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The Multiple Antibiotic Resistance Regulator Family (MarR) are transcriptional regulators, many of which forms a dimer. Transcriptional regulation provides bacteria a stabilized responding system to ensure the bacteria is able to efficiently adapt to different environmental conditions. The main function of the MarR family is to create multiple antibiotic resistance

The Multiple Antibiotic Resistance Regulator Family (MarR) are transcriptional regulators, many of which forms a dimer. Transcriptional regulation provides bacteria a stabilized responding system to ensure the bacteria is able to efficiently adapt to different environmental conditions. The main function of the MarR family is to create multiple antibiotic resistance from a mutated protein; this process occurs when the MarR regulates an operon. We hypothesized that different transcriptional regulator genes have interactions with each other. It is known that Salmonella pagC transcription is activated by three regulators, i.e., SlyA, MprA, and PhoP. Bacterial Adenylate Cyclase-based Two-Hybrid (BACTH) system was used to research the protein-protein interactions in SlyA, MprA, and PhoP as heterodimers and homodimers in vivo. Two fragments, T25 and T18, that lack endogenous adenylate cyclase activity, were used for construction of chimeric proteins and reconstruction of adenylate cyclase activity was tested. The significant adenylate cyclase activities has proved that SlyA is able to form homodimers. However, weak adenylate cyclase activities in this study has proved that MprA and PhoP are not likely to form homodimers, and no protein-protein interactions were detected in between SlyA, MprA and PhoP, which no heterodimers have formed in between three transcriptional regulators.
ContributorsTao, Zenan (Author) / Shi, Yixin (Thesis advisor) / Wang, Xuan (Committee member) / Bean, Heather (Committee member) / Arizona State University (Publisher)
Created2018
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Description
Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, is the 10th leading cause of death, worldwide. The prevalence of drug-resistant clinical isolates and the paucity of newly-approved antituberculosis drugs impedes the successful eradication of Mtb. Bacteria commonly use two-component systems (TCS) to sense their environment and genetically modulate adaptive responses.

Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis, is the 10th leading cause of death, worldwide. The prevalence of drug-resistant clinical isolates and the paucity of newly-approved antituberculosis drugs impedes the successful eradication of Mtb. Bacteria commonly use two-component systems (TCS) to sense their environment and genetically modulate adaptive responses. The prrAB TCS is essential in Mtb, thus representing an auspicious drug target; however, the inability to generate an Mtb ΔprrAB mutant complicates investigating how this TCS contributes to pathogenesis. Mycobacterium smegmatis, a commonly used M. tuberculosis genetic surrogate was used here. This work shows that prrAB is not essential in M. smegmatis. During ammonium stress, the ΔprrAB mutant excessively accumulates triacylglycerol lipids, a phenotype associated with M. tuberculosis dormancy and chronic infection. Additionally, triacylglycerol biosynthetic genes were induced in the ΔprrAB mutant relative to the wild-type and complementation strains during ammonium stress. Next, RNA-seq was used to define the M. smegmatis PrrAB regulon. PrrAB regulates genes participating in respiration, metabolism, redox balance, and oxidative phosphorylation. The M. smegmatis ΔprrAB mutant is compromised for growth under hypoxia, is hypersensitive to cyanide, and fails to induce high-affinity respiratory genes during hypoxia. Furthermore, PrrAB positively regulates the hypoxia-responsive dosR TCS response regulator, potentially explaining the hypoxia-mediated growth defects in the ΔprrAB mutant. Despite inducing genes encoding the F1F0 ATP synthase, the ΔprrAB mutant accumulates significantly less ATP during aerobic, exponential growth compared to the wild-type and complementation strains. Finally, the M. smegmatis ΔprrAB mutant exhibited growth impairment in media containing gluconeogenic carbon sources. M. tuberculosis mutants unable to utilize these substrates fail to establish chronic infection, suggesting that PrrAB may regulate Mtb central carbon metabolism in response to chronic infection. In conclusion, 1) prrAB is not universally essential in mycobacteria; 2) M. smegmatis PrrAB regulates genetic responsiveness to nutrient and oxygen stress; and 3) PrrAB may provide feed-forward control of the DosRS TCS and dormancy phenotypes. The data generated in these studies provide insight into the mycobacterial PrrAB TCS transcriptional regulon, PrrAB essentiality in Mtb, and how PrrAB may mediate stresses encountered by Mtb during the transition to chronic infection.
ContributorsMaarsingh, Jason (Author) / Haydel, Shelley E (Thesis advisor) / Roland, Kenneth (Committee member) / Sandrin, Todd (Committee member) / Bean, Heather (Committee member) / Arizona State University (Publisher)
Created2019
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Description
The purpose of this study was to observe the effectiveness of the phenylalanyl arginine β-naphthylamide dihydrochloride inhibitor and Tween 20 when combined with an antibiotic against Escherichia. coli. As antibiotic resistance becomes more and more prevalent it is necessary to think outside the box and do more than just increase

The purpose of this study was to observe the effectiveness of the phenylalanyl arginine β-naphthylamide dihydrochloride inhibitor and Tween 20 when combined with an antibiotic against Escherichia. coli. As antibiotic resistance becomes more and more prevalent it is necessary to think outside the box and do more than just increase the dosage of currently prescribed antibiotics. This study attempted to combat two forms of antibiotic resistance. The first is the AcrAB efflux pump which is able to pump antibiotics out of the cell. The second is the biofilms that E. coli can form. By using an inhibitor, the pump should be unable to rid itself of an antibiotic. On the other hand, using Tween allows for biofilm formation to either be disrupted or for the biofilm to be dissolved. By combining these two chemicals with an antibiotic that the efflux pump is known to expel, low concentrations of each chemical should result in an equivalent or greater effect on bacteria compared to any one chemical in higher concentrations. To test this hypothesis a 96 well plate BEC screen test was performed. A range of antibiotics were used at various concentrations and with varying concentrations of both Tween and the inhibitor to find a starting point. Following this, Erythromycin and Ciprofloxacin were picked as the best candidates and the optimum range of the antibiotic, Tween, and inhibitor were established. Finally, all three chemicals were combined to observe the effects they had together as opposed to individually or paired together. From the results of this experiment several conclusions were made. First, the inhibitor did in fact increase the effectiveness of the antibiotic as less antibiotic was needed if the inhibitor was present. Second, Tween showed an ability to prevent recovery in the MBEC reading, showing that it has the ability to disrupt or dissolve biofilms. However, Tween also showed a noticeable decrease in effectiveness in the overall treatment. This negative interaction was unable to be compensated for when using the inhibitor and so the hypothesis was proven false as combining the three chemicals led to a less effective treatment method.
ContributorsPetrovich Flynn, Chandler James (Author) / Misra, Rajeev (Thesis director) / Bean, Heather (Committee member) / Perkins, Kim (Committee member) / Mechanical and Aerospace Engineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
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Description
Social-emotional learning (SEL) methods are beginning to receive global attention in primary school education, yet the dominant emphasis on implementing these curricula is in high-income, urbanized areas. Consequently, the unique features of developing and integrating such methods in middle- or low-income rural areas are unclear. Past studies suggest that students

Social-emotional learning (SEL) methods are beginning to receive global attention in primary school education, yet the dominant emphasis on implementing these curricula is in high-income, urbanized areas. Consequently, the unique features of developing and integrating such methods in middle- or low-income rural areas are unclear. Past studies suggest that students exposed to SEL programs show an increase in academic performance, improved ability to cope with stress, and better attitudes about themselves, others, and school, but these curricula are designed with an urban focus. The purpose of this study was to conduct a needs-based analysis to investigate components specific to a SEL curriculum contextualized to rural primary schools. A promising organization committed to rural educational development is Barefoot College, located in Tilonia, Rajasthan, India. In partnership with Barefoot, we designed an ethnographic study to identify and describe what teachers and school leaders consider the highest needs related to their students' social and emotional education. To do so, we interviewed 14 teachers and school leaders individually or in a focus group to explore their present understanding of “social-emotional learning” and the perception of their students’ social and emotional intelligence. Analysis of this data uncovered common themes among classroom behaviors and prevalent opportunities to address social and emotional well-being among students. These themes translated into the three overarching topics and eight sub-topics explored throughout the curriculum, and these opportunities guided the creation of the 21 modules within it. Through a design-based research methodology, we developed a 40-hour curriculum by implementing its various modules within seven Barefoot classrooms alongside continuous reiteration based on teacher feedback and participant observation. Through this process, we found that student engagement increased during contextualized SEL lessons as opposed to traditional methods. In addition, we found that teachers and students preferred and performed better with an activities-based approach. These findings suggest that rural educators must employ particular teaching strategies when addressing SEL, including localized content and an experiential-learning approach. Teachers reported that as their approach to SEL shifted, they began to unlock the potential to build self-aware, globally-minded students. This study concludes that social and emotional education cannot be treated in a generalized manner, as curriculum development is central to the teaching-learning process.
ContributorsBucker, Delaney Sue (Author) / Carrese, Susan (Thesis director) / Barab, Sasha (Committee member) / School of Life Sciences (Contributor, Contributor) / School of Civic & Economic Thought and Leadership (Contributor) / School of International Letters and Cultures (Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
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Description
Calcium is the only ion capable of triggering electrical and chemical reactions in cells which are part of essential biomolecular processes, such as gene transcription and ion flux. Calcium homeostasis, the control of concentration levels, is therefore crucial for the proper functioning of cells. For example, cardiomyocytes, the cells that

Calcium is the only ion capable of triggering electrical and chemical reactions in cells which are part of essential biomolecular processes, such as gene transcription and ion flux. Calcium homeostasis, the control of concentration levels, is therefore crucial for the proper functioning of cells. For example, cardiomyocytes, the cells that form cardiac muscle, rely on calcium transfer process to produce muscle contraction.
The purpose of this work is to study aspects of calcium homeostasis in the model organism Saccharomyces cerevisiae, common yeast. Using luminometric techniques, the response of the yeast was monitored against a set of changes in the environment calcium abundance. The results indicate a complex response as both increase and decreases of external calcium induce elevations in cytosolic calcium concentrations.
Calcium is transferred across compartments by means of channels. In Saccharomyces cerevisiae, many of them have been identified; Cch1p-Mid1p, Vcx1p, Pmc1p, Pmr1p, and Yvc1p. Their participation in calcium homeostasis is well established. Observations of cytosolic calcium increase after a hypertonic shock are mainly associated with influx of ions from the environment though the Cch1p-Mid1p. This process is generally considered as driven by calcium concentration gradients. However, recent studies have suggested that the plasma membrane channel, Cch1p-Mid1p, may possess more sophisticated regulation and sensory mechanisms. The results of our experiments support these ideas.
We carried out experiments that subjected yeast to multiple shocks: a hypertonic shock followed by either a second hypertonic shock, a hypotonic shock, or a yeast dilution pulse where the solution volume increases by the calcium concentration has only a small change. The cytosolic calcium concentration of a yeast population was monitored via luminometry.
The main result of this study is the observation of an unexpected response to the combination of hypertonic and hypotonic shocks. In this case it was observed that the cytosolic calcium concentration increased after both shocks. This indicates that cytosolic calcium increases are not solely driven by the presence of concentration gradients. The response after the hypotonic pulse arises from more complex mechanisms that may include sensor activity at the membrane channels and the release of calcium from internal storages.
ContributorsMintz, David Anthony (Co-author) / Parker, Augustus (Co-author) / Solis, Francisco (Thesis director) / Marshall, Pamela (Committee member) / School of Mathematical and Natural Sciences (Contributor, Contributor) / Barrett, The Honors College (Contributor)
Created2020-05
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Description
Space microbiology, or the study of microorganisms in space, has significant applications for both human spaceflight and Earth-based medicine. This thesis traces the evolution of the field of space microbiology since its creation in 1935. Beginning with simple studies to determine if terrestrial life could survive spaceflight, the field of

Space microbiology, or the study of microorganisms in space, has significant applications for both human spaceflight and Earth-based medicine. This thesis traces the evolution of the field of space microbiology since its creation in 1935. Beginning with simple studies to determine if terrestrial life could survive spaceflight, the field of space microbiology has grown to encompass a substantial body of work that is now recognized as an essential component of NASA' research endeavors. Part one provides an overview of the early period of space microbiology, from high-altitude balloon and rocket studies to work conducted during the Apollo program. Part two summarizes the current state of the field, with a specific focus on the revolutionary contributions made by the Nickerson lab at the Biodesign Institute at ASU using the NASA-designed Rotating Wall Vessel (RWV) Bioreactor. Finally, part three highlights the research I've conducted in the Nickerson lab, as well as continuing studies within the field of space microbiology.
ContributorsMcCarthy, Breanne E. (Author) / Lynch, John (Thesis director) / Foy, Joseph (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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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
The mechanisms of extracellular respiration in Geobacter sulfurreducens, commonly considered to be a model organism for anode respiration, are yet to be completely understood. The interplay between electron and proton transport especially could be a key to gaining further insights. One way to investigate the mechanisms of extracellular respiration under

The mechanisms of extracellular respiration in Geobacter sulfurreducens, commonly considered to be a model organism for anode respiration, are yet to be completely understood. The interplay between electron and proton transport especially could be a key to gaining further insights. One way to investigate the mechanisms of extracellular respiration under varying environmental conditions is by analyzing the electrochemical response of the biofilm with respect to pH, buffer concentrations, and acetate concentrations. I seek to increase the understanding of the electrochemical response of the G. sulfurreducens biofilm through electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) techniques in concert with chronoamperometry. I used Geobacter sulfurreducens PCA biofilms in single-chamber electrochemical cells (approximately 100 mL volume) with a small gold working electrode (3.14 mm2). I observed limitations in the initial methods used for media replacement. I tracked changes in the CV data, such as EKA (midpoint potential), as a function of pH and buffer concentration. The media replacement method developed demonstrates success in pH experiments that will be transferrable to other environmental conditions to study electron transport. The experiments revealed that the clarity of data collected is dependent on the quality of the biofilm. A high quality biofilm is characterized by a high current density and normal growth behavior. The general trends seen in these experiments are that as pH increases the potential decreases, and as buffer concentration increases the potential decreases and pH increases. Acetate-free conditions in the reactor were unable to be achieved as characterized by non-zero current densities in the acetate-free experiments.
ContributorsHolzer, Denton Gene (Author) / Torres, Cesar (Thesis director) / Popat, Sudeep (Committee member) / Yoho, Rachel (Committee member) / Chemical Engineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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Description
The goal of our study is to identify socio-economic risk factors for depressive disorder and poor mental health by statistically analyzing survey data from the CDC. The identification of risk groups in a particular demographic could aid in the development of targeted interventions to improve overall quality of mental health

The goal of our study is to identify socio-economic risk factors for depressive disorder and poor mental health by statistically analyzing survey data from the CDC. The identification of risk groups in a particular demographic could aid in the development of targeted interventions to improve overall quality of mental health in the United States. In our analysis, we studied the influences and correlations of socioeconomic factors that regulate the risk of developing Depressive Disorders and overall poor mental health. Using the statistical software STATA, we ran a regression model of selected independent socio-economic variables with the dependent mental health variables. The independent variables of the statistical model include Income, Race, State, Age, Marital Status, Sex, Education, BMI, Smoker Status, and Alcohol Consumption. Once the regression coefficients were found, we illustrated the data in graphs and heat maps to qualitatively provide visuals of the prevalence of depression in the U.S. demography. Our study indicates that the low-income and under-educated populations who are everyday smokers, obese, and/or are in divorced or separated relationships should be of main concern. A suggestion for mental health organizations would be to support counseling and therapeutic efforts as secondary care for those in smoking cessation programs, weight management programs, marriage counseling, or divorce assistance group. General improvement in alleviating poverty and increasing education could additionally show progress in counter-acting the prevalence of depressive disorder and also improve overall mental health. The identification of these target groups and socio-economic risk factors are critical in developing future preventative measures.
ContributorsGrassel, Samuel (Co-author) / Choueiri, Alexi (Co-author) / Choueiri, Robert (Co-author) / Goegan, Brian (Thesis director) / Holter, Michael (Committee member) / Sandra Day O'Connor College of Law (Contributor) / School of Molecular Sciences (Contributor) / School of Politics and Global Studies (Contributor) / Economics Program in CLAS (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
Description
This thesis examines youth with mental health disorders in the juvenile justice system, and when their stories intersect with media coverage. In addition to exploring the history of juvenile justice in the United States, it reviews the relationship with youths who have a mental health disorder within the system, criticism

This thesis examines youth with mental health disorders in the juvenile justice system, and when their stories intersect with media coverage. In addition to exploring the history of juvenile justice in the United States, it reviews the relationship with youths who have a mental health disorder within the system, criticism of media’s coverage on these topics, and expert opinion on how this can be improved. Typically, both print and broadcast media have utilized these stories for sensationalism: slapping these crimes across the front page of the paper, or leading in a broadcast news show. Yet the journalistic responsibility of educating the community is tragically over looked, with these stories adding to the stigma of mental health. With this research, I aim to gather insight on how word choices, details, and story structure can improve daily reporting in a world of tight deadlines and competing interests—moving into a higher-quality news product.
Created2016-05