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Transient Receptor Potential (TRP) ion channels are a diverse family of nonselective, polymodal sensors in uni- and multicellular eukaryotes that are implicated in an assortment of biological contexts and human disease. The cold-activated TRP Melastatin-8 (TRPM8) channel, also recognized as the human body's primary cold sensor, is among the few

Transient Receptor Potential (TRP) ion channels are a diverse family of nonselective, polymodal sensors in uni- and multicellular eukaryotes that are implicated in an assortment of biological contexts and human disease. The cold-activated TRP Melastatin-8 (TRPM8) channel, also recognized as the human body's primary cold sensor, is among the few TRP channels responsible for thermosensing. Despite sustained interest in the channel, the mechanisms underlying TRPM8 activation, modulation, and gating have proved challenging to study and remain poorly understood. In this thesis, I offer data collected on various expression, extraction, and purification conditions tested in E. Coli expression systems with the aim to optimize the generation of a structurally stable and functional human TRPM8 pore domain (S5 and S6) construct for application in structural biology studies. These studies, including the biophysical technique nuclear magnetic spectroscopy (NMR), among others, will be essential for elucidating the role of the TRPM8 pore domain in in regulating ligand binding, channel gating, ion selectively, and thermal sensitivity. Moreover, in the second half of this thesis, I discuss the ligation-independent megaprimer PCR of whole-plasmids (MEGAWHOP PCR) cloning technique, and how it was used to generate chimeras between TRPM8 and its nearest analog TRPM2. I review steps taken to optimize the efficiency of MEGAWHOP PCR and the implications and unique applications of this novel methodology for advancing recombinant DNA technology. I lastly present preliminary electrophysiological data on the chimeras, employed to isolate and study the functional contributions of each individual transmembrane helix (S1-S6) to TRPM8 menthol activation. These studies show the utility of the TRPM8\u2014TRPM2 chimeras for dissecting function of TRP channels. The average current traces analyzed thus far indicate that the S2 and S3 helices appear to play an important role in TRPM8 menthol modulation because the TRPM8[M2S2] and TRPM8[M2S3] chimeras significantly reduce channel conductance in the presence of menthol. The TRPM8[M2S4] chimera, oppositely, increases channel conductance, implying that the S4 helix in native TRPM8 may suppress menthol modulation. Overall, these findings show that there is promise in the techniques chosen to identify specific regions of TRPM8 crucial to menthol activation, though the methods chosen to study the TRPM8 pore independent from the whole channel may need to be reevaluated. Further experiments will be necessary to refine TRPM8 pore solubilization and purification before structural studies can proceed, and the electrophysiology traces observed for the chimeras will need to be further verified and evaluated for consistency and physiological significance.
ContributorsWaris, Maryam Siddika (Author) / Van Horn, Wade (Thesis director) / Redding, Kevin (Committee member) / School of Molecular Sciences (Contributor) / Department of English (Contributor) / Barrett, The Honors College (Contributor)
Created2016-05
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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
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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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Description
Home Base Initiative is a student-led venture project co-founded by Madison Sutton and Sonia Sabrowsky in January 2018. As an organization, Home Base Initiative addresses the problem of teen suicide by educating parents, teachers, and students about the research-backed mental health resources currently available to them and by implementing peer-based

Home Base Initiative is a student-led venture project co-founded by Madison Sutton and Sonia Sabrowsky in January 2018. As an organization, Home Base Initiative addresses the problem of teen suicide by educating parents, teachers, and students about the research-backed mental health resources currently available to them and by implementing peer-based support programs in local high schools. With the belief that positive mental health habits are for everyone, not just individuals with a clinical diagnosis, Home Base Initiative aims to encourage positive conversations about mental health and to increase social and emotional resilience among adolescents to help them navigate the challenges in their lives. In addition to identifying the community problem our organization aims to solve, this document outlines the initial conception, development, and future outlook Home Base Initiative by describing the methods by which the organization has researched other like-minded programs, formed strategic partnerships with community members, piloted its peer-based program at a local high school, and established a foundation for future success as a student organization at Arizona State University. Currently, the Home Base Initiative team consists of 10 undergraduate students at ASU with diverse backgrounds and academic interests as well as credible mentors who are involved in the ASU Tillman Scholars Program, ASU Counseling Services, and The Courage Lab at ASU. We are united by our passion for supporting others’ mental health, and we are dedicated to playing an active role in the healthy development of our fellow community members through mental health advocacy and the facilitation of positive peer-to-peer interactions.
ContributorsSabrowsky, Sonia (Co-author, Co-author) / Sutton, Madison (Co-author) / Mokwa, Michael (Thesis director) / Eaton, John (Committee member) / School of Molecular Sciences (Contributor) / Department of Psychology (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
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Description
Structure is a critical component in drug development. This project supports antibody- facilitated structure determination for the following eleven membrane proteins: the human histamine and dopamine G protein-coupled receptors (HRH4 and DRD2) involved in a wide variety of pathologies such as allergies, inflammation, asthma, pain along with Parkinson's and schizophrenia

Structure is a critical component in drug development. This project supports antibody- facilitated structure determination for the following eleven membrane proteins: the human histamine and dopamine G protein-coupled receptors (HRH4 and DRD2) involved in a wide variety of pathologies such as allergies, inflammation, asthma, pain along with Parkinson's and schizophrenia respectively, the human cystic fibrosis transmembrane conductance regulator (CFTR), the human NaV1.8 voltage-gated sodium ion channel, the human TPC2 two-pore channel, the SARS virus proteins 3a, E and M, the MERS virus protein E and M, and the malarial chloroquine resistance transporter (PfCRT). Serum antibodies against these proteins were generated by genetic immunization, and both in vitro and in vivo expressed membrane proteins were created to characterize the serum antibodies. Plasmid clones were generated for genetic immunization, in vitro protein expression, and in vivo expression (HEK293T transfection). Serum antibodies were generated by genetic immunization of mice by gene gun. Genetic immunization promotes an immune response that allows for the generation of antibodies in the absence of purified protein. In vitro expression was accomplished through the novel technique: in vitro translation with hydrophobic magnetic beads (IVT-HMB). Transfections were performed using the HEK293T cell line to express the protein in vivo. The generated protein was then used in gel electrophoresis and silver stain and/or Western blot analyses to identify and visualize the proteins. These expressed proteins will allow for forthcoming characterization of the generated antibodies. The resulting antibodies will in turn enable structure determination of these important membrane proteins by co-crystallization.
ContributorsDrotar, Beniamin (Author) / Fromme, Petra (Thesis director) / Hansen, Debra T. (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05
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Description
G protein-coupled receptors, or GPCRs, are receptors located within the membrane of cells that elicit a wide array of cellular responses through their interactions with G proteins. Recent advances in the use of lipid cubic phase (LCP) for the crystallization of GPCRs, as well as increased knowledge of techniques to

G protein-coupled receptors, or GPCRs, are receptors located within the membrane of cells that elicit a wide array of cellular responses through their interactions with G proteins. Recent advances in the use of lipid cubic phase (LCP) for the crystallization of GPCRs, as well as increased knowledge of techniques to improve receptor stability, have led to a large increase in the number of available GPCR structures, despite historic difficulties. This project is focused on the histamine family of receptors, which are Class A GPCRs that are involved in the body’s allergic and inflammatory responses. In particular, the goal of this project was to design, express, and purify histamine receptors with the ultimate goal of crystallization. Successive rounds of optimization included the use of recombinant DNA techniques in E.coli to truncate sections of the proteins and the insertion of several fusion partner proteins to improve receptor expression and stability. All constructs were expressed in a Bac-to-Bac baculovirus expression system using Sf9 insect cells, solubilized using n-Dodecyl-β-D-Maltoside (DDM), and purified using immobilized metal affinity chromatography. Constructs were then analyzed by SDS-Page, Western blot, and size-exclusion chromatography to determine their presence, purity, and homogeneity. Along with their expression data from insect cells, the most stable and homogeneous construct from each round was used to design successive optimizations. After 3 rounds of construct design for each receptor, much work remains to produce a stable sample that has the potential to crystallize. Future work includes further optimization of the insertion site of the fusion proteins, ligand screening for co-crystallization, optimization of purification conditions, and screening of potential thermostabilizing point mutations. Success in solving a structure will allow for a more detailed understanding of the receptor function in addition to its vital use in rational drug discovery.
ContributorsCosgrove, Steven Andrew (Author) / Liu, Wei (Thesis director) / Mills, Jeremy (Committee member) / Mazor, Yuval (Committee member) / W. P. Carey School of Business (Contributor) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2016-12
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Description
The purpose of the study was to examine the associations of protective (ethic identity, parent-child closeness) and risk (perceived discrimination, parent-child role reversal) factors with mental and behavioral health in 2nd generation Cambodian American (CA) young adults. A total of 66 participants who identified as being 2nd generation CA young

The purpose of the study was to examine the associations of protective (ethic identity, parent-child closeness) and risk (perceived discrimination, parent-child role reversal) factors with mental and behavioral health in 2nd generation Cambodian American (CA) young adults. A total of 66 participants who identified as being 2nd generation CA young adults aged 18-25 years old were recruited to participate in this cross-sectional. Reliable and valid measures were used to assess protective and risk factors and mental (depressive, anxiety, somatic symptoms) and behavioral health outcomes (alcohol and drug use). We used descriptive statistics to describe sample characteristics and study variables and conducted multiple regression analysis to examine the associations of factors with each of the 5 health outcomes. The findings suggested that peer discrimination was positively and significantly associated with depressive (β = 0.42, p = 0.023; R2 = 0.397) and somatic symptoms (β = 0.63, p = 0.000, R2 = 0.554). Father role-reversal was also found to be negatively and significantly associated with predicting CA young adults’ anxiety symptoms (β = -0.32, p = 0.005, R2 = 0.456).

Majority of the CA young adults have perceived racial/ethnic discrimination in the community. Furthermore, perceived discrimination has been positively associated with their depressive and somatic symptoms, suggesting a need to address racial/ethnic discrimination issues to promote positive mental health in this population. It is important for school/work personnel and healthcare providers to assess CA young adults’ discrimination experiences, and have the sufficient resources (e.g., education, support groups) to prevent negative consequences associated with discrimination.
ContributorsOu, Jason Heng (Author) / Chen, Angela Chia-Chia (Thesis director) / Sangalang, Cindy (Committee member) / Miroballi, Barbara (Committee member) / School of Molecular Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2015-12
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Description

My research aims to determine the effectiveness of meditation and sleep applications (apps) on the reduction of anxiety and stress in college students, with a focus on sedative piano music. Results showed a significant reduction of stress and anxiety levels in college students when listening to sedative piano music versus

My research aims to determine the effectiveness of meditation and sleep applications (apps) on the reduction of anxiety and stress in college students, with a focus on sedative piano music. Results showed a significant reduction of stress and anxiety levels in college students when listening to sedative piano music versus non-sedative piano music. Music along with other therapy modalities in meditation and sleep apps show promise in reducing students’ anxiety and stress and promoting their successes.

ContributorsPantha, Bidur (Author) / Brian, Jennifer (Thesis director) / Patten, Kristopher (Committee member) / School of Molecular Sciences (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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Description

Health and Wealthness is a podcast where your hosts, Emily Weigel and Hanaa Khan discuss pressing and trending topics about health and wealth that everyone should know about. Our first four episodes focus on the opioid crisis. Both the science and healthcare sides. We then go on to talk about

Health and Wealthness is a podcast where your hosts, Emily Weigel and Hanaa Khan discuss pressing and trending topics about health and wealth that everyone should know about. Our first four episodes focus on the opioid crisis. Both the science and healthcare sides. We then go on to talk about burnout and mental health in a conversational episode.

ContributorsKhan, Hanaa S (Co-author) / Weigel, Emily (Co-author) / Olive, Foster (Thesis director) / Bonfiglio, Thomas (Committee member) / Department of Psychology (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2021-05
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There is an increase in the prevalence of mental health problems in the United States. Healthy People 2020’s leading mental health indicator is to increase the delivery of care to those with mental health issues and lower the number of youth who experience a major depressive disorder. Teachers and non-teaching

There is an increase in the prevalence of mental health problems in the United States. Healthy People 2020’s leading mental health indicator is to increase the delivery of care to those with mental health issues and lower the number of youth who experience a major depressive disorder. Teachers and non-teaching staff are well placed in the community to identify youth undergoing emotional distress and facilitate early interventions, yet do not receive adequate training in mental health.

A project was undertaken to determine if a mental health training intervention affected the community youth mentors knowledge, attitude and self-efficacy towards helping youth with mental health issues. Three instruments with good validity and reliability namely Mental Health Literacy Scale (MHLS), Attitudes to Severe Mental Illness (ASMI) scale, and Gatekeeper Behavior Scale were used in pre intervention, immediately post intervention and two weeks post intervention questionnaires. The Wilcoxon Signed Ranks test indicated changes in the pre and post intervention scores as significant in knowledge, and attitude between pre intervention and immediately post intervention time periods. Cohen’s effect size value suggested large, medium, small, and minimum clinical significance in the variables over period of time.

Mental health literacy narrows the gap between symptom onset and intervention. Numerous mental health trainings are currently available worldwide. Schools and after school clubs in collaboration with hospital mental health and other community agencies are better equipped to bridge the gap. School staff report better confidence in addressing mental health and behavioral health issues among youth when equipped with additional resources within the school in the form of psychologists, social workers, and counselors.

ContributorsMazhuvencherry, Elizabeth (Author) / Chen, Angela (Thesis advisor)
Created2020-05-03