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Membrane proteins are very important for all living cells, being involved in respiration, photosynthesis, cellular uptake and signal transduction, amongst other vital functions. However, less than 300 unique membrane protein structures have been determined to date, often due to difficulties associated with the growth of sufficiently large and well-ordered crystals.

Membrane proteins are very important for all living cells, being involved in respiration, photosynthesis, cellular uptake and signal transduction, amongst other vital functions. However, less than 300 unique membrane protein structures have been determined to date, often due to difficulties associated with the growth of sufficiently large and well-ordered crystals. This work has been focused on showing the first proof of concept for using membrane protein nanocrystals and microcrystals for high-resolution structure determination. Upon determining that crystals of the membrane protein Photosystem I, which is the largest and most complex membrane protein crystallized to date, exist with only a hundred unit cells with sizes of less than 200 nm on an edge, work was done to develop a technique that could exploit the growth of the Photosystem I nanocrystals and microcrystals. Femtosecond X-ray protein nanocrystallography was developed for use at the first high-energy X-ray free electron laser, the LCLS at SLAC National Accelerator Laboratory, in which a liquid jet would bring fully hydrated Photosystem I nanocrystals into the interaction region of the pulsed X-ray source. Diffraction patterns were recorded from millions of individual PSI nanocrystals and data from thousands of different, randomly oriented crystallites were integrated using Monte Carlo integration of the peak intensities. The short pulses ( 70 fs) provided by the LCLS allowed the possibility to collect the diffraction data before the onset of radiation damage, exploiting the diffract-before-destroy principle. At the initial experiments at the AMO beamline using 6.9- Å wavelength, Bragg peaks were recorded to 8.5- Å resolution, and an electron-density map was determined that did not show any effects of X-ray-induced radiation damage. Recently, femtosecond X-ray protein nanocrystallography experiments were done at the CXI beamline of the LCLS using 1.3- Å wavelength, and Bragg reflections were recorded to 3- Å resolution; the data are currently being processed. Many additional techniques still need to be developed to explore the femtosecond nanocrystallography technique for experimental phasing and time-resolved X-ray crystallography experiments. The first proof-of-principle results for the femtosecond nanocrystallography technique indicate the incredible potential of the technique to offer a new route to the structure determination of membrane proteins.
ContributorsHunter, Mark (Author) / Fromme, Petra (Thesis advisor) / Wolf, George (Committee member) / Levitus, Marcia (Committee member) / Arizona State University (Publisher)
Created2011
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Description
Molybdenum (Mo) is a key trace nutrient for biological assimilation of nitrogen, either as nitrogen gas (N2) or nitrate (NO3-). Although Mo is the most abundant metal in seawater (105 nM), its concentration is low (<5 nM) in most freshwaters today, and it was scarce in the ocean before 600

Molybdenum (Mo) is a key trace nutrient for biological assimilation of nitrogen, either as nitrogen gas (N2) or nitrate (NO3-). Although Mo is the most abundant metal in seawater (105 nM), its concentration is low (<5 nM) in most freshwaters today, and it was scarce in the ocean before 600 million years ago. The use of Mo for nitrogen assimilation can be understood in terms of the changing Mo availability through time; for instance, the higher Mo content of eukaryotic vs. prokaryotic nitrate reductase may have stalled proliferation of eukaryotes in low-Mo Proterozoic oceans. Field and laboratory experiments were performed to study Mo requirements for NO3- assimilation and N2 fixation, respectively. Molybdenum-nitrate addition experiments at Castle Lake, California revealed interannual and depth variability in plankton community response, perhaps resulting from differences in species composition and/or ammonium availability. Furthermore, lake sediments were elevated in Mo compared to soils and bedrock in the watershed. Box modeling suggested that the largest source of Mo to the lake was particulate matter from the watershed. Month-long laboratory experiments with heterocystous cyanobacteria (HC) showed that <1 nM Mo led to low N2 fixation rates, while 10 nM Mo was sufficient for optimal rates. At 1500 nM Mo, freshwater HC hyperaccumulated Mo intercellularly, whereas coastal HC did not. These differences in storage capacity were likely due to the presence in freshwater HC of the small molybdate-binding protein, Mop, and its absence in coastal and marine cyanobacterial species. Expression of the mop gene was regulated by Mo availability in the freshwater HC species Nostoc sp. PCC 7120. Under low Mo (<1 nM) conditions, mop gene expression was up-regulated compared to higher Mo (150 and 3000 nM) treatments, but the subunit composition of the Mop protein changed, suggesting that Mop does not bind Mo in the same manner at <1 nM Mo that it can at higher Mo concentrations. These findings support a role for Mop as a Mo storage protein in HC and suggest that freshwater HC control Mo cellular homeostasis at the post-translational level. Mop's widespread distribution in prokaryotes lends support to the theory that it may be an ancient protein inherited from low-Mo Precambrian oceans.
ContributorsGlass, Jennifer (Author) / Anbar, Ariel D (Thesis advisor) / Shock, Everett L (Committee member) / Jones, Anne K (Committee member) / Hartnett, Hilairy E (Committee member) / Elser, James J (Committee member) / Fromme, Petra (Committee member) / Arizona State University (Publisher)
Created2011
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Description
Anti-retroviral drugs and AIDS prevention programs have helped to decrease the rate of new HIV-1 infections in some communities, however, a prophylactic vaccine is still needed to control the epidemic world-wide. Despite over two decades of research, a vaccine against HIV-1 remains elusive, although recent clinical trials have shown promising

Anti-retroviral drugs and AIDS prevention programs have helped to decrease the rate of new HIV-1 infections in some communities, however, a prophylactic vaccine is still needed to control the epidemic world-wide. Despite over two decades of research, a vaccine against HIV-1 remains elusive, although recent clinical trials have shown promising results. Recent successes have focused on highly conserved, mucosally-targeted antigens within HIV-1 such as the membrane proximal external region (MPER) of the envelope protein, gp41. MPER has been shown to play critical roles in the viral mucosal transmission, though this peptide is not immunogenic on its own. Gag is a structural protein configuring the enveloped virus particles, and has been suggested to constitute a target of the cellular immunity potentially controlling the viral load. It was hypothesized that HIV-1 enveloped virus-like particles (VLPs) consisting of Gag and a deconstructed form of gp41 comprising the MPER, transmembrane, and cytoplasmic domains (dgp41) could be expressed in plants. Plant-optimized HIV-1 genes were constructed and expressed in Nicotiana benthamiana by stable transformation, or transiently using a tobacco mosaic virus-based expression system or a combination of both. Results of biophysical, biochemical and electron microscopy characterization demonstrated that plant cells could support not only the formation of HIV-1 Gag VLPs, but also the accumulation of VLPs that incorporated dgp41. These particles were purified and utilized in mice immunization experiments. Prime-boost strategies combining systemic and mucosal priming with systemic boosting using two different vaccine candidates (VLPs and CTB-MPR - a fusion of MPER and the B-subunit of cholera toxin) were administered to BALB/c mice. Serum antibody responses against both the Gag and gp41 antigens could be elicited in mice systemically primed with VLPs and these responses could be recalled following systemic boosting with VLPs. In addition, mucosal priming with VLPs allowed for a robust boosting response against Gag and gp41 when boosted with either candidate. Functional assays of these antibodies are in progress to test the antibodies' effectiveness in neutralizing and preventing mucosal transmission of HIV-1. This immunogenicity of plant-based Gag/dgp41 VLPs represents an important milestone on the road towards a broadly-efficacious and inexpensive subunit vaccine against HIV-1.
ContributorsKessans, Sarah (Author) / Mor, Tsafrir S (Thesis advisor) / Matoba, Nobuyuki (Committee member) / Mason, Hugh (Committee member) / Hogue, Brenda (Committee member) / Fromme, Petra (Committee member) / Arizona State University (Publisher)
Created2011
Description

Consider Steven Cryos’ words, “When disaster strikes, the time to prepare has passed.” Witnessing domestic water insecurity in events such as Hurricane Katrina, the instability in Flint, Michigan, and most recently the winter storms affecting millions across Texas, we decided to take action. The period between a water supply’s disruption

Consider Steven Cryos’ words, “When disaster strikes, the time to prepare has passed.” Witnessing domestic water insecurity in events such as Hurricane Katrina, the instability in Flint, Michigan, and most recently the winter storms affecting millions across Texas, we decided to take action. The period between a water supply’s disruption and restoration is filled with anxiety, uncertainty, and distress -- particularly since there is no clear indication of when, exactly, restoration comes. It is for this reason that Water Works now exists. As a team of students from diverse backgrounds, what started as an honors project with the Founders Lab at Arizona State University became the seed that will continue to mature into an economically sustainable business model supporting the optimistic visions and tenants of humanitarianism. By having conversations with community members, conducting market research, competing for funding and fostering progress amid the COVID-19 pandemic, our team’s problem-solving traverses the disciplines. The purpose of this paper is to educate our readers about a unique solution to emerging issues of water insecurity that are nested across and within systems who could benefit from the introduction of a personal water reclamation system, showcase our team’s entrepreneurial journey, and propose future directions that will this once pedagogical exercise to continue fulfilling its mission: To heal, to hydrate and to help bring safe water to everyone.

ContributorsReitzel, Gage Alexander (Co-author) / Filipek, Marina (Co-author) / Sadiasa, Aira (Co-author) / Byrne, Jared (Thesis director) / Sebold, Brent (Committee member) / Historical, Philosophical & Religious Studies (Contributor) / School of Human Evolution & Social Change (Contributor, Contributor) / Historical, Philosophical & Religious Studies, Sch (Contributor) / Department of Psychology (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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Description

Reducing the amount of error and introduced data variability increases the accuracy of Western blot results. In this study, different methods of normalization for loading differences and data alignment were explored with respect to their impact on Western blot results. GAPDH was compared to the LI-COR Revert total protein stain

Reducing the amount of error and introduced data variability increases the accuracy of Western blot results. In this study, different methods of normalization for loading differences and data alignment were explored with respect to their impact on Western blot results. GAPDH was compared to the LI-COR Revert total protein stain as a loading control. The impact of normalizing data to a control condition, which is commonly done to align Western blot data distributed over several immunoblots, was also investigated. Specifically, this study addressed whether normalization to a small subset of distinct controls on each immunoblot increases pooled data variability compared to a larger set of controls. Protein expression data for NOX-2 and SOD-2 from a study investigating the protective role of the bradykinin type 1 receptor in angiotensin-II induced left ventricle remodeling were used to address these questions but are also discussed in the context of the original study. The comparison of GAPDH and Revert total protein stain as a loading control was done by assessing their correlation and comparing how they affected protein expression results. Additionally, the impact of treatment on GAPDH was investigated. To assess how normalization to different combinations of controls influences data variability, protein data were normalized to the average of 5 controls, the average of 2 controls, or an average vehicle and the results by treatment were compared. The results of this study demonstrated that GAPDH expression is not affected by angiotensin-II or bradykinin type 1 receptor antagonist R-954 and is a less sensitive loading control compared to Revert total protein stain. Normalization to the average of 5 controls tended to reduce pooled data variability compared to 2 controls. Lastly, the results of this study provided preliminary evidence that R-954 does not alter the expression of NOX-2 or SOD-2 to an expression profile that would be expected to explain the protection it confers against Ang-II induced left ventricle remodeling.

ContributorsSiegel, Matthew Marat (Author) / Jeremy, Mills (Thesis director) / Sweazea, Karen (Committee member) / Hale, Taben (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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Description

In this creative thesis project I use digital “scrolleytelling” (an interactive scroll-based storytelling) to investigate diversity & inclusion at big tech companies. I wanted to know why diversity numbers were flatlining at Facebook, Apple, Amazon, Microsoft and Google, and took a data journalism approach to explore the relationship between what

In this creative thesis project I use digital “scrolleytelling” (an interactive scroll-based storytelling) to investigate diversity & inclusion at big tech companies. I wanted to know why diversity numbers were flatlining at Facebook, Apple, Amazon, Microsoft and Google, and took a data journalism approach to explore the relationship between what corporations were saying versus what they were doing. Finally, I critiqued diversity and inclusion by giving examples of how the current way we are addressing D&I is not fixing the problem.

ContributorsBrust, Jiaying Eliza (Author) / Coleman, Grisha (Thesis director) / Tinapple, David (Committee member) / Arts, Media and Engineering Sch T (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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The Electoral College, the current electoral system in the U.S., operates on a Winner-Take-All or First Past the Post (FPTP) principle, where the candidate with the most votes wins. Despite the Electoral College being the current system, it is problematic. According to Lani Guinier in Tyranny of the Majority, “the

The Electoral College, the current electoral system in the U.S., operates on a Winner-Take-All or First Past the Post (FPTP) principle, where the candidate with the most votes wins. Despite the Electoral College being the current system, it is problematic. According to Lani Guinier in Tyranny of the Majority, “the winner-take-all principle invariably wastes some votes” (121). This means that the majority group gets all of the power in an election while the votes of the minority groups are completely wasted and hold little to no significance. Additionally, FPTP systems reinforce a two-party system in which neither candidate could satisfy the majority of the electorate’s needs and issues, yet forces them to choose between the two dominant parties. Moreover, voting for a third party candidate only hurts the voter since it takes votes away from the party they might otherwise support and gives the victory to the party they prefer the least, ensuring that the two party system is inescapable. Therefore, a winner-take-all system does not provide the electorate with fair or proportional representation and creates voter disenfranchisement: it offers them very few choices that appeal to their needs and forces them to choose a candidate they dislike. There are, however, alternative voting systems that remedy these issues, such as a Ranked voting system, in which voters can rank their candidate choices in the order they prefer them, or a Proportional voting system, in which a political party acquires a number of seats based on the proportion of votes they receive from the voter base. Given these alternatives, we will implement a software simulation of one of these systems to demonstrate how they work in contrast to FPTP systems, and therefore provide evidence of how these alternative systems could work in practice and in place of the current electoral system.

ContributorsSummers, Jack Gillespie (Co-author) / Martin, Autumn (Co-author) / Burger, Kevin (Thesis director) / Voorhees, Matthew (Committee member) / Computer Science and Engineering Program (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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Partisan politics has created an increasingly polarized political climate in the United States. Despite the divisive political climate, women’s representation in politics has also increased drastically over the years. I began this project to see if there is a partisan rivalry between women in politics or a sense of shared

Partisan politics has created an increasingly polarized political climate in the United States. Despite the divisive political climate, women’s representation in politics has also increased drastically over the years. I began this project to see if there is a partisan rivalry between women in politics or a sense of shared “womanhood.” This thesis explores the role political parties play for women in office by examining how they vote on bills, what type of bills they propose, and whether or not they work collaboratively with their female counterparts at the Arizona State Legislature. My main goals for this project are to see how strong or weak political parties are in shaping political behavior at the Arizona State Legislature and to determine if there is a sense of “womanhood” despite different political affiliations. I also explore the role party affiliation plays within women legislators at the Arizona State Legislature.

ContributorsSanson, Claudia Maria (Author) / Lennon, Tara (Thesis director) / Woodall, Gina (Committee member) / School of Public Affairs (Contributor) / Department of English (Contributor) / School of Politics and Global Studies (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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Description

This 15-week long course is designed to introduce students, specifically in Arizona, to basic sustainability and conservation principles in the context of local reptile wildlife. Throughout the course, the students work on identifying the problem, creating visions for the desired future, and finally developing a strategy to help with reptile

This 15-week long course is designed to introduce students, specifically in Arizona, to basic sustainability and conservation principles in the context of local reptile wildlife. Throughout the course, the students work on identifying the problem, creating visions for the desired future, and finally developing a strategy to help with reptile species survival in the valley. Research shows that animals in the classroom have led to improved academic success for students. Thus, through creating this course I was able to combine conservation and sustainability curriculum with real-life animals whose survival is directly being affected in the valley. My hope is that this course will help students identify a newfound passion and call to action to protect native wildlife. The more awareness and actionable knowledge which can be brought to students in Arizona about challenges to species survival the more likely we are to see a change in the future and a stronger sense of urgency for protecting wildlife. In order to accomplish these goals, the curriculum was developed to begin with basic concepts of species needs such as food and shelter and basic principles of sustainability. As the course progresses the students analyze current challenges reptile wildlife faces, like urban sprawl, and explore options to address these challenges. The course concludes with a pilot pitch where students present their solution projects to the school.

ContributorsGoethe, Emma Rae (Author) / Brundiers, Katja (Thesis director) / Bouges, Olivia (Committee member) / School of Sustainability (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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System and software verification is a vital component in the development and reliability of cyber-physical systems - especially in critical domains where the margin of error is minimal. In the case of autonomous driving systems (ADS), the vision perception subsystem is a necessity to ensure correct maneuvering of the environment

System and software verification is a vital component in the development and reliability of cyber-physical systems - especially in critical domains where the margin of error is minimal. In the case of autonomous driving systems (ADS), the vision perception subsystem is a necessity to ensure correct maneuvering of the environment and identification of objects. The challenge posed in perception systems involves verifying the accuracy and rigidity of detections. The use of Spatio-Temporal Perception Logic (STPL) enables the user to express requirements for the perception system to verify, validate, and ensure its behavior; however, a drawback to STPL involves its accessibility. It is limited to individuals with an expert or higher-level knowledge of temporal and spatial logics, and the formal-written requirements become quite verbose with more restrictions imposed. In this thesis, I propose a domain-specific language (DSL) catered to Spatio-Temporal Perception Logic to enable non-expert users the ability to capture requirements for perception subsystems while reducing the necessity to have an experienced background in said logic. The domain-specific language for the Spatio-Temporal Perception Logic is built upon the formal language with two abstractions. The main abstraction captures simple programming statements that are translated to a lower-level STPL expression accepted by the testing monitor. The STPL DSL provides a seamless interface to writing formal expressions while maintaining the power and expressiveness of STPL. These translated equivalent expressions are capable of directing a standard for perception systems to ensure the safety and reduce the risks involved in ill-formed detections.

ContributorsAnderson, Jacob (Author) / Fainekos, Georgios (Thesis director) / Yezhou, Yang (Committee member) / Computer Science and Engineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05