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- All Subjects: Biochemistry
- All Subjects: English
- Creators: School of Molecular Sciences
- Creators: Department of English
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- Status: Published
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 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
Description
"The Problem of Hope: Literary Tragedy in Mid-Twentieth Century American Fiction" examines Arthur Miller's Death of a Salesman, Ralph Ellison's Invisible Man, and Sylvia Plath's The Bell Jar through the lens of tragedy. This thesis delves into how conflicts between internal and external identities can create a tragic individual, what kinds of success count toward achievement of the "American Dream," and whether the tragic "common man" is the socially normative one or the socially disenfranchised one. It raises a three-dimensional theoretical approach to American tragedy and, most importantly, considers the significance of tragic hope for American literature. This paper questions the construction of American identities across class, race, and gender according to social scripts. It seeks to uncover what forces these scripts exert on American cultural myths and rereads those myths through tragedy to explore Miller's idea of a noble common man. By moving from Miller to Ellison to Plath, this thesis traces the undercurrents of tragedy through some of the most identity-focused novels of mid-twentieth century American fiction to see how the overarching American narrative changed from 1940 to 1969 as the US underwent significant social changes domestically and image changes abroad. Ultimately, this paper concludes that tragedy in mid-twentieth century American fiction points toward a new idea of American success as a success that occurs beyond social scripts.
ContributorsMedeiros, Amy Marie (Author) / Holbo, Christine (Thesis director) / Sadowski-Smith, Claudia (Committee member) / Department of English (Contributor) / W. P. Carey School of Business (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
This project focuses on techniques contemporary American poets use in their work. Ten different poetry collections are analyzed for dominant writing styles and techniques, which I then apply to my own poems, concentrating on modeling that particular poet. I then reflect on those poems through an evaluation of my writing process, how those techniques were implemented, and how they affected the poem. In addition to these reviews and reflections, I also wrote three articles about the literary community and what I've learned from my interactions in that community. All these materials are organized into a website, which shows the connections between the different writings via links and menus. Creating this website brings all the materials together to demonstrate my growth as a poet, writer, and designer. This heavy focus on poetry and analysis has helped sharpen my critical thinking skills and has better prepared me for a career in design and journalism.
ContributorsHansen, Elizabeth Sarah (Author) / Murphy, Patricia (Thesis director) / Savard, Jeannine (Committee member) / Barrett, The Honors College (Contributor) / Department of English (Contributor) / Hugh Downs School of Human Communication (Contributor) / Walter Cronkite School of Journalism and Mass Communication (Contributor)
Created2015-05
Description
This honors thesis features a translation of Andrés Bello’s “Historia físicia y política de Chile por Claudio Gay” that had never before been reproduced in English, as well as a discussion of translation theories and a biographical sketch of Andrés Bello, a prolific Latin American author and philosopher. The goals of this thesis include promoting Latin American literature, bringing awareness to Bello’s contributions to Chile’s history, and promoting translation as a creative form of education.
ContributorsDominick, Alicia Lorae (Author) / Humphrey, Ted (Thesis director) / Martinez, Maria Teresa (Committee member) / Burke, Janet (Committee member) / Barrett, The Honors College (Contributor) / School of International Letters and Cultures (Contributor) / Department of English (Contributor)
Created2015-05
Description
This paper explores how marginalist economics defines and inevitably constrains Victorian sensation fiction's content and composition. I argue that economic intuition implies that sensationalist heroes and antagonists, writers and readers all pursued a fundamental, "rational" aim: the attainment of pleasure. So although "sensationalism" took on connotations of moral impropriety in the Victorian age, sensation fiction primarily involves experiences of pain on the page that excite the reader's pleasure. As such, sensationalism as a whole can be seen as a conformist product, one which mirrors the effects of all commodities on the market, rather than as a rebellious one. Indeed, contrary to modern and contemporary critics' assumptions, sensation fiction may not be as scandalous as it seems.
ContributorsFischer, Brett Andrew (Author) / Bivona, Daniel (Thesis director) / Looser, Devoney (Committee member) / Barrett, The Honors College (Contributor) / School of Mathematical and Statistical Sciences (Contributor) / Economics Program in CLAS (Contributor) / School of Politics and Global Studies (Contributor) / Department of English (Contributor)
Created2014-12
Description
In my thesis paper, I examine the gothic elements found in classical gothic fairy tales from European and Japanese tradition, particularly those works by the Brothers Grimm and Yei Theodora Ozaki. By examining the principle gothic elements that are unique to both stories, and further analyzing the commonalities of story, plot, and other major tropes, a better understanding of the message meant to be imparted and other cultural nuances can be ascertained. Gothic literature creates an atmosphere of gloom and suspense, toying with concepts of dread and darkness by employing Gothic elements such as shadows, the supernatural, sinister buildings, and strong-willed villains, all of which affect the rational mind in an irrational way. Fairytales freely use such tropes to their advantage, playing with the many fears of children, while simultaneously painting an idealistic fantasy world. The degree of usage and the application of gothic elements is closely examined in the Grimm works, "Hansel and Gretel," and "The Robber Bridegroom," as well as the Japanese tales, "The Goblin of Adachigahra,""Kintaro the Golden Boy" and "The Monkey and the Crab." These stories have been chosen due for their usage of animal tricksters, themes of control, and aspects of isolation, supernatural entities, and substantial gothic imagery. The gothic elements of death, sinister older women, the supernatural, fears of abandonment, and cunning animals are akin to both Western and Eastern tales, while the concept of gothic setting and the type of monsters prepared to feast on men is significantly different for both cultures, similar lessons are intended to be gleaned by children from these tales, with the intention of generally producing positive results \u2014 while the means differ, the message is strikingly similar, yet there remain cultural differences in terms of central themes and character traits.The effect of re-introducing the darker, gothic elements of traditional fairy tales into modern literature and retellings of the original narratives has been profound.Today, whether it has been at the bequest of the public or simply a new-age movement by modern cinema audience for the "gritty and realistic," fairy tales are returning to their former gothic forms. "Snow White and The Huntsman" is one example of a film which has gone this route, opting for a more gothic, classic telling rather than the chip, cheery, rosy cheeked Disney versions. There is a tendency for most media nowadays to be far less censored and fantastical, aiming for a more realistic, grittier approach \u2014 this bleeds into film and literature likewise, and thus children are impacted by this shift as well. Children seem to be able to handle more, perhaps desensitized at younger and younger ages by the products of our widely consumerist society, or perhaps due to parents raising their children in such a way so that the darkness that tinges these tales doesn't disturb and derail but rather, emphasizes their meaning of teaching certain lessons. Tales such as these are still valuable, and will continue to be so long as we seek a reality greater than our own, where the evil of the world is wiped away, and we all live happily ever after.
ContributorsMoschonas, Jerasimos Theodore (Author) / Ellis, Lawrence (Thesis director) / Hoyt, Heather (Committee member) / Barrett, The Honors College (Contributor) / Department of English (Contributor)
Created2015-05
Description
The purpose of this study is to determine the types of classroom instructional activities commonly used in teaching literature. Data were collected at ASU Preparatory High School. The study determined that literature-based lessons and activities fall under three categories: reading, writing, and discussion. Classroom observations revealed that reading, writing, and discursive activities were designed to promote higher-ordering thinking. These activities included silent reading, annotating text, reading aloud, keeping reading response journals, practicing essay writing, and participating in Socratic discussion. The teachers at ASU Prep used the listed activities with the intent to challenge their English students to engage in active learning, to improve reading, writing, and discursive skills, and promote critical thinking skills.
ContributorsSarik, Vivian Roathany (Author) / Blasingame, James (Thesis director) / Ingram-Waters, Mary (Committee member) / Barrett, The Honors College (Contributor) / School of International Letters and Cultures (Contributor) / Department of English (Contributor)
Created2015-05
Description
The primary objective of this project is to further the knowledge about SCL26 family of anion transporters. The goals of the experiment were to find the lowest sulfate concentration where the yeast without Sulp1 and Sulp2 is able to grow, but it grows very slowly, and to find a higher sulfate concentration where the yeast grows quickly, with or without the sulfate transporters. The lowest sulfate concentration where the yeast without the sulfate transporters is able to grow was determined to be 2-4 mM, however, this range can likely be refined by more quantitative analytical methods. At a sulfate concentration of 20 mM sulfate or higher, the yeast is able to grow quickly without high-affinity sulfate transporters. The next step in the project is to re-introduce the Sulp1 and Sulp2 genes into the yeast, so that growth in low and high sulfate conditions can be compared with and without the Sulp1 and Sulp2 proteins. The long-term goals of the project are to bring experience with yeast to Dr. Nannenga’s structural discovery lab, to determine if yeast sulfate transporters respond in the same way to drug candidates as human sulfate transporters, and to determine the structure of the proteins using cryo-electron microscopy.
ContributorsCall, Nicolas I (Author) / Nannenga, Brent (Thesis director) / Wang, Xuan (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
The devastating 2014 Ebola virus outbreak in Western Africa demonstrated the lack of therapeutic approaches available for the virus. Although monoclonal antibodies (mAb) and other molecules have been developed that bind the virus, no therapeutic has shown the efficacy needed for FDA approval. Here, a library of 50 peptide based ligands that bind the glycoprotein of the Zaire Ebola virus (GP) were developed. Using whole virus screening of vesicular stomatitis virus pseudotyped with GP, low affinity peptides were identified for ligand construction. In depth analysis showed that two of the peptide based molecules bound the Zaire GP with <100 nM KD. One of these two ligands was blocked by a known neutralizing mAb, 2G4, and showed cross-reactivity to the Sudan GP. This work presents ligands with promise for therapeutic applications across multiple variants of the Ebola virus.
ContributorsRabinowitz, Joshua Avraam (Author) / Diehnelt, Chris (Thesis director) / Johnston, Stephen (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-12
Description
The parameters of microwave-assisted acid hydrolysis (MAAH) and 1H NMR highly affect the quantitative analysis of protein hydrolysates. Microwave-induction source, NMR spectral resolution, and data analysis are key parameters in the nuclear magnetic resonance – amino acid analysis (NMR-AAA) workflow where errors accrue due to lack of an optimized protocol. Hen egg white lysozyme was hydrolyzed using an 800W domestic microwave oven for varying time points between 10-25 minutes, showing minimal protein hydrolysis after extended time periods. Studies on paramagnetic doping with varying amounts of gadolinium chloride for increased NMR resolution resulted in little T1 reduction in a majority of amino acids and resulted in significant line broadening in concentrations above 1µM. The use of the BAYESIL analysis tool with HOD suppressed 1H-NMR spectra resulted in misplaced template peaks and errors greater than 1% for 10 of 13 profiled amino acids with the highest error being 7.6% (Thr). Comparatively, Chenomx NMR Suite (v7.1) analysis resulted in errors of less than 1% for 9 of 13 profiled amino acids with a highest error value of 3.6% (Lys). Using the optimized protocol, hen egg white lysozyme C was identified at rank 1 with a score of 64 in a Gallus gallus species wide AACompIdent search. This technique reduces error associated with sample handling relative to previously used amino acid analysis (AAA) protocols and requires no derivatization or additional processing of the sample prior to analysis.
ContributorsJordan, Jacob Smith (Author) / Yarger, Jeffery (Thesis director) / Van Horn, Wade (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05