Immigration law is developed and enforced through cooperation between the state and federal government. There are times when the state legislature creates laws that supplement federal policies in order to conduct immigration enforcement unique to their situation. A state’s public perception regarding immigrants can result in the legislature drafting pro-immigrant or anti-immigrant policies. Previous research by Merkowitz and Yoder (2009) discusses how the U.S. national legislature responds to public opinion about immigrants. They find legislatures are influenced by demographics of their districts. This research will examine the state legislature of Idaho and its response to public perceptions about immigrants. I will be testing to see if immigration policy is a function of the following variables: party affiliation, state demographics, and regional industries. Each variable will be tested independently while controlling for other potential explanatory factors. I will get data for this analysis this summer by conducting a survey using a sample of Idaho state legislators, advocacy groups, and special interests that represent the state’s participants on immigration policy making. In addition to the survey, I will make an analysis on how legislators vote on immigration related legislation from the years 2000 to 2010. Finally, I will create an index that will determine if the responses from the survey and a legislator’s votes are pro-immigrant or anti-immigrant. I will define if a legislators pro-immigrant or anti-immigrant by how far their proposals deviates from current federal immigration policy. I expect the results will show that the state legislature responds more to the state’s demography and economy than the legislator’s party affiliation. The results from this research will reveal who are the primary actors in Idaho’s immigration legislation process. The results will show if there is a difference between the national legislature’s reaction to public pressure and how Idaho’s state legislature reacts.

Opiates have long been a useful tool in medicine due to their potent analgesic properties. Although there has been a significant increase in prescribed use of opioids in recent years there has also been a large increase in deaths related to prescription opioids that may involve abnormal control of ventilation. Opiate effects on ventilation have largely been considered to be due to depression of respiratory chemosensitivity (sensitivity to carbon dioxide and/or acid-base balance), but another effect may include decreased oxygen consumption and therefore decreased production of carbon dioxide. In this study, we are using a whole body plethysmograph and a respirometer to measure the rate of oxygen consumption levels in juvenile rats 18-23 days) during sleep - before and after administration of morphine at varying dosages (5mg/kg to 45mg/kg). Our initial findings show no decrease in oxygen consumption and surprisingly instead demonstrate increases up to 50% following opioid administration. These unexpected findings illustrate the need for additional investigation on possible causes of prescription opiate associated deaths. Increased oxygen consumption (and therefore increased carbon dioxide production) in the presence of drug induced decreased sensitivity to carbon dioxide may interact to lead to dangerous compromises in the control of ventilation in patients taking a broad range of opiates doses in the community.

For an animal whose brain only contains about 100,000 neurons, the fruit fly is capable of striking behavioral complexity. The goal of our work is to identify neuronal circuits that process sensory cues and transform them into changes in motor activities. Our work relies on the Gal4-UAS (Upstream Activation Sequence) system, a commonly used genetic tool that allows us to label and manipulate small subsets of neurons. The system takes advantage of different enhancer elements to drive expression of the Gal4 transcription factor in various tissues. When Gal4 flies are bred with UAS flies that contain GFP (green fluorescent protein) or RFP (red fluorescent protein), the Gal4 will bind to the UAS, and enable the production of GFP or RFP. This allows the desired cells to fluoresce, which can be tracked using a microscope. With this, we can identify Gal4 lines with interesting neuronal expression patterns and test the function of these neurons in Drosophila. Expressing the ion channel UAS-TrpA1, for example, activates neurons when the temperature is shifted from 22 °C to 27 °C. I crossed different Gal4 drivers to UAS-TrpA1. After performing a temperature shift on the progeny, I observed that various Gal4 flies became twitchy while others collapsed. These behaviors were reversible when the temperature shift ended. Fluorescing the Gal4 lines that allow for this behavior would allow insight as to the activated neurons in the brain and thoracic ganglia. The confocal images would allow us to see the neuronal expression. Through this model organism we can hope to comprehend and apply the knowledge in understanding complex systems, for example, what neurons coordinate behaviors such as navigation and locomotion.

There are currently very few effective treatments to repair damage resulting from spinal cord injury (SCI). The project I have been working on in the Moritz Lab involves the exploration of Intraspinal Microstimulation (ISMS) and its ability to promote functional recovery after SCI in a rodent animal model. Our group employs a unilateral incomplete spinal cord contusion injury using the Ohio State University contusion device at the level of the fourth lamina in the cervical spine, or C4. We then place stimulating wires in the spinal cord, specifically in the ventral gray matter caudal to the injury at C6-C8, in order to deliver ISMS to the motor pools innervating the triceps and other forelimb muscles. My independent project in the lab is a histological analysis of triceps and forelimb muscles taken from the SCI models to determine whether the use of ISMS preferentially recruits fatigue resistant (FR) muscle fibers in the affected forelimb of these rats. After paresis resulting from SCI, muscles atrophy and undergo a "slow-to-fast" transition in the myosin heavy chain (MHC) isoform content. My goal is to examine whether ISMS can slow or even reverse this slow-to-fast conversion after SCI. I am using immunohistochemistry to differentiate muscle fiber types in the triceps muscle. I stain for the unique MHC isoform that is expressed by the various slow and fast type fibers. My hypothesis is that there will be a higher population of type I and type IIA FR fibers in the stimulated muscles of ISMS group than in the control group, which will mostly consist of fast fatigable types IIB and type IID. The long-term goal of my project is to determine the benefits of using ISMS as a neural prosthesis and regenerative therapy for individuals who have suffered from spinal cord contusion injuries.

Muscle contraction begins when Ca2+ binds to sTnC, which removes the inhibition of myosin binding to actin, allowing force development or shortening. Our laboratory has engineered sTnC variants with differential Ca2+ binding affinities to investigate how it affects healthy and diseased muscle. Chief among these variants are I60Q and M80Q, which have lower and higher Ca2+ binding affinities than the native protein, respectively. Expression of these variants in diseased or damaged muscle as a therapeutic treatment will be accomplished by viral transduction with cDNA. This method does not achieve 100% replacement of the native protein. Thus, my project is to determine the minimum variant level that needs to be replaced to have a signficant effect on contraction to help design treatment studies. Replacing native sTnC with I60Q or M80Q is also expected to gradually produce reduced or greater contraction when exposed to higher [Ca2+] prior to sTnC extraction and following reconstitution with variant:WT mixtures to determine force outputs. pCa50 is compared between pre-extraction, and reconstitution. Variants used in this study also contained polyhistidine tags, allowing us to quantify the amount of reconstitued protein incorporated into the fiber. These data show us the the effect of a particular reconstitution ratio and the amount of protein necessary to produce this effect. My preliminary data suggest that there is a progressive effect of increasing I60Q in muscle fibers to reduce the calcium sensitivity of force, thus reducing contractile strength. This variant may therefore be effective in reducing contraction in muscular dystrophy, thus maintaining muscle integrity and function.

Cell transplantation has been shown to improve cardiac function following myocardial infarction (MI). The mechanisms behind improved function are not well-understood, but are correlated with changes in myofilament protein phosphorylation, which likely affect Ca2+- and length-dependent (preload) changes in force. This project was designed to test the hypotheses that 1) responsiveness to ventricular filling (Frank-Starling relationship) will decrease following MI, and 2) neonatal rat cardiomyocyte (NRC) transplantation after infarction will ameliorate this loss of function. One week after induction of MI, fluorescently-labeled NRCs were injected into the area of infarction of some of the animals. Sham-operated rats served as control. After 4-6 weeks, cardiac performance for all groups was preliminarily assessed using echocardiography. Then, whole working heart preparations were used measure cardiac power output, testing the heart’s ability to follow the Frank-Starling relationship. NRC-injected hearts exhibited higher average power and response to ventricular filling than sham (media-only) injected hearts, indicating that NRC-engraftment significantly ameliorates MI-induced decreases in preload responsiveness. Force and Ca2+ sensitivity (pCa50) of demembranated myocardial strips from these hearts were then assessed. NRC-injected hearts strips had a sarcomere length associated change in pCa50 that was similar to uninfarcted hearts (0.07 ± .02 vs. 0.10 ± .03, respectively), which was significantly improved as compared to strips from untreated, infarcted animals (0.0 ± .03). These results indicate that loss of preload dependence following MI is, at least in part, due to altered myofilament function and this is reversed following NRC engraftment. Current work involves investigating myofilament phosphorylation profiles to assess the mechanism of myocardial function changes. Preliminary results show decreased phosphorylation levels of cardiac troponin T (cTnT) in NRC-injected hearts compared to sham-operated and infarcted hearts. These changes in phosphorylation may be associated with the changes in force production following cell therapy.

In recent years, increasing numbers of single amino acid mutations in cardiac troponin C (cTnC) have been identified in patients with cardiomyopathies. The troponin protein complex regulates cardiac muscle, as Ca2+ binding to cTnC initiates contraction. Previous studies of the mutations associated with hypertrophic cardiomyopathy (HCM) patients show increased Ca2+ sensitivity and of those with dilated cardiomyopathy (DCM) patients, decreased Ca2+ sensitivity. However, it is still unclear how a change in Ca2+ signaling might precipitate the disease, and unknown why a myofilament protein variant affects the Ca2+ transient, which is regulated by calcium handling proteins. This project focuses on the I61Q cTnC variant, which previous studies have shown decreases fraction shortening (produce weaker contractions) and Ca2+ binding of myofilaments. Determining how or if abundance and activity of sarcoplasmic reticulum (SR) proteins vary will shed light on the mechanism behind the change in Ca2+ transient. All cardiomyocytes were isolated from healthy adult rats and transfected by adenoviruses containing the I61Q gene. Transduced cardiomyocytes were cultured for 48 hours. Then, Western blotting determined abundance of Ca2+ handling proteins in the SR, such as phospholamban and SERCA-2.  SDS-Page gels were used to determine protein phosphorylation. Using imaging software to analyze the bands in the gel will result in a quantitative comparison of protein abundance and phosphorylation. The difference in protein expression and regulation provide insights for understanding the mechanism behind the change in Ca2+ sensitivity, and potentially allow I61Q to be delivered through gene therapy in patients with HCM to restore normal Ca2+ sensitivity. Past contractile measurements were performed at 25°C, but since the heart normally operates at 37°C, future experiments will be performed at this temperature for transduced cardiomyocytes. Contractions will be induced by electrical stimulation so that cell lengths, rates of sarcomere contraction/relaxation, and Ca2+ transients can be measured simultaneously using IonOptix video microscopy.

The current geographical configuration of the Titlow Beach area in Tacoma, Washington has its origins in the 1903 purchase of the northern half of the W.B. Wilton Land Claim by Aaron R. Titlow. It now includes a city park, a state marine preserve, and a residential neighborhood with a few businesses. Our project is to research and delineate the geographical changes over time to this area, beginning with the arrival of Europeans to its shores. The word “arrival” is key to our project, as the various modes of transportation which brought Europeans and then Americans to the area will be used to convey each major shift in the land’s use. The three modes of transportation most responsible for these geographical changes are boat, railroad, and automobile. Boats brought the first European settlers to the area; the railroad helped fulfill Aaron Titlow’s dream of turning the area into a vacation destination; and the invention and subsequent proliferation of the automobile effectively put an end to his dream, transforming the area into a residential suburb. Our research includes comparing the area’s current features to its original Wilton land surveys of 1868; analyzing the methods used to make the area accessible by boat, then railroad, and then automobile; interviewing local long-time residents; and researching state and city data pertaining to the area’s respective marine preserve and park. Of particular interest to us is the ecological impact Titlow’s hotel had on the area, including his litigious influence on the Northern Pacific Railroad’s specific route through his tourist destination, and the three-story hotel’s subsequent transformation to a one-story lodge. We also hope to uncover the ecological impact on the beach that has resulted from the oil-tar-creosote-coated derelict pilings that are still standing after the dismantling of the beach’s ferry terminal.

Storm water runoff has been a growing issue in recent years. Many of us know that the increase of land cover by impermeable surfaces, such as concrete and home shingles, has caused much of this issue. Puyallup was once a vast valley of fertile soil, but is now seen as a fast growing suburban area. The transition between these two points in time explains a lot about the storm water runoff problem the Puyallup valley is now facing. Deforestation was the first step to the storm water runoff problem; before the early settlers of the Puyallup valley could start utilizing the fertile soil of the valley for agriculture, they had to rid of most of the trees in the area. Mono crop agriculture was the second step, depleting the soil of its natural ability to hold itself together. Urbanization, paving and building in particular, are the most recent and relevant to the issue of storm water runoff in the Puyallup valley. I have examined each of these historical events in detail to explain how the use of land in Puyallup corresponds to storm water runoff. My research includes primary source information from some of the first settlers of the Puyallup valley and land surveyors’ reports over time. I have also used GIS maps of the Puyallup area and information from peer reviewed scientific research articles on storm water runoff. My research shows that the change in land use over time has brought us to the storm water runoff issue we face today. The progression through time shows that deforestation, mono crop agriculture, and development shaped the issue of storm water runoff we face today.

Polychlorinated Biphenyls (PCBs) are a group of man-made chemical species that were manufactured as refrigerants the mid 1900’s. Though now banned, PCBs are highly resistant to degradation and persist for many decades in the environment where they are known to have adverse affects on the health of organisms, and are a suspected carcinogen in humans. My research examines the impact of tidal cycles on the re-suspension and transport of PCBs adsorbed on sediments in the Duwamish River estuary. Aquatic systems contaminated with PCBs may lead to accumulation of the toxins in the tissues of aquatic organisms, such as shellfish and endangered Chinook salmon. Consumption of these valuable food resources poses a threat to human safety, and therefore it is critical to understand the spread of PCBs due to tidal oscillations in estuarine systems. In the Lower Duwamish Waterway, a salt wedge estuary with a sharp boundary between fresh and salt water, turbulence created by the relative motion of these bodies of water can move contaminated sediments to new areas. By testing a variety of water quality parameters, including total and dissolved organic carbon content, suspended solid content, salinity, pH, and PCB concentration, my work evaluates the likelihood that elevated PCB concentrations correspond to periods of maximum tidal turbulence and re-suspension of contaminated sediment from the river bottom. Aqueous and solid samples were collected from the estuary at different depths during various stages in the tidal cycle. I have measured key aforementioned water quality parameters according to standard methods, and developed a combined microwave-accelerated solvent and solid-phase extraction procedure for PCB recovery in particulate and aqueous phases. Using these methods, I will quantify the samples’ PCB concentrations using gas chromatography/mass spectrometry to determine sample composition, and attempt to correlate these measurements with suspended sediment and organic carbon concentrations.

Opiates are considered an effective treatment of severe pain for a long time. However, repeated exposure to opiates induces analgesic tolerance, which has been correlated with an increase in opiate associated deaths due to respiratory depression. This project aims to compare and contrast tolerance to opiate-induced analgesia and tolerance to opiate-induced respiratory depression. We hypothesized that the rate of development of the former tolerance phenomenon occurred faster than that of the later tolerance phenomenon. In such a case, patients may be more likely to develop respiratory depression as the dose of opiate is escalated in an attempt to provide continued analgesia. To date, animal tests have been conducted to look at effects on nociception and ventilation produced by various doses of opiates in opiate naive and opiate experienced rats. The nociception testing was conducted through paw withdraw experiments, where the time for rats to move their paws under a hot light indicated analgesic effects, and the ventilation testing was conducted using whole body plethysmography, where ventilation signals of rats were conveniently and accurately measured and recorded. In the ventilation testing, rats’ responses to different ambient carbon dioxide concentrations up to 5% were measured. From the collected data, a potential therapeutic window for morphine was found to be between 5 mg/kg to 15 mg/kg, under which no obvious respiratory depression was observed while above which severe respiratory depression happened and almost no respiratory response to carbon dioxide was noticed. Future results may indicate the relative large discrepancy between the magnitudes of tolerance to opiate-induced analgesia and that of tolerance to opiate-induced respiratory depression. This may further indicate the different mechanisms mediating these two opiate-induced tolerance phenomena, a better understanding of which may allow treatments that make opiates safer for patients with chronic pain.

Photolysis of chlorine under sunlight results in generation of hydroxyl radical. Hydroxyl radical can in turn sensitize chlorine-resistant forms microorganisms such as Bacillus subtilis spores or Cryptospordium spp. occysts, and can make those microorganisms more vulnerable to chlorine. If the photolysis rate of chlorine and quantum yield of hydroxyl radical are known, microorganisms’ exposure to hydroxyl radical and free chlorine during a photochemically-enhanced chlorination process can in turn be derived. It is known that the decay of chlorine follows a first-order decomposition reaction under sunlight. Therefore, the reaction constant and half-life of chlorine can be computed according to first-order reaction equations. Understanding how pH and temperature affect half-life of chlorine can improve accuracy of prediction on required chlorine dosage in order to achieve a certain extent of disinfection in this photochemically enhanced chlorination process. The photolysis rate of chlorine under sunlight was investigated by determining chlorine’s half-life under various temperature and pH. Reaction solutions were irradiated by a Newport-Oriel solar simulator. Chlorine samples were taken at different time points and residual chlorine concentration was measured by DPD method. The results to be reported here showed that at the same temperature, the decay of chlorine was slower in a more acidic solution, which was due to the increasing ratio of [HOCl]/[OCl-]. Temperature also influenced chlorine’s photolysis rate: chlorine decayed more slowly in 5 °C than it did in 25°C. These findings will help to optimize the application of this disinfection process for inactivation of chlorine-resistant microorganisms.

Kaposi’s sarcoma-associated herpesvirus (KSHV) is the etiological agent of Kaposi’s sarcoma (KS), a proliferative and angiogenic tumor commonly associated with AIDS. The predominant cell type in KS tumors is the latently infected spindle cell, which expresses markers of lymphatic and blood endothelium. KSHV infection dramatically alters a number of host pathways, including angiogenesis and lymphangiogenesis. KSHV induces angiogenesis, but the viral gene or genes responsible are unknown. Also, KSHV infection of blood endothelium induces expression of lymphatic endothelium markers. Our data indicate that the viral gene vIL-6 is sufficient, but not necessary, to induce blood to lymphatic endothelial cell reprogramming. The goal of my project is to identify the latent viral gene or genes responsible for both the induction of the angiogenesis and blood to lymphatic endothelial cell reprogramming. My first aim was to determine if any single latent gene is sufficient to induce angiogenesis, through increased expression of the integrin alphaVbeta3 expression, or induce blood to lymphatic differentiation, as indicated by expression of prox-1, the master regulator of lymphatic endothelium. The results indicated that no single latent gene was sufficient to induce either change in expression. My second aim is to express the entire latency locus in endothelial cells to determine whether it is sufficient to induce angiogenesis-related phenotypes and/or lymphatic reprogramming. The KSHV latency locus will be cloned into an adenoviral vector and transfected into blood endothelial cells, which will be evaluated for alphaVbeta3 and prox-1 expression. If the latency locus is sufficient for induction of prox-1 or increased expression of alphaVbeta3, I will next silence each latent gene using siRNA to determine if any of the latent genes are necessary for the phenotypic alteration. My studies will help elucidate mechanisms by which KSHV induces activation of endothelium, and ultimately causes KS.

Antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB) have been detected in many natural and engineered water systems, such as estuaries, surface waters, and drinking and waste water treatment plants. Although many ARB are likely to be inactivated during water treatment, their ARGs may survive treatment by means of protection with cell debris or adsorption on to particulate matter. This is a potential risk for human and animal health, as certain clinically-relevant bacterial populations can undergo horizontal gene transfer by means of natural transformation, which requires only intact, free DNA and a live recipient cell (no donor cell required). These risks can possibly be mitigated through optimization of disinfection processes. However, the current treatment processes focus on deactivating the viable microbes, not the ARGs. Our study focuses on the deactivation and degradation of ARGs in disinfection processes. Our data suggests that current CT (= oxidant concentration in mg/L * time of exposure in min) values for the use of free chlorine (10⁰ to 10¹) and ozone (10^-1 to 10⁰) in disinfection are sufficient to deactivate in vitro ARGs; however the CT values for chloramines (10² to 10³) are likely not able to deactivate the in vitro ARGs. In the case of in vivo ARGs, even higher CT values are likely to required, as cell debris can protect the genes. For example, our data show that a CT value of 90 mg/l*min of chlorine, which is a very high CT value, can deactivate all the cells, but has no effect on the genes inside.

We are studying Earth’s upper atmosphere, one of the least understood atmospheric regions, wind and temperature fields to determine stimuli behind the movement of neutral particles (pushed, heated, interacted with charged particles, etc.). This project focuses on these regions in the Southern Hemisphere, which are essential to understanding movement in all areas of our atmosphere. The climate, long term trends, of the MLT (Mesosphere-Lower Thermosphere) will be studied via instrumentation at the Mount John Observatory (MJO) in New Zealand. The optical observations will provide characteristics, such as wind, kinetic temperature, and emission rates, for groups of species of molecules, which act as tracers for the ambient gas. The time series of motions and temperature from MJO, which provides an understanding of upper atmospheric behavior, extends slightly over one solar cycle. This extensive amount of data can provide us with a broader view into the effects of Global Climate Change. My role is to alter the spectrometer design to enhance the light collection section enabling improved measurements of the atmospheric changes and increased accuracy regarding wind/temperature. Future efforts consist of investigating effects of the lower on the upper atmosphere, and studying relative importance of the processes in action.

Attention Deficit Hyperactivity Disorder (ADHD) is a pediatric disorder of higher executive functioning which may persist into adulthood. This disorder is estimated to affect 5.4 million children aged 4 to 17. Neuroimaging evidence suggests the underlying neuropathology is characterized by a delayed, but otherwise typical, brain maturation process. A previous study analyzed electroencephalograms (EEG) recorded from children (aged 10 to 13) while they were actively anticipating visual stimuli. They found under-connectivity (a) within the frontal lobes and (b) between the frontal and parietal lobes in the ADHD group. These data were consistent with a delay in cortical maturation in the ADHD group. This project builds upon the results of the previous study by examining the eyes closed, resting state-EEG recorded from the same children (42 ADHD and 21 controls) at the same recording session. Cortical maturation is assessed using dense array EEG coherence spectra to characterize functional connectivity differences between groups. We are investigating if the previously demonstrated connectivity differences are static in respect to behavioral state or if these differences are a function of cognitive demands.

Why do smokers with epilepsy suffer from cooking and not cigatte-induced burns? 21% of our patients smoke, yet cigarette-induced burns are uncommon. We studied whether nicotine might change seizure thresholds in patients undergoing video EEG telemetry (VEEG). We tested NP effects on duration of stay, rate of seizure aquisition and number of seizures collected in patients undergoing VEEG. 762 consecutive patients charts were screened for proven epilepsy (EPI) and NP use during their VEEG hospitalization (n=33). Randomized controls, EPI patients without NP (n=33), were also selected. Outcome variables included duration of stay number of seizures collected on day 2, and total number of seizures. T-test comparisons were done. Patients with psychogenic non-epileptic events (PNES) with NP (n=52) were compared to a randomized NP-free PNES group (n=52). The duration of VEEG in EPI patients with NP was 5.42 days (s.d=1.66) vs. 4.33 (s.d=1.74) days in patients without NP (p= 0.01). EPI patients with NP had a total of 3.88 (s.d.=8.82) seizures, while those without had 7.58 (s.d.=1.44, p=0.21). On day 2, 0.94 (s.d.=1.44) events were captured in patients with NP, and 2.18 (s.d.=5.38) events in patients without (p=0.21). PNES patients with NP spent 3.73 days for VEEG, those without 3.98 (p=0.46). No relationships were seen in PNES patients for day 2 or for total events captured. No differences in provider-specific stay durations were seen. In epilepsy subgroups, partial-onset EPI patients without NP stayed 4.41 days, those with NP 5.56 days (p < 0.05). Generalized EPI patients showed no differences. In conclusion, NP use during VEEG may prolong length of hospital stay in EPI patients, but not for those with PNES, this effect may be more notable in patients with partial onset epilepsy.

Reactive halogen species (RHS, e.g., X·, X2·- and HOX), which can be generated during solar irradiation of sea water, can abiotically produce organohalogen compounds by reacting directly with natural organic matter (NOM). In order to obtain a better understanding of the mechanism involved in these systems, a novel technique that allows monitoring RHS production in sea water is required. For this purpose, I have performed a variety of tests to aid in the development of a highly-sensitive method for measuring RHS yields using the probe compound 3,5-dimethylpyrazole (DMPZ), which can rapidly and selectively form monohalogenated products by interacting with certain RHS. In this work, samples of artificial sea water containing DMPZ were irradiated in a 1700-W Xe arc lamp solar simulator for 48 hours. The concentrations of the main photochemical reaction byproducts (4-bromo-3,5-dimethylpyrazole and 4-iodo-3,5-dimethylpyrazole), which are formed in proportion to the concentration of RHS generated during irradiation, were determined by liquid chromatography with mass spectrometry (LC-MS). From this work, we have demonstrated that DMPZ is suitable for use as a probe of RHS. In addition, we have used this method to study the influence different chemical species (NO2-, NO3- , Fe3+, I- and natural organic matter) present in the water matrix on yields of halogenated DMPZ. Results of these experiments will be presented, and discussed with respect to likely relevance for natural environmental systems.

It is well known that mammals contain several types of phospholipase A2. The cytosolic phospholipases A2 (cPLA₂s) are one type, and are composed of six enzymes: cPLA₂alpha, cPLA₂beta, cPLA₂gamma, cPLA₂delta, cPLA₂epsilon, and cPLA₂zeta. There has been significant interest in the cPLA₂alpha isoform because of the enzyme's 10 fold preference for the hydrolysis at the sn-2 position of the glycerol backbone in phospholipids, resulting in the liberation of arachidonic acid. Arachidonic acid serves as a precursor for several highly regulated inflammatory mediators that play an important role in asthma, atherosclerosis, and arthritis. To address these problems, inhibitors that target cPLA₂alpha have been developed to serve as anti-inflammatory therapeutics. Wyeth pharmaceuticals has released a class of indole inhibitors that inhibit cPLA₂zeta at <10 nM IC₅₀. However, a recent study has shown that in cPLA₂alpha^-/- stimulated lung fibroblasts arachidonic acid production is lessened but still pressent. cPLA₂zeta has been identified as the other enzyme involved in the release of arachidonic acid. Consequently, my research project is to develop a selective and potent inhibitor of the cPLA₂zeta isoform. Our synthetic strategy is to modify the scaffold of Wyeth's cPLA₂alpha inhibitor. Analysis of the structure-activity relationships and 3D molecular modeling has revealed 4 different places on the scaffold to introduce substitutions. To date, we have generated and assayed over 20 inhibitors, with many more in progress. These inhibitors display low nanomolar potency, but are not selective for cPLA₂zeta. Our main focus is to increase the selectivity of our inhibitors towards cPLA₂zeta in order to further regulate and understand cPLA₂zeta's role in arachidonic acid production. 

Nitrogen, a limiting nutrient, controls oceanic primary production which directly affects the global carbon cycle. Consequently the nitrogen cycle has great implications for how the oceans will react to climate change. Oxygen Minimum Zones (OMZs) are areas of the ocean characterized by low oxygen levels and as areas of nitrogen removal. OMZs are expected to grow in the future, possibly increasing nitrogen removal from the ocean. Because nitrogen is requirement for primary productivity, the growth of OMZs has potential to slow the biological pump. The Eastern Tropical Pacific (ETNP) OMZ is the largest OMZ in the world, running from 0-25 ° N with a core reaching depths from 280m to 850m.  Anammox (anaerobic ammonium oxidizing) bacteria are of particular interest in the ETNP because their ability to convert fixed nitrogen (ammonium and nitrite) into the form of N₂ gas, which is biologically unavailable. Anammox bacteria produce N₂ autotrophically by oxidizing ammonium in low oxygen environments using nitrite as an electron acceptor. Understanding the presence of anammox bacteria through the water column is crucial to gaining a clear understanding of the nitrogen cycle. Presence/ absence data of anammox bacteria has been constructed using PCR with primers that target the anammox specific Hzo gene. Samples from 5 vertical profiles were collected while aboard the RV Thomas G. Thompson during the oceanography senior thesis cruise from March 16^th to 27^th. Samples were taken at every layer in the water column, including the oxycline, the OMZ core and bottom of the OMZ. During the cruise denitrification rates and oxygen concentrations were determined. Comparing denitrification rates to presence absence data of anammox bacteria will help uncover the role of anammox bacteria as denitrifyers in the ETNP.

Diatoms are unicellular photosynthetic algae, or phytoplankton, and account for approximately 20% of global primary production. The diatom Pseudo-nitzschia can produce a neurotoxin called domoic acid (DA) that builds up in the tissues of shellfish when this diatom blooms. DA poisoning causes life-threatening conditions in mammals and humans when these shellfish are ingested. Parameters such as shellfish exposure length and bloom toxicity can be found by understanding what regulates diatom communities. One mechanism of bloom regulation that we know little about is that of viral infection, despite viruses being the most abundant predator in the ocean. My lab is isolating and characterizing viruses to develop a model system. After a toxic Pseudo-nitzschia bloom at Sunset Beach, Oregon was sampled in 2009, the first Pseudo-nitzschia infecting virus was isolated by infecting the host P. multiseries Clnn-16. This virus was named PmDNAV and was characterized by its burst size, latent period, host range, morphology, and nucleic acid content. I hypothesize that there are many different viruses that can infect diatoms of the genus Pseudo-nitzschia in addition to the PmDNAV. My project focuses on isolating and characterizing more viruses, in order to compare them to the PmDNAV. I have samples from 4 different blooms and 43 different hosts in culture to test these lysates on. I found that the host Clnn-16 dies when inoculated with lysates from the Sunset Beach sampling. I repeated this experiment to prove that my lysates could infect a new healthy host. I am working to prove that this death is due to a virus by looking for small pinpricks of light on a slide after staining the nucleic acids of the putative viruses. After that, I will make the virus clonal and work to characterize this virus, later repeating this process with as many viruses as possible.

Thanks to advances in safety equipment and emergency care, most spinal cord injuries that occur in people today are incomplete, leaving a portion of the spinal cord neural tissue intact. Further, recent research has shown that there is plasticity and axonal sprouting following injury, expanding the possibilities for rehabilitation from what was once considered a permanent and unrecoverable injury. Our goal is to enhance this process and work towards functional recovery. Dopamine, a powerful neurotransmitter involved in processes such as learning, memory and motivation, plays an integral role in brain plasticity. We are currently testing whether pairing dopamine release with functional movements can increase plasticity of the remaining active pathways and lead to increased functional recovery following an injury in the same way that it can form powerful associations between behaviors and reward sensations. Recently concluded tests followed the recoveries of two rats which received dopamine-releasing electrical brain stimulation paired with functional movements of an impaired limb following identical unilateral cervical spinal cord injuries. These rats activated the stimulation by using their impaired forelimb to push a lever which activated their medial forebrain bundle, a key pathway in the mesocorticolimbic dopamine system. One of these animals showed a significant recovery, nearly to full pre-injury movement capability, and much greater than control animals that received no dopamine stimulation (results in the second animal were inconclusive). Ongoing experiments will compare animals receiving dopamine-releasing stimulation paired with movement of the injured limb to control animals where phasic dopamine release is uncorrelated with movement. These experiments may lead directly to novel methods for promoting nervous system recovery from injury by using dopaminergic pathways to promote and reinforce neural plasticity. This may lead to therapies which allow people with incomplete spinal cord injuries to improve functional motor skills.

The Earth’s magnetic field deflects most of the energetic particles contained in solar wind, but some are captured and trapped in regions called radiation belts. Sometimes, high-energy particles migrate from the radiation belts into Earth’s upper atmosphere, where they interact with matter to produce X-rays. Our project focuses on attempting to understand radiation belt loss processes by analyzing the X-ray data. Professor Michael McCarthy and I modeled an existing X-ray detector on the computer and simulated electron populations (having various energy distributions) as they move through the upper atmosphere. We recorded the energy distribution of X-rays captured in the detector. We then compared this distribution with the actual X-ray energy distribution recorded by real experiments. This method of working backwards limits what the energy distribution of the original charged particles may have been. This is significant, because knowing the energy distribution of charged particles in radiation belts and understanding how they move into the upper atmosphere will allow engineers to design spacecraft and satellites that are more resistant to damage from high-energy particles and X-rays. It is also an interesting research question that will teach us about the radiation belts of other planets in the universe that have magnetic fields. Our project will be applied to more experiments; in December 2012, the research team will return to Antarctica to release more balloons equipped with X-ray detectors into the upper atmosphere. Another extension of this work is to model the effects of high-energy protons from solar wind on Earth’s upper atmosphere.

Vashon-Maury Island has always had a fascinating history, involving both its people and the land. As Vashon-Maury Island became more populated around the 1880s, citizens began to recognize the true benefits that the island was able to offer. Industries began to bloom, creating an inordinate economic growth for the community which led to a greater recognition of the island. One of the consequences of the economic boom was that industries were situated throughout the island, especially around Quartermaster Harbor. Research various documents and data have been compiled to understand the relationship between the population impacts on the environment especially around the island. Studies that were being conducted for this experiment included examining literature reviews from previous researchers, and compilation reviews from environmental journals. The conclusion from researchers state that the pollution of bacteria from the waters was not only accumulating directly from industries but also the homes and recreational activities. In order to understand the holistic impact of changes in relation to water pollution scientists took samples of soil and other marine organisms at Quartermaster Harbor and it has been revealed that the water has been significantly polluted over the past 100 years. Bacteria such as Escherichia coli have been recorded as having an immense population growth due to decades of activities. Remarkably, it appears that outdoor activities ranging from boating, picnicking, swimming and other recreational events have led to massive inputs of pollution. By understanding these conditions, future and present researchers are able to educate and inform the public on ways they can prevent future contamination to the harbor. With the right education and deliverance of information the community has the potential to increase the health of the environment on land and water.

There are numerous applications of porous ceramics, including electrodes for solid oxide fuel cell, membranes and filters, and thermal insulation.  Most of these applications utilize uniform isotropic porous ceramics. Directional freeze-casting offers a novel way to make porous ceramics with tailored microstructures. Using this process, followed by partial sintering, we are able to fabricate ceramics with directionally aligned and hierarchical porosity. Freeze-casting involves directional freezing of a liquid suspension and then sublimation of the solvent by freeze drying in order to achieve uni-directional pores. Overall porosity and pore size and alignment can be controlled by several variables including: solvent to powder ratio, freezing rate, and sintering temperature. Our research focuses on two different applications of anisotropic porous zirconia ceramics. One project focuses on creating filters for catalytic hydrogen peroxide decomposition. Hydrogen peroxide (H₂O₂) is an undesirable byproduct and hindrance of a new process to produce chlorine using solar radiation. Porous ceramics provide the means of developing a filter with sufficient surface area and structure for photochemical reactor solutions to be continuously scrubbed of H₂O₂; thereby enabling unhindered chlorine production by this photochemical process at levels sufficient for water purification. The other project focuses on developing a glass-infiltrated reusable ablative ceramic surface for high temperature aeronautical applications. Porous yttrium stabilized zirconia is used as a substrate through which glass can be both initially infiltrated and subsequently drawn to a high temperature surface through capillary action for ablative purposes. At high use temperatures, the glass rises in the capillaries reaching the hot surface where it is ablated. The heating required for ablation reduces the surface temperature of the system.

No commercial mining has been attempted in the glacier basin area surrounding the ghost town of Monte Cristo, Washington since 1920. My research investigates the ecological history of mining in the Monte Cristo area and the impacts which are still evident today. Mining in Monte Cristo has/had three direct impacts on the ecology of its surroundings; runoff, tailings at each mining site, and the discarded “waste” from the mineral concentrator. The ore concentrator is long gone and had not been in operation for nearly a century. However the tailings at the mines remain and water continues to run from the scars ripped into the hillside, infiltrating the flow with pollutants. I did an in-depth historical analysis of newspaper articles along with interpretation of historical photographs to create a clear image of the indirect impacts mining had to the glacier basin area. The ancillary impacts included logging, grading and tunnel building from the railroad construction and building of a complete township. My analysis of historical photographic evidence and the contrasting photos from my recent visit to the area show that the wilderness has regained a foothold by the regrown thick forest and destruction of most of the man-made structures in what was once a booming mining town. Ongoing research shows that the ecological recovery of the Monte Cristo mines is a tenacious battle. Much physical evidence remains yet the environmental disturbances of the Monte Cristo mineral mines are being alleviated by nature.

The Dickinson lab studies the neurobiology of fruit flies. Particularly, we tested the effects of the neurotransmitter octopamine on a fly's ability to respond to visual stimuli while flying and walking. Evidence shows that a set of visually sensitive cells that are thought to be involved in stabilizing reflexes during flight have different responsive properties with and without octopamine. To further investigate the role of octopamine in this system, we looked for optomotor response deficits in flies that do not produce octopamine. We used mutant Drosophila melanogaster flies that no longer have the gene for enzyme tyramine-beta-hydroxylase, which converts tyramine to octopamine. As a result, these flies build up an excess of tyramine and are unable to produce octopamine. We tethered a fly to a metal pin, and placed it an LED light arena, where we presented it with different moving stripe patterns. We placed an infrared light above the fly, and an infrared sensor below it. Infrared light was used because it is outside the visual range of the fly and would not interfere with our intended visual stimuli. The fly cast a shadow on the sensor, from which we computed the wingbeat frequency and amplitude. We used the left and right wingbeat amplitudes to determine the fly's torque, which indicates its turning response. We analyzed this turning response to measure the fly's ability to respond to the visual stimuli. Lastly, we compared the octopamine-null fly's response to wild type flies. These experiments will further our understanding of the role of octopamine in the optomotor response system of fruit flies.

Heart disease is the leading cause of death in the US. Current therapies are limited, and most slow progression of the disease but do not address restoring function. Dr. Regnier’s laboratory is pursuing a novel heart disease treatment. The lab discovered that a nucleotide analog of ATP, 2-deoxy-ATP (dATP), which is produced in heart cells (cardiomyocytes), greatly improves cardiac performance by increasing myofilament contraction. We have established a transgenic mouse line that overexpresses ribonucleotide reductase (RR), the enzyme that converts ATP to dATP. In addition, we have developed viral vectors to overexpress RR specifically in the heart. Both methods of RR overexpression elevate intracellular dATP, improving contractile strength of cardiomyocytes and cardiac function. An added benefit is increased rate of relaxation by dATP, as is the decay of the calcium transient. The calcium transient behavior is primarily governed by proteins of the sarcoplasmic reticulum (SR), which is the storage site for intracellular calcium. I am studying what causes the improved rate of calcium decay. I measure calcium handling protein expression and activity modification via western blots, and measure contraction and relaxation of isolated cardiomyocytes. I am comparing healthy and infarcted hearts from transgenic and viral RR overexpression mice. Mechanical measurements demonstrate the frequency dependence of the amplitude and kinetics of contraction, relaxation, and the calcium transient. Western blots reveal if faster calcium transient decay is due to SR changes in relation to RR overexpression. The infarct hearts have loss of function due to the death of muscle tissue, but we predict that RR overexpression and subsequent increased concentration of dATP will increase myofilament calcium sensitivity, thus rescuing cardiac function and restoring damaged cells to regular contractile strength. RR gene therapy may be a breakthrough treatment of heart trauma if damaged cells are rescued and normal contraction is achieved.

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 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-beta2), an anti-angiogenic cytokine. In addition to six viral genes, KSHV also encodes 17 recently discovered microRNAs (miRNAs) during latent infection that appear to directly contribute to pathogenesis and angiogenesis by targeting host gene expression.  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. We have screened the miRNAs of KSHV to determine if any are involved in the downregulation of TGF-beta2 by KSHV. We found that two KSHV miRNAs, miK-K3 and miR-K8, are sufficient to downregulate TGF-beta2. To determine if these miRNAs act directly on the mRNA of TGF-beta2 as opposed to altering genes that affect its expression, I proposed two aims: (1) use quantitative RT-PCR to determine the levels of TGF-beta2 expression during KSHV infection when miR-K3 and miR-K8 are inhibited by antagomirs, small antisense inhibitors of miRNAs, and (2) determine if miK-K3 and miR-K8 directly bind to the 3ʼ UTR of TGF-beta2 mRNA using luciferase reporter constructs. By identifying specific targets of KSHVʼs miRNAs such as TGF-beta2, we can improve our understanding of how KSHV is able to maintain latent infection and disrupt the normal regulation of cell proliferation. These studies are the first steps to better understanding KSHV pathogenesis and to developing targets that disrupt KSHV replication.