Childhood apraxia of speech (CAS) is a proposed motor speech disorder. At present, there is no universally accepted catalogue of diagnostic criteria for CAS. Peter & Raskind described a motor-based familial speech disorder in two multigenerational families where one or more children had a diagnosis of CAS. The current study attempted a qualitative analysis of errors made by adult members of five multigenerational families (N = 22), including the two families with the previously described motor-based speech disorder subtype, on real word and nonword repetition tasks, rapid automatic naming tasks, and measures of reading. In one family, three biologically related individuals showed slowed alternating, but not repetitive, oral movements and deficits in sequencing during multisyllabic word and nonword repetition and reading, but unimpaired naming speeds. Results are consistent with a familial CAS subtype characterized by multimodal sequencing deficits, which has implications for future diagnosis and treatment of CAS.

Adolescent use of alcohol, tobacco, and illicit drugs is on the rise in the United States. According to the Centers for Disease Control's (CDC) annual National Youth Risk Behavior Survey (NYRBS), nearly fifty percent of all teens have smoked cigarettes at some point and nearly seventy-five percent have tried alcohol. Research has established several factors known to put adolescents at risk for negative behaviors, including family problems, socioeconomic status, and lack of parental supervision. However, fewer studies have examined what factors prevent at-risk adolescent populations from using alcohol, tobacco, and illicit drugs. Many potential factors have been identified as buffers for at-risk adolescent populations; some these are creativity, service learning, and community involvement. But research investigating these factors is still limited in scope. The present study examines relationships between protective factors and risky health behaviors of at-risk adolescents. First, we pilot-tested our measures on emerging adult college students and found that students who reported higher levels of family cohesion reported lower levels of cigarette use, illicit substance use, and drunk driving. However, contrary to our hypothesis, the higher the level of reported creativity, the more students reported illegal drug use. Based on our preliminary findings, we are revising our survey and plan to survey adolescents at a Boise, Idaho community center about their use of alcohol, tobacco, and illicit drugs using the NYRBS. In addition, factors thought to serve as buffers against these risky health behaviors, including the five characteristics of Positive Youth Development, competence, confidence, connection, character, and caring using the Positive Youth Development Inventory (PYDI). We hypothesize that adolescents who possess these five characteristics will be less likely to engage in risky behaviors. We will test this hypothesis using Pearson's r correlations. Using our research findings, we hope to suggest future prevention program offerings to the community center.

Cognitive theorists argue that mental states, including disorders, are generated and maintained by personal, subjective beliefs, and that events can only be appropriately labeled by the individual experiencing them. Additionally, pervasive negative thoughts are a part of both depression and anxiety. Whereas with depression a person is likely to believe they are incompetent, or that life is hopeless, with anxiety a person is likely to have negative thoughts about what might happen or about threats in the future. Additionally, high levels of self-hate are associated with bulimia and high levels of self-control are associated with anorexia. Current research indicates a connection between depression, anxiety, and self-talk (ST). However, studies have yet to examine whether ST in men suffering from depression, anxiety, or disordered eating (DE) differs from that of women. The purpose of this study is to explore potential gender differences in ST (as measured by the automatic thoughts questionnaire-revised and the anxious self-statements questionnaire) with student’s scores on the Beck Depression Inventory, the State-Trait Anxiety Inventory, and the Eating Disorders Inventory. These differences, if found, may help to explain the disparity between the prevalence of depression, anxiety, and DE between genders. Women are twice as likely as men to be diagnosed with depression or anxiety and ten times as likely to be diagnosed with an eating disorder. We hypothesize that there will be differences in ST between men and women, and that these differences in ST will mediate the relation between gender and depression, anxiety, and disordered eating.

Past studies have documented that musical tones separated by an octave interval are perceived as highly similar (“octave equivalence”). A recent study of children showed that they downshifted their fundamental frequency (F0) during a nonword imitation task to match the target, which was below their vocal ranges, at an octave above the target. This was the case for stressed syllables in all children and for unstressed syllables as well in the children without speech sound disorder (SSD). This finding was interpreted as implying that children utilize the octave equivalence effect to enhance their imitations when the target is below their vocal ranges. Here, we expand on this finding by investigating the hypothesis that octave-shifted pitch matching can also be observed in adults. Specifically, we ask whether men match a target spoken by a female adult at an octave below the target, and whether women and children match targets spoken by a male adult an octave above the target. Participants were not asked to match the pitch of the target; they were simply instructed to imitate the nonwords as they heard them. For each participant, imitation/target ratios were calculated and averaged. As in the 2008 study, the role of lexical stress and speech sound disorder (SSD) is addressed as well. Initial results show octave shifting occurred most often with the adult male participants. One woman directly pitch-matched the male target. During the remainder of this project, we will evaluate the role of lexical stress in each participant group and the role of an SSD history in the adults. We will also compare conversational F0 for each participant, drawing on the results from a companion study. Evidence of pitch matching at an octave interval implies the presence of octave based perceptual similarity in speech sounds.

Decision making and risk factors such as media, peers/friends, and family, represent the two main areas that researchers have examined in an effort to understand the cause of disordered eating. Research on decision making has identified a strong tendency toward self-gratifying or disadvantageous decision making among disordered eating populations. Furthermore, persons with disordered eating demonstrate sensitivity towards risk factors as a source for self-perception which in turn plays a large role in binge eating symptomology. However, research on the interaction of these two components with regards to disordered eating is yet unexplored. This gap in understanding was the source of inquiry for my study. Binge Eating Disorder (BED) is the most prevalent of the eating disorders. Being that I did not have access to a sizable BED group, I focused on subthreshold BED (SBED). I hypothesized that (a) SBED participants will demonstrate disadvantageous decision making; (b) SBED participants will place high value on media, peers, and family as a source self-perception; (c) SBED participants’ high sensitivity to media, peers and family promotes self-gratifying DM; (d) Gender differences will be significant with regards to external risk factors, but no difference in decision making. Participants were recruited from introductory psychology classes. An online survey was administered to measure eating attitudes, decision making pattern, and influence of media, peers, and family on binge eating symptomology. I anticipate finding that SBED participants demonstrate decision making impairments and sensitivities to risk factors. Additionally, I anticipate differences in the way risk factors are perceived between genders. Understanding the interaction between decision making and risk factors could lead to possible therapies for and preventive measures against disordered eating. A more in-depth study of clinical and subthreshold disordered eating is needed to ascertain a clearer understanding of the interplay between these two key components of disordered eating.

Learning and memory require that neural activity be flexibly organized. Neurons must sometimes fire together in ensembles so that important signals are not lost in the complex activity of the brain, but these ensembles must also be capable of forming, breaking apart, or reforming at a moment’s notice. Such transient synchrony of neural activity may be achieved by means of oscillatory firing. Oscillatory firing at various frequencies is commonly found in electrophysiological recordings from the brain and may underlie the dynamic organization of neural activity. We investigated oscillatory activity in a brain region critical to learning and memory, the hippocampus. The power of oscillatory activity in the hippocampus at several frequency bands including gamma (30-80 hz) has been linked to successful encoding and recall in hippocampus-dependent memory tasks. Prior work has also shown that inputs from the ventral tegmental area are necessary for normal hippocampal function and for spatial learning and memory. These inputs occur in two forms: tonic baseline firing and intermittent phasic bursts. We tested genetic knockout mice that lack phasic but not tonic inputs from the ventral tegmental area while animals foraged for food on a spatial navigation task. We simultaneously recorded local field potentials in the hippocampus, which represent the net inputs to a region of the brain and allow us to assess the power of oscillatory activity. In the preliminary data, knockout mice demonstrated more rapid learning in response to successive manipulations of the reward location compared to controls. Knockouts also showed higher gamma-frequency but not theta-frequency (4-10 hz) power. These results suggest that phasic inputs from the ventral tegmental area selectively inhibit gamma-frequency oscillations in the hippocampus. This role may be behaviorally relevant, reflecting the possibility that these oscillations play a role in organizing learning and memory.

Traditionally, nonword imitations have been used as a measure of phonological awareness. In a recent study of nonword imitations by children where the targets were below the children’s vocal pitch ranges, it was surprising to find that the children matched the low vocal pitch one octave above the targets (Peter et al., 2008). This was interpreted as a covert awareness of the perceptual similarities between octave-shifted vocal pitches. It was unknown whether children and adults would match pitches directly when the target is within their vocal range. This question is addressed in my current project, which also evaluates the role of lexical stress and speech sound disorder (SSD). SSD occurs when articulation and phonological mistakes continue beyond the point when the child should have mastered the sound through normal development. In a dataset collected for a multigenerational family genetics study, men, women, and children imitated nonwords produced by men and women. Each family in the study had a history of SSD represented in several members of the family. Data analysis is complete for all participants in the study. This consisted of determining the fundamental frequency for each vowel in the non-word for two different imitation tasks (Syllable Repetition Task and Tennessee Test of Rhythm and Intonation Patterns). The participant’s fundamental frequency is compared to the fundamental frequency of the target. Results indicated that the participants showed strong evidence for direct pitch matching in these tasks. I am currently working on determining conversational pitch for each individual to show that the participant is changing his/her pitch to match that of the target voice. In addition, I am determining what effect SSD has on the direct pitch-matching task. Finally, I will also consider the role of stressed verses unstressed syllables has on the expected pitch matching.

Mast cells regulate both inflammatory responses and tissue repair in human diseases but there are conflicting reports on the role of these cells in the pathogenesis of various kidney diseases. Interleukin-6 (IL-6) is an interleukin that acts as both a pro-inflammatory and anti-inflammatory cytokine. From our preliminary studies we have demonstrated that IL-6 and its receptor gp130 are pro-fibrotic in renal fibrogenesis. Here we measure mast cell function in unilateral urethral obstruction, a well-characterized model of renal fibrosis, using mice in which Cre recombinase is expressed under the control of a segment of the carboxypeptidase A3 (Cpa3) promoter bred to IL-6 flox/flox mice. Mast cells from the C57BL/6 Cpa3-IL-6 (fl/fl) mice do not express IL-6. We have groups of 10 for each of the following strains: control1; Cpa3-Cre, control2; IL-6fl/fl and experimental mice; Cpa3-IL-6. Mice under general anesthesia had the ureter ligated at 2 sites distally from the kidney and sacrificed after 14d. We measured the level of fibrosis by measuring hydroxyproline colorimetrically and by measuring area of Picro Sirius red in stained kidney sections, and quantified by pixel analysis. Our preliminary results indicate that the absence of IL-6 in mast cells is important in renal fibrosis. When IL-6 was ablated from mast cells the level of fibrosis measured by collagen and the expansion of extracellular matrix was reduced. These preliminary results suggest that mast cell-derived IL-6 promotes renal fibrosis after unilateral urethral obstruction.

Many occurrences of glomerulonephritis present recurrent acute inflammation episodes. With time these conditions often lead to progressive fibrosis and chronic renal failure. My project studies the role of Interleukin-6 (IL-6), one of the most proinflammatory cytokines, using the nephrotoxic serum (NTS) nephritis mouse model. The ubiquitous expression of IL-6 makes it a challenge to study this interleukin. We have genetically engineered mice with ablated IL-6 in the F4/80+ macrophages and the Ytgax (CD11c+) dendritic cells. We hypothesize a greater level of glomerular damage and disease in the wild type mice compared to the IL-6 depleted mice. IL-6 +/+ (IL6WT) and -/- mice (IL6KO) were injected with NTS in the tail vein and placed in metabolic cages for seven days. Urine was collected to measure renal failure and progression of the disease. Mice were sacrificed at 7 days after the injection and both kidneys harvested and sectioned. Extraction, quantification and quality analysis (RNA integrity number) from the IL6WT and IL6KO frozen kidney samples were conducted and later cDNA synthesized. Quantitative PCR (qPCR) reactions were carried out to analyze gene expression of IL-6 family genes and pathological analysis of frozen or paraffin embedded tissue was done to assess glomerular and interstitial damage. Preliminary results from histopathology analysis and qPCR suggest a significant role played by IL-6 in glomerular damage. IL6WT diseased mice show upregulated expression of IL-6 and greater degree of disease relative to the IL6KO. We plan to continue gene expression quantifications and perform immunostaining to compile more data and validate our results. These findings will help to establish the efficacy of IL-6 neutralization as a therapeutic option for kidney disease.

The Greek sovereign debt crisis has required controversial bailouts, austerity measures that have caused Greek citizens to riot in the streets, and tense political negotiating in the Eurozone. However, it is only one part of a larger problem of economic stability and political unity facing the European Union (EU) today. My research, then, seeks to answer three main questions: (1) what were the causes of the Greek sovereign debt crisis, (2) what are the potential policy solutions to the Greek debt problem, and (3) what are the implications of these policy proposals on the future of the European Union as a whole? Using an institutionalist approach, which emphasizes the role of institutions in determining the rational decisions of political actors, I analyze both the policies of the EU (in bailing out and financially strengthening Greece) and the institutional defects of the Eurozone (i.e., the European Central Bank has limited power and banking regulations are still national). I evaluate proposed solutions according to which would present the greatest benefits for the fewest costs, in terms of democratic legitimacy, economic competitiveness and efficiency, and political feasibility. My research finds that the Greek debt crisis was caused largely by incentives created by the European political environment and by the institutional structure of the EU’s Economic Monetary Union. Thus, to move forward, the EU must take an active role in restructuring its institutions to promote both economic and political convergence. Correspondingly, it must develop strong political leadership, a polity that identifies with the European project, and a democratic process that provides democratic legitimacy without forgoing its technocratic efficiency.

Quorum sensing is cell density-dependent communication that regulates many functions in bacteria. Signal synthases (I) produce acyl-homoserine lactones, which diffuse into the environment and activate signal receptors (R) when they reach a critical concentration. Synthases and receptors often form cognate pairs. The soil bacterium Burkholderia thailandensis encodes three I-R pairs. The I-R1 system controls cell-cell aggregation and activates many putative aggregation factors including the bce genes that likely encode an exopolysaccharide. The I-R2 system controls antibiotic production. The antibiotics are members of the bactobolin family and are effective against both Gram-positive and Gram-negative bacteria. Both aggregation and antibiotics are believed to promote competitiveness in the diverse soil communities where B. thailandensis is found. We have developed genetic screens to learn more about quorum-controlled antibiotics and aggregation in B. thailandensis. The first screen identified bactobolin-resistant isolates of the soil bacterium Bacillis subtilis, which is normally sensitive to bactobolin. We submitted these mutants for whole genome sequencing to investigate the mutations that confer resistance, and with our results we hope to gain insight into how bactobolin acts on target cells. The second screen was designed to isolate variants that overproduce aggregation factors. Quorum sensing mutants do not wrinkle but variants arise that produce aggregation factors by other means. We are isolating such variants and identifying the underlying mutations to understand how they compensate for the loss of quorum sensing regulation. This may reveal the elements involved in regulating aggregation. These variants will also be useful to learn about the advantages of aggregation in different contexts. Through our two screens we hope to better understand quorum-controlled aggregation and antibiotic phenotypes. Our findings will be useful in further studies of the importance of each of these in competition.

Chronic inflammation is a silent killer, contributing to death in patients with chronic kidney disease (CKD) and other chronic disorders, such as diabetes and obesity. Although the specific molecular pathways linking inflammation contribute to CKD pathogenesis still remain unclear, interleukin-6 (IL-6), the prototypic inflammatory biomarker and a potential mediator, is believed to be able to help us understand the mechanism that leads to CKD. We have been investigating the role of IL-6 family of genes using different animal models, such as the Unilateral Ureteral Obstruction (UUO), to induce interstitial fibrosis. The results from our studies have demonstrated that IL-6 and gp130 are profibrotic molecules. Therefore, our goal was to understand the mechanisms of IL-6 in myofibroblasts, the major contributors of collagen during fibrosis. For these studies, we used wild-type mice, IL6 (+/+), IL6 (-/-) and αSMA-Cre-IL-6(fl/fl), in the UUO model of interstitial fibrosis. We hypothesized that IL-6 regulates the crosslinking of collagen by regulating the lysyl-oxidase gene. Therefore, we assessed the Extracellular matrix expansion and collagen accumulation by measuring the changes in gene expression of Col-1, 3, and 4, IL-6, Lysyl-oxidase, Fli-1 and Hells at different time points during renal fibrogenesis. We consider that the change in collagen crosslinking is an important factor in the turnover rate of collagen and it changes its stability to be degraded/accumulated. The examination of the role of IL-6 could potentially provide a new therapeutic breakthrough for treatment and better detection/assessment of renal fibrosis.

The purpose of this study was to investigate one explanation for why religious people have been found to be happier than non-religious people. Given that gratitude has been shown to be an important aspect of psychological well-being, we theorize that an individual’s gratefulness towards God should mediate their happiness due to an increased availability of gratitude expression as well as creating a larger pool of events to be grateful for. We expected this mediation to manifest in the form of a participant memory bias as well as significant contributions to well-being after accounting for their gratitude disposition. In order to test this hypothesis, we analyzed the reported religiosity, well-being, and recalled significant events taking place in the past month of 133 college students. The results from this study seem to indicate that gratefulness towards God may not explain the happiness of religious people beyond their general gratitude.

Gluten intolerance disorders such as Celiac sprue elicit a painful immunological response in the intestines of patients who consume gluten. Gluten is a polypeptide found in wheat, barley, and rye, and contains repeating Pro-Gln-Leu-Pro (PQLP) motifs. Currently the only treatment for gluten intolerance is elimination of gluten from the diet. Our goal is to design an enzyme therapeutic that can be taken as a drug therapeutic in pill form. The engineered enzyme will break down the immunogenic PQLP motifs before gluten reaches the small intestine and can elicit an inflammatory response. Kumamolisin-As, a protein isolated from Alicyclobacillus sendaiensis, demonstrates promising proteolysis on PQ residues, and works in the low pH of the stomach. Using the protein-engineering program “FoldIt,” we designed novel mutations to Kumamolisin to improve its activity on PQLP. Our best version of the enzyme, dubbed KumaMax, degrades gluten over 700 times faster at pH 4 than SC-PEP, an enzyme currently in clinical trials. After further protein characterization, we hope to start clinical trials on KumaMax with the future goal of creating an effective treatment for gluten intolerance.

This project was conducted as part of a larger research endeavor called the Triadic Eye Gaze (TEG) project, a 5-year study funded by the National Institutes of Health. The TEG study is an ongoing investigation into the efficacy of one type of communication intervention for children 10 to 25 months of age with moderate-to-severe motor impairments and delayed expressive communication. Our project's purpose was to compare the routine interventions these children received for early signals of communication to standard clinical practices as recommended in the literature. We specifically wanted to know how much treatment the children were receiving and from which professionals, how much of that early intervention focused on communication, and how parents were involved in the treatment sessions. The population for this study was a convenience sample from the larger TEG study, comprised of 15 children with a range of disorders including cerebral palsy, several genetic syndromes, and general developmental delay. Intervention documents, including Individualized Family Service Plans, daily notes, and attendance records, were obtained for each participant from the appropriate birth-to-three center. Quantitative data reduction methods were used to determine the average number of treatment sessions per month, average minutes of treatment per month, and the proportion of total treatment time that each professional provided. Qualitative methodology was used to determine how much of the intervention focused on early communication signals and how parents were involved in this intervention. Results will be compared to clinical practices recommended in the literature. If we find inconsistencies, it will suggest that the typical treatment these children receive is not sufficient. It will be imperative to conduct further research with a larger, more in-depth study to expand upon these findings.

Homo floresiensis exhibits lower limb morphology that is different from that of H. sapiens, leading some to conclude that gait would be different between the two species. For instance, the ratio of foot to fibular length in H. floresiensis is smaller than the ratio in modern humans, even after differences in body size are accounted. This increased foot length in H. floresiensis is thought to lead to a reduction in energy efficiency. In this study we artificially lengthened the foot of 8 women aged 18-30 years so that their ratio of foot to fibular length was similar to that of H. floresiensis. This was accomplished by creating a foot extension device (FED). Participants walked unshod on a treadmill at five self-selected and randomized velocities. Each velocity was maintained for five minutes, with a four-minute rest period between velocities while the volumetric rate of oxygen consumption (VO2) was assessed with a SensorMedics Vmax29 metabolic cart. We then increased the foot length with the FED and the participants walk at three self-selected velocities. The cost of transport (CoT) curve for each participant was calculated from their normal and FED walking trials. Walking with the FED did not change CoT (p > 0.05). From this result, we conclude that lengthening the foot to the proportion seen in H. floresiensis does not change walking energetics.

Women have been steadily increasing their representation in many careers that have been traditionally male-dominated; however, women continue to be underrepresented in computer science. If women learn that their gender is underrepresented, that could lower their interest or deter them from joining because they believe they do not belong in this field. On the other hand, it could increase their interest if they feel they are wanted. This study examined whether learning about the underrepresentation of women in computer science would increase women's interest in this field. We predicted that women's interest would increase especially if they see it as an opportunity to stand out and be unique. Female students in the psychology subject pool (PSP) at the University of Washington voluntarily signed up to participate in the study. There were two conditions: in the control condition, participants were given an article that stated computer science had high demand and that less than 30% of job openings are filled by college graduates; in the underrepresentation condition, participants were given an article that stated computer science had high demand for women and that less than 30% of college degrees are earned by women. After participants read the article they answered questions about their interest in computer science. Results were statistically significant: women who were told that women are underrepresented in computer science showed more interest and belonging in the field than those who were not told. These results imply that an intervention to raise awareness of underrepresentation of women in computer science could help more females become interested and likely to join this field.

Women who are targets of a positive stereotype (i.e., women are nurturing) assume that the stereotyper also holds negative stereotypes about them (e.g., women are bad at math). The present research investigates whether being the target of a positive stereotype leads women to underperform on a math test because they fear confirming assumed negative perceptions of their group. In this study, female participants were randomly assigned to either the positive stereotype condition (i.e., “why women are generally more nurturing than men”) or the no positive stereotype condition (i.e., “why some men and women are generally more nurturing than others”). Participants then proceeded to take a math test. Results showed that women in the positive stereotype condition underperformed on the math test compared to women in the no positive stereotype condition. This study suggests that praise in the form of a positive stereotype may be counterproductive, resulting in negative consequences for people who are targets of a positive stereotype. Moreover, targets of a positive stereotype may perceive negative stereotypes as being implicitly imposed onto them. Therefore, positive stereotypes may lead women to not only underperform on a math test but also accept associated negative perceptions of their group.

‘Gaydar’ is the phenomenon by which people are able to determine a person’s sexual orientation based on mere observation. Recent research has shown that people can judge sexual orientation with above-chance accuracy from facial photographs. The present research investigated if smiling is a cue that impacts judgments of men’s sexual orientation. Smiling is associated with femininity, and as gay men are generally stereotyped as feminine, we hypothesized that a man smiling would be rated as appearing more gay (vs. straight) than a man who was not smiling. We also predicted that because smiling, a femininity cue, may be less expected on more masculine faces, the impact of smiling will be greater in straight (vs. gay) men’s faces. Participants judged the sexual orientation of a total of 64 gay and straight men’s faces with smiles or neutral facial expressions (using a scale of 1=very gay to 7=very straight). As expected, men were judged as looking more gay when smiling (vs. neutral), and this effect was greater among straight (vs. gay) men’s faces. The findings of this research suggest that subtle gender cues can be conveyed by emotional facial expressions and influence perceptions of sexual orientation.

Asian Americans respond negatively to being the target of a positive stereotype (e.g., Asians are smart) even though these stereotypes are often attempts at praising their group (Siy & Cheryan, under review). The current work focuses on how responses to positive stereotypes vary depending on the race of the person stating the positive stereotype. We hypothesize that Asian Americans will respond more positively to other Asians who state positive stereotypes about their race than Whites who state positive stereotypes about Asians, because Asians are assumed to have "cultural credentials" about their own race. In this case, cultural credentials are defined as the perception that a person has sufficient knowledge about a group to enable them to make comments about that group that outsiders cannot. In this study, Asian American participants imagined a situation in which they were positively stereotyped by either a White classmate or an Asian classmate. Results showed that participants responded more positively to the positive stereotype when the stereotyper was Asian, compared to when the stereotyper was White. Positive responses to Asians who state positive stereotypes about their own race were explained by the belief that Asian stereotypers had stronger cultural credentials. This study shows that taking the time to establish cultural credentials may be a useful strategy for times when one desires to praise a group for their accomplishments.

The Ronald E. McNair Scholars Program’s goal is to introduce first-generation, low-income, under-represented group college students to effective strategies for succeeding in graduate programs. One way to explore program effectiveness is to ask the McNair graduates themselves. This interview study will explore McNair graduates understandings of issues they face in adjusting to graduate school and how McNair participation prepared them for addressing these issues. Typically McNair program evaluations emphasize the collection and analysis of quantitative data – e.g. academic performance and degree attainment; however, little qualitative research has been conducted on graduate’s perceptions of the impact of program participation on graduate school adjustment and success. Using Bourdieu’s concept of cultural capital, along with Sociology-based ideas of role-as-resource and role mastery, this study will explore students’ perceptions of the McNair program’s effectiveness in regards to helping them understand the “graduate student” role and to use that role to succeed in graduate school. 

DNA benefits from a system of self-maintenance, containing genes that code for the repair of its damage. The base-excision repair pathway serves as the primary means of preventing mutations attributed to oxidative stress, such as 8-oxoguanine (8-oxoG), within both the nuclear and mitochondrial DNA. Initiated by the 8-oxogunine DNA glycosylase (OGG1), the removal of 8-oxoG is known to prevent genomic instability and neoplasia. The current study presents new data indicating a role for OGG1 in the maintenance of energy homeostasis at both cellular and whole-body levels. Oxidative stress introduced via high-fat diet (HFD) yielded increased adiposity and hepatic steatosis within the OGG1 knockout (OGG1-/-) mouse model. The HFD-fed OGG1-/- mice were also prone to increased levels of plasma insulin and impaired glucose tolerance. The same mice exhibited a higher resting VCO₂, indicating a preference for carbohydrate metabolism versus fat oxidation. Analysis of expression levels in the livers of OGG1-/- mice revealed significant downregulation of the metabolism-regulating transcriptional coactivator PGC1 and other genes integral to fatty acid oxidation. Consistent with a reduction in PGC1a, multiple genes involved in the TCA cycle were significantly downregulated. Also noted was a reduction in glycogen stores within the OGG1-/- livers, and fasting plasma ketones were significantly reduced. We conclude that OGG1 deficiency alters cellular substrate metabolism, fosters a fat-sparing phenotype, and renders the OGG1-/- mouse highly susceptible to obesity and related diseases. An understanding of OGG1 and its role in energy homeostasis will contribute to the direction of research within the realm of disease initiation due to oxidative stress.

In the United States, there are approximately 50,000 people newly infected with HIV each year. Fear of rejection and other factors related to stigma affect how and when people communicate their HIV status. Research has shown that HIV disclosure through non-face-to-face modes, for example with e-mail and chat, afford benefits that include helping both parties control and process emotions and lessening the chance of violence. To help facilitate the process of an HIV-positive person communicating her or his HIV status in an effective, informative, and non-face-to-face manner, we designed an e-disclosure service called PositiveMagnetism. PositiveMagnetism is a web-supported animated e-card service, allowing the sender to input personalized text and video to convey a message to the recipient. The personalized message is incorporated into an educational video of evidence-based facts and preventative tips regarding HIV - all conveyed in an animated info-graphic form. PositiveMagnetism consists of three parts: a website allowing a person to upload or webcam a personalized video message; an animated e-card video incorporating HIV facts and prevention strategies with the personalized video message; and an invitational e-mail welcoming the recipient of the e-card to view the e-card video. We assembled and collaborated with an advisory board consisting of HIV/AIDS researchers, people living with HIV, and other related professionals to help us throughout the design process. In addition, we conducted two surveys targeting participants who have experienced HIV disclosure. The contributions of this work are design artifacts consisting of a service website, an animated infographic of core supportive information, and a video prototype demonstrating two users (sender and receiver) interacting and utilizing the PositiveMagnetism service. This work contributes to increasing knowledge, awareness, and education regarding HIV to help facilitate HIV disclosure between potential serodiscordant partners.

The formation of amyloid ß (Aß) plaques, resulting from the inappropriate aggregation of the protein beta-amyloid, is a major pathology associated with Alzheimer's disease (AD). One potential therapy for Alzheimer's disease is to reduce cerebral Aß plaque deposition through stem cell therapies. We hypothesize that myeloablative therapy, using lethal irradiation, as well as non-myeloablative bone marrow transplantation, using non-lethal irradiation, will result in a reduction in Aß plaque accumulation in the cerebrum. To test the therapeutic potential of myeloablative and non-myeloablative bone marrow transplantation, we transplanted bone marrow into either lethally irradiated AD (APPswePSIdeltae9) mouse model recipients (myeloablative therapy) or non-lethally irradiated AD mouse model recipients (non-myeloablative therapy). The number of plaques present and the total plaque area are quantified using ImageJ software. Initial immunofluorescence quantification has shown that non-myeloablative bone marrow transplantation reduces total Aß plaque area in the cortex by 33% when compared to non-treatment controls. To further assess the therapeutic potential of both myeloablative and non-myeloablative bone marrow transplantation therapies, additional immunofluorescence, as well as immunohistochemistry, quantification of Aß plaque deposition will be performed. These preliminary results suggest non-myeloablative bone marrow transplantation is a potentially effective stem cell therapy in treating an Alzheimer's related pathology.

Organic matter (OM) production and decomposition is important to wetland health as it serves to cycle nutrients throughout the ecosystem. Current wetland monitoring programs in the Laurentian Great Lakes focus on structural indicators, such as fish, bird, and vegetation communities but lack the use of functional metrics such as organic matter processing. In this study we aim to create a new suite of functional indicators that focus on OM production and decomposition. Decomposition rates were measured using mesh litter bags containing dried bulrush tissue (Schoenoplectus acutus) as well as by the use of cotton strips incubated for 8 weeks at 20 coastal wetlands of Lakes Michigan and Huron. Nutrient limitation of decomposition was also measured using nutrient diffusing bags containing nitrogen, phosphorus, and a combination of nitrogen and phosphorus fertilizers. OM production rates were measured by periphyton biomass accrual on acrylic rods over an 8-week period. In addition, nutrient diffusing substrates were deployed to test the effects of nutrient limitation on algae growth. We predicted that decomposition rates would be positively associated with higher nutrient concentrations. We also predicted that algae growth rates would be greatest on substrates containing high nutrient concentrations. When developed, functional indicators could be used alongside current structural indicators to provide a better overall assessment of wetland health.

Transcranial magnetic stimulation (TMS), the use of electromagnetic induction to generate an electrical current beneath a conductive coil, has proven to be a non-invasive technique that can be applied to cortical areas in the brain to produce evoked action potentials. The behavioral effects of TMS are subject to the frequency, duration, and intensity of the magnetic stimulus being applied, and either long-term potentiation or long-term depression of neurons can be produced. Low-frequency repetitive transcranial magnetic stimulation (rTMS) employs the use of repeated stimulation at 1 Hz, and has been shown to cause a short-term depression of cells. A newly developed protocol, continuous theta-burst stimulation (cTBS), applies three pulses (50Hz) in bursts of triplets, and has been reported to produce stronger long-term depressive effects than low-frequency rTMS. While the effects of both low-frequency rTMS and cTBS have been extensively studied in regards to their effect on motor cortex function, few analyses have been conducted on their effect on language. In the current experiment, we compare the effects of cTBS and low-frequency rTMS on two tests of language comprehension abilities including a sentence comprehension task and a naming task. If language processing can be modified using TMS, one would predict that the response times for the tests of language comprehension would increase, with greater response time increases following cTBS than following low-frequency rTMS. Initial results from this experiment show that pre-post TMS response times for a sentence-comprehension task after low-frequency rTMS is applied to their left hemisphere is decreased, but is increased after cTBS is administered. Opposite results are produced when applied to the right hemisphere, with rTMS increasing response time, and cTBS decreasing it, respectively. Although data collection is still under way, our goal is to determine whether rTMS and cTBS produce significantly different effects on sentence reading and word naming paradigms.

Traditional classical guitar making heavily relies on the use of precious wood and animal materials, many of which are endangered. These materials are illegal to trade under the CITES Treaty (Convention on International Trade in Endangered Species of Wild Flora and Fauna) and the U.S. Lacey Act which have unintentionally shifted material demand to countries of weak legal enforcement, resulting in environmental degradation. I would like to examine different factors involved through a production function, which comprises organization, technology, human capital, physical capital, labor, and natural resources to suggest a better institutional arrangement for classical guitar making. I will also gather historical data on Brazilian Rosewood, an essential material in guitar making. This will include its relative price to alternatives, its users, and the tree population to examine what is at stake. I predict that a better-designed international legal institution will facilitate the process of conservation while redressing the issue of resource limitation. Next, the formation of an international de facto standardizing institution will emerge to facilitate certification and ecolabeling programs, and incentivize sustainable forestry and business through market force. Furthermore, an increase in demand will lead to acoustic and construction technology advancement to remedy resource deficit. Through examining guitar making, I hope to propose a model for sustainable business and to tackle challenges in resource management for an ever increasingly populated world.

According to American cultural tradition, domestic and work responsibilities are typically split between the spouses. The wife satisfies the domestic role and stays at home to care for the house and children while the husband goes out into industry to earn money. But with the recent economic decline, there has been an increase in the number of husbands enacting the domestic role as stay-at-home dads. Like many emerging social profiles, stay-at-home dads are struggling to establish a sense of community and individual identity given their limited networking resources and experience in the domestic domain. Transitioning from a role defined by the status of their job and their level of income, new stay-at-home dads may find it difficult to adjust to their new role. The primary scope of this project is twofold: 1) to create a profile of usability needs for the emerging audience of stay-at-home dads, and 2) to develop an online space for both new and current stay-at-home dads so they can network and share their experiences in the domestic domain. While our contextual inquires and competitive analysis of preexisting stay-at-home dad forums demonstrate a growing audience, our individual interviews with stay-at-home dads confirm the high demand for a user-friendly online space to stay connected with other stay-at-home dads. We report initial findings from research activities focused on designing an online space that meets the expectations of personal and social growth for stay-at-home dads.

When water is placed in contact with a hydrophilic surface and light, a zone clear of all solutes measuring several micrometers long forms at the area of contact. The water in this exclusion zone (EZ) differs from bulk water in that EZ water is negatively charged, more viscous, and highly stabilized with the water molecules being further aligned and constrained. The energy required to generate this zone comes from ultraviolet, visible, and near-infrared wavelengths of light, and while the specific mechanism for their production is unknown, it is hypothesized that light increases hydrogen bonding from the hydrophilic surface into the water, producing the EZ. Metals also contain the necessary properties for generating EZs, and have the potential of producing exclusion zones on a large scale. Current results show that EZs form only when the interface between copper and water is exposed to sunlight, suggesting that exposure to light specifically at the interface might be responsible for producing a photoelectric effect involved with separating the interfacial water into EZ water. Repetitions will be needed to confirm the trend that sunlight light is responsible for producing EZs along copper. A positive result will have significant influence on developing water filtration technologies using just sunlight and copper by harvesting the pure EZ water, fit for drinking. The potential also exists to engineer solar panels using copper and sunlight, and harvesting the stored negative charge in the EZ as a source of energy.

Huntington’s disease (HD) is a genetic neurodegenerative disease caused by a polyglutamine expansion in the protein huntingtin. The age of symptom onset has been correlated with the length of this repeat. Adults with a repeat length of greater than 35 polyglutamines typically present symptoms between the ages of 40 and 50. Greater than 60 polyglutamine repeats causes a rapidly progressing early onset of the disease in adolescents. While both types of HD result in cognitive deficits and neuron loss, 25-35% of early-onset HD patients exhibit epileptic seizures that become less responsive to anti-epileptics over time. One of the earliest molecular changes in HD is the loss of the cannabinoid receptor 1 (CB1) before symptom onset. Activation of CB1 inhibits the release of neurotransmitters and protects neurons from cell death, whereas genetic deletion in HD models exacerbates motor symptoms. Additionally, activation of CB1 prevents the onset of seizures. The goal of this study was to determine if loss of CB1 activation increased epileptic activity, decreased survival, and whether elevating endocannabinoid (CB1 agonists) levels reversed these effects in early-onset HD. R6/2 mice (a rapidly progressing mouse model of early-onset HD) were treated with SR141716A (a CB1 antagonist), WWL123 (elevates the level of endocannabinoids), or vehicle (a control) and the frequency and severity of spontaneous and handling-induced seizures were analyzed. Preliminary results demonstrated that R6/2 mice treated with SR141716A had a decreased lifespan and more frequent seizures whereas WWL123 decreased seizure severity. This suggests that the endocannabinoid system has a regulatory effect on seizures associated with early-onset HD. Future experiments in the lab will further characterize the role of endocannabinoids and CB1 in early-onset HD seizures as a novel therapeutic pathway.

The vestibular system, a complex system involving the inner ear, is responsible for maintaining eye stability and posture during head movements.  Rotational and linear accelerations of the head are sensed by the end organs of the vestibular labyrinth, and are used to initiate and modulate reflexes.  Dysfunction of the vestibular system can produce symptoms ranging from mild dizziness to debilitating attacks of vertigo.  Our group has developed a vestibular prosthesis to replace vestibular function through the use of electrical stimulation. The device has three electrodes which are surgically implanted into the three semicircular canals of vestibular labyrinth to allow for direct electrical stimulation of the end organ. Our ultimate goal is to artificially restore vestibular function in patients, in part to accurately control eye movements.  To do this, we need to understand the relationship between eye position and eye movements during stimulation with the prosthesis. Previous studies in our laboratory have shown that transient eye velocities elicited by short stimulation are linearly related to the initial position of the eye in the orbit.  We need to understand if during prolonged stimulation eye position also has a significant effect on eye velocity. In this study, we characterize the effects of eye position on the maintained velocities elicited by a long stimulus train. These observations will allow us to more accurately control eye movements to stabilize the eye with electrical stimulation of varying durations, or at different frequencies of head rotation. We have trained one rhesus monkey to track a moving visual target.  A long duration stimulus was administered to each of the three semicircular canals in that animal.  All visual stimuli were removed during the stimulations.  Preliminary results suggest that there is a relationship between eye position and eye velocity elicited by long duration stimuli.

Historically, if an individual entered the U.S. without authorization it was not considered a criminal offense. Today the term “illegal immigrant” is commonplace; with many simply assuming this is how it has always been. "Crimmigration”refers to the phenomenon of increasing overlap between criminal and immigration law, procedures, and enforcement mechanisms.Using newspaper articles, this research identifies the percentage share of crimmigration discussion in several major news outlets as a proxy for the climate of public opinion.This study analyzes the changing rhetoric of immigration since WWII. Through an investigation of newspaper articles, this research examines whether economic downturns have contributed to the criminalization of immigration debates in the public sphere. I expect to find that economic factors are important predictors of the prevalance of crimmigration-related articles, after controlling for the other explanations suggested by past research, such as changes in immigration law.

The human body is roughly two-thirds water, much of which interacts with intracellular hydrophilic surfaces, materials that exhibit water-attracting properties. Near any hydrophilic surface such as biological tissue, a zone of water extending a few hundred micrometers from the surface called the “exclusion zone (EZ)” exhibits exclusive and stunning properties far different from those in normal, bulk water. For instance, EZ water does not exist as a solid, liquid, or gas but instead forms a unique, ordered liquid crystalline state that is solute-free, highly organized, and electrically polarized. When EZs are exposed to light, their sizes increase as they absorb and store energy. Cells could utilize this potential energy through growth or other cellular activities. However, it is hypothesized that EZs absorb not only light energy, but also other energy sources such as acoustic energy. To measure the effects of sound on EZs, sheets of hydrated Nafion, a hydrophilic polymer that generates EZs, were submerged in water and exposed to various frequencies and intensities of audible sound produced from an underwater speaker. Microspheres, small spherical particles made from synthetic material, were used to visualize the EZs. Because EZs exclude solutes, the EZs were represented by the areas void of microspheres. During and after exposure to sound, the EZ sizes were monitored over time. Recent results showed that during exposure to multiple audible frequencies (1-8 kHz), the sizes of the EZs decreased transiently. After the sound was removed, in some cases, the size recovered to its original value. These results may suggest enormous biological implications. Everyday sounds produced in our environment could have a large impact on cellular behavior.

Pelvic morphology varies between quadrupedal and bipedal primates due to locomotor behavior.  Flat, posteriorly oriented ilial blades and narrow bi-acetabular distances characterize terrerstrial quadrupeds, while bipeds have curved blades with wider acetabular distance.  While both groups exhibit an orthograde body plan, an ancestal characteristic for arboreality, humans as bipeds have modified this for a terrestrial environment.  Due to the constraints of parturition, human females have been shown to have wider bi-acetabular distances than males, but whether or not their interspinous distance or bi-iliac breadth is different remains to be determined.  The main objective of this study was to evaluate bi-iliac, interspinous, and bi-acetabular breadth in human males and females, using macaques, as the quadrupedal comparison group. Using Osirix imaging software, these pelvic dimensions were measured from the radiographs of 10 human males and females, and 11 adult macaques.  Since macaques only show sexual size dimorphism in the pelvis, we did not distinguish between sexes.  From the average dimensions, three ratios were calculated (bi-iliac breadth/interspinous distance (BI/IS), bi-iliac breadth/bi-acetabular distance (BI/BA), interspinous distance/bi-acetabular distance (IS/BA)). Sex and species differences were examined statistically using Stata (StataCorp, College Station, Texas).  As expected, the ratios in macaques were significantly different than humans (all p’s < 0.001). While human bi-iliac breadth and interspinous distance was greater than bi-acetabular distance (all p’s < 0.001), macaques showed nearly identical proportions in the three measurements.  These findings reflect different locomotor patterns morphologically shown by the transition between vertically oriented iliac blades to more anteriorly wrapped iliac blades.  Between males and females, females differed from men in bi-iliac/interspinous ratio (p = 0.038).  Unexpectedly, the sexes did not differ in the other two ratios.  Like macaques, humans did not show evident sexual dimorphism in relative pelvic proportions.  

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.

The Short-tailed Albatross (Phoebastria albatrus) is listed as endangered under the US Endangered Species Act (ESA) due to small population size, and limited breeding range. The Short-tailed Albatross (STAL) breeds on Torishima and Minami-kojima Islands off Japan and migrates to Alaskan waters during the summer. The NOAA Alaska Fisheries Science Center’s Observer Program, in collaboration with the USFWS, instructs observers to carry out seabird monitoring activities in addition to their recording of bycatch during fishing operations. Using Observer Program seabird sightings and vessel-bird interaction data from 1993-2009, we examined potential vessel interaction threats to the recovery of the population, as well as the frequency and geographic expansion of Short-tailed Albatross sightings. Our data indicate that Short-tailed Albatross sightings increased more than the Short-tailed Albatross population during this time period (6.5% population growth per year), with observer days remaining relatively stable. We also found that incidences of discard feeding and/or vessel following are not increasing disproportionately to population size as the population continues to grow, thus it is unlikely that vessel interactions threaten the STAL population recovery. Lastly, we found that the sightings of Short-tailed Albatross show an eastward expansion since 1993, thus suggesting an expansion of this species within their historic range.

The San Juan Islands of Washington are a unique habitat for both humans and wildlife. Although geographically close to many urban cities, these islands remain relatively rural and have maintained close connections to wildlife. Marine and terrestrial wildlife are very different in their ecology and how humans perceive them. Yet, there are few distinctions between how marine and terrestrial wildlife is generally presented in educational materials for the public. The San Juan Islands are also home to a large, seasonal ecotourism industry. Consequently, there tends to be a high density of tourists in the area, particularly during the summer. I worked with Wolf Hollow Wildlife Rehabilitation Center in Friday Harbor, WA on San Juan Island to evaluate human interactions and attitudes regarding wildlife. For ten weeks I volunteered and used participant observation methods at the facility. I conducted eight semi-structured interviews ranging from 30 to 120 minutes to examine differences in human actions and attitudes towards terrestrial and marine wildlife, as well as any differences in attitude between visitors and residents of the islands. Initial results indicate that, unlike visitors, residents are more likely to view all wildlife as a nuisance. Moreover, marine animals tend to be more appreciated and less intentionally harmed by humans. Based on these findings I evaluated the current outreach materials used by Wolf Hollow and made recommendations for improvements, which included implementing new methods and materials in order to more effectively educate the public about how to live harmoniously with all types of wildlife.

Vascular calcification (VC) is an indicator of atherosclerotic plaque burden and is a strong, independent predictor for future cardiovascular complications, significantly increasing the risk for cardiovascular morbidity and mortality. Type II diabetes (T2D), the 7th leading cause of death in the US, is associated with increased incidence of VC clinically, as well as in our recent animal study. Although VC was considered a degenerative process involving passive accumulation of calcium-phosphate salts associated with tissue necrosis, numerous studies have indicated that VC is an active, cell-mediated process that resembles embryonic bone formation and remodeling. Smooth muscle cells (SMCs) have been implicated in giving rise to bone- and cartilage-like cells in calcifying blood vessels by undergoing lineage reprogramming in response to local environmental cues. We investigated the association of S100A11, a calcium binding protein, with a multi-ligand receptor, the receptor for advanced glycation end products (RAGE), as a possible signaling mechanism involved in the lineage reprogramming of SMCs under T2D settings. We used a genetic fate mapping strategy to trace SM-derived cells in the model T2D mice that developed vascular calcification. Immunofluorescent dual staining for S100A11 and RAGE, along with immunohistochemical staining for Runx2/Cbfa1, an early osteochondrogenic marker, was performed on aortic sections of the T2D mice. The results revealed that S100A11-RAGE signaling is most likely to be involved in cartilage lesion development. This is supported by co-localization of S100A11-RAGE markers within cells of chondrocyte morphology in cartilaginous and calcific areas of blood vessels. Interestingly, almost all S100A11-RAGE positive cells were derived from SMCs that have been labeled by specific transgenes. These results highlight the significance of S100A11-RAGE signaling in VC and possibly provide for the development of preventative or therapeutic targets against T2D-related VC.

Latinos are the largest and fastest-growing majority in Washington State, but over the past thirty years they have been systematically underrepresented in local office. This research focuses on city council and school board races across the ten counties which have the largest Latino populations, and the resulting report is the first to document the severe history of Latino underrepresentation in Washington. Through the same surname analysis used by the Department of Justice in its investigations of racially polarized voting, we analyzed the precinct-by-precint electoral results and found that Latino candidates are routinely defeated by non-Latino candidates. Previous studies have demonstrated that at-large elections - which are used in more than 90% of Washington's local races - tend to dilute minority votes and depress minority representation when minority candidates run against majority candidates. We found that Latinos win far less than half of their races against non-Latinos, suggesting that the extensive use of at-large elections has been combined with racially motivated voting patterns to systematically deny Latino candidates local office. Through interviews with Latino leaders and prominent members of various communities in Yakima County, we also found that Latino communites face more than just structural barriers to effective political participation and proportional representation. Our findings strongly suggest that the Revised Code of Washington ought to be amended to allow local jurisdictions to move away from at-large elections, that local jurisdictions should strive to provide more bilingual services, and that community organizations ought to prioritize the cultivation of local Latino leaders. This report was written as part of the State of the State for Washington Latinos project, which is an ongoing community-based research program at Whitman College. Our research was conducted in partnership with the National Voting Rights Advocacy Initiative.

Decision-making is a complex behavior where one must weigh cost and benefit to maximize the likelihood of the best possible outcome. A common example of this is a decision between a small yet certain reward (e.g. $1 or 1 piece of candy), and a larger uncertain reward (e.g. $10 or 10 pieces of candy 50% of the time). It has been suggested that context and dopamine levels play a critical role in the willingness of decision makers to take risks. We tested this hypothesis by assessing behavioral and neural differences between groups of rats trained on an equivalent risk-based paradigm conducted in different contexts. Rats started training in an operant chamber or a decision maze, and then switched contexts (i.e. tasks). These tasks involved the rat making a decision between two levers or doors, one of which was associated with a certain small reward (1 sugar pellet), and the other which is associated with a large (4 sugar pellets) reward that appeared with decreasing probability (100%, 50%, 25%, 12.5%). To test the influence that dopamine has on risk taking behavior, we used a dopamine antagonist (flupenthixol) to inhibit dopamine in both the maze and operant chamber contexts. Our results suggest that when analyzed as a group, rats are less willing to take risks in the maze context. We hypothesize this is because risk taking on the maze is a more complex behavior, perhaps because of a longer delay between a choice and reward delivery. Our pharmacological findings of high numbers of trials in which levers were not pressed in the chamber, or trials not run on the maze, suggest that blocking dopamine interferes with the ability of rats to make risky decisions. Together, these results support the idea that risk-based decision-making is influenced by many factors, including context and dopamine.

 The vestibular system is composed of the semicircular canal system, which detects rotational movements, and the otolith organ system, which senses linear accelerations and tilt with respect to gravity. The two systems can interact in a complex way during movement. The vestibular-ocular reflex (VOR) stabilizes a retinal image during head movement by commanding the eyes to move in a direction opposite to that of the head movement. Under some conditions, both the otolith organs and the semicircular canals contribute to the VOR. Unilateral or bilateral loss of vestibular function often results in vertigo and oscillopsia, which are debilitating and could be life-threatening because they result in falls or injury. In principle, a vestibular prosthesis could treat a functional loss of inner ear vestibular function by sending electrical stimulation into the three semicircular canals via electrodes to help maintain balance and stable vision. However, if the otolith organs are still functional, the signals from the prosthesis might be interpreted incorrectly in different head orientations, limiting the efficacy of the treatment. The interaction between the output from the otolith organs and artificial electrical stimulations from the implant is not known. In this experiment, we oriented a head-fixed rhesus monkey in different static positions along either of the roll and pitch axes prior to electrically inducing a VOR using the implanted vestibular prosthesis. Results showed that stimulation in the posterior canal induced a higher eye velocity in backward-tilted position than in the upright position, while stimulation in the anterior canal induced a slower eye velocity in the forward position than in the upright position. This project demonstrates that the performance of the implant changes under different body orientations. The prosthesis provides a unique experimental tool allowing us to disambiguate canal and otolith inputs in the intact organism.

Illustrations of the Irish dating from the Victorian era were often layered with political commentary, making it important to differentiate between the art of caricature and that of cartoons. A defining characteristic of caricature is the emphasis on the grotesque, whereas in cartoons it is associated with comedic content. This research will examine the hidden nature of British political cartoons that demeaned the character of the Irish by veiling caricature under the guise of comedy. Through analysis of political caricatures from the mid to late-nineteenth century British publication Punch magazine, I will trace the emergence of the use of crude simian imagery to portray the Irish. The horrific appearance of such caricature had its basis in political suppression and was implemented in order to rationalize British sovereignty over Ireland. The art of caricature was a common tool for cartoonists, such as John Tenniel, to promote stereotypes and false sciences regarding the Irish. These caricatures also have far reaching implications, which went beyond the confines of the United Kingdom. Emigrating along with Irish immigrants, who fled their nation during the Great Famine, the stereotypical depictions of the Irish extended into the United States and even impacted the Kennedy administration. Caricature in America perpetuated the second-class status of Irish immigrants and created obstacles on the path toward assimilation.

Research on resilience (ability to overcome obstacles in life) has shown that early childhood socio-emotional competence is a strong predictor of future resilience as an adolescent and adult. Research shows that resilience is not a stable quality of individuals, but rather a changeable, dynamic pattern of functioning that is influenced by protective factors such as positive relationships, strong attachment bonds, and self-regulation skills. Early childhood intervention programs have been found to make lasting effects on the functioning of individuals into adulthood. Children who have attended preschool programs are less likely to be incarcerated, drop out of school, and placed into special education programs. (Barnett, 2002) However, research on preschool intervention has focused mostly on cognitive and adult outcomes, rather than socio-emotional competence in childhood. This study seeks to learn if preschool programs targeting disadvantaged children, is related to outcomes of protective factors and behavioral concerns, as measured the the DECA-C, an assessment tool becoming widely used to study resilient functioning in children ages 2 to 5 years. The design of this study involves two full-day, demographically-similar preschool classrooms. In addition to comparing children’s scores on the DECA instrument, teachers were audio-taped and observed for 20 hours to identify the quality of teacher-child relationships, a factor known to be very influential to child self-efficacy. It is hypothesized that children that participate in high-quality preschool intervention programs will show higher DECA scores measured over the course of one year. This research has implications for teacher-training, curriculum design, and family programs that have a stronger emphasis on socio-emotional development in low-income early childhood classrooms. This research is also specifically interested in how early childhood intervention can promote resilience to future adverse life experiences for underprivileged children.

The American Farmland Trust estimates that we are losing more than an acre of farm and ranch land every minute. This rapid conversion of farmland to residential and commercial development presents an urgent and dangerous problem, as it severely compromises our ability to grow our own food, and raises serious issues of food security, resiliency, and environmental degradation. However, there are numerous tools that counties can employ to reverse this trend and protect local farmland. Counties are in the driver’s seat when it comes to farmland preservation, and this research looks critically at what counties can do to prevent conversion using the tools of regulation, development rights, tax incentives, and economic development, while highlighting the Puget Sound Basin as a case study for what is working and what needs improvement at the county level. I gathered most of this information in my internship with the American Farmland Trust, where I worked with a graduate student to create a scorecard using the above criteria, and used it to evaluate the farmland protection programs of the 12 counties in the Puget Sound Basin. Among our findings are that total farmland acreage in the Puget Sound is dwindling, that no county reaches the ideal in farmland protection programs, and that farmland across the Sound remains vulnerable. Fortunately, there are a number of counties that are using certain tools very effectively, and counties can draw on one another’s strengths and examples to shape better farmland protection programs across the region.

Neurons contain two highly specialized compartments: the dendrite, which receives synaptic or sensory inputs, and the axon, which relays these signals to other cells. Different types of neurons often have unique, type-specific dendrite morphologies and proper dendrite patterning is important for normal neuronal function. Underscoring the importance of dendrite morphology to neuronal function, defects in dendrite patterning likely contribute to numerous neurological disease states and the cognitive decline that accompanies normal aging. However, the many genes that contribute to the complex dendrite patterns remain largely uncharacterized. To identify genes involved in this process, we conducted a genetic screen for mutants that alter the patterning of sensory neuron dendrites that innervate the body wall of fruit fly larvae. Briefly, we mutagenized fruit flies with EMS, a chemical mutagen, and screened for late-larval stage lethal mutations that perturb dendrite patterning using a fluorescent protein that allowed us to visualize a single functional class of sensory neurons in living larvae. Here, we report our analysis of a novel mutant, messi. Using live cell imaging and quantitative analysis, we found that messi leads to exuberant dendrite branching and intermingling of dendrites in sensory neurons. This latter defect is of particular interest since “self-avoidance” may represent a general mechanism for organizing sensory dendrites and little is known about the genetic basis for this phenomenon. Using time-lapse microscopy, we found that messi causes progressive dendrite defects: the dendrite patterning of messi mutants is indistinguishable from wild type early in development, but becomes abnormal later in development. Similarly, dendrite defects in many neurological disorders are progressive, therefore further analysis of messi may contribute to our understanding of how dendrite maintenance is deregulated in diseased states. Currently, we are working towards identifying the gene affected in messi mutants and determining whether messi affects dendrite patterning in other functional classes of sensory neurons.

At a time in the United States when religious participation by youth has decreased (in a recent Pew Forum report, those born in 1981 or later hold the lowest church attendance) there is a rapidly growing church with roots in Washington: Mars Hill. Practicing a unique brand of evangelism, Mars Hill employs a variety of methods, some of them technological, to attract increasing numbers both in their campuses and online. In order to explore what these methods were, I began attending services and researching the current state of religious participation in American Protestants. I then approached my conception of Mars Hill through the works of various philosophers, including Paul Tillich and Emmanuel Levinas. The third aspect of my work is a journal-based account of my experience throughout this project. My project will reflect the above methods in three main sections: The Numbers, The Theory, and The Experience. I plan to present what it is about the reality of Mars Hill that is fulfilling and attractive for today’s religious population. The reality Mars Hill creates uses novel modes of delivery for its message, which has a high-production value and can be found on Facebook, Twitter, and an Iphone app, among a larger web presence. In addition, Mars Hill offers an experience of community that substantiates strict doctrine on a weekly basis and provides unique accountability for members. This doctrine argues against a conception of Christianity as a “religion”, which moves Mars Hill into a space of new orthodoxy where meaning is born from contradiction in the potentialities embedded in language. My telling of my experience there will demonstrate the difficulty I experienced regarding the dichotomy between participant and observer roles, resulting in a difficult ultimate decision. To uncover the difference in Mars Hill, I started with numbers, continued with theory, and end in experience.

 In 1943 Hanford, Washington became home to the first full scale nuclear production complex and is now the most contaminated nuclear Superfund site. How have the lived experiences, local knowledge, health indicators and perceptions of risk in disparately-impacted communities shaped the complex processes of planning and decision-making of the clean-up at Hanford? Community Based Action Research can provide critical insight into the socio-cultural dimensions of the Hanford site needed to understand the complex processes that shape clean-up efforts, and how those decisions affect the lives of primary stakeholders. This methodology can also be utilized to critically examine how those decisions impact the experiences and health of the larger community. Archival research, participant observation and interviewing have been utilized to identify stakeholder groups and create accessible collaborative descriptive accounts of the Hanford site. This research will add to our collective understanding of the importance and use of cultural resources found on the Federal reservation, the contemporary economic importance of the super fund site, as well as how Hanford has and continues to affect a diverse array of stakeholders. This research has the potential to contribute to understandings of environmental justice policies as well as increase involvement of marginalized stakeholders and facilitate the implementation of collaborative, sustainable solutions to mitigate threats to population health and safety.

The Paul Yager Lab in the University of Washington’s Bioengineering Department has been researching the dynamics of fluids on two dimensional paper networks of nitrocellulose. Knowledge of this science will allow for the development of distributed diagnostics, akin to pregnancy tests, that can be used in remote settings without access to electricity or medical supplies. By testing the storage and flow of various proteins and carbohydrates on nitrocellulose, we are developing lateral flow enzyme-linked immunosorbent assays (ELISAs) for the diagnosis of various diseases. My research involves the control of proteins once they are already on the nitrocellulose membrane. By printing proteins and carbohydrates in various conformations onto the nitrocellulose, we were able to test the effectiveness of our methods for delaying and mixing protein flow. We showed that you can delay protein flow by printing a wall of sugar around the protein that needs to dissolve before the protein can flow with the liquid. We tested the flow patterns of two consecutive patterns of proteins and found that we can predict the time and efficiency with which they arrive at the downstream mixing site. We also developed ways to effectively load up to 1µl of solution onto the nitrocellulose strips. This research will be useful in the eventual development of a working ELISA, when combined with other results from researchers in the lab.

Aminoglycoside antibiotics such as kanamycin and neomycin have been known to cause severe hearing loss in some patients. The Rubel Lab in the UW Department of Otolaryngology has recently reported a molecule, termed PROTO1, which has been shown to protect cochlear hair cells from aminoglycoside induced degradation. However, due to the poor vascularization of the cochlea, it is difficult to deliver ototherapeutic drugs systemically, and thus other methods of delivery must be explored. This study reports on the development of a PROTO1-loaded hydrogel based on the amphiphilic triblock copolymer poly(ethylene oxide)-poly[(r)-3-hydroxybutyrate]-poly(ethylene oxide) (PEO-PHB-PEO). In this study, two cross linking mechanisms were explored. The first uses the oligosaccharide α-cyclodextrin to thread α-cyclodextrin onto the terminal PEO chains of the micelles resulting in a supramolecular structure termed Pluronic/aCD pseudorotaxanes. Hydrogels formulated in this manner showed controlled linear release of PROTO1 over 25 days but are difficult to manipulate and requires sonication and heating to achieve gelation. An ideal hydrogel would be able to undergo in situ gelation, in which the hydrogel components remain liquid prior to being introduced to the active site, upon which they immediately cross-link to form a hydrogel. In this work, we synthesize materials for an alternative approach that is based on crosslinked cyclodextrin-modified polymers. The synthesis and characterization of the polymer components are reported.

The 2007 financial crisis and subsequent recession in the United States has brought the political economy of unsustainable debt to the forefront of the minds of people everywhere in a way not seen in decades.  This paper will delve into the puzzle of why neither dominant political party has been able to institute the recommendations of the four highly regarded debt and deficit reduction commissions in light of their general consensus regarding goals that need to be met in order to avoid a currency crisis.  We ask the question: Why has the Congress of the United States failed to enact measures to reverse the trajectory of the triple deficits of the United States (fiscal, trade, unfunded liabilities) and thereby avoid this crisis?  Our hypothesis is that decades of continued entrenchment of budgetary, trade, and social policies led to gridlock.  We will investigate the coalitions that have formed around the aging baby boomer population, national health care, and national defense.  First, we will discuss the recommendations of the various commissions.  Second, we will review the literature regarding the interests and coalitions within representative bodies contending with looming fiscal imbalances. Third, we will analyze the interest groups, coalitions, and beneficiaries of inaction at this particular time and relate this to the previous literature review.  Finally we speculate as to what would be necessary for congress to act, and what are the likely scenarios if congress fails to act. Having done so, it is our hope that we can provide policy makers and constituents with a deeper understanding of the political-economic problem that the country faces.

 Galaxy formation theories call for all galaxies to be embedded in massive extended dark matter halos. Ideally, we would use thousands to millions of tracer particles to map the dynamical environment around galaxies to constrain the properties of dark matter. Reality limits us to using only the dynamical tracers that are luminous enough to actually be observed. The spiral galaxy NGC 4258 has numerous small galaxies surrounding it that appear to be satellite galaxies, and can therefore be used as dynamical tracers. To determine if these smaller galaxies are indeed satellites, we examine their redshifts by combining spectroscopic observations from the APO 3.5-meter telescope with SDSS spectra. Of the 33 observed objects, 22 appear to be members of the system. Using the projected line-of-sight velocities of the satellite galaxies and a Jeans equation approximation, we estimate the mass of NGC 4258 out to a radius of 260kpc to be 6x10¹² solar masses with an error of 50%. When utilizing a more rigorous set of criteria to determine which candidate galaxies are actually satellites, we are left with only 8 true satellites and derive a new mass to be 1.3x10¹² solar masses out to a radius of 230kpc. Further observations are expected to yield around 20 to 40 more satellite velocity measurements, which will improve our mass estimate and significantly reduce our error. By searching the SDSS database, we believe we can find other galaxy systems that host enough satellites to repeat this technique.

M dwarf stars, which are smaller, dimmer, and redder than our Sun, are the most common type of star in our galaxy. Like our Sun, these stars exhibit activity due to inherently strong magnetic fields at their surfaces. In M dwarfs, this activity is often classified by the emission line strength of the first transition of the hydrogen Balmer series (H alpha). We examine a sample of five equal-mass M dwarf wide binaries; these “twin” stars should have nearly identical stellar properties and therefore have very similar spectral features. We hypothesize that these stars should exhibit nearly identical properties in H alpha. If our hypothesis proves false, it means there are unexpected dynamics present in the binary system. In initial observations to classify these objects, we found that the M dwarfs in each binary, which were twins in every other way, showed very different levels of H alpha emission. To examine the range of H alpha line strengths we obtained a time series of optical spectra using the Astrophysical Research Consortium’s 3.5-meter telescope at Apache Point Observatory in New Mexico. In each binary, the slightly less massive star exhibits stronger mean H alpha, consistent with large survey results. Using data from 70 hours over 14 nights, we determined our stars have a characteristic variability timescale of around 2 hours; indicating our sample is active on shorter timescales than the typical rotational period.

Exoplanets are planets outside our solar system. As they do not emit light of their own, these objects have historically been much harder to find and study than stars. One detection method exploits the slight dimming of a star caused by a planet passing across its face. In astronomy, these events are termed transits, and they cause distinct u-shaped dips in the lightcurve—a plot of observed brightness versus time—of their host star. In recent years, the Kepler Space Telescope has provided lightcurve data on a fixed group of stars that is unprecedented in precision. Our research is primarily concerned with searching released Kepler data for previously undiscovered exoplanets. Due to the vast quantity of information, we have developed a pipeline of computer code to expedite the search process. The code, written in python, detrends variations in quiescent star brightness to normalize each lightcurve around a fixed, flat value. This readies the data for input to a Quasi-Periodic Automated Transit Search algorithm (QATS). Thus far, we have identified a handful of planet candidates as well as a number of undocumented binary star systems. Current and future tasks involve automating QATS output classification to further improve the efficiency of the planet finding process.

The effects of amphetamine on learning and memory are important areas of research due to the widespread use of amphetamine among people, ranging from prescription to recreational. It has been shown in animals that repeated treatment of amphetamine will result in sensitization to the drug. Sensitization is a process through which treated subjects develop an elevated response to a drug. Long term potentiation (LTP) is a physical change strengthening the connection between synapses that is thought to underlie learning and memory. Enhancement of LTP in the striatum has been shown to occur as a result of amphetamine use. The striatum is a brain region important in mediating a large number of behaviors including motor control and learning. For this reason it is possible that sensitization to amphetamine may affect learning in mice. To investigate this possibility, wild-type mice sensitized to amphetamine were tested in behavioral paradigms designed to measure learning ability. It was found that they do not show enhanced performance in motor learning, appetitive learning, or spatial learning, but they may show an improvement over controls when they are required to alter their strategy to escape from a water-maze. In order to explore the neurological pathway through which sensitization to amphetamine may affect learning, we also tested knockout mice that lack N-methyl-D–aspartate-type glutamate receptors (NMDARs) in medium spiny neurons, the primary neuron type in the striatum. In many brain regions, NMDARs are important for various forms of learning and memory, and it has been shown that these knockout mice display impaired motor and appetitive learning and yet are still able to sensitize to amphetamine. Using the same learning paradigms as with our wild-type mice, we found that knockout mice did not show any enhancement in motor or appetitive learning as a result of amphetamine sensitization.

All cells including bacteria exhibit an intricate spatial organization that is essential for cellular function. The mechanisms by which this ultra structural organization is established and maintained are not yet understood. The resolution limit of conventional wide-field fluorescence microscopy is set to 250nm by the diffraction limit, however the building blocks cellular ultra-structure, protein, are typically a few nanometers in size. Therefore, in order to probe the cell-machinery that gives rise to the organization of Escherichia coli chromosome, I use a super resolution fluorescence microscopy technique called PhotoActivation Localization Microscopy (PALM). I am able to resolve DNA structures with a resolution near ~20nm by labeling short sections of E. coli chromosome with a photoactivatable fluorescent protein. I then relate the measured size of chromosome sections to known physical parameters of DNA in order to asses the viability of various polymer models of DNA, which will lead to a better understanding of the physical factors that lead to structure in E. coli.

Bacterial conjugation is a form of horizontal gene transfer that serves as an important source of genetic plasticity and diversity in both Gram positive and Gram negative bacteria. It has also been proved to be advantageous for dissemination of antibiotic resistance. Conjugation is a plasmid-mediated mechanism that allows single stranded DNA to be transferred across cell membranes from a donor to a recipient cell. In Escherichia coli, the conjugative F (fertility) plasmid encodes all necessary proteins for assembling the transfer apparatus thus allowing transfer and propagation of DNA into a recipient bacterial cell. Previous conflicting studies of bacterial conjugation in E. coli have shown DNA transfer to occur between donor and recipient pairs in intimate cell-cell contact or between distant donor and recipient pairs, connected only by the pilus, a filamentous protein extended from the donor cell. Using various fluorescent protein constructs to differentially mark donor and recipient cells and F plasmid DNA itself, we visualize conjugation in real time with single-cell resolution by fluorescent microscopy. Here we show that conjugation occurs only when donor and recipient cells are in intimate and close physical contact with each other. In addition, there appears to be no preference for cellular orientation for transfer. Furthermore, we propose a simple rate equation to model DNA transfer between bacteria and measure the rate of DNA transfer per unit time per cell contact. This quantitative model is a tool to elucidate conjugation at a single-cell scale and will provide a deeper understanding into the mechanism of bacterial conjugation.

Centromeres in many species are found to reside near origins of replication that are activated early in S-phase, yet the biological significance of this conserved early replication of centromeres is unknown. Previous research has shown that the activation time of an origin is affected by its chromosomal location. A recent study has found that centromeres in the yeast Saccharomyces cerevisiae advance the activation time of nearby origins, and that this effect depends on the centromere’s ability to establish a functional kinetochore. These observations lead to the hypothesis that centromeres need to be replicated early to allow sufficient time for the assembly of functional kinetochores. If kinetochores fail to be established in a timely fashion, chromosomes may not be correctly bi-orient at mitosis. Daughter cells therefore may inherit the incorrect number of chromosomes creating conditions of aneuploidy such as those found in Down syndrome and many cancers. I will test the importance of early centromere replication by deleting origins immediately flanking a yeast centromere to delay that centromere’s replication. I will then conduct an assay to test for the rate of chromosome mis-segregation events in the mutant strain compared to the wild-type strain. I anticipate that the mutant strain may have a higher rate of loss for the modified chromosome, suggesting that early replication of centromeres is important for maintaining genome stability. This project will also provide insights into the regulation of the temporal program of replication origin activation and provide the first functional test for the consequences of disrupting the temporal program of chromosome replication.

Vitamin A, whose active form in our body is all-trans-retinoic acid (atRA), is essential for normal embryonic development and many other biological functions. CYP26A1 is an isoform of cytochrome P450 enzyme which is responsible for atRA metabolism and elimination. Since we cannot synthesize vitamin A on our own, humans must consume vitamin A as nutrients. Because both too much and too little intake of the vitamin are detrimental, a tightly regulated feedback system controls the level of vitamin A circulating in the body. CYP26A1, which is shown to be strongly inducible by atRA, takes a major role in modifying atRA into its inactive forms. Common aryl hydrocarbon environmental toxins are also known to induce the activity of CYP26A1 by binding to aryl hydrocarbon receptor (AHR). My current work aims to increase the understanding of the effects of environmental toxin on CYP26A1 in comparison to its other isoforms. I treated HepG2 cell line with varying concentration of 3-Methylcholanthrene (3MC) for 24 hours and quantified the amount of CYP26A1 mRNA. The results show that 3MC induces CYP26A1 transcription up to a certain toxic concentration. With better understanding of how common environmental toxins affect the health of an individual at a molecular level, it is possible to mediate the effects of the environmental toxins.

Ubiquitin is a small post-translational signaling molecule. Its roles in repairing damaged DNA, apoptosis and protein degradation make it critical for tumor suppression. Genetic mutations affecting proteins in the ubiquitination pathway have been implicated in multiple inherited human cancers. The ubiquitination pathway attaches the protein ubiquitin (Ub) onto a substrate molecule with the help of three enzymes, ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2), and ubiquitin ligase (E3). Transfer of Ub onto the substrate requires the formation of a thioester bond between Ub and E2, denoted as E2~Ub. A catalytically active E3 ligase which can bind E2~Ub, but never directly binds Ub, is also necessary. E3’s catalytic mechanism is not fully understood. One function of E3 is to bring E2~Ub and substrate in close enough proximity to facilitate a reaction. However, E3 enhances the release of Ub from E2~Ub in the absence of substrate molecules. This suggests E3 plays a role in priming E2~Ub for transfer of Ub. Using site directed mutagenesis and protein-protein interaction assays visualized through western blotting, I have investigated what electrostatic properties of Ub are important for E3’s catalytic activity. Informative Ub mutations don’t stop formation of E2~Ub, but disrupt the catalytic activity of E3. I propose that polar/charged Ub residues in the contact region between Ub and E2 affect E2~Ub binding modes, promoting inhabitance of E2~Ub conformations favorable to Ub transfer upon binding of E3.

MicroRNAs (miRNAs) are small noncoding RNA molecules that post-transcriptionally regulate gene expression. MiRNA biogenesis involves two key ribonucleases, Drosha and Dicer. I am investigating the Drosha enzyme. Previous research from the Ruohola-Baker lab reveals that reducing levels of Drosha in stem cells significantly reduces levels of some miRNAs, while only slightly reducing levels of another group of miRNAs. This surprising result led us to conduct a bioinformatics analysis to identify characteristics that differentiate these two groups of miRNAs. We discovered structural differences, specifically that bulges in the central region of miRNAs were more frequent in heavily reduced miRNAs than in miRNAs that were not affected. My mentor, Henrik Sperber and I propose a model explaining why miRNAs with mismatches (bulges) in the central region of the sequence are more sensitive to reduced levels of Drosha than rigid miRNAs with no mismatches in that region. To test whether a bulge causes differential miRNA processing when Drosha is the limiting factor, I am adding and removing bulges to primary miRNA transcripts, overexpressing them in cells with low Drosha levels, and comparing their mature levels using qPCR. In addition, I am testing whether in nature cells vary Drosha levels as a mode of regulation. A previous study shows that T cells at specific stages of differentiation reduce Drosha expression by as much as 10 fold. My own bioinformatics analysis reveals that certain tissues in mice have an 8-fold reduction of Drosha. To see whether reduction of Drosha levels correlates with differential microRNA processing, I am using microarray data to find subsets of microRNAs that have dramatically reduced levels when Drosha expression is significantly low. With my research, I hope to discover a potentially novel mechanism of regulation in which cells alter Drosha levels to selectively process a subgroup of miRNAs.

Schizophrenia is a devastating neurodevelopmental disorder whose genetic influences continue to be elusive. Rare, gene-disrupting genomic deletions and duplications – called copy number variants (CNVs) – have been implicated in schizophrenia; however, much remains to be understood about which genes are causative, as well as the cellular mechanisms involved. Biological follow up of individual CNVs may give insight to the origin of schizophrenia. We focused on a CNV present in a patient with early onset schizophrenia. This mutation duplicates the 5' ends of two genes that lie head-to-head on chromosome 11q22: DCUN1D5, a previously uncharacterized gene predicted to be involved in cullin neddylation of ubiquitin ligase complexes, and DYNC2H1, a dynein active in cilia. Both are expressed in brain and are plausible candidate genes for schizophrenia. Using RNAseq, we detected novel DCUN1D5 transcripts in the patient’s lymphoblasts that we predict will result in aberrant, truncated DCUN1D5 protein. To test the conformational stability of these aberrant proteins, we then expressed epitope-tagged full length and truncated DCUN1D5 in HEK293 cell lines. These transfected cells also allow us to study the functional capabilities of the aberrant proteins in the cullin neddylation pathway, and the specific cullin binding partners of DCUN1D5. Investigation of these aberrant proteins and their function may shed light on pathways that contribute to schizophrenia, potentially guiding the search for new candidate genes and the development of novel treatment strategies.

   The stereoscopic image has risen and fallen in commercial popularity since its concepton in the late 19th century, but recent advances in computing and image technology have extended the medium's accessibility and popularity once more. At the UW, in the DXARTS department, I have spent the last year and a half examining the technical and aesthetic properties in stereoscopic pictures. My research and experiments have approached issues in projection surfaces, methods of compositing, graphics, and editing, and technical solutions for live-action stereo-photography. In addition to these practical issues, I also aim to fit my practice into a longer lineage of art-tradition, citing artists who have found the medium to be a fertile ground of expression. Today, my own efforts culminate into a single project as I plan a new work in stereo, and I hope to share video and still images of my experiments and completed works with the students at the University.

A profound irony underscored the Cold War-era U.S. government’s clandestine support of American dance and film; the main innovators in these forms were Russian immigrants themselves. One of these was Maya Deren, a largely unacknowledged avant-garde filmmaker, theorist, and creator of the “dance for camera” genre. She was born Eleanora Derenkowsky in Kiev, Ukraine and immigrated to the U.S. in 1922. Defying mainstream Hollywood trends, which followed linear, cause-and-effect story arcs, Deren created unique “dance for camera” films that emphasized the movements of the dancer and the camera. Her films created dances that could not exist in a live performance by utilizing film’s ability to cut instantly between scenes and play footage in reverse. Deren viewed film as “an art which makes its statement in terms of time and movement. This medium is not so much concerned with any single instant as with the change from instant to instant.” A dancer would begin a movement in one scene and complete the same movement in a different space entirely. In a single moment, the dancer could traverse space and time. By using such film tools, Deren expanded the limits of the human body.  Though many films today reflect her influence, mainstream film theory histories ignore Deren’s contributions. By analyzing Deren’s A Study in Choreography for the Camera (1945) using her own writings as well as contemporary film theory, I attempt to recuperate Deren to film theory history.

Little is known about the 19th Century painter Francois Leupalle and his 1832, oil on canvas painting, Marie Taglioni and Joseph Mazilier in the First Scene of La Sylphide. Despite Leupalle’s obscurity, many Romantic Ballet history books reproduce the painter’s La Sylphide as visual evidence for Romantic Ballet. Leupalle’s painting, then, represents not only La Sylphide but also the Romantic ballerina. In the 19th Century, the ballerina usurped the danseur and reached celebrity status. Through her pointe work, Taglioni helped to facilitate this radical shift. If her pointe work symbolized this sea change, why then does Leupalle illustrate Taglioni barefoot? Lithographs of this period predominantly portray dancing ballerinas, framing their pointe work; yet Leupalle’s barefoot ballerina kneels beside a kilted Scotsman, who slumbers in an armchair, surrounded by a vista of mist, light and foliage. This paper examines how Leupalle’s choice to portray Taglioni as barefoot affects scholars’ use of the painting as visual evidence for Romantic Ballet. Through analysis of the painting and archival documents, this research critiques the use of this image as visual evidence of the Romantic ballerina using male gaze and visual pleasure.

Do we embody our social identities? It is well known that men and women are taught to speak differently, but they also learn to move differently. Like in spoken language, differences exist between both individuals and groups of people in how they communicate non-verbally. Men and women have generally distinct sex differences which result from acculturation; how much is a result of biological factors and how do these overlap? I will undertake research that examines how sex is perceived based on movement alone. Using Kinect technology to document subjects performing movement prompts in the form of gender-ambiguous animations, I will solicit responses from viewers of the animations to determine how accurately the viewer judges the sex of the mover. I will measure the accuracy of their perceptions against several factors, which might influence gender performance including: the dance-experience of the mover, the music played for the mover, the movement prompt, and whether the mover is actively trying to embody a particular gender. This research will contribute to research on non-verbal expression of identity and inform representations of gender in the performing arts.

Somali women immigrants have significant trauma histories related to their experiences of torture, rape, and displacement. They commonly present with somatic complaints for which there is no "medical explanation" and few acceptable or helpful treatment options. A community health program, Daryel, was started 3 years ago by a nurse at Harborview Medical Center to address somatic pain among Somali women through yoga and massage. Daryel primarily serves women over 50 years of age who are not fluent in English. This project consists of preparatory work for the development of a culturally congruent training manual for Health Care Practitioners working with Somali women. The following three methods will guide training manual development: 1) Conducting a literature review using databases such as ETHNOMED, CINHAL, PUBMED and gathering background on Somali traditional medicine, beliefs, and the cultural practices of Muslim women with regard to health; 2) Participating in individual conversations with Somali women and the Health Care Practitioners who serve them; and 3) Outreach to massage therapists who have stopped volunteering at Daryel with a set of questions on their volunteer experience. The results: 1) Findings based on the literature review show that taking time to establish rapport, understanding Somali culture and traditions, listening to their explanatory model of illness, and working skillfully with interpreters are important factors that will help gain success with Somali women patients; 2) Conversations with Somali women in Seattle confirm that a program such as Daryel is helpful for Somali women immigrants; 3) A survey of past and current volunteer Massage Therapists showed they want more training on Somali culture and language. In conclusion, a culturally congruent training manual will aid Health Care Practitioners in providing essential quality care to Somali women immigrants.

 Postcards, as the modern Western world knows them, function as tools for communication, providing evidence of a supposedly successful touristic experience. Acting as an essentialized idea of a location, postcards are bought as an artifact by tourists to qualify their experience yet are inherently misleading in not only their apparent sense of direct contact with the peoples of a different location but with their supposed depiction of photographic reality.By extensively analyzing modern literature regarding postcards, anthropological literature on tourist motivations,and hundreds of individual postcards, this research examines how the work of 20th century British photographer John Hinde depicts Ireland as a fantasy, thereby transcending the nature of the already illusory postcard to altogether loftier heights of deception. By making deliberate, significant enhancements and manipulations in composition, color, and subject, Hinde created postcards of an idyllic Ireland that amplified the unreal in the name of both economy and pleasure. Hinde’s postcards were especially problematic because of the ultimate illusion they constructed. This imagined fantasy as constructed by Hinde further disconnects the tourist from the reality of the place they are visiting, allowing their use of the false images as evidence, ultimately reflecting the inevitable failure of the tourist: to really understand the place they are visiting. The significance of this research lies in examining the futility of being a tourist and how visual representation can further complicate cross-cultural experiences. It is important for all people, tourists in particular, to question and become aware of how images can influence their perceptions and motivations.

The diatoms are a prolific and productive group of phytoplankton belonging to the Phylum Heterokontophyta. They are responsible for ~23.5% of global primary production and their high photosynthetic capabilities depend on their abilities to synthesize chlorophyll. A key step in the biosynthesis of chlorophyll is catalyzed by the enzyme 'light-dependent protochlorophyllide oxidoreductase’ (POR). Early in the evolution of diatoms, the gene encoding this enzyme (por) was duplicated, resulting in two homologous but later divergent genes: por1 and por2. We aim to create a comprehensive heterokont por gene phylogeny to determine whether the gene duplication event occurred before diatoms diverged from sister taxa in the Heterokontophyta, early in diatom evolution, or in only one diatom lineage. Additionally, this phylogeny will allow us to determine the effects of natural selection in shaping the evolution of these genes. Preliminary studies suggest that positive selection acted in the past on both por1 and por2, though these protein sequences are now conserved within diatoms. To create this por gene phylogeny, we will sequence the por genes of various organisms representative of the major orders within the diatom class, Bacillariophyceae. Additionally, we have successfully sequenced numerous representatives of other Heterokont classes that will serve as the background to the Bacillariophyceaen gene phylogeny. These studies require the cultivation of various strains of algae, DNA extraction, PCR with por specific primers, and gene sequencing.

The volume of glaciers and ice sheets controls global sea level and plays a major role in climate change. However, it is difficult to reconstruct the history of glaciers and ice sheets from their deposits because these deposits are commonly overrun and re-worked in each successive glaciation. An alternative approach is to measure cosmic-ray-produced nuclides, such as isotopes Beryllium-10, Aluminium-26, and Chlorine-36 (in order of decreasing half-life) in bedrock. These nuclide concentrations build-up due to cosmic ray exposure during interglacial periods when glaciers are at their smallest extent. The present day concentrations are the cumulative result of exposure during ice-free periods and radioactive decay during periods of ice cover. By working with nuclides of different half-lives, we can obtain information about the long-term glacial history. We are measuring these nuclides in samples from elevation transects on mountains beside Scott Glacier and Beardmore Glacier in Antarctica. Concentrations of all three increase with elevation. Summit samples have Be-10 and Al-26 concentrations close to saturation levels, but lower Cl-36 concentrations. This suggests that ice cover during glacial periods was thicker and more prolonged in the last 0.5 – 1 Myr than the 2-5 Myr average. In contrast, samples from the base of these mountains have lower and more uniform concentrations of the three nuclides, which is consistent with glacial erosion of the surfaces. Measuring depth profiles in the Beardmore Glacier samples will allow us to test this interpretation.

Motivated by recent interest in developing compact, inexpensive, and reliable eye-safe lasers, the effect of quantum confinement on a saturable absorber material, cobalt-doped zinc selenide (Co²⁺:ZnSe), was explored in colloidal quantum dots. Co²⁺:ZnSe diluted magnetic semiconductor quantum dots (DMS-QDs) are semiconducting nanocrystals with structural lattices that include substitutional magnetic ions in addition to the primary metal cations. While previous work has been conducted on the saturability of optical absorption in bulk Co²⁺:ZnSe, the research presented here focuses entirely on quantum-confined colloidal DMS-QDs. The ability of these particular DMS-QDs to behave as saturable absorbers was explored in depth. This property is directly related to the long lived ⁴T₂(F) excited state of the Co²⁺ dopants, which gives rise to bleaching of the Co²⁺ ligand field transitions, leading to increased transmission of light upon irradiation at high powers. Specifically, absorbance features associated with excitation of ground-state electrons localized in the d-orbitals of the dopants are lost. To examine the saturable absorber behavior of Co²⁺:ZnSe, we looked at the optical transmittance of the quantum dots as a function of photoexcitation power. This was done by slowly increasing the power of a pump laser exciting the sample and observing changes in the absorbance spectrum at characteristic Co²⁺ ligand-field transition features. The bleaching of the ligand field Co²⁺:ZnSe in its nanocrystal form demonstrates that these colloidal materials are candidates for saturable absorbers in Q-switched eye-safe lasers. From this point, it should also be possible to explore excitation of the electrons from the first excited state (⁴T₂(F)) into higher energy states (excited-state absorption).

The longevity of an organism is influenced both by internal (genetic) and external (environmental) factors. With respect to internal factors, a significant effort is being made to identify novel pharmacological agents that extend lifespan by targeting genetic pathways with a defined role in the aging process. On the external side, the molecular mechanisms responsible for the positive influence of environmental interventions, such as dietary restriction and mild stress, are being widely explored. The environment experienced by humans in modern societies already contains countless compounds that may influence longevity. Understanding the role that the most prevalent of these compounds play in the aging process will be important to predicting and interpreting the outcome of introducing new interventions. Caffeine is the most widely used psychoactive drug worldwide. Prior studies indicate that caffeine may positively impact age-associated neurodegenerative diseases, such as Alzheimer’s. In the nematode, Caenorhabditis elegans, we observe that low concentrations of caffeine extend lifespan in worms in a temperature-dependent manner. Initial evidence from experiments combining caffeine with previously identified aging interventions suggests caffeine may increase life span by reducing signaling through the insulin/IGF pathway. We have also shown that caffeine delays pathology in a worm model of polyglutamine disease. Additionally, caffeine and dietary restriction by bacterial deprivation show non-additive life span extension, suggesting that they impact aging through a similar mechanism. Our initial effort is focused on characterizing the effect of caffeine on longevity and health span, and exploring the interplay between caffeine and other aging interventions.

The Cordilleran Ice Sheet filled Puget Sound during the last ice age. Its maximum extent is defined by glacial deposits, including large "erratic" boulders, near Olympia in the southern Puget Lowland. The age of maximum glaciation can be determined using the build-up of cosmic ray produced nuclides in these rocks. We sampled two large granite erratics and separated quartz for analysis of cosmic ray produced Be-10. After dissolution, beryllium was extracted and purified, and the Be-10/Be-9 ratio measured by accelerator mass spectrometry. The Be-10 concentrations (approximately 75,000 atom/g) are used to calculate the time the boulders have been exposed to cosmic rays, and thus date the retreat of the Puget Lobe of the Cordilleran Ice Sheet from its maximum extent.

Our world is becoming increasingly integrated with motion sensor technology. However, the usefulness of motion sensor data is still vague and difficult to understand. Several applications using sensor infrastructure in the home, often referred to as smart homes, aim to support independence for older adults. The goal of this project is to try to understand smart homes for aging in place in general and complex motion sensor data within independent retirement communities. Our focus is in trying to understand how aggregated motion sensor data can be useful for clinicians, patients and family members, how to present aggregated motion sensor data, and how to proactively manage health to support aging in place. Our group is implementing ubiquitous motion sensors in independent retirement communities to assess mobility. Various parameters that influence mobility, including cognitive ability, psychosocial activity, and environmental characteristics, are noted and integrated into the data. The projects goal is to evaluate mobility in older adults and consider actions or treatments to prevent disease onset or discomfort. Preliminary findings from the ongoing longitudinal feasibility study provide strength to the argument that motion sensors may provide a supporting role for aging in place.

Recent advances in remote-controlled multi-rotor helicopters have propelled them to become a uniquely capable means of aerial cinematography. The video camera can now chart paths never before possible, weaving around and over objects, and bringing us along as it moves quickly through a space. The implications for art are largely unexplored. I will investigate ways this camera movement’s affect on our sense of space in film, and how these techniques, when used over a large area, may refigure our understanding of the city. My research requires constructing an experimental remote-controlled helicopter, building off the most current research this a dynamically evolving field. I will then employ my system to explore and capture footage the suburban environment, from which I will compose a non-narrative short film. The goal of this project is to see what compositional structures follow from these new techniques, and ultimately to see how they might lead to a reimagining of neighborhoods. Portions of this film and documentation of the process involved will be presented.

The nuclear transport pathway performs the fundamental function of moving cargo between the cytoplasm and nucleus in all eukaryotes. Nuclear transport is carried out through a highly conserved mechanism across all eukaryotes. Yet, in Drosophila, several components of the nuclear transport apparatus evolve rapidly under positive selection. Genetic conflict with selfish elements has been suggested as a possible cause for this pattern of rapid evolution. Here, we present a comprehensive phylogenomic analysis of importin gene evolution in Drosophila. Importins are adapter molecules that directly mediate the transport of cargo into the nucleus. Our analysis reveals a recurrent pattern of gain and loss of importin paralogs across independent lineages of Drosophila. Interestingly, we discovered that almost all new copies of importins have acquired a testes-specific expression pattern since their birth through gene duplication. This pattern of repeated gains of testes-specific copies of importins and signatures of episodic lineage-specific positive selection suggests a function in suppressing genetic conlifcts such as segregation distortion in the male germline.

A spherical domed ceiling has been a proven artistic space since the era of Antonio da Correggio’s famous dome fresco, The Assumption of the Virgin. Mix that together with the University of Washington’s planetarium and an extremely fertile space for artistic collaboration and exploration is revealed. Today, brand-new digital projection technology at University of Washington’s planetarium provides a unique space for digital artwork in comparison to more classical, non-digital domes. However, because of a planetarium’s contextual history, a significantly greater amount of time has been spent continuing planetarium-like experiences rather than attempting more artistic and experimental explorations. Over the past year, I have worked in collaboration with the Astronomy and DXARTS departments to create a fully immersive and experimental film for fulldome projection. Through research, development, and testing, I have also produced a robust framework for future artists to create their own works in fulldome spaces, especially at the University of Washington. As Correggio has shown, being creative with illusionistic perspective on a dome can lead to impressive results. Even in the 21st century, his techniques can be applied towards scientific and artistic experiences as well as open up other unique avenues of exploration within a fulldome space.

Zebrafish have a defined striped pattern that results from pigment cell differentiation and migration during development, and this developmental process provides a good model for studying the process of boundary formation. Elucidating the mechanisms by which boundaries are formed is critical for understanding vertebrate development, and will give insight into what might cause them to malfunction during metastasis where cancer cells invade other tissue types, thus breaking the boundaries that have formed. While zebrafish have these defined patterns, a closely related species, the pearl danio, lacks stripes and instead has different pigment cell types intermingled. We hypothesize that the pearl danios have lost the mechanism for pigment cell boundary formation found in zebrafish and my goal is to identify genetic differences between the two species that could have caused this change pattern development. Preliminary RNA sequencing has identified candidate genes that appear to be differentially expressed. I am using in situ hybridization to compare the expression patterns of candidate genes between the two species. This is a technique where whole larval fish are incubated with RNA probes which are complimentary to the mRNAs of the genes I am screening for. Wherever the candidate genes are transcribed into mRNA, and thus expressed, the probe will hybridize. I am able to visualize this through labeling of the probe and staining with antibodies. I expect to find that some of the candidate genes will have different patterns of expression between the two species. This result will indicate that these genes are involved in the pigment pattern formation process, although it will not explain their specific function. However, once I know which genes are involved I will be able to manipulate their expression through knockdown or over-expression, which will provide further insight on the mechanisms of boundary formation in pigment patterns.

Paleontological evidence has shown that during large scale extinction events, hydrogen sulfide (H2S) rises to abnormal levels in Earth’s atmosphere. To survive these extinction events, an organism would be required to counter-act the negative effects of H2S by evolving unique adaptations. It can be hypothesized that many of these unique adaptations still exist in present-day lineages. Modern plants, for example, may have developed a method of utilizing H2S in order to survive, and possibly thrive, in what would be considered a highly toxic atmosphere. Through experiments involving species Phaseolus vulgaris and Pisum sativum, it was discovered that applications of H2S to seedlings increased growth at rates of up to 200% within 1 week. In addition to the increases in growth, increases in plant mass and photosynthetic activity were also observed. In order to better understand the effects of H2S on P. vulgaris and P. sativum, seeds of the species were also exposed to H2S, under the same controlled conditions. The effects of the seed experiment were similar to the initial seedling tests. Time to germination decreased with exposure, and the overall rates of germination increased. Large scale implications of these findings present benefits such as greater crop yields and perhaps a more efficient method of bio-fuel production. Continuing research of H2S and its effects on plant life could someday create a powerful means to deal with our ever-expanding human population…and the ever-expanding human footprint that goes along with it.

More than 3000 landslides have been identified as occurring as a direct result of the March 11, 2011, Tohuko Earthquake, using Google Earth disaster satellite imagery (1 m resolution). Landslides were categorized into 5 types: disrupted, lateral spread, flow, rock fall, and block slides. These types were analyzed using ArcMap, a type of Geographical Information System (GIS) to map spatial patterns of landslides in relation to surficial geology, peak ground acceleration (PGA), distance to epicenter, and topography. The areal size versus frequency of the Japan landslides was also compared to distributions of landslides inventoried in previous studies. The Japan distribution was found to follow an inverse power law function; this is an extremely useful relationship because it allows us to “fill in” an incomplete inventory, and can have predictive value for landslides in areas that were not covered by the disaster imagery. In knowing how many landslides and where the susceptible areas are likely to occur, potential damage and injury can be avoided. Landslides can be dangerous and the disruption they cause in distribution of supplies especially aid after an event, this research can strengthen the ability for governments to plan for and find ways to diminish problems associated with such blockages. Further work can be done to determine how roadway systems and infrastructure can be safeguarded against failures to ensure life and property damage does not occur.

The glass transition temperature is a feature of polymer systems that dictates the transition between glassy and molten-like states in amorphous solids. Glassy polymers have many advantageous properties and are highly desirable for commercial and industrial uses. However, the conditions that lead to glass formation are usually well above room temperature, making implementation of most polymer glasses unfeasible. In a recent discovery, ultra-stable poly(methyl methacrylate) (PMMA) glasses were formed using a technique known as matrix assisted pulsed laser evaporation (MAPLE). These results showed a significant decrease in the polymer’s density, while maintaining chemical stability under normal conditions. The exciting experimental discoveries hold great promise for future technological developments. However, there is much to understand about the underling atomic scale features that arise from MAPLE deposited glassy polymer films. To help answer some of these questions, molecular dynamics (MD) computer simulations were used to understand the mechanisms behind the ultra-stable polymer formation. Previous studies include observing the collapse of the polymer PMMA from an extended to a globular-like state under various solvent and temperature conditions. This poster will show how we track formation of the globular state using different order parameters (e.g., radius of gyration) and also show our generalized strategy for quickly building new polymer simulations. Understanding the fundamentals behind ultra-stable glass formation can unlock discoveries in the development and implementation of new materials.

What is the effect of corporate environmental performance (EP) and social performance (SP) on financial performance (FP)? Conducting research on the effects of environmental performance and social performance on financial performance could encourage corporations to engage in more sustainable business operations. The potential benefits of better environmental performance and social performance range from a cleaner environment to healthier societies. Therefore, the goal of this research is to investigate the relationships among SP, EP, and FP. Freeman’s stakeholder theory (1984), defined as integrating (through who and what, processes, and allocating resources) multiple stakeholders, who affect and are affected by the firm on multiple issues (i.e. their “stakes”), is the basis for testing the following hypotheses. H1: Better environmental performance leads to higher financial performance. H2: Better social performance leads to higher financial performance. Our sample, which includes 90 Fortune 500 publically traded companies from the US, Europe, and Asia, represents a variety of industries. Environmental, social responsibility, and financial performance data were extracted from 2008 and 2009 annual financial and sustainability reports. We analyzed the data with descriptive statistics, exploratory factor analysis, and linear regression. If research findings reveal positive correlations among environmental, social responsibility, and financial performance, corporations will find it beneficial to be environmentally and socially responsible. The current study may contribute to the literature and managerial practices in two ways. First, reduced pollution, waste, and energy used can lower costs to corporations, thereby increasing profits, while benefiting the planet. Second, better relations with stakeholders can enhance the firm’s reputation thus financial performance.

Clean water reserves have become difficult to obtain for much of the world’s population, due to drought as well as heavy use by certain industries in some regions. There are multiple solutions to this issue but they all require high costs that are only affordable to large corporate institutions. Therefore this project proposes a solution that attempts to be financially accessible and environmentally conscientious utilizing the analysis and critique of previous work. Building on the previous work my research suggest that the sun and seawater can work together to produce a purified form of water by using a magnifying glass to evaporate seawater and condense it into a container. I think that the availability and simplicity of this device would make it financially accessible and environmentally conscientious.

This research examines the way Indigenous cultures and identities are represented at Tillicum Village, a Seattle-area tourist attraction. Recent changes in ownership and management at the site have proved controversial among the Indigenous populations represented, who perceive the focus as shifting away from cultural authenticity and toward economic profit. This project explores the issues of a corporation's responsibility in accurately and fairly representing the cultures it profits from. The concepts of staged authenticity and heterotopia are essential to the theoretical framework of this issue, as is placing Tillicum Village in the context of colonial tourism. Data are being collected using traditional ethnographic methods and historical research, and analyzed through critical theory. This research ultimately hopes to illuminate way in which Tillicum Village can better address the grievances of, and collaborate with, the Indigenous communities it portrays.

Autophagy, or “self-eating”, is a process cells use during times of stress or starvation to break down their own proteins or organelles to provide the required nutrients. I focused on the autophagy-related protein 13 (Atg13), a regulatory protein that controls the initial stages of autophagy. Specifically, I was interested in defining the distinct roles of the N and C-termini of the Atg13 protein. Using Saccharomyces cerevisiae expressing the full length, N-terminal, and C-terminal half of Atg13, I tested how these protein domains affected autophagy processing. To monitor this I looked at the autophagy dependent processing of aminopeptidase 1 (Ape1) and alkaline phosphotase 8 (pho8). Western blots were used to monitor the changes in Ape1 protein length after processing, and a colormetric phosphatase assay measured autophagy-dependent increases in pho8activity. Using these techniques I found that the two domains of Atg13 play distinct roles in the process of autophagy.

The reduction of CO₂ has become a global priority as the concerns of climate change and the adverse effects of atmospheric carbon dioxide on the environment continue to rise. One method of reducing CO₂ emissions is through conversion and utilization as a chemical feedstock. CO₂ is cheap, nontoxic, and can be recycled to produce valuable new products, making it an attractive feedstock. The increasing importance of CO₂ utilization processes requires a better understanding of the underlying mechanisms of its conversion. The industrial conversion of CO₂ is typically achieved using a heterogeneous catalyst, usually consisting of small transition metal particles supported on a high surface area metal oxide. Our project focuses on elucidating the mechanism of the reverse water-gas shift (RWGS) reaction on platinum, an extensively studied catalyst with a highly active surface. Our goals are (1) to quantitatively measure the RWGS rate under conditions of low H coverage and (2) to compare the measured rate to the rate of CO₂ dissociation (the rate-limiting step according to the surface redox mechanism) and to the theoretical rate calculated using a micro-kinetic model based on the associative mechanism. The information we gather from the investigation will clarify whether the RWGS reaction proceeds on platinum via the surface redox mechanism or the associative mechanism.

Estimation of the firing range is often critical for reconstructing gunshot fatalities, where the main measurable evidence is the gunshot residue aslo known as stippling and soot, which will be used throughout the study. In the present study, a diverse range of gunshot wounds with four different guns with their appropriate ammunition have been used to gain knowledge about the image and patterns of gunshot wounds depending on the weapon and the firing distance. There were a total of 21 target shootings, there are about 6 targets per weapon used each with varying tested distances (2, 4, 6, 8, 10, and 12 in.). By increasing the firing distance away from targets, targets demonstrated a clear decrease of soot pattern (the black powder left by the firing)  and increase of stippling (gunshot residue- burnt gunpowder) diameter. Further analysis showed a linear relationship between the diameter of stippling and the firing range. The amount of soot and stippling was highly dependent upon the gun and ammunition used. The results allowed a basis for estimating the firing range in diverse gunshot wounds, helping to determine the manner of death between suicide or homicide. The reason that the research has to be re-done various times, due to law and individual cases, but the research sets a starting point to where the detectives can initiate study.

Ultra-high vacuum (UHV) techniques make it possible to observe interactions on clean, ordered surfaces without the complexity of realistic systems.  Final construction and testing of an improved UHV adsorption microcalorimeter capable of measuring the heat of adsorption of metals on single crystalline surfaces have recently been completed.  This calorimeter is the first of its kind to employ an electron beam evaporator to create a high, stable flux of metal atoms with low source radiation, which is then pulsed onto the thin single crystalline sample. Due to the properties of the electron beam evaporator, metals with high heats of sublimation such as Pd, Pt, and Au can be evaporated while maintaining the base pressures necessary for maintaining ideal surface conditions.  The new chamber is also equipped with a sample preparation chamber in which metal oxide surfaces can be grown on single crystalline samples.  Absolute calibration of the microcalorimeter was achieved by measuring the heat of formation of Cu multilayers on a Pt(111) surface, indicating that the theory behind our electron beam evaporator is correct, and will allow for the first direct measurements of the heats of adsorption of catalytically important metals such as Pt, Pd, and Au on any surface.

Mitochondrial dysfunction and oxidative stress have been proposed to contribute to organismal aging. In the nematode C. elegans, reduction of mitochondrial function through knockdown of genes encoding components of the electron transport chain (ETC) is sufficient to increase life span. The increased longevity is not solely the by-product of decreased oxidative damage generated by the ETC, but also involves induction of the mitochondrial unfolded protein response (UPR^mt), as evidenced by the observation that life span extension requires an intact UPR^mt. The cytoplasm, endoplasmic reticulum, and mitochondria have each evolved compartment-specific signal transduction pathways to alter nuclear gene expression in response to perturbations in protein homeostasis. However, little is known about the UPR^mt and the mechanism by which unfolded proteins are recognized in the mitochondria and signal changes in gene expression. Previous studies demonstrate that knockdown of ETC components and a subset of mitochondrial proteins induce the GFP reporters for mitochondrial chaperones hsp-6 and hsp-60. To identify novel genes involved in the UPR^mt, we are performing a forward genome-wide RNAi screen using the hsp-6::GFP reporter and measuring UPR^mt activity by fluorescent microscopy. Upon completion of the screen, we expect positive hits to include mitochondrial proteins, signaling molecules, and transcription factors that regulate the UPR^mt. Consistent with previous studies, gene knockdowns that have robustly induced the UPR^mt thus far in our screen are largely components of the ETC. In addition, the screen has yielded many novel hits including genes outside of the mitochondria and of unknown function. After the screen is complete, we will validate the RNAi clones that induce the UPR^mt and perform lifespan experiments to understand how the UPR^mt is involved in the aging process. Our screen will further understanding of how the mitochondria and nucleus communicate with one another in order to maintain mitochondrial proteostasis and illuminate how mitochondrial stress contributes to aging.

Phase Change Memory (PCM) exploits the difference in resistivity between the amorphous and crystalline solid phases of chalcogenide alloys. With easier scalability, faster write speeds, and higher switch endurances, PCM is set to replace Flash Memory and lead the way to greater functionality of our everyday electronic devices. The major obstacle preventing development of this technology is the power required to transition between the two solid states. In order to switch between phases, the material must first be melted and at greater cell densities, it becomes uncertain if the memory access device can supply enough current for resistive heating. A solution for the high power consumption is to adopt a nanowire (NW) structure whose geometry lowers the melting temperature of the phase change material. Our research was focused on improving simulation codes written by Ph.D. student Jie Liu for Nanowire Phase Change Memory (NW-PCM). As the code was based on macroscopic electro-thermal equations, a significant part of the research became to determine how accurately macroscopic physics could describe the operations of NW-PCM cells with mesoscopic dimensions - where quantum mechanics is known to cause fluctuations from classical results. By simulating published experiments on NW-PCM and comparing the results, we concluded that NW-PCM with dimensions ranging from 5nm – 200nm can be adequately described by classical physics within an error margin. The conformity of results for NW-PCM dimensions as low as 5nm was unexpected as devices with dimensions smaller than 10nm have been experimentally observed to undergo ballistic electron transport. The anomaly piqued interests in the published experiment that caused it and the data analysis from our research as a whole went towards identifying how the simulation should be improved.

Synthetic biology holds great promise regarding the production of important compounds, and the degradation of harmful ones. Last summer, we harnessed the power of synthetic biology to meet the world’s needs for fuel and medicine. We constructed a strain of Escherichia coli that produces a variety of alkanes, the main constituents of diesel fuel, by introducing a pair of genes recently shown convert fatty acid synthesis intermediates into alkanes. Using this method, we were able to produce on average 160 mg of alkanes per liter. Also, we identified a protease with gluten-degradation potential, and then reengineered it to have 100 times increased gluten-degrading activity, allowing for the breakdown of gluten in the digestive track when taken in pill form. Finally, to enable next-generation cloning of standard biological parts, BioBrick vectors optimized for Gibson assembly, an emerging DNA cloning technique, were constructed and used to construct the Magnetosome Toolkit: genes for biofabrication of magnetic particles.

In 2011, over 18% of all international adoptions to the United States were from Ethiopia; an increase of over 1,000% in the past decade. Ethiopian adoptees have also shown to have better health outcomes and less behavioral issues than adoptees from other regions of the world. Even though Ethiopian adoption to the United States has become an increasingly prevalent phenomenon, virtually no research has been done to explore the complex and unique processes of trans-racial family building, attachment, and identity formation experienced by Ethiopian adoptees and their families. While adoptees are integrated into new social and cultural settings, adoptive parents are forced to navigate their family through a new social and cultural space within a heavily racialized society, and facilitate these complex processes for their children and themselves. Why has Ethiopian adoption become such a recent popular trend? How do adoptive parents of Ethiopian adoptees navigate the critical experiences of family building/attachment and identity making/re-making? What strategies and resources do they use or create to facilitate these processes? In order to explore these questions, this project has been developed to investigate and analyze the lived experience of Ethiopian adoption by Euro-American parents to identify and understand these complex and poorly understood processes. Six case studies have been generated through semi-structured interviews, narrating the adoption process from the perspective of adoptive parents living in the Seattle area who retrospectively examined the experiences they faced and continue to face as Euro-Americans adopting from Ethiopia. Key informants, such as a pediatric physician specializing in adoptive medicine, have been consulted to provide further input and direction in analysis and theory development. Data is qualitative and the analysis interpretive with a goal of understanding these complex, unique, and challenging processes of identity making/re-making, cultural reconciliation, trans-racial family building, parent-child attachment, and other critical fundamental transitions.

Despite an increasing US-born population, Asian-Americans are often regarded as perpetual foreigners and unrecognized as ‘American.’ The present research examines whether Asian-Americans that are heavier, a stereotype commonly associated with Americans, are perceived as more American than normal weight Asian-Americans. Sixty-eight students were randomly assigned to see a photo of one of three Asian-American women. The same three women were used in both conditions and photos were edited to reflect a change in weight (the weights estimated by participants differed on average by 46 pounds between conditions). Results showed that heavier Asian-Americans were rated as more likely to have been born in the US and rated as worse at math than normal weight Asian-Americans. This research suggests that weight may reduce the extent to which Asian-Americans are perceived as foreigners and increase acknowledgement of their American identity. 

Chronic infections with Pseudomonas aeruginosa are a significant cause of deterioration of lung function in patients with cystic fibrosis (CF), and have substantial clinical importance due to their marked resistance to treatment with antibiotics. Furthermore, existing laboratory methods for testing bacterial susceptibility to antibiotics are poor indicators of clinical response to therapy, which further increase the difficulty associated with treating these infections. Current research shows that phenotypically diverse P. aeruginosa subpopulations evolve within a single patient during prolonged infection, and that clinical antibiotic therapy causes some of these subpopulations to change significantly in relative abundance. Observed changes in abundance following antibiotic treatment signify varying degrees of antibiotic susceptibility within each patient’s P. aeruginosa population as a whole, with an increase in a particular subpopulation abundance indicating resistance, and a decrease in a particular subpopulation abundance indicating sensitivity. As previous studies have only evaluated P. aeruginosa antibiotic sensitivity prior to antibiotic treatment, we attempted to demonstrate the correlation between an increase subpopulation abundance following clinical treatment and the degree of resistance to antibiotics in laboratory testing. This was done by evaluating the antibiotic minimum inhibitory concentration (MIC) of a range of isolates collected from CF patients who had undergone antibiotic therapy, using antibiotics which had been given to each patient in clinical treatment. Our results demonstrate a marked lack of correlation between relative subpopulation abundance and antibiotic MIC, with clinically resistant subpopulations showing the same or lower MIC than subpopulations which were clinically sensitive treatment. We conclude that isolates resisting treatment do not have a different MIC than isolates responding to treatment, suggesting that mechanisms other than classic antibiotic resistance are responsible for treatment resistance in the CF lung. Further studies to evaluate these mechanisms, such as biofilm formation and formation of persister cells, are currently under way.

The type VI secretion system (T6SS) is a large protein complex utilized by bacteria to transport proteins into other cells. Initially implicated in virulence, the T6SS also participates in interbacterial interactions. A T6SS found in the opportunistic pathogen Pseudomonas aeruginosa, the hemolysin coregulated protein secretion island I T6SS (H1-T6SS), has been found to deliver a set of toxins to other bacteria. This provides P. aeruginosa a fitness advantage over competing bacteria. The H1-T6SS can target neighboring P. aeruginosa, which protects itself with cognate immunity proteins. Deleting these immunity proteins leaves P. aeruginosa susceptible to self-attack. However, the H1-T6SS as well as other T6SSs can target other bacteria, specifically they only target certain species. One of the goals of this project is determine the nature of such specificity. These findings would implicate new functions for the H1-T6SS. To further this, I am seeking to extend the scope in which the T6SS is viewed, both mechanistically and genetically. In doing so I am searching for new genetic loci that participate in T6SS-dependent interactions. Taken together, this work provides insights into a new aspect of type VI secretion and may change the genetic composition of T6SSs.

Rhododendron is an aesthetically valuable plant genus primarily distributed in Northern hemisphere. Previous work by collectors and taxonomists has revealed that Rhododendron taxa have a scattered pattern of distribution around the globe. Yet, the evolutionary history that led to this modern distribution pattern is not well understood. Fossil records of once extant Rhododendron populations can be a key component for inferring the diversification and migration of Rhododendron. Macrofossils such as leaves and stems have more restricted requirements in their formation, and hence are rarely discovered. On the contrary, microfossils of pollen are more widely preserved and found. In this study, I explored the potential of using pollen microfossils of Rhododendron to understand how the modern distribution pattern of Rhododendrons came about. To that end, I compared and contrasted modern Rhododendron pollen from three subgenera and twenty four subsections in order to discover the degree of consistency in pollen morphology within and unique combination of traits of each taxon. All observations were performed under light microscopy. Experimental data include both structural differences and quantitative measurements. Preliminary results demonstrate promising level of variation in measurements such as the structure of apperture edges, pollen wall sculpture, overall size, and wall thickness. These palynological features are then fitted into a Rhododendron phylogeny based on molecular data to reveal any potential taxonomic significance of variation in pollen morphology. 

In 2011, a 3 week research cruise on board the R/V Atlantis was undertaken to the Endeavor segment of the Juan de Fuca spreading ridge. The main purpose of this expedition was to determine conductive heat flow from lightly sedimented sea floor within the axial valley of the spreading center (in close proximity to an isolated hydrothermal vent system). During a series of Remotely Operated Vehicle (ROV) dives to the seafloor, hundreds of heat flow time series were measured by the deployment of 13 “thermal blanket” heat flow instruments, which will enable researchers to better constrain the spatial extent of the hydrothermal fluid recharge zone. Throughout each dive, continuous video footage was recorded from several cameras on-board the ROV, along with the geographic position, altitude, heading and three dimensional orientation of the vehicle. A geological map of the sea floor across the vent field will be created using geo-referenced and orthorectified frame grabs (“flattened” onto the topography) from ROV video. The primary goal of this project is to provide a means of quantitatively measuring the porosity of spreading ridge seafloor basalt in this area of active hydrothermal venting, through the application of a technique that has never before been attempted at this scale.  A series of trigonometric algorithms were developed to use this reference information as a means of translating a still video image from ROV footage into a series of geo-referenced images.  Geographic Informatics System (GIS) software is being used to create a mosaic-like map of volcanic, biological and geological features from the sea floor near this hydrothermal vent system from these images.  An additional data layer will be produced simultaneously to provide additional information concerning the spatial extent and distribution of geologic features on the seafloor to assist in understanding the results of the heat flow study. 

Obstetric Fistula is a childbearing injury that occurs as a result of neglected labor. Obstetric Fistulas occur in 5% of live births and accounts for 8% of all maternal deaths. It is one of the four major causes of maternal mortality and morbidity. Worldwide about 50,000 to 100,000 women are affected by obstetric fistula each year (World Health Organization, 2011). The purpose of this project was to: 1) Create greater awareness of obstetrical fistula in the community, 2) Assess if giving circles and my presentation accomplished this goal. Giving circles is a strategy used to inform and educate small groups of people about obstetric fistula. I conducted a series of giving circles, starting with family, then to a group of friends, followed by students in the UW School of Nursing Programs, and hope to set up another for UW faculty. I used PowerPoint slides and showed a clip of the 2009 Emmy award winning documentary “A Walk to Beautiful”. I asked the attendees to fill out a written evaluation at the end of each presentation as a means to evaluate the presentation itself and the impact it had on the attendees. The evaluation consisted of eight direct questions about the information and delivery of the presentation and allowed the attendees to comment and/or grade content. In total I had a response rate of about 50%. I identified major themes across all responses and sub-themes in each of the individual questions. One major theme identified was the need to spend more time explaining the anatomy of an Obstetric Fistula and the research around the social implications of having the fistula. As a result I concluded that there is a general interest in the community about Obstetric Fistula and giving circles are an effective method of raising awareness.

In the last decade, Seattle, like many other cities across the world, has tried to encourage bicycling by developing new bicycle infrastructures, cultures, and policies. In many cities, these efforts often start nearly from ‘scratch’. However, in The Netherlands, bicycling has been an important part of the transportation network for more than 100 years, with comprehensive infrastructure in place from policies developed during the 1960’s and 70’s. Our team, composed of two members, spent several months in Groningen, The Netherlands, on an exchange where we collected data on the bicycle infrastructure success that this region of the world is renowned for. Working with the Multimodal Project Development team at Seattle Department of Transportation (SDOT), we brought the lessons-learned and design of the Netherlands to our home, Seattle. We have evaluated the design of four critical Seattle intersections: 43rd Ave NE and University Way, 7th Ave NE and NE 40th St., 25th Ave NE and the Burke Gilman Trail, and 34th Ave N and N Fremont Ave. and compared them to successful infrastructure in Groningen. Through background research, interviews, and observation, we have assessed the comfort, safety, and directness of these selected intersections using quantitative and qualitative measures. Based on this data, we have suggested two alternatives for each intersection: one based on Dutch design which is novel to Seattle and very ambitious, and one of which is more practical and less expensive. The possible results of this study are far-reaching and significant. Whether our designs are adopted in some form by SDOT or not, our final designs have influenced SDOT employees. We are taking a step towards making our community a safer, healthier, and more enjoyable place for all street users.

My research examines fairness and equity in the distribution of three wastewater treatment types across seven counties in the Puget Sound region. I begin my analysis with a thorough exploration into the concepts of fairness and equity, within the context of a variety of disciplines. Drawing on this analysis, I then look into the costs of these different systems, making sure to take into account all applicable considerations, including implementation, maintenance, and repair costs. Specifically, I investigate the costs of combined-sewer systems, separated sewer systems, and onsite septic systems at the household scale. Lastly, I use Geographical Information Systems and spatial analysis to compare the costs of these systems to their spatial distribution through the subject counties, and to the household median income within their respective census tracts. This methodology allows me to uncover any inequities in the way that the costs of these treatment types are distributed, and address a specific instance of fairness within urban planning that has yet to be investigated.

Examining three of Booker prize winner Ian McEwan’s novels Atonement, At Chesil Beach and Saturday provides a passage into his exploration of changing attitudes and viewpoints regarding female sexuality during the later 20th and early 21st century. Utilizing the genre of historical fiction, McEwan explores the various sociological and cultural influences such as improving neonatal care, contraceptives and improved birth control options that severed the previously inescapable link between sexuality and motherhood. Additionally, the rise of women in higher education and sectors of the workforce beginning in the 1930s onwards contributed to the autonomy and, as these novels indicate, increasing sexual freedom of women. Each novel explores the 1930s, 1950s and 2000s respectively as critical turning points in British culture. In the 1935 pre-WWII setting of Atonement, the prevalence of women in college and independent professions challenge gender norms. Against this backdrop, McEwan examines reciprocal sexual desire against victimization and societal attitudes towards women as equally desirous individuals. At Chesil Beach explores the physical relationship of a newly married couple during the 1950s and the emergence of clinical contraceptives. Finally, Saturday examines the attitudes of a mother and daughter in a post-sexual revolution setting and explores issues of generational differences as well as the link between sexuality and victimization still present in a contemporary time period. Ultimately, critically examining the attitudes and historical contexts of each novel against other scholarly work provides a literary interpretation of the evolution of attitudes regarding female sexuality, and sub issues of autonomy, consensual desire, the role of women and victimization, spanning from a pre-WWII era to contemporary society. 

Bilinguals outperform monolinguals in a number of tasks involving executive functions or higher-order thinking processes. For bilinguals, the everyday use of more than one language creates increased demands for language selection, inhibition, and switching. Little is known, however, about which facet of the bilingual experience shapes the brain in a way that gives rise to these improvements in executive functions. Previous research on this topic has primarily focused on increased inhibitory control, but the evidence has not been consistent. To explore the possibility that the switching component of executive functions is improved in bilinguals, an fMRI task-switching paradigm, with four experimental conditions (non-switch, language-switch, semantic task-switch, and double-switch), was created. This task allows for the comparison of the neural circuits involved in language switching and task switching in bilinguals. Specifically, this research tests the hypothesis that increased practice with coordinating and executing rules in two languages shapes the general task shifting sub-component of executive functions in bilinguals. Preliminary analysis of the data suggests that behaviorally, for bilinguals, the language-switch condition was significantly easier than the task-switch condition, although the double-switch condition was not significantly more difficult than the task-switch-alone condition. Neurally, the task-switching condition recruited more parietal and inferior frontal regions, whereas, language switching involved more medial frontal regions. All switching conditions recruited the basal ganglia though. Lastly, a comparison between the double-switch condition and the combination of the two single-switch conditions showed that classic working memory regions are recruited when executing a double-switch. By understanding which of the many unique facets of the bilinguals experience shapes the brain to give rise to improved executive functions, we can simultaneously better understand the bilingual brain and gain insight into how executive function might be improved in other populations.

The University of Washington Tacoma is considering a change to the campus telephone system. The current Avaya Private Branch Exchange (PBX) switch system is aging and inadequate for future expansion. The University is considering the implementation of a Voice over Internet Protocol (VoIP) system. This work briefly examines potential risks and rewards related to the implementation of VoIP phones. Initial considerations include the risks of inaction, as opposed to the risks of a large scale VoIP implementation. The main component of this work is related to the potential pitfalls and benefits of VoIP telephony. Of the many possible issues and benefits related to implementation of VoIP, only a few can be considered risks which are directly related to replacing a traditional PBX telephone switch with VoIP. Those specific risks are evaluated qualitatively; the qualitative data is then used to create a quantitative analysis. The quantitative data is used to perform a Monte Carlo risk simulation using the software tool ModelRisk version 4.3 by Vose Software. Conclusions include recommended actions to reduce the probability and impact of potential risks which are most relevant.

Dr. Gerald Pollack and lab associates have discovered that when a polymer or gel is placed in a chamber with an aqueous suspension of polystyrene microspheres, the microspheres migrate away from the polymer or gel surface to about ~200-500 micrometers, leaving a microsphere free zone. This region is known as an “exclusion zone” (EZ) and is characterized by crystalline water. We tested the effects of changing the solution temperature on the size of the exclusion zone. A critical aspect of this experiment is what role thermal energy plays, if any, on the exclusion zone. In this experiment, we placed the polymer (Nafion) in an aluminum chamber, which was then filled with microsphere solution (500:1 deionized water to polystyrene microspheres). We then placed this chamber into a larger chamber filled with water. This larger chamber was connected to a water circulator with which we could control the temperature of the water surrounding the aluminum chamber. In our experiments, we varied the temperature of the circulating water at 15 randomly chosen temperatures between 5˚C and 75˚C, and measured the size of the EZ. We found that EZ size increases as temperature diminishes. This finding has fundamental implications for understanding basic properties of water, and particularly the EZ phase of water.

Fe₃O₄ nanoparticles have been extensively studied for biomedical applications including magnetic resonance imaging, chemotherapeutic delivery and gene delivery due to their advantageous properties of superparamagnetism, biocompatibility and biodegradability. However, ultrafine, monodisperse and sized controlled nanoparticles synthesized by thermal decomposition are commonly encapsulated by hydrophobic layers and can’t be dispersed in aqueous solutions. Besides, during the surface modification of nanoparticles, it is still difficult to obtain a highly stable and dense polymeric surface layer. To this end, a robust multi-step polyethylene glycol(PEG)ylation approach is presented to produce biocompatible multilayer surface coated magnetic iron oxide nanoparticles that are hydrophilic and display long-term stability in biological media. Ultrafine, monodisperse and 12nm hydrophobic nanoparticles were first synthesized by elevated temperature decomposition of nontoxic iron oleate complex in the presence of 1-octadecene and oleic acid. Subsequently, the oleic acid layer of nanoparticles was replaced by 3-(triethoxysilyl)propyl succinic anhydride(SSA) through ligand exchange to form SSA layer coated nanoparticles. Then, during the first step of PEGylation, to prevent the aggregation of nanoparticles, adequate amount of methyl PEG-NH₂ was attached to the part of SSA layer through the coupling reaction of functional groups. And in the second step of PEGylation, NH₂-PEG-NH₂ was attached to SSA to achieve a SSA/PEG dense multilayer surface with an adequate number of functional groups for further conjugation of therapeutics or targeting agents. The tools such as transmission electron microscopy (TEM), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FTIR) were utilized to characterize the structure and properties of surface modified nanoparticles. The excellent dispensability and stability of surface modified nanoparticles in biological media was confirmed by no significant size change after at least 30 day’s incubation in Dulbecco’s phosphate buffered saline (PBS) and dulbecco’s modified Eagle’s medium (DMEM) with 10% of fetal bovine serum (FBS), demonstrating a huge potential for biomedical applications.

Single molecule microscopy techniques make it possible to observe the fluorescence of individual dye molecules. One observation previously obscured in ensemble measurements is fluorescence intermittency, where molecules undergo prolonged non-emissive periods, as well as other intermediate emissive states. We are trying to identify the photochemical processes that cause these fluctuations in intensity. The fluorescence of dye molecules can be affected by the environment in which they are immersed, specifically the substrate they are in. The polymer polyvinylidene fluoride (PVDF) is of special interest because of its ferroelectric properties, which allows polarization of domains in the polymer. I have been exploring procedures to make PVDF films with a high degree of domain polarization. Using infrared (IR) spectroscopy, I have characterized the percentage of domain polarization in these films. Future investigation is still needed though to understand how PVDF will act as a substrate and what influences it will have on fluorescent organic dyes. An expanded knowledge of organic materials is essential for the design of photochemically stable devices. Based on organic electronics, these new devices will be greener alternatives to semiconductor technologies.

The Ihara Zeta Function (IZF) of a finite graph encodes its spectral, combinatorial and number theoretic properties. Our aim is to calculate the IZF of the Cayley graph of the modular group PSL(2,Z), which can be thought of as a special group of square matrices. Since this group plays a central role in number theory and other areas of mathematics and physics, we hypothesize that the zeta function will reflect important information about modular forms and elliptic curves. Our previous research has yielded an algorithm that computes the IZF of a sequence of finite approximations of this group, but it does not give the IZF of the entire graph. The IZF can be expressed as a product of L-functions, which encode how successive graphs in the sequence are related. Our ultimate aim is to use the L-functions of the finite approximations to compute the IZF of the modular group. We are currently exploring L-functions of simpler Cayley graphs, in the hope that the methods can be generalized to calculate the IZF of PSL(2,Z).

This research discusses healthcare accessibility issues among the Hmong ethnic group in the Twin Cities metropolitan area located in Minnesota. Research was based on the geographical access to healthcare in the Hmong community. Through the Geographical Information System (GIS), I predict that the Twin Cities’ Hmong community lacks access to healthcare. The Hmong community, when mapped, was concentrated in two particular locations in Minneapolis and Saint Paul. The use of GIS helped to project statistical analysis of different aspects concerning healthcare such as healthcare facilities, healthy and affordable food, and public transportation. These were analyzed by mapping their distances from the Hmong community. Statistical analysis was examined by scatter plots were the correlation between the Hmong community and where healthcare, food sources, and public transportation was located were weak. Statistical analysis showed that the Hmong community of the Twin Cities metropolitan area is centrally located. Given the study area at the scaled researched (1km), the Hmong were not especially isolated from healthcare, food, and public transportation. This showed that this community is isolated but isolation is not a major factor shaping healthcare outcomes. Since isolation would not be a major factor of access to healthcare other factors may contribute to the limitations in this community such as cultural and language barriers and health literacy. More studies about the Hmong would be beneficial in order to examine the gap in health and healthcare in the community.

The regulation of organismal life cycles by genetic components is a research area that has shown great promise in recent years. The process of aging is becoming more important to understand, not only due to the increasing incidence of age-related diseases, but also in order to determine what factors contribute to prolonged health and longevity. A variety of physiological pathways can regulate lifespan in C. elegans nematodes. Among these, one pathway of great interest involves dietary restriction. It is known that “stresses” on the worm, such as reduced food intake, can extend its lifespan. A mutation in the eat-2 gene affects the pharyngeal pumping mechanism whereby the worms ingest bacteria, thus creating a genetic model for dietary restriction. This research investigates the role of steroid signaling in dietary restriction-induced lifespan extension and improved stress resistance, using eat-2 mutant worms as a background and using a heat shock assay to induce stress at various points during the worms’ life cycle. By studying genetic determinants of longevity in C. elegans, my research will help to uncover more information about what can make nematodes long-lived, thus contributing to our understanding of the genetic basis for longevity regulation. More importantly, the existence of human orthologs for many longevity genes in C. elegans suggests that our findings will be valuable in identifying factors that are crucial for human longevity and health.

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.

Synechococcus is a unicellular cyanobacterium about 1µm in diameter. It is an abundant photosynthetic microorganism and a great contributor to the primary productivity of the Earth’s oceans. Synechococcus communities are composed of different phylogenetic clades that are believed to represent physiologically and genetically diverse populations. Previous data has shown two phylogenetically distinct populations of Synechococcus, clade I and clade IV, to be dominant in Puget Sound, with fluctuations in these populations during different seasons and regions. For my research, I will attempt to identify a clade IV strain. In an attempt to enhance conditions where clade IV will be most competitive, I have varied the culturing parameters such as using multiple media types. Filtered whole water samples containing mixed assemblages of Synechococcus were collected from Puget Sound and were initially cultured in natural Puget Sound seawater. These cultures were poured onto agar plates with sterilized natural Puget Sound water as well as plates with artificial seawater with different concentrations of AMP1 nutrients. The 16S rDNA of the isolates identified as clade IV will be sequenced by the use of PCR using primer sets that target specifically Synechococcus clade I or clade IV. The success, or failure, of amplification will indicate whether the Synechococcus DNA template was Synechococcus clade I or Synechococcus clade IV. Any Synechococcus DNA that results in a successful amplification using the Synechococcus clade IV primer set, I will send off to be sequenced. We have nutrient and physical measurements for these samples to help explain the environmental parameters that control distribution of these populations. With this information and ITS sequences I can compare Synechococcus clade IV populations within Puget Sound, and compare these with coastal populations that have been previously isolated. We seek to understand how these populations fluctuate seasonally and throughout different regions of the Sound.

Beginning with the industrial revolution, levels of pCO₂ in the oceans have increased which in turn decreases the pH of waters and creates a chemical change known as ocean acidification. There are many potential impacts that ocean acidification may have on marine ecosystems and biodiversity, yet little research has been conducted on the effects of low pH levels on fish, particularly temperate species. In this study, we evaluated how different pCO₂ levels affect the survivorship and behavior of larval lingcod (Ophiodon elongates) under starvation conditions. To accomplish this task, we reared lingcod eggs that we obtained from the Manchester NOAA hatchery and the resulting larvae under three different pCO₂ levels that correspond to mean present day (~550 ppm) and possible moderate and high future pCO₂ levels (1050 and 2159 ppm, respectively) in Puget Sound. For the survivorship experiments, we recorded daily mortalities, which allowed comparisons of the cumulative mortality curves and mean survival times among treatments. For the behavioral experiments, we scored several different behaviors to assess activity levels and group cohesion. Five fish were taken from each container in each treatment and placed in a 4cm tall enclosure where they were filmed for ten minutes. Initial results indicate that mortalities were highest under present day conditions (550 ppm) and lowest under predicted high future pCO₂ levels (2159 ppm). These findings suggest that low pH, high pCO₂ levels benefit some species of fish larvae, which is the opposite effect demonstrated in most published larval fish studies to date. We discuss our findings in the context of Puget Sound’s dynamic pH environment and highlight areas of future research on this important yet understudied topic.

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.

For controversial subjects like evolution, language used by experts can actually reinforce naïve conceptions held by students. For example, students interpret the word “purpose” to mean that traits evolve to serve an immediate function, but experts mean that a trait increases reproductive fitness. Students also often incorrectly assume that evolution follows a predetermined goal, that individuals choose to evolve in a certain direction, and that fit individuals are stronger, bigger, or faster. In this study, I tested the hypothesis that an upper division biology textbook includes some naïve conceptions and unintentionally reinforces others. I analyzed nine chapters of Biology of Microorganisms (13th Ed) by Brock et al., a standard text for teaching microbiology. In my preliminary analysis of these chapters, I found 15 statements that seemed to imply or promote misconceptions. For example, Brock et al. stated that “[e]volution selected for improvements and diversification of these early cells…” (34) which could be misconstrued by students to mean that cells evolve towards a predefined, yet unspecified, improvement. Students may interpret this quote as supporting the inaccurate idea that evolution has a goal in mind and that individual organisms can choose to adapt. Only four chapters discussed evolution as a process, and the others talked about evolutionary patterns. Understanding the patterns is an essential component of evolutionary thinking, for example, interpreting how different lineages are related to each other. However, the text did not explicitly describe the process through which different species diverged from their most recent common ancestor. This result confirms what has been found in other studies, that even though evolution is a central theme in biology, discussion of evolutionary process is often restricted to only a few chapters in textbooks. By being aware of potentially negative impacts behind ambiguous words, educators can address common misconceptions and clarify existing naïve conceptions.

In primates, ventral striatum contains many reinforcement sites. These sites, when stimulated with electrical current, mimic behavioral reinforcement. Monkeys trained to perform behavioral tasks will often work solely for stimulation at these sites without food reward. Using these sites, we can further understand the plasticity of the brain by altering neural activity through conditioned changes in connectivity. We implanted moveable stimulating electrodes near the nucleus accumbens. Reinforcement sites are identified by whether stimulation elicits reinforcement behavior. This is done by the use of alternating reinforced and non-reinforced trials (R/NR) while the monkey performs a simple manual force step-tracking program by controlling a cursor’s movement on a computer screen with forces generated with their wrist. Their objective is to move the cursor into different boxes representing different wrist forces. During each R period, the monkey receives electrical stimulation at the potential reinforcement site for every completed trial. During NR periods, however, the monkey has the opportunity to work but no stimulation is given for completed trials. If the number of completed trials is significantly higher during R periods than NR periods, a reinforcement site has been found. We next condition single neuron firing rates using this reinforcing stimulation. Chronically implanted micro-wires record discriminated neuron activity from primary motor cortex. We determine the cell’s baseline firing rate and trigger stimulation of the reinforcement site on action potentials that exceed this rate-threshold. Our objective is to train the monkey to increase the cell’s activity for reinforcing stimulation in set time periods. This experiment enables us to condition volitional control of brain activity to manipulate neural prosthetics and restore motor function in paralyzed patients. Since this conditioning can also be done with an autonomous brain-computer interface, this paradigm can be extended to freely behaving monkeys and has the potential to be applied to humans.

Cell labeling and tracking hold enormous potential to illuminate cellular pathway mechanisms. Because current labeling methods are either unsafe or unavailable for humans, a need exists for a safe and effective cell labeling technique suitable to track cells administered for cell therapy. The stable carbon isotope, C13, is safe for human use, incorporates into cellular DNA when taken up as C13 glucose in culture medium, and can be detected by GC mass spectrometry. We proposed to evaluate the use of C13 to label and track exogenous bone marrow derived macrophages (BMMCs) administered after myocardial infarct (MI). To confirm exogenous BMMCs can be detected in the heart, we injected green fluorescent protein (GFP) labeled BMMCs into the tail vein of mice three days following MI. Some heart sections were assessed for GFP positive cells by immunohistochemistry. Other hearts underwent dispersion and isolation of macrophages. Quantitation revealed the percentage of exogenously delivered C13 labeled GFP BMMCs in the cardiac macrophage population was 10 percent. Preliminary cell proliferation assay results suggest no considerable difference between the growth of BMMCs with or without C13 glucose. Finally, we cultured GFP BMMCs for ten days in 100% C13 glucose media for tail vein injections to mice following MI. Three days later, we excised the hearts to isolate cardiac macrophages. Cultured heart cells and macrophages were analyzed via GC mass spectrometer to quantify labeled DNA. Although we could detect C13 in in vitro labeled macrophages, increased C13 cellular label could not be detected in isolated macrophages. Unlabeled macrophages appropriately had no increase in label. Ultimately, while this study was unsuccessful in detecting a C13 cellular label in vivo with a GC mass spectrometer, in vitro successes suggest technique refinement could lead to a powerful and safe therapeutic stem cell label.

The secretion of signals or toxins is a vital aspect of bacterial communication with its environment. We recently found that type VI secretion systems (T6SSs) in Gram-negative bacteria mediate interbacterial interactions by translocating protein effectors into neighboring bacterial cells, providing a large fitness advantage. One protein exported by the T6SS is a type VI-exported (Tse) cell wall amidase that degrades peptidoglycan, and secretion of this effector, Tse1, results in target cell lysis. Bacteria can also self-intoxicate by T6S, potentially posing a problem for the survival of a given species. However, bacteria encode cognate immunity (Tsi) proteins that bind and inactivate their Tse partners. As we identify more T6 effectors, it is becoming clear that Tse–Tsi pairs are part of a widespread protein–protein recognition network underlying bacterial diversity and community structure. This study sheds light on how this network influences interbacterial interactions by uncovering the mechanism of Tsi1-dependent inhibition of Tse1 and defining the molecular determinants of this recognition. I have reconstituted the complex in vitro by using recombinant proteins to probe for direct interaction. Then, by mapping conserved residues onto the recently solved structure of Tse1, I have identified and mutationally analyzed candidate surface residues that may be involved in Tsi1 recognition. Further understanding the determinants of this interaction may allow us to predict which bacterial species are vulnerable to Tse1 or provide a template for the design of novel effector-immunity complexes that escape recognition in naturally occurring bacteria as an antibacterial strategy.

A strong understanding of the intricacies of blood coagulation is essential for proper treatment of coagulopathies such as hemophilia, hemorrhage, Von Willebrand’s disease, and numerous thrombosis related conditions. The coagulation behavior of blood is directly related to the structure of the supporting protein networks. Fibrinogen is a protein found in human blood that, upon polymerization into cross-linked fibers, forms the scaffold-like support structure of a blood clot. Based on the observation that emergency room patients with traumatic injuries experience a greater extent of hemorrhage and abnormal hemostasis, our group has investigated the effect of acidic oxidation on the structure and mechanical properties of fibrin gels. Purified fibrinogen was oxidized using hypochlorous acid to simulate the acidification of blood during trauma and shock. Rheological, x-ray scattering, and turbidity studies were performed to characterize the structural and mechanical differences between oxidized and normal fibrin gels. Preliminary results from these studies indicate the oxidation causes a delay in fibrin gelation as well as a decrease in the elastic modulus of the fiber network. Additionally, evidence from turbidity studies suggests that oxidation prevents lateral aggregation between protofibrils leading to an overall reduction in fiber diameter.

Diluted magnetic semiconductor (DMS) technology combines the properties of a semiconductor with ferromagnetism. Nanoparticles doped with magnetic elements such as Manganese offer a prime example of this type of material. Our specific task was to analyze the effects of p-type doping with nitrogen in ZnO quantum dots (QD) of 2.4 nm in diameter. p-type doping in this material involves the replacement of an O^2- ion with an N^2- ion, which has one less electron resulting in a hole due to an empty N^2- p-orbital. We hypothesize that when p-type doping is combined with paramagnetic Mn²⁺ doping, the unpaired 3d electrons of two Mn²⁺ ions separated by an N^2- ion will adopt a ferromagnetic orientation as opposed to their traditional antiferromagnetic state. Testing for this experiment was done with the development version of Gaussian. Energy calculations within this program and further data analysis reveal information such as the total energy of each state of the system, molecular orbital data, and energy diagrams concerning the semiconductor properties of these materials. Results show that the lowest energy state occurs when the Mn²⁺ ions adopt a ferromagnetic arrangement with the N^2- hole sharing this alignment. The potential for this QD and others with similar properties is extreme as many semiconductor applications could be revolutionized by DMS's. New applications such as solar cells made of DMS's are also under consideration.

All solid materials have a stress limit beyond which cracks and ultimately fractures will develop; we are analyzing the possibility of uneven solar heating as a process that causes cracking. Research at the Universities of New Mexico and North Carolina drew attention to patterns of near vertical cracks in desert rocks, which known weathering processes cannot sufficiently explain. To investigate, temperature and strain data as well as cracking events were recorded using an instrumented boulder. We created a 3D-Finite-Element-simulation of the boulder incorporating thermal radiation, heat conduction, and thermo-mechanical stress analysis. Simulation provides the link between insolation and stresses, allowing us to assess size and shape effects on tensile stresses. We are validating our model by comparing measured and simulated temperature and strain data. As an example, we are calculating the maximum and minimum strain for each surface collection point and validating them against provided data. We expect to find that uneven heating from the diurnal cycle of the sun creates tensile stresses sufficient to cause cracking in these rocks. Future research will include a comprehensive data analysis concerning stress distribution and orientation within spherical boulders of various sizes, and an assessment of potential crack growth rates and crack orientation.

Pedaliaceae (Sesame Family) are a small family of plants, about 14 genera with approximately 70 species that occur mainly in arid regions and deserts of the Old World, with their center of distribution in Southern Africa. Contained in Pedaliaceae are two widely known species: Sesamum indicum, which is cultivated and provides sesame seed and Harpagophytum procumbens, Devils Claw, an endangered species which is harvested wild and used for medicinal purposes. Evolutionary relationships (phylogeny) within the family are currently not very well understood. Previous classification systems have included the Martyniaceae, a small New World family, which occurs in similar environments and exhibits similar fruit morphology, with the Pedaliaceae but this inclusion of has since been questioned. In order to infer a phylogeny for the Pedaliaceae I am using the chloroplast loci ndhF and tnrL-tnrF which generally provide good resolution to determine the relationships between families and major lineages within a family of plants. Preliminary analyses based on these two loci indicate that the genus Sesamum is not monophyletic. To achieve resolution of the relationships of the species within Sesamum, I am sequencing two commonly used nuclear loci: ETS and ITS, the external and internal transcribed spacers of the nuclear ribosomal repeat, and one of the many loci in the pentatricopeptide repeat (PPR) which have only recently been developed for phylogenetic study in the Olmstead Lab. The goal of this project is to infer a phylogenetic tree of the Pedaliaceae on the species level, resolve their relationship to Martyniaceae and the genus Trapella, as well as the placement of these groups within the larger tree of the Lamiales.

Self-assembling silver nanoparticles have been of great interest to the scientific community due to their unique optical properties and potential use in areas such as cancer therapy and semiconductors. This research explores using a simple one-pot technique to produce self-assembling silver nanoparticles with controllable geometries based on steric interactions. The core advantage of this method is that it employs a bottom-up approach using commonly available and relatively inexpensive materials. Successive doping of long hydrophilic polyethylene glycol (PEG) chains followed by a short hydrophobic alkane-thiol renders the nanoparticles amphiphilic. Clusters of different sizes self-assemble depending on the concentration of PEG added to the silver dispersion. As PEG concentration increases, the extent of clustering decreases as the long chained PEG molecules create a corona around the silver particles and inhibit functionalization with alkane-thiol. This synthesis method is analyzed with Dynamic Light Scattering (DLS), Small Angle X-ray Scattering (SAXS), and Transmission Electron Microscopy (TEM). While the samples follow the expected clustering size trends, data indicates that monodisperse silver is critical to the reproducibility of the technique.

Oil operations that took place in the Northeast Ecuadorian Amazon between 1964 and 1992 have altered the ecological functions of thousands of acres of land by dumping crude in open pits, burying oil extraction byproducts, and burning unwanted oil without proper treatment. Many people in the area experience a number of health issues including flu, respiratory problems, skin issues, stomach aches, and headaches possibly associated with air and water contamination. My research focuses on the analysis and mapping of water quality, specifically of 5 parameters: temperature, dissolved oxygen, turbidity, conductivity, and pH. My colleagues and I collected 55 geo-referenced water samples within the Pacayacu and Sacha River watersheds, where oil operations have taken place. Preliminary data shows extremely acidic drinking water (pH<5.0) along with decreased oxygen levels means of 1.9 mg/L. My hypothesis is that sample sites such as wells and streams that are closer to oil production features such as residual pools, pumps, and oil refineries have inferior water quality to those located further away. I used geographic information systems to map and operationalize spatial variables such as path distance from the sites to oil operation features among other topographic factors. I will test the hypothesis using ordinary least square regression analysis. The results of this study will inform local organizations about the direct impacts on water quality and public health of oil operations in the area. We hope this study will help direct conservation and restoration efforts towards reversing anthropogenic environmental degradation in the area.

Model membranes are important tools for investigating physical properties of lipid bilayers. Unfortunately, current techniques for producing giant unilamellar vesicles (GUVs) with biologically and physically relevant properties (namely: inclusion of charged lipids, encapsulation of high salt buffer, and compositional asymmetry between the inner and outer leaflets) are difficult, unreliable, and/or low throughput. We examine a method called cDICE, developed by Abkarian et al., that has the potential to address all of these issues. Our results here for charged lipids in high salt buffers are promising, whereas asymmetry has proven difficult to measure.

The field of gene therapy relies on vehicles that can introduce exogenous genetic material into target cells in order to affect the production of a desired therapeutic protein. Viruses have long been used as delivery vehicles, but concerns over immunogenicity have fueled efforts to develop alternative nonviral delivery vehicles. Current polycationic polymers used in gene delivery methods generally exhibit poor transfection efficiencies and also dose-dependent cell toxicity. This project seeks to address these parameters through the synthesis of a novel copolymer of 2-hydroxypropyl methacrylate (HPMA) and a methacryated oligoarginine peptide (MaAhxR10). In phase one, the HPMA-oligoarginine copolymer was synthesized and tested for basic functionality. Transfections of HeLa cell populations demonstrated that the transfection capabilities of this polymer were comparable to those of the oligolysine standard, but the toxicity was significantly enhanced. Phase two consisted of modifying the peptide with neutral amino acid residues (such as glycine) to reduce the charge density of the polycationic peptides. Further, a more efficient approach to preparing arginine peptides was developed by chemically guanidinylating lysine residues. This approach was used to generate a lysine and a homoarginine version of each polymer. The resulting copolymers were tested for transfection efficiency and cell toxicity in cultured mammalian cells. Copolymers made from the resulting ‘spaced’ peptides (MaAhx(GK)5 and MaAhx(GhR)5) demonstrated reduced cytotoxicity compared the MaAhxR10 and PEI standards, but were still comparable in transfection efficiency to the oligolysine standard. While the delivery vehicle has not performed as expected, the project may have revealed a relationship between charge density (and therefore accessibility of charge) and cytotoxicity. The next step will be to generate another panel of polymers, this time with even more spacing between positive charge residues. This will allow for the identification of a correlation between charge density and cytotoxicity.

Gene therapy is the treatment of disease by inserting genes or altering them in a subject’s cells and tissues. The Pun lab develops cationic polymer-based delivery vehicles. Our agenda was to determine the intracellular fate of drug delivery vehicles carrying the therapeutic genes in order to better understand cellular barriers that limit efficient delivery. For this project, we applied two methods to study the intracellular trafficking of polyplexes (polymer-DNA complexes). The first approach is a quantitative method using subcellular fractionation to separate organelles and determine the distribution of polyplexes in various cellular compartments after internalization. We hypothesized that by using methods of centrifugation, organelle-specific enzyme assays, and radiolabeling, polyplex distribution could be quantitatively analyzed. We demonstrated successful separation and quantification of organelles using a combination of differential and density-gradient centrifugation. The polyplex distribution in each fraction was determined through radiolabeling. In this way, a quantitative “snapshot” was taken of the distribution of polyplexes inside the cell, and yielded information for the identification of potential bottlenecks in trafficking to the nucleus. Our second approach is to study the intracellular trafficking of drug delivery vehicles by time-lapse fluorescence imaging combined with single-particle tracking. This complementary approach reveals both the time-course and behavior of particles as they are trafficked within the cell. By fluorescently -labeling the polymer and applying the polyplexes to cells, we successfully observed their motion within a single cell. We further optimized the quantity of polyplexes delivered and the image capture techniques to robustly track polyplexes within a single cell. Our work therefore provides improved methods for determining the mechanism and efficacy of gene delivery systems and potentially any particle-based drug delivery system. For future work, we plan to investigate the process of unpackaging of the polyplexes inside cells.

Many soldiers acquire traumatic brain injuries (TBI), such as edema, hemorrhage, and traumatic axonal injury, from improvised explosive devices. Currently there are no adequate brain-imaging devices for use near the battle field. Patients with TBI may require a complete hemicraniectomy to examine the extent and location of the injury. This presents a great need for field-deployable imaging devices optimized for brain imaging with the spatial resolution of CT scans. Previous work has shown that intraoperative ultrasound, ultrasound used during surgery, can create images of brain-based tissue stiffness. For this project, rats have been given TBI using controlled cortical impact methods and their brains have been imaged using shear wave imaging to quantify the stiffness of brain tissue. The ultrasound images have shown patterns of brain stiffness that exhibit the extent and location of TBI. After the brain tissue stiffness for TBI was quantified using shear wave ultrasound, the data was used to optimize ultrasound machines for imaging moderate TBI. The Verasonics ultrasound device in the lab was optimized to image brain tissue by tracking the shear wave, generated by the transducer, as it propagates through brain tissue and correlating propagation speed with stiffness values. The images obtained from the ultrasound device were compared against histology. The success of this project would ultimately lead to increased quality of care for soldiers with TBI near the battlefield.

The Banda Islands in Indonesia have been important cultural and trade centers for centuries. With their unique native nutmeg production, they have attracted merchants from around the world including Islamic spice traders and Dutch and British merchant companies. The site of BN1 located on Banda Neira Island may contain an important archaeological record of early contact with Islam. Following the thinking that behavioral and dietary changes soon follow religious conversions, this study looks to the presence and use of pigs as a proxy for Islamic versus Animistic religious orientation. I examine the density of pig skeletal remains through time by using tradeware ceramics as chronological markers. Additionally, I investigate the site formation processes with geochemical methods and search for chemical evidence of pig presence in the form of fecal lipid biomarkers. Gas Chromatography-Mass Spectrometry of total lipid extracts from the BN1 archaeological sediments can determine with greater resolution and accuracy the presence or absence of pigs at the site based on known sterol lipid ratios derived from decomposed feces. This suite of archaeological and geochemical data will provide an insightful picture of local-level impacts of Islam in the Banda Islands.

Stroke is the degeneration of brain tissue caused by blockage of blood flow to the brain. It is one of the leading causes of adult disability and death worldwide. We used ultrasound elastography, a technique that measures local tissue deformation from ultrasound-induced shear wave propagation, to measure shear modulus, the ratio of shear stress to the shear strain, of brain tissue after ischemic infarction, loss of brain function due to low brain blood supply, to determine changes in individual hemisphere due to that injury. We imaged live mice that were 3 hours (hyperacute), 24 hours (acute) and 72 hours (subacute) after surgical occlusion of the middle cerebral artery, and measured the shear modulus of ipsilateral and contralateral brain hemispheres compared to corresponding values of control mice. The mice with hyperacute stroke showed statistically significant increase in shear modulus in both hemispheres compared with control animals. The mice with acute stroke showed a significantly decreased shear modulus in ipsilateral hemisphere and a significantly increased shear modulus in contralateral hemispheres compared to control mice. The mice with subacute stroke showed no significant difference in shear modulus compared to control mice. Significant differences were shown between ipsilateral and contralateral shear modulus values measured 24 hours and 72 hours after infarction. We hypothesize that the shear modulus differences among hyperacute, acute, and subacute stroke mice reflect the initial development of edema and reduction of cerebral blood flow in the ipsilateral hemisphere, and the initial reduction of blood flow then later development of edema in the contralateral hemisphere. Thus, ultrasound elastography may be a sensitive method to detect subtle changes in brain elasticity post stroke. Future work will include an attempt to remove the imaging artifact through the design of a new elastic imaging machine. This research will be a key step toward exploring the ultrasound parameters for imaging stroke, which will ultimately contribute to the development of a better imaging tool for potential stroke patients.

Ultrasonic neuromodulation is the process of stimulating different neural pathways in the brain through the use of transcranially delivered ultrasound in order to induce clinically relevant emotional, cognitive, physical or behavioral reactions. My research goal is to find out if it is possible to add anatomical specificity to neuromodulation through the use of modulated focused ultrasound (mFU), where 'modulated' refers to adding complex temporal structure to the waveform to optimize neuromodulation. Since mFU has a small focus relative to the brain, the acoustic power from the ultrasound can be moved within the brain; previous experiments have used unfocused ultrasound which sends ultrasound through the entire brain. We hypothesize that we can map varying physical reactions through stimulation of different parts of the brain, with our initial emphasis on the motor cortex because stimulation there produces readily observable results. With lightly anesthetized mice as our subjects, and always delivering ultrasound through the skin and skull, we began neuromodulation with unfocused transducers to transmit our ultrasound. This critical step was done to determine the optimal intensity, waveform shape and carrier frequency of ultrasound that created a robust observable effect. Next we translated these ultrasound parameters to our mFU transducer. The current stage of the process includes moving through different parts of each hemisphere and recording the associated motor response such as leg movements and tail flicks. We have already seen a change in activity between activating the quadrant of the brain containing motor cortex relative to the opposite quadrant. Going forward, we intend to see if we could produce varied motor movements as we change the point of stimulation within the brain hemisphere containing the motor cortex, along with changes in applied intensity, and duration of stimulation. Clinical implications for neuromodulation research include retraining brain function after injury.

The purpose of the study is to contribute to the development of HIV prevention efforts by determining primary sources of reproductive health information for Luo youth of Western Kenya. In 2006, there were an estimated 55,000 new HIV infections in Kenya; the majority affected youth aged 15-24. HIV prevalence is highest, 20.2%, among the Luo ethnic group. The high infection rate among youth generally, and among the Luo in particular, stresses the need for HIV prevention tailored for Luo youth, which can be informed by existing survey data. The Asembo Baseline Cross Sectional Survey (Asembo BCS) was conducted by the Kenya Medical Research Institute & Centers for Disease Control and Prevention’s HIV Research Branch in 2003-2004 in the rural, predominately Luo community of Asembo. Interviewers collected data on health information-seeking behaviors, sexuality, gender norms, and HIV prevalence and risk factors from 1822 randomly selected Asembo residents. We conducted a secondary analysis using the Asembo BCS database, focusing on Luo youth respondents ages 13-24 (n=1,283). Factors examined included identification of the most important and preferred source of sexual health information, types of sexual health issues typically discussed with parents and friends, comfort level speaking with parents about sexuality, sources of HIV/AIDS information, and whether the youth had recently received HIV/AIDS information. Using descriptive statistics we characterized the distribution, dispersion, and central tendency of these factors. Preliminary results suggest that important and preferred sources of reproductive health information include schools, media and friends. Additional analyses are being conducted to describe parent-youth communication and recent exposure to HIV/AIDS information. Findings regarding where and from whom Luo youth receive information about sexual and reproductive health will be used to inform the design of targeted HIV prevention for Luo youth.

This project will investigate the hypothesis that in utero exposure to clinically relevant doses of diagnostic ultrasound increases the risk for the manifestation of symptoms of autism. It is hypothesized that autism is caused by a triple hit of risk factors: a genetic susceptibility, a developmental time window of vulnerability, and an environmental factor. Diagnostic ultrasound is a candidate environmental factor within this “Triple Hit Hypothesis” for autism. To take the first steps toward testing our hypothesis, we propose to assay for ultrasound-associated changes in behavior and neuroanatomy of three mouse strains that have been well characterized in terms of social behavior. One strain is known for normal social behavior and the other two are known to exhibit autistic-like behaviors. Specifically, standard diagnostic ultrasound will be applied to mice at a key developmental time point during pregnancy to half of a group of mothers, and the other half will undergo the same procedure, but the ultrasound will not be turned on (sham application). At three weeks of age, the social, repetitive, and other relevant behaviors of the pups will be assayed using a battery of tests that includes a social approach test, a juvenile play test, open field observation, and several other methods. We will then analyze the data for significant differences in magnitude or occurrence of these behaviors within the three strains of mice based on their ultrasound exposure group. If the magnitude and/or occurrence of these behaviors are greater in the ultrasound-exposed group compared to the sham group in at least one of these strains, then our hypothesis will be supported and further experiments will be critical.

Sepsis is a complex, incompletely understood and often fatal disorder that is one of the most common causes of death in children worldwide. The overlapping pro- and anti-inflammatory responses, and resulting complications contribute to the complexity of sepsis and have made it difficult to find biomarkers and therapeutics targets for this disorder. Mast cells and basophils are well-known for being key players in allergic responses. However, it is less known whether these cells can influence the outcome of bacterial infections. We hypothesize that mast cells and/or basophils can limit the severity of the inflammatory response against bacterial infections to preven sepsis. We found that mast cell and basophil deficient mice (Cpa3-Cre; Mcl-1^flox/flox mice) exhibit signs of morbidity (reduced body temperature), kidney dysfunction (high blood urea nitrogen levels), and are unable to govern the inflammatory response induced by cecal ligation and puncture (CLP), a well-established model of sepsis. The inability to regulate the inflammatory response induced by CLP is shown by the increased number of neutrophils in the peritoneal cavity of the mast cell and basophil deficient mice. Our initial results indicate that Cpa3-Cre; Mcl-1^flox/flox mice that have been previously engrafted with mast cells show similar signs of morbidity, kidney dysfunction, and neutrophil numbers in the peritoneal cavity than non-engrafted mice. These results could indicate that basophils are more responsible than mast cells for the down-regulation of inflammation and reduced morbidity after CLP. Considering these new results, the next step will be to obtain basophil deficient mice to see the response after CLP and what effect basophils might have on the anti-inflammatory response during sepsis. 

The Sarmiento formation in the Gran Barranca region of Argentina is comprised of eolian, fluvial and volcanic sediment formed between twenty and forty million years ago. Climate change, sea level variation and tectonics driving subsidence were all important factors in the deposition of the Sarmiento Formation and research on the paleoenvironments of the region includes study of eolian and fluvial processes, paleoclimate and the evolution of grasslands. The sediment of the 33.581 million year old Vera section of the Sarmiento Formation is the focus of this study. To better understand the environments of deposition of this section, grain size and magnetic susceptibility were analyzed for over fifty sediment samples. A particle size analyzer was used to provide grain size distributions for each sample, which were then used to look for loess signatures in the sediment. Loess (wind blown sediment) is predominantly composed of fine sand and silt sized particles, and its detection can be evidence of arid, windy climates. Ongoing sedimentary analysis has revealed a predominance of silt sized particles, indicating a loess signature. Magnetic susceptibility analysis is a process that identifies iron-bearing minerals in sediment. The amount of iron present in the Vera sediment can be an indication of chemical weathering in Gran Barranca during the Eocene. Extent of weathering can also indicate past climate conditions. The results of the grain size and magnetic susceptibility analysis for each individual sample will be related to stratigraphic height and age so that a history of climate and environmental change for the Sarmiento Formation can be created.

Through much of the 20th century, the ASARCO copper smelter in Tacoma released heavy metal-rich ash, contaminating local soils and sediments. However, the smelter plant is not the only source of contamination: arsenate pesticides and other heavy metal sources contributed to soil contamination. Understanding the source of contamination can lead to remediation or replacement of contaminated soil. Here we use soil magnetic properties to identify minerals associated with smelter contamination. Specifically, iron bearing minerals can uniquely identify emissions generated from the smelting process. We use magnetic susceptibility (both high and low field), coercivity and magnetic remanence to produce a “magnetic fingerprint” of minerals from smelter dust. These properties are affected by grain size and mineralogy of magnetic particles. Remanance (Mr/Ms) and susceptibility/coercivity (LF/Hc) ratios are similar in all soils regardless of degree of contamination. However, high field susceptibility does appear to be related to arsenic concentrations of smelter contamination (R2= 0.4656). Further sampling and testing are being conducted to extend that relationship to other sites. Our results suggest that magnetic properties and testing can be used to efficiently and cheaply assess smelter contamination throughout the South Puget Sound region.

The neuromodulator dopamine is important for motivated behavior. Past research provides evidence that dopamine neurons encode the expected value of reward-predicting stimuli in situations with a single option. The current project investigated dopamine transmission when multiple options are present, as previous studies have arrived at conflicting conclusions. One study found that dopamine neurons encode the value of the subsequently chosen option, regardless of the expected value of the option foregone; whereas the other concluded that dopamine neurons encode the value of the greater available reward, regardless of which option was actually chosen. To address this discrepancy, we recorded changes in nucleus accumbens dopamine concentration using fast-scan cyclic voltammetry in rats performing a decision-making task in operant chambers. Sessions included single-option trials intermixed with choices between two options that differed in reward size (number of food pellets) and effort requirement (number of lever presses). When rats were presented with a low-reward/low-effort option (1 pellet for 4 presses) and a high-reward/intermediate-effort option (4 pellets for 8 presses), they preferred the high-reward/intermediate-effort option. In separate sessions, when presented with the low-reward/low-effort and a high-reward/high-effort (4 pellets for 32 presses) options, the rats preferred the low-reward/low-effort option. Preliminary voltammetry results from single option trials suggested that dopamine release was greater following the presentation of the high-reward/intermediate-effort option compared to the presentation of the low-reward/low-effort option. In the other session type, however, dopamine concentration did not differ between the high-reward/high-effort option and the low-reward/low-effort option. In two-option choice trials, dopamine release associated with each reward/effort combination was similar to that in the corresponding single-option trials. Thus far, these results are more consistent with the claim that dopamine encodes for the option subsequently chosen, but the interaction between behavioral preference and encoding of expected reward value requires further investigation.

Since political violence first broke out in 1999, Côte d’Ivoire has experienced a drastic loss of medical equipment, drugs, and skilled health personnel. Two civil wars in the past decade have posed significant challenges to outside intervention from international NGOs, and the weakening economy has provided little support for a state-funded health system. Currently, Côte d’Ivoire suffers from some of the highest maternal and neonatal mortality rates in the world, resulting from problems ranging from infection and hemorrhages to the compound effects of malaria, malnutrition, and HIV/AIDS. My research responds to these challenges by exploring what would be needed in a grant proposal to The Global Fund, a global funding agency that supports the fight against HIV/AIDS, malaria, and tuberculosis in developing countries. Of the thirteen Global Fund grants approved for Côte d’Ivoire in the past, none specifically address the problems of maternal and neonatal mortality. This project aims to understand why this gap in maternal and neonatal care persists and how it can be diminished in the context of political instability. To assess the efficacy of attempts to address health care gaps in Côte d’Ivoire, I analyzed the interventions currently in place to see if they were meeting their established goals and if not, why. Building upon this data and development theory, I created a proposal for improving maternal and neonatal mortality through a self-sufficient system that can remain in place should political violence persist. Preliminary research suggests that weak medical assistance is a leading cause of birth complications. Increased health monitoring during and immediately following pregnancy, as well as greater education for mothers on the crucial steps during this time period are vital to reducing mortality. Establishing self-sustaining health systems that are resilient in the face of violence and political uncertainty is invaluable across the developing world.

Ultracold molecules are a new frontier in atomic physics that have many possible applications which include tests of fundamental physical theories and the development of quantum information science. One method of creating such molecules is through the process of photo-association (PA), where two ultracold atoms are optically excited into a bound molecular state. We will present the design and construction of an extended cavity laser diode system that will be used for PA of ultracold Lithium (Li) and Ytterbium (Yb) atoms. The exact energies for molecule formation are not known a-priori, so the laser has to be tunable over a wide range of frequencies near the wavelength of 556nm. To maximize the “mode hope free” (MHF) range where the laser frequency varies smoothly, we will simultaneously vary the current powering the diode and the length of the laser cavity. Once the MHF range has been maximized, the laser will be used in PA spectroscopy in order to form Li-Yb molecules and characterize their energy structure.

Drug resistant strains of Staphylococcus aureus are now possibly responsible for more deaths than any other infectious disease in the United States. Phosphosignaling enzymes are thought to constitute a major class of drug targets in humans, but their potential as drug targets in bacteria are yet to be known to their full extent. In this study, we sought to develop an assay to test activity of an S. aureus signaling molecule, the tyrosine kinase CapB, which controls capsule biosynthesis. By interfering with capsule biosynthesis, we hope to interfere with virulence of S. aureus, allowing better clearance of the infection by the immune system. To assay for kinase activity, we recombinantly expressed the known kinase substrate Cap5O. Cloning of the Cap5O insert into the pQE30 vector created an expression plasmid with a 6XHis-tag on the C-terminus of Cap5O. The plasmid was grown and harvested in Rosetta cells. Purification of the protein was carried out through Metal Affinity Chromatograpy to obtain pure protein for activity assays. The results of this study will provide the starting point for an assay to screen for compounds that interfere with capsule biosynthesis. To date, no screens for the bacterial tyrosine kinases have been developed.

Stroke is one of the top causes of death in the United States; however, due to the apoptotic death of neuronal cells from lack of oxygenated blood, prognosis and recovery is not always favorable even if the event is survived. Our lab focuses on finding agents that are already clinically relevant in other areas, but might also show neuroprotective qualities in cerebral ischemic events. The compounds we have focused on in this study are progesterone and its metabolite allopregnanolone. Previous studies have shown that both compounds have neuroprotective qualities in vitro and that progesterone administration in vivo suppresses inflammatory response and subsequent edema. Our hypothesis is that both progesterone and allopregnanolone would show neuroprotective, as well as neurogenic, qualities in vivo. This is tested through a mouse stroke model using progesterone receptor knockout mice and wild type mice, both being subjected to a middle cerebral artery occlusion. Different groups are treated with different compounds or control. The brains are then perfused, fixed, and sliced to be put through immunohistochemistry, where cells are identified by certain neurogenic markers (bromodeoxyuridine incorporation and doublecortin labeling). Slides are finally analyzed through confocal microscopy to look for these markers in certain areas such as the hippocampus, striatum, and lateral cortex. Finally, positive cells are counted in both the ipsilateral (stroked) and contralateral hemispheres. We anticipate an increase in the number of bromodeoxyuridine and doublecortin labeled cells in mice subjected to progesterone and allopregnanolone, and we also expect this to be more dramatic in the ipsilateral side due to new cell migration to the injury site. Hopefully, these results will allow for a greater push in implementing progesterone and allopregnanolone in clinical use for the recovery of ischemic stroke.

Vietnam, which is experiencing a heath transition from communicable to non-communicable diseases, has an increased burden of stroke. Understanding its risk factors, including stress, are needed. The goal of this study was to evaluate the relationship of chronic stress and stroke symptoms (which are indicative of a potential stroke) as well as hypertension and diabetes in participants ages 35 and older in Da Nang, Viet Nam. We randomly identified adults, age 35 and older using voter registration lists for inclusion in this study. Interviews and health procedures were conducted either at the local health clinic or participants’ homes depending on preference. Stress was measured using the chronic stress burden scale assessing problems occurring in five domains: problems with one’s own health, with another’s health, job difficulties, financial strain, and relationship problems. Hypertensions was defined as self-reported or measured systolic blood pressure of 140 mm Hg or diastolic of 90 mm Hg and higher. Stroke was ascertained as a self-report of symptoms related to stroke or transient ischemic attack collected by the Questionnaire to Verify Stroke-Free Status (QVSFS). Multiple logistic regressions assessed the odds of increased risk of stroke related to level of stress categorized as no stress, 1 or 2 symptoms, and 3 to 5 symptoms in unadjusted and adjusted models. After adjustment for demographic, health characteristics, as well as diabetes and hypertension, individuals experiencing stress at the highest level (3-5 domains) were more than eight times as likely to report stroke symptoms as those with no stress (OR: 8.13, 4.43-14.91). Lower levels of stress (1-2 domains) also increased significantly the risk of stroke symptoms (OR: 3.27, 95% CI: 2.01-5.34). After adjusting for diabetes and hypertension, along with other potential confounding factors, we found stress to be independently associated with risk of stroke as measured by stroke symptoms.

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.

T lymphocytes protect against the universe of pathogens by generating a diverse repertoire of antigen receptors called T cell receptors (TCRs). Each individual T cell expresses TCRs with a single antigen specificity. The induction of self-tolerance is crucial to the health of the individual because it culls the T cell population of those cells that bear self-antigen reactive receptors. Such autoreactive T cells are the driving force behind autoimmune diseases. One mechanism for inducing self-tolerance is TCR revision, by which peripheral T cells replace self-reactive TCRs with newly generated, non-autoreactive TCRs. This replacement results in cell rescue and involves cleaving DNA, which requires implementation of many controls to protect genome stability. Cells undergoing TCR revision appear to be localized in the spleen and lymph nodes inside a confined microenvironment known as the germinal center (GC), and this constrainment may provide a means for controlling this process. Given the rarity of revising T cells, my project focused on potential means for increasing the frequency of cells undergoing TCR revision using as a model system TCR transgenic mice, in which the majority of T cells express TCRs of the same antigen specificity. We hypothesized that expanding the GC T cell compartment would increase the frequency of cells undergoing TCR revision. I carried out injections of lipopolysaccharide, sheep red blood cells, and anti-CD40 antibody into TCR transgenic mice in an effort to induce GC formation. I then analyzed the accumulation of post-revision and revising T cell populations by flow cytometry. Use of any of these agents to increase TCR revision would facilitate the isolation of post-revision and revising T cells and allow further investigation into the mechanistic details of this tolerance process. Our goal is to understand how the challenging balance between having a diverse T cell repertoire and avoiding autoimmunity is maintained.

Diabetes is a serious and prevalent disease affecting the lives of many people. Studies have shown that cardiovascular calcification, or calcium-phosphate mineral deposition in blood vessels, cardiac valves, and myocardium, is prominent in patients with Type II diabetes (T2D), accounting for the heightened cardiovascular morbidity and mortality in this population. Using a T2D mouse model, our laboratory has shown a positive correlation of hyperglycemia with cardiovascular calcification. A multi-ligand receptor, namely the receptor for advanced glycation-end products (RAGE), was high in cartilaginous and calcific areas of blood vessels and aortic valves and translocated to the cell membrane of the cells with osteochodrogenic (bone and cartilage precursor) and chondrocytic (cartilage) properties. To study whether RAGE is involved in the early signaling of osteochodrogenic differentiation and calcification of cardiac valves, we cultured bovine valve interstitial cells (BVICs) in procalcific medium containing normal and high glucose in a range of hyperglycemia. We found that high glucose increased calcification of BVICs by up to 3 folds. Expression levels of RAGE and its ligands are determined by reverse-transcription PCR and Western blot analysis. The biological activity of RAGE are blocked by a commercially available neutralizing antibody to see whether RAGE is required for the high glucose-associated BVIC calcification. Our results show a decrease in BVIC calcification when RAGE activity is blocked by a neutralizing antibody. For future studies, we will create a cell line with lower expression levels of RAGE via small RNA interference.

Glioblastoma multiforms (GBMs) are malignant brain tumors and are among the most lethal cancers in the United States. Current clinical treatments involve surgical resection followed by chemo radiation therapy. Temozolomide (TMZ) is a DNA methylating agent that has shown promise in post-operative chemo radiation therapy, yet its effects are mediated greatly by the DNA repair protein O6-methylguanine-DNA methyltransferase (MGMT). The addition of DNA repair inhibitor O6-benzylguanine (BG) as part of a concurrent treatment with TMZ has shown promise, but suffers from unacceptable toxicity due to BGs poor pharmacokinetics. Here we report the development of nanoparticles (NPs) loaded with a BG payload that can efficiently cross the blood brain barrier (BBB) and with the addition of targeting agents may be a more effective method for drug delivery by reducing toxicity in healthy tissue. These NPs are composed of a magnetic core and biocompatible polymer surface coating. The NPs are then conjugated with BG (NP-BG) by attachment to the polymer surface. NP-BG is further modified with chlorotoxin (NP-BG-CTX), a tumor targeting peptide, through a polyethylene glycol linker. NP-BG-CTX had a mean hydrodynamic size of 75nm with a surface charge of about +4mV and have demonstrated specific targeting in vitro of GBM cell line SF767. Finally, we have shown that NP-BG substantially reduces MGMT activity in SF767 cells in vitro. These results suggest that NP-BG-CTX have a higher specificity to tumor cells than normal healthy cells in cancer patients. This would decrease drug toxicity in patients and reduce tumor resistance to TMZ, potentially providing a more effective treatment and improve clinical outcome.

Every year, 700,000 people in the US alone suffer a stroke. Out of this, 87% of these incidences comprise of ischemic strokes. Ischemic strokes occur when normal blood flow to the brain is inhibited by some sort of blockage, thus starving the brain of oxygen and glucose. This results in neural tissue damage. HDACs (Histone Deacetylases) seem to play a major role in promoting neural tissue damage, thus using an HDAC inhibitor should protect already existing neurons from insult resulting from ischemia or any other traumatic brain injury. We believe that the recently developed HDAC inhibitor, MI-192, has the potential to show strong affinity towards a key HDAC involved in disease pathogenesis and thus confer neuroprotective effects. We have analyzed drug affinity for HDAC 3 and the effects of MI-192 on neuronal damage and on infarct size using western blotting and a variety of histological methods, respectively. MI-192 may have the potential to make it to clinical trials.

Huntington’s disease is a hereditary neurodegenerative disease that is characterized by motor and cognitive dysfunction and currently has no known cure. One of the earliest molecular events in Huntington’s disease pathogenesis is the down-regulation of the cannabinoid 1 receptor (CB₁), occurring prior to symptom onset and loss of any other receptor. Though the exact mechanism is unclear, it has been shown that CB₁ loss occurs almost exclusively in medium spiny neurons (MSNs), cells that play a key role in movement and degenerate later in disease progression. CB₁ knockouts in mouse models also exhibit exacerbated disease phenotype. Previous studies have attempted to rescue CB₁ function in mouse models by administering chronic cannabinoid agonists, but have produced inconclusive results. Our study investigates the effects of CB₁ receptor rescue from a genetic approach, by inserting flox-stop CB₁ (fsCB₁) specifically activated in striatal MSNs of R6/2 mice, the best characterized Huntington’s disease mouse model. We first confirmed the localization and functionality of the knocked-in fsCB₁ in R6/2 and CB₁ null mice (control), and then measured motor phenotype and synaptic integrity over time. Performance of R6/2 mice with and without fsCB₁ rescue was measured using rotarod, open field, novel object recognition, and clasping behavior tests biweekly from weeks 4 to 12 of age. MSN axon integrity was measured by IHC staining for caspase-6 and NPY-Y1R, markers for Wallerian-type degeneration. By observing behavior and axon integrity from pre- through late-symptomatic stages, we hope to establish a relationship between early MSN axonal loss and impaired motor phenotype in R6/2 mice, as well as link CB₁ function to axonal integrity through R6/2 and fsCB₁-R6/2 comparisons. This research will increase the present understanding of molecular mechanisms behind Huntington’s disease and provide insight into CB₁ as a therapeutic target.

In recent years, nanoscience has become a major focus: from technologies in solar cell coatings, to developments in biomolecular detection methods. Utilization of unique optical absorption properties of noble metals, such as gold or silver, opens the door for harnessing photons in new and innovative ways. For these reasons, great emphasis has been placed on the synthesis and characterization of nanostructures. The Pozzo Group has recently shown that structured gold clusters can be formed in solution by exploiting the hydrophobic and steric interactions in self-assembling nanoparticles. Hydrophilic polyethylene-glycol (PEG) chains control long-range steric interactions, while hydrophobic alkane-thiol chains induce clustering through short-range attraction. This simple method has the potential to be a cheap and viable option for nanoscale fabrication; starting from the molecular level and building up. This method of nanoclustering yields multi-particle aggregates in solution. Clusters of different geometries and sizes are formed based on the number of consistuent gold particles in each aggregate. It has been shown that the total amount of PEG adhered to the surface of the gold particle impacts the extent of clustering. At lower PEG concentrations, more diverse populations of nanoclusters are observed. In this project, size-exclusion chromatography (SEC) is being utilized for separating cluster populations. A cluster dispersion is pumped through a column that is packed with a porous stationary phase. By size-exclusion, the largest clusters exit the bottom of the column first, while the smaller clusters have a higher retention time as they navigate the pores of the matrix. The viability of SEC for controlled separation of cluster populations is being investigated on different systems, with the goal of isolating populations of similar aggregation types. Population analysis with SEC is simple and can be used to identify the fundamental parameters that lead to polydispersity in cluster size and shape.

More than 50 percent of stars are found in binary systems. A cataclysmic variable star is a binary system consisting of a white dwarf and a larger, cooler star in an orbit with a period of only a few hours. These stars are so close that the white dwarf distorts the gravitational field of the other star and mass is transferred from the cool star to the white dwarf via an accretion disk. The defining characteristic of the type of cataclysmic variable called dwarf novae is that they irregularly increase and decrease in brightness. The increases (called outbursts) are caused by an instability in the accretion disk that causes an increase in accretion. After an outburst, these stars steadily decline in brightness until they return to their original state. However, the subgroup of dwarf novae called Z Cam systems sometimes become stuck in a state called a standstill, where they stay at a brightness near outburst level, and then decline to their original level. Using data taken at Kitt Peak National Observatory and Apache Point observatory over a series of five months, we were able to analyze spectra of two unusual objects as they go through their outbursts. We can find out what state they are in by comparing their spectra to light curves from the American Association of Variable Star Observers. What is unusual about these objects is that rather than fade to their original state after standstills, they go up to another outburst. Using these spectra, we hope to find the accretion rates of the stars at standstill, and then compare to other stars of the same type to better understand why they have this peculiar behavior.

This project explores the innate immunity of the earthworm species Eisenia fetida during development in context of its symbiotic relationship with Verminephrobacter eiseniae, a known bacterial symbiont. Innate immunity, unlike adaptive, has largely unspecific properties against foreign particles yet many organisms that possess only innate immunity manage to allow select beneficial symbionts to thrive within their body while at the same time defending against pathogenic infection. In this study I begin to explore involvement of innate immunity in the relationship between E. fetida and its symbiont V. eiseniae. This project examines expression of a common innate response, oxidative stress, in early stages of V. eiseniae colonization of its host E. fetida. We hypothesize that innate immunity activity is spatially separated from colonization of bacterial symbionts. To test this hypothesis, I will stain for nitric oxide synthase (NOS), an enzyme commonly involved in oxidative stress, during the development of E. fetida in the presence of V. eiseniae. I will then proceed to compare the location of NOS to the location of V. eiseniae. If the hypothesis is correct, then oxidative stress response in the worm eliminates foreign organisms everywhere except where the symbionts reside. On the other hand, if oxidative stress is at the same location as the known symbionts at the same time during development, then the oxidative stress may eliminate other foreign organisms except the symbionts, which have developed a mechanism for overcoming the stress. The results of this research can reveal how innate immunity overcomes the dilemma of accommodating symbionts. In addition, if E. fetida is particularly easy to work with for this type of experimentation, then E. fetida could prove to be an ideal model organism for studying innate immunity. Furthermore, insights into innate immunity may hold the key to new innovative ways of combating disease.

Stellar flares are the explosive release of energy caused by the reconnection of magnetic fields on stars. Flares release large amounts of UV and X-ray radiation, and may affect the possibility of life on exoplanets, planets orbiting stars other than our own Sun. A star's mass and age greatly affect the frequency, duration, and energy of flares. Therefore, it is important to understand these aspects for a variety of masses and ages. The age of a star is related to the rate of its rotation as well as the rate at which it flares; older stars spin much more slowly and also flare less frequently than younger stars. M dwarfs, stars less than half the mass of the Sun, are commonly used in stellar flare research because they make up the majority of stars in the Milky Way galaxy. To conduct our research, we used two months of data from the NASA Kepler spacecraft to study the light curves of one young and three old M dwarfs. We were able to identify flares in these light curves and measured their durations and energies. We found that the vast majority of flares on all four stars have a short rise time and a long decay time. We also found that flares with long durations have larger total energies, and that older stars have fewer large energy flares compared to younger stars. Through using the light curves, larger, more prominent flares were easily detectable in comparison to smaller flares. Further research would concentrate on more accurate ways toward detecting such small flares.

MRI is by far the most sensitive technique for screening high-risk women and assessing the extent of breast cancer. However, dynamic contrast-enhanced imaging (DCE-MRI), the standard breast MRI method, produces many false positives. As the American Cancer Society now recommends annual MRI screening for high-risk woman, estimated to affect up to 1.8 million women in the US, there is a quickly growing need to reduce the false-positive rate of breast MRI in order to reduce the number of unnecessary biopsies that could result. Our research explores the diagnostic potential of diffusion-weighted imaging (DWI), a MRI technique that depicts the diffusivity of water molecules in the tissue as a way to characterize the cellularity of breast lesions. Research has shown that DWI has the potential to increase the diagnostic accuracy of breast cancer, serving as a promising adjunct tool to DCE-MRI for reducing false positives. Our goal is to evaluate the ability of DWI for non-invasively discriminating benign and malignant breast lesions that were identified as suspicious on conventional breast MRI. Specifically, we will investigate image registration to improve alignment of DWI images, the effects of the contrast agent used in DCE-MRI on DWI measures, and the optimal diffusion sensitization of DWI in discriminating benign and malignant breast lesions. These characterizations of DWI can potentially help to optimize the acquisition and processing of DWI scans for future clinical implementation and help guide the appropriate use of DWI as an adjunct tool in radiological diagnosis of breast cancer.

In eukaryotic organisms, ubiquitin is a small protein modifier that is attached to proteins and targets them for proteasomal degradation. In an analogous fashion, in Mycobacterium tuberculosis (Mtb), the prokaryotic ubiquitin-like protein (Pup) is conjugated to lysine side-chains of substrate proteins and targets them for degradation by the Mtb 20S proteasome. This pathway has been shown to be essential for Mtb infection in mice, and is a potential target for theraputic reseach. The enzyme PafA is responsible for attaching Pup to its substrates and has previously been shown to be essential for lethal infections by Mtb in mice. We have developed a fluorescent activity-based probe to monitor PafA activity in vitro by conjugating Fluorescein isothiocyanate (FITC) to free lysine, a known substrate of PafA. FITC-Lys demonstrates a high degree of specificity for PafA, even in the presence of cell lysates. This probe was employed to visualize PafA activity by in-gel fluorescence. It has allowed us to identify a minimal sequence of Pup, the C-terminal 26 amino acids, that is recognized by PafA. Alanine mutants of the 25th and 26th C-terminal residues demonstrate that leucines at these positions are critical for PafA interaction.

This project is aimed to examine the ways gender stereotypes are reinforced in sports. Particularly I wanted to investigate the impact this has on transgendered, intersex or otherwise gender nonconforming student athletes. This connects to current research published in the National Transgender Discrimination Study, examples of homophobia in women’s college basketball, the impact of genetic testing on athletes competing in the Olympic Games, the publication of the NCAA Inclusion of Transgender Student-Athletes, and the recent critical analysis of transgender legislation politics by Spade. For this I implemented the following methods: Literature review, focused analysis of data collected by Grant et al., social media analysis of internet blogs and websites, photo voice, critical discourse analysis and review of documentary films. These different approaches were intended to help me in the investigation of my research question by approaching the subject from multiple angles. With my research I hope to constructively contribute to the ongoing dispute of lingering discrimination based upon sexual orientation, gender politics and human rights in the United States and around the globe. Furthermore I aim to show how the struggle for acceptance of gender nonconforming students is still complicated by barriers raised on the grounds of biases and unfounded stereotypes. This project therefore shows how, despite the claims of policies aimed to end discrimination on the basis of race, creed, ethnicity, nationality, gender, sexual orientation or ability, not all students are treated equally. Finally it also connects to my undergraduate honors research project where I examine the transgender experience and quality of life in connection to gender reassignment surgery in the United States and Germany.

The hippocampus is well known for its role in spatial memory. A better understanding of how the hippocampus functions will provide us with insight into how memories are made and stored. This knowledge may ultimately lead to improved preventative and treatment strategies under conditions in which the hippocampus is altered, such as after traumatic brain injury or in cases of age-related memory impairment. Within the hippocampus there are cells commonly known as “place cells”. These cells are most likely to fire when a navigating organism is in a certain space in an environment, known as that cell’s “place field”. The activity of place cells may be modulated by dopamine input, which can be released tonically or phasically. For example, when overall dopamine input from the ventral tegmental area is disrupted, the place fields of cells within the CA1 subregion of the hippocampus are disrupted and spatial memory deficits occur. To test the idea that phasic dopamine, in particular, is necessary for normal place cell activity and spatial memory performance, we tested spatial working memory while recording electrophysiological activity of neurons in the hippocampus in normal mice and genetically engineered ‘knockout’ mice whose phasic dopamine signaling was selectively disrupted. Preliminary electrophysiological results show differences between the groups, including a nearly significant increase in the spatial selectivity that corresponds to a significant decrease in the size of place fields from the knockout mice when compared to controls. Behaviorally, knockout mice adapt more quickly than control mice following a change in the location of a reward, indicating that phasic dopamine input is not required to perform this task. Rather, removal of the phasic firing pattern may have unmasked a potential facilitating role of tonic dopamine release in flexible goal directed behaviors.

Over the past ten years we have analyzed a variety of plants that produce volatile oils grown in our Chemical Garden on The Evergreen State College campus. These plants lend themselves well to extraction by steam or ethereal solvents and are easily analyzed through Gas-Chromatography Mass Spectrometry (GC/MS). The plants we have focused on are garlic, St. John's Wort, hops, mints, and lavender. This poster reports on our progress towards isolating and identifying individual constituents of Devil's Club (Oplopanax horridus), a local plant with a long history of use as a medicine. We have been curious as to the best time of year to harvest the plant, as there is some debate in the herbalism community on this point. The Devil's Club was distilled at different times of the year and analyzed using GC/MS. The non volatile constituents were extracted using a number of different solvents and analyzed using Thin Layer Chromatography (TLC). This research will provide information on the effects of harvest time on constituent levels, as well as providing knowledge of the chemical make up of this plant.

Bacterial conjugation is the process of transferring genetic material from one cell to another. In the case of F-plasmid of Escherichia coli, the conjugation system is composed of inter-membrane proteins as well as hairlike appendages called pili encoded by the plasmid. Current interest in the characterization of F-plasmid lies with its ability to not only transfer itself to other cells but also its potential to mobilize auxiliary genes, such as antibiotic resistance, that yield some selective advantage. However, many basic details about the F-plasmid system, such as how donor cells recognize recipients and the function of the protruding pili, remain unknown. Through the use of an optical tweezer setup, I am able to isolate individual, fluorescently labeled E. coli cells in close proximity to each other. Once in close proximity, I will monitor the forces applied by donor cells on recipient cells and compare those forces to the forces potentially applied by donor cells on other donor cells. In addition, by utilizing the same optical tweezer setup, I will explore the statistical behavior of F-plasmid transfer as a function of cell-to-cell separation to investigate the potential function of the pili as transfer conduits.

Despite the numerous studies of the bacteria E. Coli, the basic motion of macromolecules confined to the cellular cytoplasm, such as mRNA, remains uncharacterized. I employ a technique called fluorescent correlation microscopy, which consists of binding synthesized mRNA with fluorescent proteins, imaging the cell throughout its lifetime, and analyzing the images using statistical autocorrelation. This technique allows me to determine the diffusion constant (D) as a function of cellular location while bypassing the error caused by higher order analysis. We predict that D is primarily dependent on the molecular crowding at its position, and there exists a tendency for mRNA to gather in the less crowded spaces in the cell, e.g. at the poles.

The spinal cord represents a key site of convergence for sensory feedback and cortical commands, but determining how each contributes to movement has not been uncovered.  Understanding how spinal circuits are structured and utilized to generate complex movements will be necessary to understand how the nervous system processes multiple inputs.  One approach to understanding motor circuits is to know how the output effects of neurons change between tasks.  Here we investigate the state of the spinal cord by stimulating at specific sites in the cord and recording electromyographic activity (EMG) in the forelimb muscles of a non-human primate.  By comparing the output effects of stimulation between two behaviors, a flexion/extension wrist task and a reaching task, we hope to understand the effects of cortical and sensory interactions in shaping motor output.  In addition, by creating a spinal map of stimulus triggered EMG activity, we hope to reveal the functional organization of spinal outputs during the behaviors.  In this experiment, we compare the effects of stimulus triggered averages of EMG between these two tasks at various sties in the cord.  Our results show a change in the effects on EMG activity dependent on the behavior performed in roughly 2/3 of sites.  Indicating that the configuration of the circuit that our electrode stimulates is dependent on the task performed.  When the non-human primate performs a wrist task, cortical input and sensory feedback recruit specific spinal pathways to activity certain muscles in order to perform the task.  When the task is altered, different cortical inputs and sensory feedback may inhibit or further excite that circuit, changing EMG activity.  Understanding the circuits responsible for the control of arm movements and how these circuits change with varying tasks, may allow us to utilize and exploit these spinal circuits to treat patients with spinal cord injury.

Manipulation of inflation reports is a rarely known tool that policymakers use to generate temporary growth and more revenue than they would with an explicit monetary expansion. The Argentine government has intervened in its Statistic Department since early 2007. Several private estimates and scholarly studies have shown that the real inflation rate is in fact between two or three times the officially reported inflation rate. However, the current studies have only produced price indexes from December 2007 to present, which does not include the crucial period when the manipulation in inflation reports was first implemented. In this project, I will create an alternative price index from January 2006 to January 2009, which will bridge the gap from when the Argentine Statistic Department was still respectable to when its was widely known of misreporting inflation rate. I will assess the validity of my index by comparison with the published studies for 2008, the year that already has an alternative price index. From my index, I will be able to determine an accurate date when the misreporting inflation date started, and by also having the data for that crucial period, I will contribute to the study on the cost of misreporting inflation. I will be using the monthly average prices of 60 representative goods to construct an alternative price index comparable to the officially reported one. This valuable dataset includes the publications of the Consumer Action League and receipt records in a supermarket’s archives. Both database are rare to find because the Argentine government started to censor and punish domestic independent studies on inflation with heavy fines beginning on 2011. The preliminary results show that the manipulation of reported inflation started in February 2007.

In Acute Myeloid Leukemia (AML) the adhesion interaction between the AML blast cells and the bone marrow microenvironment confers resistance of the leukemia cells to the chemotherapy drugs. One possible receptor that might facilitate this adhesion is the VLA-6 integrin. Our laboratory has been found it to be highly expressed in the AML stem cells, the cells that are believed to give rise to leukemic relapse. To investigate the role of the VLA-6 receptor, I performed assays to observe the effects of blocking this receptor on cell adhesion as well as the chemotherapy resistance. The adhesion assay consists of measuring AML patient cell binding to laminin-coated wells, as laminin is the ligand for the VLA-6 adhesion receptor. Adhesion mediated by VLA-6 is assessed by measuring binding in the presence or absence of the anti-VLA-6 antibody. The chemotherapy resistance assay involved the same conditions as the adhesion assay. We expected that blocking the VLA- 6 receptor would prevent the cells from adhering to the laminin and that this would also lead to lessened drug resistance. However, for the cells from one patient studied so far, there has not been a significant difference between the amount of adhered cells in these conditions. Additional experiments are in progress with more patient samples to elucidate the significance of the VLA-6 receptor in AML physiology and survival.

Measurements of the rotation of galaxies indicate that visible matter cannot account for all of a galaxy's mass. The prevailing theory is that each galaxy is surrounded by a halo of dark matter. Dark matter halo mass can be determined by measuring the velocities of dwarf satellite galaxies orbiting a central galaxy. We used data from the Sloan Digital Sky Survey (SDSS) DR8 and HyperLeda databases to identify massive, isolated galaxies including M94, M101, and NGC 2841 and prioritize their satellites for observation. Next we will make spectrographic observations to determine satellite velocities and use these results to infer the mass of the central galaxy. By comparing this value to the amount of visible mass in the galaxy, we will determine the mass of the dark matter halo.