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Description
Pathogenic Gram-negative bacteria employ a variety of molecular mechanisms to combat host defenses. Two-component regulatory systems (TCR systems) are the most ubiquitous signal transduction systems which regulate many genes required for virulence and survival of bacteria. In this study, I analyzed different TCR systems in two clinically-relevant Gram-negative bacteria, i.e., oral pathogen Porphyromonas gingivalis and enterobacterial Escherichia coli. P. gingivalis is a major causative agent of periodontal disease as well as systemic illnesses, like cardiovascular disease. A microarray study found that the putative PorY-PorX TCR system controls the secretion and maturation of virulence factors, as well as loci involved in the PorSS secretion system, which secretes proteinases, i.e., gingipains, responsible for periodontal disease. Proteomic analysis (SILAC) was used to improve the microarray data, reverse-transcription PCR to verify the proteomic data, and primer extension assay to determine the promoter regions of specific PorX regulated loci. I was able to characterize multiple genetic loci regulated by this TCR system, many of which play an essential role in hemagglutination and host-cell adhesion, and likely contribute to virulence in this bacterium. Enteric Gram-negative bacteria must withstand many host defenses such as digestive enzymes, low pH, and antimicrobial peptides (AMPs). The CpxR-CpxA TCR system of E. coli has been extensively characterized and shown to be required for protection against AMPs. Most recently, this TCR system has been shown to up-regulate the rfe-rff operon which encodes genes involved in the production of enterobacterial common antigen (ECA), and confers protection against a variety of AMPs. In this study, I utilized primer extension and DNase I footprinting to determine how CpxR regulates the ECA operon. My findings suggest that CpxR modulates transcription by directly binding to the rfe promoter. Multiple genetic and biochemical approaches were used to demonstrate that specific TCR systems contribute to regulation of virulence factors and resistance to host defenses in P. gingivalis and E. coli, respectively. Understanding these genetic circuits provides insight into strategies for pathogenesis and resistance to host defenses in Gram negative bacterial pathogens. Finally, these data provide compelling potential molecular targets for therapeutics to treat P. gingivalis and E. coli infections.
ContributorsLeonetti, Cori (Author) / Shi, Yixin (Thesis advisor) / Stout, Valerie (Committee member) / Nickerson, Cheryl (Committee member) / Sandrin, Todd (Committee member) / Arizona State University (Publisher)
Created2013
Description
This study was devised to elucidate key information concerning the potential risk posed by Legionella in reclaimed water. A series of biological experiments and a recharge basin soil column study were conducted to examine the survival, growth, and transport of L. pneumophila through engineered reclaimed water systems. A pilot-scale, column study was set up to measure Legionella transport in the columns under Arizona recharge basin conditions. Two columns, A and B, were packed to a depth of 122 cm with a loamy sand media collected from a recharge basin in Mesa, Arizona. The grain size distribution of Column A differed from that of Column B by the removal of fines passing the #200 sieve. The different soil profiles represented by column A and B allowed for further investigation of soil attributes which influence the microbial transport mechanism. Both clear PVC columns stand at a height of 1.83 m with an inner diameter of 6.35 cm. Sampling ports were drilled into the column at the soil depths 15, 30, 60, 92, 122 cm. Both columns were acclimated with tertiary treated waste water and set to a flow rate of approximately 1.5 m/d. The columns were used to assess the transport of a bacterial indicator, E. coli, in addition to assessing the study's primary pathogen of concern, Legionella. Approximately, 〖10〗^7 to 〖10〗^9 E. coli cells or 〖10〗^6 to 〖10〗^7Legionella cells were spiked into the columns' head waters for each experiment. Periodically, samples were collected from each column's sampling ports, until a minimum of three pore volume passed through the columns.
The pilot-scale, column study produced novel results which demonstrated the mechanism for Legionella to be transported through recharge basin soil. E. coli was transported, through 122 cm of the media in under 6 hours, whereas, Legionella was transported, through the same distance, in under 30 hours. Legionella has been shown to survive in low nutrient conditions for over a year. Given the novel results of this proof of concept study, a claim can be made for the transport of Legionella into groundwater aquifers through engineering recharge basin conditions, in Central Arizona.
The pilot-scale, column study produced novel results which demonstrated the mechanism for Legionella to be transported through recharge basin soil. E. coli was transported, through 122 cm of the media in under 6 hours, whereas, Legionella was transported, through the same distance, in under 30 hours. Legionella has been shown to survive in low nutrient conditions for over a year. Given the novel results of this proof of concept study, a claim can be made for the transport of Legionella into groundwater aquifers through engineering recharge basin conditions, in Central Arizona.
ContributorsMcBurnett, Lauren Rae (Author) / Abbaszadegan, Morteza (Thesis advisor) / Alum, Absar (Committee member) / Fox, Peter (Committee member) / Arizona State University (Publisher)
Created2014
Description
With the aid of metabolic pathways engineering, microbes are finding increased use as biocatalysts to convert renewable biomass resources into fine chemicals, pharmaceuticals and other valuable compounds. These alternative, bio-based production routes offer distinct advantages over traditional synthesis methods, including lower energy requirements, rendering them as more "green" and "eco-friendly". Escherichia coli has recently been engineered to produce the aromatic chemicals (S)-styrene oxide and phenol directly from renewable glucose. Several factors, however, limit the viability of this approach, including low titers caused by product inhibition and/or low metabolic flux through the engineered pathways. This thesis focuses on addressing these concerns using magnetic mesoporous carbon powders as adsorbents for continuous, in-situ product removal as a means to alleviate such limitations. Using process engineering as a means to troubleshoot metabolic pathways by continuously removing products, increased yields are achieved from both pathways. By performing case studies in product toxicity and reaction equilibrium it was concluded that each step of a metabolic pathway can be optimized by the strategic use of in-situ adsorption as a process engineering tool.
ContributorsVasudevan, Anirudh (Author) / Nielsen, David R (Thesis advisor) / Torres, César I (Committee member) / Wang, Xuan (Committee member) / Arizona State University (Publisher)
Created2014
Description
In this thesis, I present a lab-on-a-chip (LOC) that can separate and detect Escherichia Coli (E. coli) in simulated urine samples for Urinary Tract Infection (UTI) diagnosis. The LOC consists of two (concentration and sensing) chambers connected in series and an integrated impedance detector. The two-chamber approach is designed to reduce the non-specific absorption of proteins, e.g. albumin, that potentially co-exist with E. coli in urine. I directly separate E. coli K-12 from a urine cocktail in a concentration chamber containing micro-sized magnetic beads (5 µm in diameter) conjugated with anti-E. coli antibodies. The immobilized E. coli are transferred to a sensing chamber for the impedance measurement. The measurement at the concentration chamber suffers from non-specific absorption of albumin on the gold electrode, which may lead to a false positive response. By contrast, the measured impedance at the sensing chamber shows ~60 kÙ impedance change between 6.4x104 and 6.4x105 CFU/mL, covering the threshold of UTI (105 CFU/mL). The sensitivity of the LOC for detecting E. coli is characterized to be at least 3.4x104 CFU/mL. I also characterized the LOC for different age groups and white blood cell spiked samples. These preliminary data show promising potential for application in portable LOC devices for UTI detection.
ContributorsKim, Sangpyeong (Author) / Chae, Junseok (Thesis advisor) / Phillips, Stephen M. (Committee member) / Blain Christen, Jennifer M. (Committee member) / Arizona State University (Publisher)
Created2011
Description
The diversity of industrially important chemicals that can be produced biocatalytically from renewable resources continues to expand with the aid of metabolic and pathway engineering. In addition to biofuels, these chemicals also include a number of monomers with utility in conventional and novel plastic materials production. Monomers used for polyamide production are no exception, as evidenced by the recent engineering of microbial biocatalysts to produce cadaverine, putrescine, and succinate. In this thesis the repertoire and depth of these renewable polyamide precursors is expanded upon through the engineering of a novel pathway that enables Escherichia coli to produce, as individual products, both δ-aminovaleric acid (AMV) and glutaric acid when grown in glucose mineral salt medium. δ-Aminovaleric acid is the monomeric subunit of nylon-5 homopolymer, whereas glutaric acid is a dicarboxylic acid used to produce copolymers such as nylon-5,5. These feats were achieved by increasing endogenous production of the required pathway precursor, L-lysine. E. coli was engineered for L-lysine over-production through the introduction and expression of metabolically deregulated pathway genes, namely aspartate kinase III and dihydrodipicolinate synthase, encoded by the feedback resistant mutants lysCfbr and dapAfbr, respectively. After deleting a natural L-lysine decarboxylase, up to 1.6 g/L L-lysine could be produced from glucose in shake flasks as a result. The natural L-lysine degradation pathway of numerous Pseudomonas sp., which passes from L-lysine through both δ-aminovaleric acid and glutaric acid, was then functionally reconstructed in a piecewise manner in the E. coli L-lysine over-producer. Expression of davBA alone resulted in the production of over 0.86 g/L AMV in 48 h. Expression of davBADT resulted in the production of over 0.82 g/L glutaric acid under the same conditions. These production titers were achieved with yields of 69.5 and 68.4 mmol/mol of AMV and glutarate, respectively. Future improvements to the ability to synthesize both products will likely come from the ability to eliminate cadaverine by-product formation through the deletion of cadA and ldcC, genes involved in E. coli's native lysine degradation pathway. Nevertheless, through metabolic and pathway engineering, it is now possible produce the polyamide monomers of δ-aminovaleric acid and glutaric acid from renewable resources.
ContributorsAdkins, Jake M (Author) / Nielsen, David R. (Thesis advisor) / Caplan, Michael (Committee member) / Torres, Cesar (Committee member) / Arizona State University (Publisher)
Created2012
Description
How do you convey what’s interesting and important to you as an artist in a digital world of constantly shifting attentions? For many young creatives, the answer is original characters, or OCs. An OC is a character that an artist creates for personal enjoyment, whether based on an already existing story or world, or completely from their own imagination.
As creations made for purely personal interests, OCs are an excellent elevator pitch to talk one creative to another, opening up opportunities for connection in a world where communication is at our fingertips but personal connection is increasingly harder to make. OCs encourage meaningful interaction by offering themselves as muses, avatars, and story pieces, and so much more, where artists can have their characters interact with other creatives through many different avenues such as art-making, table top games, or word of mouth.
In this thesis, I explore the worlds and aesthetics of many creators and their original characters through qualitative research and collaborative art-making. I begin with a short survey of my creative peers, asking general questions about their characters and thoughts on OCs, then move to sketching characters from various creators. I focus my research to a group of seven core creators and their characters, whom I interview and work closely with in order to create a series of seven final paintings of their original characters.
As creations made for purely personal interests, OCs are an excellent elevator pitch to talk one creative to another, opening up opportunities for connection in a world where communication is at our fingertips but personal connection is increasingly harder to make. OCs encourage meaningful interaction by offering themselves as muses, avatars, and story pieces, and so much more, where artists can have their characters interact with other creatives through many different avenues such as art-making, table top games, or word of mouth.
In this thesis, I explore the worlds and aesthetics of many creators and their original characters through qualitative research and collaborative art-making. I begin with a short survey of my creative peers, asking general questions about their characters and thoughts on OCs, then move to sketching characters from various creators. I focus my research to a group of seven core creators and their characters, whom I interview and work closely with in order to create a series of seven final paintings of their original characters.
ContributorsCote, Jacqueline (Author) / Button, Melissa M (Thesis director) / Dove-Viebahn, Aviva (Committee member) / School of Art (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
In the past ten years, the United States’ sound recording industries have experienced significant decreases in employment opportunities for aspiring audio engineers from economic imbalances in the music industry’s digital streaming era and reductions in government funding for career and technical education (CTE). The Recording Industry Association of America reports promises of music industry sustainability based on increasing annual revenues in paid streaming services and artists’ high creative demand. The rate of new audio engineer entries in the sound recording subsection of the music industry is not viable to support streaming artists’ high demand to engineer new music recordings. Offering CTE programs in secondary education is rare for aspiring engineers with insufficient accessibility to pursue a post-secondary or vocational education because of financial and academic limitations. These aspiring engineers seek alternatives for receiving an informal education in audio engineering on the Internet using video sharing services like YouTube to search for tutorials and improve their engineering skills. The shortage of accessible educational materials on the Internet restricts engineers from advancing their own audio engineering education, reducing opportunities to enter a desperate job market in need of independent, home studio-based engineers. Content creators on YouTube take advantage of this situation and commercialize their own video tutorial series for free and selling paid subscriptions to exclusive content. This is misleading for newer engineers because these tutorials omit important understandings of fundamental engineering concepts. Instead, content creators teach inflexible engineering methodologies that are mostly beneficial to their own way of thinking. Content creators do not often assess the incompatibility of teaching their own methodologies to potential entrants in a profession that demands critical thinking skills requiring applied fundamental audio engineering concepts and techniques. This project analyzes potential solutions to resolve the deficiencies in online audio engineering education and experiments with structuring simple, deliverable, accessible educational content and materials to new entries in audio engineering. Designing clear, easy to follow material to these new entries in audio engineering is essential for developing a strong understanding for the application of fundamental concepts in future engineers’ careers. Approaches to creating and designing educational content requires translating complex engineering concepts through simplified mediums that reduce limitations in learning for future audio engineers.
ContributorsBurns, Triston Connor (Author) / Tobias, Evan (Thesis director) / Libman, Jeff (Committee member) / Department of Information Systems (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
Description
There has been a recent push for queer fiction, especially in the young adult genre, whose focus is gay and lesbian relationships. This growth is much needed in terms of visibility and the furthering of acceptance, but there are still subjects within the LGBTQ+ community that need to be addressed, including bisexual, asexual, and non-binary erasure. There are many people who claim that these identities do not exist, are labels used as a stepping stone on one's journey to discovering that they are homosexual, or are invented excuses for overtly promiscuous or prudish behavior. The existence of negative stereotypes, particularly those of non-binary individuals, is largely due to a lack of visibility and respectful representation within media and popular culture. However, there is still a dearth of non-binary content in popular literature outside of young adult fiction. Can You See Me? aims to fill the gap in bisexual, asexual, and non-binary representation in adult literature. Each of the four stories that make up this collection deals with an aspect of gender and/or sexuality that has been erased, ignored, or denied visibility in American popular culture. The first story, "We'll Grow Lemon Trees," examines bisexual erasure through the lens of sociolinguistics. A bisexual Romanian woman emigrates to Los Angeles in 1989 and must navigate a new culture, learn new languages, and try to move on from her past life under a dictatorship where speaking up could mean imprisonment or death. The second story "Up, Down, All Around," is about a young genderqueer child and their parents dealing with microaggressions, examining gender norms, and exploring personal identity through imaginary scenarios, each involving an encounter with an unknown entity and a colander. The third story, "Aces High," follows two asexual characters from the day they're born to when they are 28 years old, as they find themselves in pop culture. The two endure identity crises, gender discrimination, erasure, individual obsessions, and prejudice as they learn to accept themselves and embrace who they are. In the fourth and final story, "Mile Marker 72," a gay Mexican man must hide in plain sight as he deals with the death of his partner and coming out to his best friend, whose brother is his partner's murderer.
ContributorsOchser, Jordyn M. (Author) / Bell, Matt (Thesis director) / Free, Melissa (Committee member) / Department of English (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
Pandora is a play exploring our relationship with gendered technology through the lens of artificial intelligence. Can women be subjective under patriarchy? Do robots who look like women have subjectivity? Hoping to create a better version of ourselves, The Engineer must navigate the loss of her creation, and Pandora must navigate their new world. The original premiere run was March 27-28, 2018, original cast: Caitlin Andelora, Rikki Tremblay, and Michael Tristano Jr.
ContributorsToye, Abigail Elizabeth (Author) / Linde, Jennifer (Thesis director) / Abele, Kelsey (Committee member) / Department of Information Systems (Contributor) / Economics Program in CLAS (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
After freelancing on my own for the past year and a half, I have realized that one of the biggest obstacles to college entrepreneurs is a fear or apprehension to sales. As a computer science major trying to sell my services, I discovered very quickly that I had not been prepared for the difficulty of learning sales. Sales get a bad rap and very often is the last thing that young entrepreneurs want to try, but the reality is that sales is oxygen to a company and a required skill for an entrepreneur. Due to this, I compiled all of my knowledge into an e-book for young entrepreneurs starting out to learn how to open up a conversation with a prospect all the way to closing them on the phone. Instead of starting from scratch like I did, college entrepreneurs can learn the bare basics of selling their own services, even if they are terrified of sales and what it entails. In this e-book, there are tips that I have learned to deal with my anxiety about sales such as taking the pressure off of yourself and prioritizing listening more than pitching. Instead of trying to teach sales expecting people to be natural sales people, this e-book takes the approach of helping entrepreneurs that are terrified of sales and show them how they can cope with this fear and still close a client. In the future, I hope young entrepreneurs will have access to more resources that handle this fear and make it much easier for them to learn it by themselves. This e-book is the first step.
ContributorsMead, Kevin Tyler (Author) / Sebold, Brent (Thesis director) / Kruse, Gabriel (Committee member) / Computer Science and Engineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05