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Description
Transient Receptor Potential (TRP) ion channels are a diverse family of nonselective, polymodal sensors in uni- and multicellular eukaryotes that are implicated in an assortment of biological contexts and human disease. The cold-activated TRP Melastatin-8 (TRPM8) channel, also recognized as the human body's primary cold sensor, is among the few

Transient Receptor Potential (TRP) ion channels are a diverse family of nonselective, polymodal sensors in uni- and multicellular eukaryotes that are implicated in an assortment of biological contexts and human disease. The cold-activated TRP Melastatin-8 (TRPM8) channel, also recognized as the human body's primary cold sensor, is among the few TRP channels responsible for thermosensing. Despite sustained interest in the channel, the mechanisms underlying TRPM8 activation, modulation, and gating have proved challenging to study and remain poorly understood. In this thesis, I offer data collected on various expression, extraction, and purification conditions tested in E. Coli expression systems with the aim to optimize the generation of a structurally stable and functional human TRPM8 pore domain (S5 and S6) construct for application in structural biology studies. These studies, including the biophysical technique nuclear magnetic spectroscopy (NMR), among others, will be essential for elucidating the role of the TRPM8 pore domain in in regulating ligand binding, channel gating, ion selectively, and thermal sensitivity. Moreover, in the second half of this thesis, I discuss the ligation-independent megaprimer PCR of whole-plasmids (MEGAWHOP PCR) cloning technique, and how it was used to generate chimeras between TRPM8 and its nearest analog TRPM2. I review steps taken to optimize the efficiency of MEGAWHOP PCR and the implications and unique applications of this novel methodology for advancing recombinant DNA technology. I lastly present preliminary electrophysiological data on the chimeras, employed to isolate and study the functional contributions of each individual transmembrane helix (S1-S6) to TRPM8 menthol activation. These studies show the utility of the TRPM8\u2014TRPM2 chimeras for dissecting function of TRP channels. The average current traces analyzed thus far indicate that the S2 and S3 helices appear to play an important role in TRPM8 menthol modulation because the TRPM8[M2S2] and TRPM8[M2S3] chimeras significantly reduce channel conductance in the presence of menthol. The TRPM8[M2S4] chimera, oppositely, increases channel conductance, implying that the S4 helix in native TRPM8 may suppress menthol modulation. Overall, these findings show that there is promise in the techniques chosen to identify specific regions of TRPM8 crucial to menthol activation, though the methods chosen to study the TRPM8 pore independent from the whole channel may need to be reevaluated. Further experiments will be necessary to refine TRPM8 pore solubilization and purification before structural studies can proceed, and the electrophysiology traces observed for the chimeras will need to be further verified and evaluated for consistency and physiological significance.
ContributorsWaris, Maryam Siddika (Author) / Van Horn, Wade (Thesis director) / Redding, Kevin (Committee member) / School of Molecular Sciences (Contributor) / Department of English (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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Description
Musical interpretation is challenging when one's goal is to evoke an emotional response from an audience. In order to develop a well-informed interpretation of Mozart's Fantasia in D minor K. 397, a study was conducted on the historical background of the piece and various performances by well-regarded performers. Fantasias are

Musical interpretation is challenging when one's goal is to evoke an emotional response from an audience. In order to develop a well-informed interpretation of Mozart's Fantasia in D minor K. 397, a study was conducted on the historical background of the piece and various performances by well-regarded performers. Fantasias are written works, but improvisatory by nature. Mozart's fantasias were influenced by C. P. E. Bach's, which included sudden changes in emotion. An Emil Gilels performance provided a classically trained approach, while Mitsuko Uchida's performance provided an emotional approach. Colin Tilney and John Irving performances elucidated the sound of the instruments that Mozart would have been composing with. Altogether, the research culminated in an interpretation of the D minor Fantasia that endeavored to capture the essence of fantasy, improvisation and emotion.
ContributorsMo, Gina Nan (Author) / Emmery, Laura (Thesis director) / Creviston, Hannah (Committee member) / Department of Psychology (Contributor) / School of International Letters and Cultures (Contributor) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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Description
Volunteer motivation and satisfaction were assessed at Project CURE, a nonprofit that collects, sorts, tests, and ships donated medical supplies and equipment to healthcare facilities in developing countries. This research was the result of a yearlong partnership between the Community Action Research Experiences (CARE) program and Project CURE. Volunteers at

Volunteer motivation and satisfaction were assessed at Project CURE, a nonprofit that collects, sorts, tests, and ships donated medical supplies and equipment to healthcare facilities in developing countries. This research was the result of a yearlong partnership between the Community Action Research Experiences (CARE) program and Project CURE. Volunteers at Project CURE were surveyed (N=147) after completing a volunteer session to assess their motivation for volunteering, satisfaction with their experience, and any recommendations they had for improving the volunteer program. Five categories of motivating factors were assessed and it was found that the Values and Understanding categories were the strongest motivating factors. Overall, volunteers rated their experience highly, but the results indicated a number of small changes that Project CURE could make to better meet volunteers' needs, and better communicate the impact of volunteers' work, which could pave the way to increases in the numbers of volunteer hours Project CURE receives and increased quality of volunteer work.
ContributorsStepanek, Rachel (Author) / Reesing, Amy (Thesis director) / Dumka, Larry (Committee member) / School of Molecular Sciences (Contributor) / School of Human Evolution and Social Change (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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Description
The purpose of this study was to create an evaluation plan for the Preventing Abusive Conduct (PAC/Peace) program, within the community-based organization Chrysalis. PAC/Peace is a program that provides treatment for domestic abuse offenders through psycho-educational counseling groups. This study analyzed evaluation methods used by similar programs to determine which

The purpose of this study was to create an evaluation plan for the Preventing Abusive Conduct (PAC/Peace) program, within the community-based organization Chrysalis. PAC/Peace is a program that provides treatment for domestic abuse offenders through psycho-educational counseling groups. This study analyzed evaluation methods used by similar programs to determine which methods were successful, as well as what criteria were used to evaluate these programs. A literature review was conducted on similar studies to find which measures should be used to determine whether a treatment program met the desired goals for its participants. The literature review, along with interviews with Chrysalis staff were used as resources to create a cost-effective evaluation plan for Chrysalis to measure the effectiveness of their offender treatment program. The results indicated a two part evaluation plan would be the most beneficial for Chrysalis at this time. The first part consists of official reports of recidivism collected at one year, and three years post-graduation from the program. The second is utilization of The Revised Conflict Tactics Scale as an offender self-reporting measure, administered at program intake and exit, as well as one year post-graduation. When implemented, the results of this evaluation will give Chrysalis insight to which aspects of the treatment are working, and which aspects might be improved upon. The results acquired from the evaluation may also help PAC/Peace to become an evidence-based program.
ContributorsScouras, Natasha Elexa (Author) / Low, Sabina (Thesis director) / Dumka, Larry (Committee member) / School of Social Transformation (Contributor) / School of Molecular Sciences (Contributor) / School of Human Evolution and Social Change (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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Description
We hypothesized that recurrent exposure to a temporal discounting task would habitize participants, so that they become insensitive to framing effects. Temporal discounting is a behavioral trend which describes how people discount the value of a reward dependent on the time until receipt. Participants completed a temporal discounting task weekly

We hypothesized that recurrent exposure to a temporal discounting task would habitize participants, so that they become insensitive to framing effects. Temporal discounting is a behavioral trend which describes how people discount the value of a reward dependent on the time until receipt. Participants completed a temporal discounting task weekly for five weeks, to promote formation of a habitual decision strategy. Concomitant with this, we expected that people would shift their decision process from a deliberate, goal-oriented approach that is sensitive to changes in reward outcomes and environmental context, to a simplified, automatic approach that minimizes cognitive effort. We expected that this shift in decision strategy would be evident in a reduced influence of contextual effects on choice outcomes. We tested this hypothesis by leveraging two framing effects \u2014 the date/delay effect and the decimal effect. Consistent with our hypothesis, we find that the date/delay effect is significant on week 1, shows significant changes in week 1 to week 5, and is no longer significant on week 5. The results for the decimal effects were not significant. We discuss these results with respect to the cognitive processes that underlie temporal discounting and self-control.
ContributorsSt Amand, Jesse Dean (Author) / McClure, Samuel (Thesis director) / Sanabria, Federico (Committee member) / School of Molecular Sciences (Contributor) / Department of Physics (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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Description
Orbiting space debris is an active issue that affects the capability of space launch for future satellites, probes, and space shuttles, and it will become a nearly insurmountable problem without action. Debris of varying sizes and speeds orbit the Earth at a range of heights above the atmosphere and need

Orbiting space debris is an active issue that affects the capability of space launch for future satellites, probes, and space shuttles, and it will become a nearly insurmountable problem without action. Debris of varying sizes and speeds orbit the Earth at a range of heights above the atmosphere and need to be removed to avoid damage to crucial equipment of active orbiting satellites including the International Space Station. Finding a feasible solution to space debris removal requires that several facets be covered to become a reality; these include being aware of the problem in magnitude and source. This literature assessment covers the magnitude of space debris in low-earth and geosynchronous orbit as well as collision events which have increased the amount of space debris. There have been efforts made by several space agencies to control the amount of space debris added to orbit by current and future launches over the last decade \u2014 serving as a temporary fix before removal can be executed. This paper explores known removal efforts through mitigation, projects conceived and tested by DARPA, related space policies and laws, CubeSat technology, and the cataloguing of known space debris. To make space debris removal a reality, roadblocks need to be removed to acquire permission from states or countries for space missions. For example, these restrictions are in place to protect the assets of several countries and organizations. Guidelines set to curb the growth of space debris fail to prevent the growth due to the restrictions for ownership rights making them not as effective. This paper covers space policy and laws, the economy, satellite ownership, international conflict, status of space debris, and the overall feasibility of space debris removal. It will then discuss currently proposed solutions for the removal of space debris. Finally, this paper attempts to weight the advantages and disadvantages of the idea that space debris removal should include the opportunity to recycle materials. For example, defunct satellites and other discarded space crafts could be used for future launches. It will conclude with a personal exploration of what materials can be recycled, what chemical processes can be used to break down materials, and how to combine recycling and chemical processes for space-based recycling stations between Earth and the moon. The overall question that drives the search for making space debris removal a reality is whether it is feasible in multiple areas including technologically, legally, monetarily, and physically.
ContributorsBreden, Elizabeth Catherine (Author) / Foy, Joseph (Thesis director) / Thoesen, Andrew (Committee member) / Maximon, Leonard (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05
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Description
Brown adipose tissue (BAT) is thought to be important in combating obesity as it can expend energy in the form of heat, e.g. thermogenesis. The goal of this study was to study the effect of injected norepinephrine (NE) on the activation of BAT in rats that were fed a high

Brown adipose tissue (BAT) is thought to be important in combating obesity as it can expend energy in the form of heat, e.g. thermogenesis. The goal of this study was to study the effect of injected norepinephrine (NE) on the activation of BAT in rats that were fed a high fat diet (HFD). A dose of 0.25 mg/kg NE was used to elicit a temperature response that was measured using transponders inserted subcutaneously over the BAT and lower back and intraperitoneally to measure the core temperature. The results found that the thermic effect of the BAT increased after the transition from low fat diet to a high fat diet (LFD) yet, after prolonged exposure to the HFD, the effects resembled levels found with the LFD. This suggests that while a HFD may stimulate the effect of BAT, long term exposure may have adverse effects on BAT activity. This may be due to internal factors that will need to be examined further.
ContributorsSion, Paul William (Author) / Herman, Richard (Thesis director) / Borges, Chad (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05
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Description
Cystic Fibrosis (CF) is a genetic disorder that disrupts the hydration of mucous of the lungs, which promotes opportunistic bacterial infections that begin in the affected person’s childhood, and persist into adulthood. One of the bacteria that infect the CF lung is Pseudomonas aeruginosa. This gram-negative bacterium is acquired from

Cystic Fibrosis (CF) is a genetic disorder that disrupts the hydration of mucous of the lungs, which promotes opportunistic bacterial infections that begin in the affected person’s childhood, and persist into adulthood. One of the bacteria that infect the CF lung is Pseudomonas aeruginosa. This gram-negative bacterium is acquired from the environment of the CF lung, changing the expression of phenotypes over the course of the infection. As P. aeruginosa infections become chronic, some phenotype changes are known to be linked with negative patient outcomes. An important exoproduct phenotype is rhamnolipid production, which is a glycolipid that P. aeruginosa produces as a surfactant for surface-mediated travel. Over time, the expression of this phenotype decreases in expression in the CF lung.
The objective of this investigation is to evaluate how environmental changes that are related to the growth environment in the CF lung alters rhamnolipid production. Thirty-five P. aeruginosa isolates from Dartmouth College and Seattle Children’s Hospital were selected to observe the impact of temperature, presence of Staphylococcus aureus metabolites, and oxygen availability on rhamnolipid production. It was found that the rhamnolipid production significantly decreased for 30C versus 37C, but not at 40C. The addition of S. aureus spent media, in any of the tested conditions, did not influence rhamnolipid production. Finally, the change in oxygen concentration from normoxia to hypoxia significantly reduced rhamnolipid production. These results were compared to swarming assay data to understand how changes in rhamnolipid production impact surface-mediated motility.
ContributorsKiermayr, Jonathan Patrick (Author) / Bean, Heather (Thesis director) / Misra, Rajeev (Committee member) / Haydel, Shelley (Committee member) / School of International Letters and Cultures (Contributor) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05
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Description
Almost every form of cancer deregulates the expression and activity of anabolic glycosyltransferase (GT) enzymes, which incorporate particular monosaccharides in a donor acceptor as well as linkage- and anomer-specific manner to assemble complex and diverse glycans that significantly affect numerous cellular events, including tumorigenesis and metastasis. Because glycosylation is not

Almost every form of cancer deregulates the expression and activity of anabolic glycosyltransferase (GT) enzymes, which incorporate particular monosaccharides in a donor acceptor as well as linkage- and anomer-specific manner to assemble complex and diverse glycans that significantly affect numerous cellular events, including tumorigenesis and metastasis. Because glycosylation is not template-driven, GT deregulation yields heterogeneous arrays of aberrant intact glycan products, some in undetectable quantities in clinical bio-fluids (e.g., blood plasma). Numerous glycan features (e.g., 6 sialylation, β-1,6-branching, and core fucosylation) stem from approximately 25 glycan “nodes:” unique linkage specific monosaccharides at particular glycan branch points that collectively confer distinguishing features upon glycan products. For each node, changes in normalized abundance (Figure 1) may serve as nearly 1:1 surrogate measure of activity for culpable GTs and may correlate with particular stages of carcinogenesis. Complementary to traditional top down glycomics, the novel bottom-up technique applied herein condenses each glycan node and feature into a single analytical signal, quantified by two GC-MS instruments: GCT (time-of-flight analyzer) and GCMSD (transmission quadrupole analyzers). Bottom-up analysis of stage 3 and 4 breast cancer cases revealed better overall precision for GCMSD yet comparable clinical performance of both GC MS instruments and identified two downregulated glycan nodes as excellent breast cancer biomarker candidates: t-Gal and 4,6-GlcNAc (ROC AUC ≈ 0.80, p < 0.05). Resulting from the activity of multiple GTs, t-Gal had the highest ROC AUC (0.88) and lowest ROC p‑value (0.001) among all analyzed nodes. Representing core-fucosylation, glycan node 4,6-GlcNAc is a nearly 1:1 molecular surrogate for the activity of α-(1,6)-fucosyltransferase—a potential target for cancer therapy. To validate these results, future projects can analyze larger sample sets, find correlations between breast cancer stage and changes in t-Gal and 4,6-GlcNAc levels, gauge the specificity of these nodes for breast cancer and their potential role in other cancer types, and develop clinical tests for reliable breast cancer diagnosis and treatment monitoring based on t-Gal and 4,6-GlcNAc.
ContributorsZaare, Sahba (Author) / Borges, Chad (Thesis director) / LaBaer, Joshua (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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Description
Popular Culture of today, particularly books and movies have begun to influence the way individ- uals and society as a whole, views specific concepts. In this case, the fairly recent phenomenon of the Sci- ence Fiction Drug Niche has produced significant thought among audiences as to both the benefits and

Popular Culture of today, particularly books and movies have begun to influence the way individ- uals and society as a whole, views specific concepts. In this case, the fairly recent phenomenon of the Sci- ence Fiction Drug Niche has produced significant thought among audiences as to both the benefits and costs of cognitive enhancers in our world. Through the use of both a thorough analysis of modern films and novels on the topic as well as focus groups of the average college students this study analyzes the influence that this niche has had on the perceptions that students have towards the use of such cognitive enhancements. Small groups of students were shown the same film: Limitless, and discussion after the film displayed the students thoughts and attitudes towards the ideas shown in the film. Limitless itself falls into this Science Fiction drug niche and discusses both benefits and harms of chemical cognitive enhancement. The study indicates that audiences have thought not only about the issues that may arise with the presence of cognitive enhancement in our world but also the possible benefits of this enhancement. The results go even further to preliminarily show that there are common thoughts that arise in such situations. These common ideas that arise show, at least on a very basic level, that the presence of these Science Fiction Drug-inspired works are influencing the way audiences perceive the use of cognitive enhancement as well as influencing what doubts, questions, hopes, and fears arise from these pharmaceuticals. This preliminary study could use further research to ana- lyze the effects of popular culture on perceptions of cognitive enhancement and pharmaceuticals to alter consciousness.
ContributorsSyed, Mariha Batool (Author) / Zachary, Gregg (Thesis director) / Hurlbut, Ben (Committee member) / School of Historical, Philosophical and Religious Studies (Contributor) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05