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Description
Recombinases are powerful tools for genome engineering and synthetic biology, however recombinases are limited by a lack of user-programmability and often require complex directed-evolution experiments to retarget specificity. Conversely, CRISPR systems have extreme versatility yet can induce off-target mutations and karyotypic destabilization. To address these constraints we developed an RNA-guided

Recombinases are powerful tools for genome engineering and synthetic biology, however recombinases are limited by a lack of user-programmability and often require complex directed-evolution experiments to retarget specificity. Conversely, CRISPR systems have extreme versatility yet can induce off-target mutations and karyotypic destabilization. To address these constraints we developed an RNA-guided recombinase protein by fusing a hyperactive mutant resolvase from transposon TN3 to catalytically inactive Cas9. We validated recombinase-Cas9 (rCas9) function in model eukaryote Saccharomyces cerevisiae using a chromosomally integrated fluorescent reporter. Moreover, we demonstrated cooperative targeting by CRISPR RNAs at spacings of 22 or 40bps is necessary for directing recombination. Using PCR and Sanger sequencing, we confirmed rCas9 targets DNA recombination. With further development we envision rCas9 becoming useful in the development of RNA-programmed genetic circuitry as well as high-specificity genome engineering.
ContributorsStandage-Beier, Kylie S (Author) / Wang, Xiao (Thesis advisor) / Brafman, David A (Committee member) / Tian, Xiao-jun (Committee member) / Arizona State University (Publisher)
Created2018
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Description
Synthetic biology is an emerging field which melds genetics, molecular biology, network theory, and mathematical systems to understand, build, and predict gene network behavior. As an engineering discipline, developing a mathematical understanding of the genetic circuits being studied is of fundamental importance. In this dissertation, mathematical concepts for understanding, predicting,

Synthetic biology is an emerging field which melds genetics, molecular biology, network theory, and mathematical systems to understand, build, and predict gene network behavior. As an engineering discipline, developing a mathematical understanding of the genetic circuits being studied is of fundamental importance. In this dissertation, mathematical concepts for understanding, predicting, and controlling gene transcriptional networks are presented and applied to two synthetic gene network contexts. First, this engineering approach is used to improve the function of the guide ribonucleic acid (gRNA)-targeted, dCas9-regulated transcriptional cascades through analysis and targeted modification of the RNA transcript. In so doing, a fluorescent guide RNA (fgRNA) is developed to more clearly observe gRNA dynamics and aid design. It is shown that through careful optimization, RNA Polymerase II (Pol II) driven gRNA transcripts can be strong enough to exhibit measurable cascading behavior, previously only shown in RNA Polymerase III (Pol III) circuits. Second, inherent gene expression noise is used to achieve precise fractional differentiation of a population. Mathematical methods are employed to predict and understand the observed behavior, and metrics for analyzing and quantifying similar differentiation kinetics are presented. Through careful mathematical analysis and simulation, coupled with experimental data, two methods for achieving ratio control are presented, with the optimal schema for any application being dependent on the noisiness of the system under study. Together, these studies push the boundaries of gene network control, with potential applications in stem cell differentiation, therapeutics, and bio-production.
ContributorsMenn, David J (Author) / Wang, Xiao (Thesis advisor) / Kiani, Samira (Committee member) / Haynes, Karmella (Committee member) / Nielsen, David (Committee member) / Marshall, Pamela (Committee member) / Arizona State University (Publisher)
Created2018
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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
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
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Description
This study investigates how the patient-provider relationship between lesbian, gay, and bisexual women and their healthcare providers influences their access to, utilization of, and experiences within healthcare environments. Nineteen participants, ages 18 to 34, were recruited using convenience and snowball sampling. Interviews were conducted inquiring about their health history and

This study investigates how the patient-provider relationship between lesbian, gay, and bisexual women and their healthcare providers influences their access to, utilization of, and experiences within healthcare environments. Nineteen participants, ages 18 to 34, were recruited using convenience and snowball sampling. Interviews were conducted inquiring about their health history and their experiences within the healthcare system in the context of their sexual orientation. The data collected from these interviews was used to create an analysis of the healthcare experiences of those who identify as queer. Although the original intention of the project was to chronicle the experiences of LGB women specifically, there were four non-binary gender respondents who contributed interviews. In an effort to not privilege any orientation over another, the respondents were collectively referred to as queer, given the inclusive and an encompassing nature of the term. The general conclusion of this study is that respondents most often experienced heterosexism rather than outright homophobia when accessing healthcare. If heterosexism was present within the healthcare setting, it made respondents feel uncomfortable with their providers and less likely to inform them of their sexuality even if it was medically relevant to their health outcomes. Gender, race, and,socioeconomic differences also had an effect on the patient-provider relationship. Non-binary respondents acknowledged the need for inclusion of more gender options outside of male or female on the reporting forms often seen in medical offices. By doing so, medical professionals are acknowledging their awareness and knowledge of people outside of the binary gender system, thus improving the experience of these patients. While race and socioeconomic status were less relevant to the context of this study, it was found that these factors have an affect on the patient-provider relationship. There are many suggestions for providers to improve the experiences of queer patients within the healthcare setting. This includes nonverbal indications of acknowledgement and acceptance, such as signs in the office that indicate it to be a queer friendly space. This will help in eliminating the fear and miscommunication that can often happen when a queer patient sees a practitioner for the first time. In addition, better education on medically relevant topics to queer patients, is necessary in order to eliminate disparities in health outcomes. This is particularly evident in trans health, where specialized education is necessary in order to decrease poor health outcomes in trans patients. Future directions of this study necessitate a closer look on how race and socioeconomic status have an effect on a queer patient's relationship with their provider.
Created2016-05
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Previous research has found improvements in motor and cognitive measures following Assisted Cycle Therapy (AC) in adolescence with Down syndrome (DS). Our study investigated whether we would find improvements in mental health in older adults with DS as measured from the Adapted Behavior Dementia Questionnaire (ABDQ), Physical Activity Self Efficacy

Previous research has found improvements in motor and cognitive measures following Assisted Cycle Therapy (AC) in adolescence with Down syndrome (DS). Our study investigated whether we would find improvements in mental health in older adults with DS as measured from the Adapted Behavior Dementia Questionnaire (ABDQ), Physical Activity Self Efficacy Scales (PACES), Children's Depressive inventory, which are early indicators of Alzheimer's disease (AD) in persons with Down syndrome. This study consisted of seven participants with Down syndrome between the ages of 31 and 54, inclusive, that cycled for 30 minutes 3 x/week for eight weeks either at their voluntary cycling rate (VC) or approximately 35% faster with the help of a mechanical motor (ACT). Our results were consistent with our prediction that self efficacy improved following ACT, but not VC. However, our results were not consistent with our prediction that dementia and depression were improved following ACT more than VC. These results were interpreted with respect to the effects of exercise in older adults with DS. Future research should focus on recruiting more participants, especially those with deficits in mental health.
ContributorsPandya, Sachin (Author) / Ringenbach, Shannon (Thesis director) / Coon, David (Committee member) / School of Nutrition and Health Promotion (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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Description
Exercise is a great boon to both the health of individuals and the national as a whole. Exercise provides a wide range of significant and well established benefits to both physical and psychological health, with many benefits that are still being discovered. The effects of exercise on health are somewhat

Exercise is a great boon to both the health of individuals and the national as a whole. Exercise provides a wide range of significant and well established benefits to both physical and psychological health, with many benefits that are still being discovered. The effects of exercise on health are somewhat unique, as exercise is one of a limited number of ways to improve health that is not harm a reduction strategy, but instead increases health through direct benefit, rather than increasing health by decreasing damage and harm. Support is also given to the proposal that individuals are in best position to determine the intensity of exercise and to choose primary activities to participate in, in order to provide near maximum physical and psychological benefit, with the understanding that frequency of exercise is of the upmost importance for benefit. The accessibility of exercise and the tremendous health benefit of exercise, makes exercise a huge asset in reducing the exorbitant health care spending and improving mediocre health outcomes in this country; a reasonable goal as numerous countries have better health the United States, even though the United States spends the more than any other country on health.
ContributorsRael, Ashur Scott (Author) / Kashiwagi, Dean (Thesis director) / Kashiwagi, Jacob (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor) / School of Film, Dance and Theatre (Contributor)
Created2015-05
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Description
Bacteria have been shown to possess a large array of regulatory mechanisms to not just respond to a diverse array of environmental stresses, but to injurious artificial proteins as well. A previous investigation introduced DX, a man-made ATP sequestering protein into Escherichia coli (E. coli) which resulted in the formation

Bacteria have been shown to possess a large array of regulatory mechanisms to not just respond to a diverse array of environmental stresses, but to injurious artificial proteins as well. A previous investigation introduced DX, a man-made ATP sequestering protein into Escherichia coli (E. coli) which resulted in the formation of novel endoliposome structures and induced a viable but non-culturable state (VBNC) that was not easily reversed. It was hypothesized that the broadly conserved bacterial stringent response pathway may have been responsible for the observed phenotypic changes. With the goal of unveiling the molecular mechanism behind this novel response, changes in cellular morphology and physiology upon DX expression were assessed in a population of E. coli encoding a dysfunctional relA gene, one of the two genes controlling the induction of the stringent response. It was ultimately shown that RelA directly contributed to cellular filamentation, endoliposome structure formation, and the induction of a VBNC state. While the stringent response has been extensively shown to induce a VBNC state, to our knowledge, relA has not yet been shown to induce filamentation or coordinate the formation of endoliposome structures in bacteria. As the stringent response has been shown to be increasingly involved in antibiotic tolerance, this study provided an exciting opportunity to further characterize this adaptive response pathway to aid in the future development of novel therapeutics. In addition to this, this study continued to highlight that the DX protein may serve one of the first tools to allow for the direct selection of bacteria in a VBNC state by morphologically distinguishing non-culturable cells through cellular filamentation.
ContributorsFrost, Fredrick Charles (Author) / Chaput, John (Thesis director) / Wachter, Rebekka (Committee member) / Korch, Shaleen (Committee member) / Barrett, The Honors College (Contributor) / Department of Chemistry and Biochemistry (Contributor)
Created2014-12
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This paper explains what factors influence mental health issues and what type of care is provided in various countries. The countries in this study will include the United States, Japan, Ethiopia and South Africa, all of which have varying degrees of ethnic diversity, economic status and understanding of mental health

This paper explains what factors influence mental health issues and what type of care is provided in various countries. The countries in this study will include the United States, Japan, Ethiopia and South Africa, all of which have varying degrees of ethnic diversity, economic status and understanding of mental health issues. It discusses the specific healthcare systems in each country, as well as the attitudes and problems associated with depression and schizophrenia, two prevalent mental health disorders. This paper examines the different ways that a diagnosis is reached for schizophrenia and major depression in these different countries, as well as what methods are used for treating individuals with these disorders. It will also examine the prominent notion that schizophrenia has better outcomes in developing countries than in places that have wider medical care available. It then discusses what treatments are available in each country, as well as social constructs that exist regarding those treatments in order to understand the ways that treatments can be expanded to improve outcomes. This paper will then examine the different outcomes of these mental health disorders that are common in each country, and conclude with ideas on how to make global mental health a reality.
ContributorsOlsen, Rachel Lindsay (Author) / Gaughan, Monica (Thesis director) / Wood, Reed (Committee member) / Barrett, The Honors College (Contributor) / School of Politics and Global Studies (Contributor) / School of Human Evolution and Social Change (Contributor) / Department of Psychology (Contributor)
Created2015-05