading to fatal respiratory and cardiac complications. It is caused by loss of functional dystrophin through mutation of the dystrophin gene. Possible treatments could use Adeno-associated virus (AAV) to introduce genes into muscle cells. This strategy requires muscle specific regulatory cassettes containing sequences specific for transcribing the therapeutic gene in muscle cells. My studies seek to increase the activity of one such cassette, MHCK7. Since AAV vectors can package only ~5000 basepairs of DNA, the dystrophin cDNA and regulatory cassette must be miniaturized. I am modifying MHCK7 by adding a 95 bp intronic enhancer (SIE) from the muscle creatine kinase (MCK) gene that is important for MCK expression in slow muscle fibers and cardiac muscle. The SIE has been placed ~1kb 3’ of the MHCK7 luciferase reporter cDNA to mimic its endogenous genomic location. SIE addition was performed by introducing a HindIII site 3’ of the luciferase polyA sequence by site directed mutagenesis, and the subsequent opening of DNA at that site. The SIE was transferred from a previously made plasmid and ligated into the opened MHCK7. Restriction digests and gel electrophoresis were used to assess successful introduction of HindIII and to confirm digests for opening the DNA and isolating the SIE. New recombinant MHCK7-3’-SIE will be transfected into skeletal and cardiac muscle cultures, and its activity will be assessed through luciferase activity. The SIE’s contribution to expression will be compared to that of the original MHCK7 plasmid as well as to MHCK7 with the SIE located immediately 5’ of the cassette. If the new regulatory cassette is more active than MHCK7 and 5’-SIE-MHCK7, further studies will test the cassette in conjunction with a microdystrophin cDNA. If activity is not increased, I will test alternative locations of the SIE.

Historically, Islamist leaders — from the Muslim Brotherhood in Egypt to the Jamaat e Islami in Pakistan and Bangladesh — have insisted that Sharia law is applicable to the modern nation state without political molding. Upon obtaining political power, however, they have always significantly moderated their conservative religious politics and adjusted their ideology to pragmatic realities. In my research, I ask what happens when an Islamist group does not moderate their politics to match political realities. I argue that any Islamist group which does not align their ideology with pragmatic realities after achieving power is destined to become a failed revolution. To test this theory, I analyzed the Afghan Taliban, a group which made no attempt at altering Sharia law to meet political demands domestically and internationally. Ultimately, this led to their deterioration as a political party. Through this, I conclude that it was the direct implementation of politically unmodified Sharia law which undermined their once robust popular support and lead the regime on a path to collapse. In turn, this paper implies that a government truly based upon Qu’ranic law is not sustainable in the modern world. In my research I relied entirely on an extensive literature review. I reviewed the premier literature on Islamist movements in and out of Afghanistan, as well as extensively on the history and ideology of the Taliban revolution in Afghanistan. Additionally, I reviewed Theda Skocpol’s theory on revolution to place the failed Taliban revolution in the frame-work of contemporary social science.

As hundreds of thousands Pakistanis inundated the cities of Lahore and Karachi to hear the rising cricketer-turned-politician Imran Khan speak, commentators and politicians alike were left baffled: how did the peripheral figure and his respective party become a formidable voice in Pakistan’s politics within a few months? This paper will argue that the ascension of Imran Khan’s party, the Tehreek-e-Insaaf, can be attributed to its adoption of Post-Islamist rhetoric of religiosity and rights, which came to be publicly appealing following the formation and failure of the Islamist conglomerate the Muttahida Majles-e-Amal. Due to Postcolonial Pakistan’s institutional divide between State and society, as well as Islamism’s failure to bridge such a gap, Pakistan is experiencing a Post-Islamist turn, where a novel relationship between political institutions and the public sphere is being negotiated. In conclusion, this paper will contend that Post-Islamism may prevail given Pakistan’s pre-existing democratic institutions.

In the wake of the Ottoman Empire, Mustapha Kemal sought to modernize the republic of Turkey, ushering it head-first into the 20th century. His manifested ideas resulted in a huge departure from the country’s Ottoman past. An officer in the Ottoman military, Kemal viewed the empire as decadent and corrupt, embarking on a journey of reformation that altered the future of Turkey forever. Independence and democracy were at the forefront of Kemal’s mission, and in Kemal’s mind these pursuits were inseparable from secularism and nationalism. Kemal claimed that politics corrupted religion and that only secularism could restore purity to Islam in a post-Ottoman world. However, the 1970’s saw the rise of the National Order Party, an Islamist political group. Unfortunately for the ambitious party, unrest and a failing economy framed their rise to power and in 1980 the military took control of the government, claiming that the Islam in government was unconstitutional. The military used Kemalism to legitimate that, and several future coups even though Kemal himself stated that the military should also be separate from politics. The resulting struggles between Islamic groups and the Kemalists saw the rise of the AKP, a group composed of Islamist actors. The AKP has straddled the fence between secularism and Islam, promoting social, democratic, and economic growth. The argument proposed shows how Muslim leaders have moderated themselves and become the heirs to Kemal’s Revolution by following Kemalism's tenets for modernization: republicanism, statism, nationalism, popularism, revolutionism, and secularism. By examining the current economic and political academic discourses concerning Turkey's historic and current events I am able to contradict the debated notion that assumes that Islam and democracy are not compatible while taking it a step further to assert that a party with Islamic roots has continued one of the most successful revolutions in history.

Since the attacks on September 11, 2001 the international eye has fixed on the effects of political Islam and the impacts that transnational Islamic organizations such as al Qaeda have on global politics. A critical impact of transnational Islamic organizations is their involvement and impact on nationalist struggles. By using Chechnya as a case study this paper analyzes the effect of transnational Islamic organizations, in particular al Qaeda, on a nationalist struggle. This project is the result of my extensive research and a peer review process in an attempt to understand how nationalist struggles are changed by transnational Islamic organizations. Chechnya has been fighting for independence from Russia since the collapse of the Soviet Union in two separate wars. Al Qaeda started to become involved in Chechnya after the first Chechen War of Independence and their presence has caused dramatic changes. During the Second War for Chechen Independence there was a dramatic shift in both ideology and practices by the Chechens. New organizations, such as the Islamic International Brigade (IIB) appeared and preached a hard line of fundamentalist Islam in addition to carrying out violent attacks in the name of Islam and Chechnya. Their tactics shifted from local guerilla warfare to grandiose terrorist plots within Russia proper, including taking over a school in Beslan which resulted in the death 300 Russians. In addition to the large scale attacks suicide bombing became a technique the Chechens used, though there is debate on whether this is the result of al Qaeda. My research studies how the battle for Chechnya changed from nationalism to jihad, and how this case study can shed light on the question of how transnational Islamist organizations impact nationalist struggles.

Urban women’s boycotting of the 2005 elections in Iran cleared the way for hardliner Mahmoud Ahmadinejad’s ascension to the presidency and a subsequent regression in women’s rights. This serves as an entry point to an understanding of the intricacies of a feminist movement in which women subjected themselves to an ideologically conservative president as part of a conscientious and contentious strategy for achieving equal democratic rights. If not through formal politics, what articulations and organizational capacities are available to and employed by women to demand and influence reforms within an Islamic Republic? This paper will examine three facets of the women’s movement in Iran; first, the role of women in mobilizing support for and successfully electing the reform candidate Mohammad Khatami to the presidency in 1997, and how the successes and failures of reform politicians and women in government reflected the limits of formal politics to elevate Iranian women’s rights through legislation from 1997-2005. Second, the paper will address how Iranian feminists have employed an Islamic justification for women’s rights that is both compatible with the prevailing Iranian politico-religious context, and useful in reconciling secular and Islamic feminist perspectives under a common cause. Finally, the implications of various feminist approaches to engaging in “informal” politics will be explored; specifically, the organizational structures supported through the use of new media that mobilizes feminist solidarities and collective action, pressuring the Iranian regime from within and without. The efforts of Iranian women reveal a movement that is not content with gradually reforming within, or conforming to, a restrictive legislative process; rather, Iranian feminists have adopted an Islamic modernist position focused on the removal of religious authority from the state, based on the recognition that the achievement of a fully democratic political space is fundamental to the adaptability and sustainability of the women’s movement.

This project focuses on using microbiota to determine environmental changes in Puget Sound, specifically benthic foraminifera. Benthic foraminifera are single celled organisms that produce a shell and live at the bottom of the ocean. Because foraminifera have characteristic ecologic requirements, they can be used to document environmental changes over time. In addition, foraminifera’s short reproductive cycle (weeks to months) and species specific reactions to environmental changes enable high resolution studies of ecosystem health and reaction to mitigation efforts. By examining the density, diversity and assemblage composition of foraminifera in samples of sediment several determinations can be made about environment in the local region at the time the sediment was deposited. This study focuses on stations in Bellingham Bay, WA using 11 sediment samples taken between 1997 and 2011 by the Washington state Department of Ecology. From 2002 to 2006 a clear drop in foraminiferal density is observed in Bellingham Bay. This indicates a serious change in environment for the area during this period; changes that may include an increase or decrease in pollution, an increase in pH, or changing climatic conditions. In the Bellingham Bay area a very low species diversity is also observed. This lack of diversity can also indicate various climatic or other environmental conditions. For example, the most dominant species in these samples, Eggerella advena, is known to colonize areas of sewage pollution in other bays. By comparing the Bellingham Bay stations to others in Puget Sound, it will be possible to document environmental shifts over the last 15 years.

The harmful bloom-forming alga, Heterosigma akashiwo, has a dual-stage life cycle, transitioning between a vegetative, motile stage in the water column and a resting, non-motile stage in the sediments. Previous observations have revealed a large shift in neutral lipid content as cells transition between vegetative and resting life stages. The relative neutral lipid content per cell decreases as cells enter the resting stage, followed by a rapid synthesis of neutral lipids once cells emerge from the resting stage. Neutral lipid bodies are thought to be a source of cell energy reserves. The purpose of my research is to study how fatty acid profiles shift in H. akashiwo during these life cycle transitions. Understanding changes in fatty acid profiles is important in understanding how cells respond to changes in environmental conditions and how lipids are used in metabolic processes. In this study, two strains of H. akashiwo (CCMP 452 and UWC 13.03) are signaled to enter the resting stage by putting cell cultures into the dark at 10 °C for 18 days. Resting cells are activated by exposing the cultures to growth supporting conditions of light and increased temperature (12 °C). Cell viability and relative neutral lipid content is determined by staining with the SYTOX-Green DNA stain and the BODIPY505/515 neutral lipid stain, respectively, using flow cytometry. Fatty acid profiles are determined using an established gas chromatography- mass spectrometry method developed in the Cattolico laboratory. I will report on the outcomes of the fatty acid profiling for H. akashiwo over the alga’s dual-stage life cycle. I anticipate distinct change in the fatty acid profiles between these life stages.

Puget Sound is home to countless aquatic species, all of whom are dependent on healthy water conditions. At the bottom of the trophic web are foraminifera—single celled organisms that directly reflect the environment they live in, because each species has a very narrow range of acceptable habitats. Foraminifera can be used to assess local conditions on a very fine scale, and to assess the large-scale health of the body of water. This type of study has been done on numerous bodies of water throughout the world and is well proven, but has never been applied to Puget Sound. As a small part of this overarching project, I focus on identifying and analyzing foraminifera from Port Madison, a part of Puget Sound relatively sheltered from anthropogenic impact. Port Madison thus acts as a control with which to compare samples from other locations in Puget Sound. Preliminary results indicate that Port Madison has a very high diversity of foraminifera with at least 12 different species immediately apparent, but a relatively low abundance of about 28 foraminifera per gram of sediment. In comparison with other locations throughout Puget Sound, this shows that anthropogenic conditions decrease the diversity of foraminifera, and areas like Port Madison where little anthropogenic activity is present yield higher diversity. Data from sheltered areas help build a knowledge base for a healthy Puget Sound, which in turn gives us an idea of what to work toward in future attempts to promote a healthier Puget Sound.

With the rise of Islamist parties to power in Egypt, the question of how women will be affected by the Islamist agenda has gained new urgency. This research project examines the rise of Islamic health services as well as several Islamist political movements in Egypt in order to assess how women are affected by these phenomena. Governmental Egyptian health services have declined in both quality and quantity since the establishment of the Nasserist welfare state during the 1960s. Services affecting women, in particular, have varied in coincidence with the state’s pattern of repression, supervision and cooption of healthcare organizations in the Parallel Islamic Sector. Islamic hospitals and clinics, some affiliated with Islamist political organizations such as the Muslim Brotherhood, have grown in popularity and influence during the past several decades. I discovered through my research that Islamic and Islamist health clinics consistently out-perform the state in the provision of health services to many Egyptians, and generally provide high quality, modern and Western medical care. This trend towards religiously influenced healthcare provision raises several important concerns for women’s health in Egypt, as Islamist organizations promote female domesticity and oppose reforms to personal status laws seeking to improve protections for women within marriage bonds. My exploration of these diverse aspects of Islamic healthcare in Egypt will illuminate how these parallel institutions have substituted for and subsequently challenged the legitimacy of the state in providing healthcare to Egyptian women.

Conversation analysis has shown that certain linguistic factors (avoidance of pronouns, increased negative words, e.g.) are affected by the act of deception. Studies have proven that the act of deception increases cognitive load and results in behavioral and linguistic indicators. Little research has been done on the potential cognitive strain that could result from speaking in a second language and none has been done to explore the effect that deception and second language production would have when done simultaneously. Second language discourse has also been proven to increase cognitive load. Therefore we expect that the added cognitive strain resulting from the use of a second language while participating in an act of deception will further exaggerate the well known linguistic indicators of deception. We believe our findings will show that the increased cognitive effort caused by a speaker using a second language while participating in deception will exaggerate the linguistic factors affected by deception, making it easier to identify in discourse analysis.

Preventing a child from accessing a self-defense weapon in the parents closet when home alone is difficult if the parent does not even know that the child has tried to acquire it. Designing a home monitoring system that can alert parents when children access restricted areas around the home even when they are away from the home could help prevent the child from harming himself or others. To achieve this goal, an Internet connected microcontroller and sensors were used in congruence to detect the presence of a child in a restricted room. The system consists of an Arduino microcontroller connected to a PIR(passive infrared) sensor that could detect when there was motion. Using an internet connection, the microcontroller can send a twitter post to a dedicated twitter account. The twitter account can be linked to a smart phone and alert parents even when they are not around the house. A smart phone could be configured to automatically retrieve the post within minutes of the child’s breaching into the restricted area, notifying parents of the breach.  A system like this would be a great way to monitor a child’s activities at home and help parents control what their child is doing.

Warming temperatures and the declining extent of sea ice are encouraging economic development in the Russian Arctic. The Russian Federation’s implementation of a foreign policy which places priority on sustainable socio-economic development, and protection of indigenous peoples and environment, surprised many within the international community. The strengthening of the indigenous voice in Arctic governance is another significant recent transition. Indigenous peoples have gained the status of Permanent Participants in the Arctic Council, which is a high level intergovernmental forum for the Arctic region. My scholarly research aims to examine achievements by the indigenous peoples to ensure sustainable socio-economic development in the Russian Arctic. My work highlights ways in which indigenous peoples are utilizing customary international law and diplomacy to shape state policies. This research project uses four indigenous organizations as case studies to illustrate the evolving role of indigenous peoples in governance of the Russian Arctic. By examining the high degree of participation and political agency wielded by these non-traditional actors, it is clear that a paradigm shift in governance is taking place. This shift is one which marks the end of the strictly state-centric model of international relations towards a more cooperative and multi-lateral global political arrangement.

Deuterostome adult body plans greatly differ between echinoderms, chordates and hemichordates. Genetic analysis has helped to understand the evolution of deuterostomes and the relationships between species. In this project, we have sequenced the Hox genes in Ptychodera flava, an indirect developer of the hemichordates, which has larvae that are morphologically similar to echinoderms. This information, along with gene expression data, gives insight to the similarities in developmental patterns between phyla as well as insight to their evolution. Once the genomic organization of Ptychodera flava Hox genes and Hox gene expression is complete, we can begin to deduce the developmental pattern of the common ancestor of hemichordates and echinoderms. Since the Hemichordate phyla contains both indirect and direct developers that are equally related to echinoderms, which are indirect developing, a deep comparison between gene expression and genomic organization will help us understand their common ancestor. We approached this by sequencing Bacterial Artificial Chromosomes (BACs) that contain the P. flava Hox complex and by examining gene expression via in situ hybridization. We are specifically interested in whether the Hox complex is rearranged, as in echinoderms, or co-linear, as in chordates and where the genes are expressed during different stages of development. Results show that P. flava Hox 5 through Hox 11/13c are collinear on the chromosome, but the arrangement of Hox 1 through Hox 4 is still unknown.

Cooperative interactions are observed at all levels of biological organization. Cooperation facilitates major transitions in evolution such as from unicellularity to multicellularity. How a cooperative system evolves from its incipient state is unknown. I have used a synthetic cooperative system termed CoSMO (Cooperation that is Synthetic and Mutually Obligatory) as a model for incipient cooperation. CoSMO is an engineered yeast system consisting of two non-mating strains, differentially labeled with fluorescence. It relies on direct exchange of distinct metabolites between the two cooperators. Evolved CoSMO cocultures exhibit an improvement in viability, defined as an increased ability to survive reductions in population density. I have determined that this viability improvement is heritable, by measuring the viability of reconstituted evolved CoSMO pregrown in rich media which allow cells to grow in the absence of cooperation. Furthermore, by mixing an evolved strain with its ancestral partner, I determined that a single evolved cooperator was responsible for community-level viability improvement. I am characterizing phenotypic changes in evolved cooperators that contribute to increased viability. The phenotypic changes can potentially be categorized as either “self-helping”, such as through increased starvation tolerance and higher affinity for metabolites, or “partner-helping”, such as through increased metabolite release. In all cases, I will identify the molecular basis for these adaptations through genome-wide re-sequencing. Furthermore, genomic analysis on entire evolved CoSMO cocultures will be performed to reveal the diversity and frequency of relevant mutations in the population over time. Lastly, I will perform similar analyses on replicate populations. If similar adaptive changes that result in improvement in system viability are observed, a deterministic process to improve viability is implied. Otherwise, diverse mechanisms can achieve improved fitness. In both cases, we will gain insights into how cooperative systems evolve from inception.

Cancer vaccines offer a promising method of manipulating the immune system to seek and destroy malignant cells. However, there have been many limitations on their ability to enhance stimulation and persistence of CD4+ Type 1 helper (Th1) and CD8+ cytotoxic T lymphocytes (CTL) necessary to promote adequate tumor regression. The long-term goal of this research is to use polymeric carriers to deliver antigens and immunomodulatory siRNA to antigen presenting cells (APCs), in order to promote anti-tumor cellular immunity. We have designed pH-responsive, endosomolytic, “smart” polymers that are capable of being conjugated to antigens and complexed to siRNA. Also, we have demonstrated the ability of these polymers to enhance class I antigen presentation by APCs. In addition, we have shown that polymeric delivery of siRNA promotes knock down of a model gene, glyceraldehyde 3-phosphate dehydrogenase, in dendritic cells. We plan to demonstrate similar results with other immune cells, both immortalized and primary, and genes of interest, as well as extend this technology to relevant tumor-associated antigens. These data point to the modular nature of our polymeric delivery systems, as they provide a promising way of approaching challenges in cancer immunotherapy through multiple delivery modalities.

Duchenne Muscular Dystrophy (DMD) patients have mutated dystrophin genes and suffer from muscular degeneration, which leads to cardiac, respiratory and general skeletal muscle disabilities. Potential treatment of DMD patients could involve gene therapy in which Adeno-Associated Viruses (AAV) deliver functional dystrophin cDNA ligated to muscle specific regulatory cassettes that express dystrophin exclusively in striated muscle tissues. The Hauschka lab has been designing regulatory cassettes (e.g. MHCK7) that contain control regions from genes that are highly active in skeletal muscles such as the muscle Creatine Kinase (MCK) and α-Myosin Heavy Chain (α-MHC) genes. Due to the limited size of DNA that can fit into AAV capsids, the cassettes need to be as compact as possible. My goal is to test whether adding a small intronic enhancer (SIE) from the MCK gene, that is known to have high activity in skeletal muscle in vitro, will improve transcriptional activity in cardiac and skeletal muscle cultures compared to MHCK7. The SIE was isolated and cloned immediately 5’ of MHCK7. After sequencing to confirm the SIE orientation, we carried out a pilot study in which 5’-SIE-MHCK7 was transiently transfected into skeletal muscle cultures. 5’-SIE-MHCK7 activity, as measured by the reporter gene firefly luciferase, was several fold higher than MHCK7. This suggests that the addition of the SIE to two known muscle specific enhancers increases transcriptional activity in skeletal muscle cells. Future studies will focus on confirming this result, and comparing 5’-SIE-MHCK7 activity to aMHCK7-SIE cassette in which the SIE is located 3’ of the luciferase reporter, so as to mimic its normal location in the MCK gene. The 5’-SIE-MHCK7 cassette will also be tested in cardiomyocyte cultures, and smaller cassette versions will be designed that eliminates additional non-conserved DNA sequences from the SIE and MHCK7 components.

The Institute of Medicine‘s (IOM) report on health disparities, “Unequal Treatment”, identified the medical system’s inability to distinguish minority groups with meaningful specificity as a primary barrier to recognizing critical disparities in health care and outcomes. A recent follow-up report on “Race, Ethnicity, and Language Data” called for more detailed demographic data collection. This study evaluated the validity of self-reported primary language to detect disparities in diabetes care across language groups of first generation, mostly non-English speaking outpatients at an international clinic. The REALL (Race, Ethnicity and Language proficiency Level) study is a cross-sectional pilot study, surveying a convenience sample of immigrant and refugee patients receiving primary care at an urban, county hospital. Participants were eligible for this study if they had diabetes based on medical record data and completed the REALL survey question about how many and what languages they speak, their reading ability in those languages, and the culture they most identify with. Survey data were linked with medical record data that included hemoglobin A1C (HA1C), low density lipoprotein (LDL) and systolic blood pressure (SBP). Diabetes control was evaluated across the 6 largest language groups. 250 patients were eligible for the study. The survey data were 90.84% concordant with registration data for primary language. Mean HA1Cs were lowest among Vietnamese patients, significantly lower than Somali patients and Cambodian patients. While 52% of Vietnamese patients had HA1C<7, only 26% of Somali patients had HA1C<7. There were no statistically significant differences across language groups in mean LDL level or SBP. Results suggest that there is variability in diabetic outcomes across language groups in first generation immigrant communities. Recent refugee communities are potentially more stressed due to lack of acculturation and community establishment. Further inquiry into their duration in the US, the clinical care system, and their dietary practices are advised.

Ocean Research College Academy (ORCA) students partake in monthly State of Possession Sound (SOPS) cruises to collect data from various sites in the Snohomish River estuary near Everett. The focus of this SOPS research is on dissolved oxygen fluctuations in Possession Sound and the Snohomish River. Large fresh water inputs into Possession Sound bring sediment, organic matter, nutrients, and pollutants from both natural and human influences, which may impact the dissolved oxygen (DO) levels of the water. In addition, the tides affect the amount of DO in the Snohomish River during flood tide and ebb tide. The hypothesis formulated for this research was that less populated areas upriver would have a higher DO content rather than more populated areas downriver. Measurements were taken at four different locations along the Snohomish River and at the delta during ebb tide to negate the tidal effects. The parameters collected with a YSI 85 were DO, temperature, and salinity in order to find the influences that determine levels of DO in the areas of interest. Future temporal studies based off of this research could directly link anthropogenic and seasonal influences on specific respiring marine organisms in the ecosystems of Possession Sound.