Matching Items (189)
Description
Effectively modeling Alzheimer’s disease will lend to a more comprehensive
understanding of the disease pathology, more efficacious drug development and
regenerative medicine as a form of treatment. There are limitations with current
transgenic mouse models of Alzheimer’s disease and the study of post mortem brain tissue of Alzheimer’s diseases patients. Stem cell models can overcome the lack of clinical relevance and impracticality associated with current models. Ideally, the use of stem cell models provides the foundation to study the biochemical and physiological aspects of Alzheimer’s disease, but at the cellular level. Moreover, the future of drug development and disease modeling can be improved by developing a reproducible and well-characterized model of AD that can be scaled up to meet requirements for basic and translational applications. Characterization and analysis of a heterogenic neuronal culture developed from induced pluripotent stem cells calls for the understanding of single cell identity and cell viability. A method to analyze RNA following intracellular sorting was developed in order to analyze single cell identity of a heterogenic population
of human induced pluripotent stem cells and neural progenitor cells. The population was intracellularly stained and sorted for Oct4. RNA was isolated and analyzed with qPCR, which demonstrated expected expression profiles for Oct4+ and Oct4- cells. In addition, a protocol to label cells with pO2 sensing nanoprobes was developed to assess cell viability. Non-destructive nanoprobe up-take by neural progenitor cells was assessed with fluorescent imaging and flow cytometry. Nanoprobe labeled neurons were cultured long-term and continued to fluoresce at day 28. The proof of concept experiments demonstrated will be further expanded upon and utilized in developing a more clinically relevant and cost-effective model of Alzheimer’s disease with downstream applications
in drug development and regenerative medicine.
understanding of the disease pathology, more efficacious drug development and
regenerative medicine as a form of treatment. There are limitations with current
transgenic mouse models of Alzheimer’s disease and the study of post mortem brain tissue of Alzheimer’s diseases patients. Stem cell models can overcome the lack of clinical relevance and impracticality associated with current models. Ideally, the use of stem cell models provides the foundation to study the biochemical and physiological aspects of Alzheimer’s disease, but at the cellular level. Moreover, the future of drug development and disease modeling can be improved by developing a reproducible and well-characterized model of AD that can be scaled up to meet requirements for basic and translational applications. Characterization and analysis of a heterogenic neuronal culture developed from induced pluripotent stem cells calls for the understanding of single cell identity and cell viability. A method to analyze RNA following intracellular sorting was developed in order to analyze single cell identity of a heterogenic population
of human induced pluripotent stem cells and neural progenitor cells. The population was intracellularly stained and sorted for Oct4. RNA was isolated and analyzed with qPCR, which demonstrated expected expression profiles for Oct4+ and Oct4- cells. In addition, a protocol to label cells with pO2 sensing nanoprobes was developed to assess cell viability. Non-destructive nanoprobe up-take by neural progenitor cells was assessed with fluorescent imaging and flow cytometry. Nanoprobe labeled neurons were cultured long-term and continued to fluoresce at day 28. The proof of concept experiments demonstrated will be further expanded upon and utilized in developing a more clinically relevant and cost-effective model of Alzheimer’s disease with downstream applications
in drug development and regenerative medicine.
ContributorsKnittel, Jacob James (Author) / Brafman, David (Thesis director) / Salvatore, Oddo (Committee member) / School of Life Sciences (Contributor) / Dean, W.P. Carey School of Business (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Cell fate is a complex and dynamic process with many genetic components. It has often been likened to “multistable” mathematical systems because of the numerous possible “stable” states, or cell types, that cells may end up in. Due to its complexity, understanding the process of cell fate and differentiation has proven challenging. A better understanding of cell differentiation has applications in regenerative stem cell therapies, disease pathologies, and gene regulatory networks.
A variety of different genes have been associated with cell fate. For example, the Nanog/Oct-4/Sox2 network forms the core interaction of a gene network that maintains stem cell pluripotency, and Oct-4 and Sox2 also play a role in the tissue types that stem cells eventually differentiate into. Using the CRISPR/cas9 based homology independent targeted integration (HITI) method developed by Suzuki et al., we can integrate fluorescent tags behind genes with reasonable efficiency via the non-homologous end joining (NHEJ) DNA repair pathway. With human embryonic kidney (HEK) 293T cells, which can be transfected with high efficiencies, we aim to create a three-parameter reporter cell line with fluorescent tags for three different genes related to cell fate. This cell line would provide several advantages for the study of cell fate, including the ability to quantitatively measure cell state, observe expression heterogeneity among a population of genetically identical cells, and easily monitor fluctuations in expression patterns.
The project is partially complete at this time. This report discusses progress thus far, as well as the challenges faced and the future steps for completing the reporter line.
A variety of different genes have been associated with cell fate. For example, the Nanog/Oct-4/Sox2 network forms the core interaction of a gene network that maintains stem cell pluripotency, and Oct-4 and Sox2 also play a role in the tissue types that stem cells eventually differentiate into. Using the CRISPR/cas9 based homology independent targeted integration (HITI) method developed by Suzuki et al., we can integrate fluorescent tags behind genes with reasonable efficiency via the non-homologous end joining (NHEJ) DNA repair pathway. With human embryonic kidney (HEK) 293T cells, which can be transfected with high efficiencies, we aim to create a three-parameter reporter cell line with fluorescent tags for three different genes related to cell fate. This cell line would provide several advantages for the study of cell fate, including the ability to quantitatively measure cell state, observe expression heterogeneity among a population of genetically identical cells, and easily monitor fluctuations in expression patterns.
The project is partially complete at this time. This report discusses progress thus far, as well as the challenges faced and the future steps for completing the reporter line.
ContributorsLoveday, Tristan Andre (Author) / Wang, Xiao (Thesis director) / Brafman, David (Committee member) / Harrington Bioengineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
Chronic Traumatic Encephalopathy (CTE) is a neurodegenerative brain disease that results from repetitive brain trauma causing brain structure, personality, behavioral, and cognitive changes. CTE is currently undiagnosable and untreatable in living patients. This thesis investigates research surrounding CTE and presents a comparative discussion of the advantages and disadvantages of current diagnostic methods used for other neurodegenerative diseases that may be useful for the diagnosis of CTE.
ContributorsBlair, Sierra (Co-author) / Blair, Taylor (Co-author) / Brafman, David (Thesis director) / Stabenfeldt, Sarah (Committee member) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2019-05
Description
The importance of second language learning in today’s ever-increasing globalized world is becoming ever more paramount. Despite seeming trends which indicate aversion to globalization, the international phenomenon which describes the connection made between peoples and cultures, will only increase its influence in coming years. With the advances globalization has made, it is becoming more important to learn and study foreign languages in order to keep abreast of this trend and not be left behind by globalization. Why electronic translation is not so viable in the long run (at least currently) is that culture and syntax are not things which can be simply ascertained via mediums such as application use. Due to the fact that advanced language proficiency is considered to be an integral piece towards stronger sentiments of “integration” (i.e. Syrian refugees integrating into EU and US) it is of more importance that increasing second language proficiency receives the adequate study and implementation to reflect a more cohesive globalized world. Accompanying this necessity is the simple fact that adult second language learners often struggle to overcome difficulty to the challenging yet rewarding task of learning and eventually mastering a second language.
To truly understand the difficulty some adult second language learners have with learning a second language it can be helpful to compare second language acquisition to how one naturally, and seemingly effortlessly in many cases, acquires their native language. How can a comparative analysis of how native speakers and adult second language learners each learn their first and second language respectively be successfully converted to a specific means of assisting adult second language learners achieve the highest level of possible fluency? In order to more accurately propose a viable solution to the overarching question, the following three questions have been proposed as a means to springboard into better understanding the nature of the main topic.
The points to consider while analyzing the main question throughout this analysis are as follows: How would it be best for an adult second language learner to achieve the same level off proficiency as a native speaker of a given language whom has been exposed to all of its intricacies since birth? At what point exactly is someone considered to have the same level of proficiency which a native speaker of a given language would have and how does that differ from being a heritage speaker? With the final supporting question being: What type of learning would be best suited in helping a heritage speaker (someone who learns a language in the home by virtue of their heritage) or adult second language learner to become highly proficient in a second language?
In order to propose a wide variety of integration between these questions with the conjoined purpose of answering the inquiry of this thesis, many different sources supporting each of the above questions will contain certain overlap, providing a clear basis for constructing a tri-fold argument in answering the thesis question as acutely as possible. In regards to the first question, the question of “proficiency” will be a subsection committed to understanding the nature of how language proficiency works and at what point (if ever) one can ever be considered “highly proficient” in a second language.
All three exploratory questions are compatible with a theory known as Critical Period Theory, a theory in linguistics proposed by Montreal neurologist Wilder Penfield, which states, “There is a critical age, before puberty, that one must learn language. If one has not learned to speak before puberty it is much more difficult, and sometimes impossible, to learn language and speak in a meaningful way.” The overlap which this method binds to Universal Grammar is a rather close-knit relationship. Research composed by certain linguists suggest that “children are born with a certain universal grammar wired into their brains.” This will be compared and cross-examined to a higher degree in a later section of the paper.
The importance of Universal Grammar in relation to Critical Period Theory cannot be overstated. Universal Grammar in relation to the second language Critical Period Theory will help explain at which point, someone is considered to be a “native speaker” of a given language. The third question posed of how would it be best for heritage and second language learners to increase their proficiency in a second language really touches on both of these theories in regards to at which age someone is exposed to a specific language in addition to how Universal Grammar affects the development of second language acquisition. In the realm of perpetually working towards mitigating an answer to this analysis’s thesis, the connecting thread or “roter Faden”, as it is said in German, will be the integrative domain the above questions will have on arriving to a clearer understanding of the nature of how a comparative analysis of second language learners and mother-tongue speakers can expedite the language learning process of second language learners, using techniques of native speakers which they inherently pick-up.
To truly understand the difficulty some adult second language learners have with learning a second language it can be helpful to compare second language acquisition to how one naturally, and seemingly effortlessly in many cases, acquires their native language. How can a comparative analysis of how native speakers and adult second language learners each learn their first and second language respectively be successfully converted to a specific means of assisting adult second language learners achieve the highest level of possible fluency? In order to more accurately propose a viable solution to the overarching question, the following three questions have been proposed as a means to springboard into better understanding the nature of the main topic.
The points to consider while analyzing the main question throughout this analysis are as follows: How would it be best for an adult second language learner to achieve the same level off proficiency as a native speaker of a given language whom has been exposed to all of its intricacies since birth? At what point exactly is someone considered to have the same level of proficiency which a native speaker of a given language would have and how does that differ from being a heritage speaker? With the final supporting question being: What type of learning would be best suited in helping a heritage speaker (someone who learns a language in the home by virtue of their heritage) or adult second language learner to become highly proficient in a second language?
In order to propose a wide variety of integration between these questions with the conjoined purpose of answering the inquiry of this thesis, many different sources supporting each of the above questions will contain certain overlap, providing a clear basis for constructing a tri-fold argument in answering the thesis question as acutely as possible. In regards to the first question, the question of “proficiency” will be a subsection committed to understanding the nature of how language proficiency works and at what point (if ever) one can ever be considered “highly proficient” in a second language.
All three exploratory questions are compatible with a theory known as Critical Period Theory, a theory in linguistics proposed by Montreal neurologist Wilder Penfield, which states, “There is a critical age, before puberty, that one must learn language. If one has not learned to speak before puberty it is much more difficult, and sometimes impossible, to learn language and speak in a meaningful way.” The overlap which this method binds to Universal Grammar is a rather close-knit relationship. Research composed by certain linguists suggest that “children are born with a certain universal grammar wired into their brains.” This will be compared and cross-examined to a higher degree in a later section of the paper.
The importance of Universal Grammar in relation to Critical Period Theory cannot be overstated. Universal Grammar in relation to the second language Critical Period Theory will help explain at which point, someone is considered to be a “native speaker” of a given language. The third question posed of how would it be best for heritage and second language learners to increase their proficiency in a second language really touches on both of these theories in regards to at which age someone is exposed to a specific language in addition to how Universal Grammar affects the development of second language acquisition. In the realm of perpetually working towards mitigating an answer to this analysis’s thesis, the connecting thread or “roter Faden”, as it is said in German, will be the integrative domain the above questions will have on arriving to a clearer understanding of the nature of how a comparative analysis of second language learners and mother-tongue speakers can expedite the language learning process of second language learners, using techniques of native speakers which they inherently pick-up.
ContributorsRoselius, Thomas Jacob (Author) / Lee, Sara (Thesis director) / Reves, Christiane (Committee member) / College of Public Service and Community Solutions (Contributor) / College of Integrative Sciences and Arts (Contributor) / School of International Letters and Cultures (Contributor) / Barrett, The Honors College (Contributor)
Created2018-12
Description
Alzheimer’s Disease (AD) affects over 5 million individuals in the U.S. and has a direct cost estimated in excess of $200 billion per year. Broadly speaking, there are two forms of AD—early-onset, familial AD (FAD) and late-onset-sporadic AD (SAD). Animal models of AD, which rely on the overexpression of FAD-related mutations, have provided important insights into the disease. However, these models do not display important disease-related pathologies and have been limited in their ability to model the complex genetics associated with SAD.
Advances in cellular reprogramming, have enabled the generation of in vitro disease models that can be used to dissect disease mechanisms and evaluate potential therapeutics. To that end, efforts by many groups, including the Brafman laboratory, to generated patient-specific hiPSCs have demonstrated the promise of studying AD in a simplified and accessible system. However, neurons generated from these hiPSCs have shown some, but not all, of the early molecular and cellular hallmarks associated with the disease. Additionally, phenotypes and pathological hallmarks associated with later stages of the human disease have not been observed with current hiPSC-based systems. Further, disease relevant phenotypes in neurons generated from SAD hiPSCs have been highly variable or largely absent. Finally, the reprogramming process erases phenotypes associated with cellular aging and, as a result, iPSC-derived neurons more closely resemble fetal brain rather than adult brain.
It is well-established that in vivo cells reside within a complex 3-D microenvironment that plays a significant role in regulating cell behavior. Signaling and other cellular functions, such as gene expression and differentiation potential, differ in 3-D cultures compared with 2-D substrates. Nonetheless, previous studies using AD hiPSCs have relied on 2-D neuronal culture models that do not reflect the 3-D complexity of native brain tissue, and therefore, are unable to replicate all aspects of AD pathogenesis. Further, the reprogramming process erases cellular aging phenotypes. To address these limitations, this project aimed to develop bioengineering methods for the generation of 3-D organoid-based cultures that mimic in vivo cortical tissue, and to generate an inducible gene repression system to recapitulate cellular aging hallmarks.
Advances in cellular reprogramming, have enabled the generation of in vitro disease models that can be used to dissect disease mechanisms and evaluate potential therapeutics. To that end, efforts by many groups, including the Brafman laboratory, to generated patient-specific hiPSCs have demonstrated the promise of studying AD in a simplified and accessible system. However, neurons generated from these hiPSCs have shown some, but not all, of the early molecular and cellular hallmarks associated with the disease. Additionally, phenotypes and pathological hallmarks associated with later stages of the human disease have not been observed with current hiPSC-based systems. Further, disease relevant phenotypes in neurons generated from SAD hiPSCs have been highly variable or largely absent. Finally, the reprogramming process erases phenotypes associated with cellular aging and, as a result, iPSC-derived neurons more closely resemble fetal brain rather than adult brain.
It is well-established that in vivo cells reside within a complex 3-D microenvironment that plays a significant role in regulating cell behavior. Signaling and other cellular functions, such as gene expression and differentiation potential, differ in 3-D cultures compared with 2-D substrates. Nonetheless, previous studies using AD hiPSCs have relied on 2-D neuronal culture models that do not reflect the 3-D complexity of native brain tissue, and therefore, are unable to replicate all aspects of AD pathogenesis. Further, the reprogramming process erases cellular aging phenotypes. To address these limitations, this project aimed to develop bioengineering methods for the generation of 3-D organoid-based cultures that mimic in vivo cortical tissue, and to generate an inducible gene repression system to recapitulate cellular aging hallmarks.
ContributorsBounds, Lexi Rose (Author) / Brafman, David (Thesis director) / Wang, Xiao (Committee member) / Harrington Bioengineering Program (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
This study examined the type and frequency of questions asked by attorneys in cases of children alleging sexual abuse. Of interest was whether child age would affect the questions asked. The participants included 25 child witnesses testifying in criminal trials in Maricopa County over a recent ten-year period. Children were placed into two groups: younger (five to seven-year-olds) and older (eight to nine-year-olds). Every question asked, and answer provided, during children's testimony, was systematically and reliably coded for the content of the interaction. Attorneys exhibited developmental sensitivity, varying the amount of question they asked across content areas by the age of the child. In addition, attorneys varied in what they asked about: the prosecution focused more on the plausibility of abuse, whereas the defense focused more on how others may have suggestively influenced the child's report. Both attorneys were equally concerned about the consistency of narratives. The findings from the present study have direct policy implications for how attorneys structure their arguments, both in an attempt to establish, and question, children's credibility in these important cases. Keywords: children, age, suggestibility, consistency, inconsistency, plausibility
ContributorsHogan, Caitlyn Rose (Author) / Stolzenberg, Stacia (Thesis director) / Fradella, Hank (Committee member) / College of Public Service and Community Solutions (Contributor) / School of Criminology and Criminal Justice (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
Given the prominence of alcohol in the Korean adult's daily life, further investigation into the negative effects of harmful alcohol consumption and the treatment of and culture surrounding alcohol use disorders is needed. An investigation into the status and treatment of alcohol use disorders in South Korea was chosen due to the significant cultural differences from the United States with regards to alcohol consumption, mental health, and healthcare. The investigation used academic and grey literature, news reports, and current healthcare and public health policy to evaluate South Korea's weak areas in addressing alcohol use disorders. Greater investment into mental health research and healthcare delivery, as well as further development of the continuum of care to help patients transition from treatment of the acute symptoms of an alcohol use disorder to lifetime management are needed. Lastly, increased alcohol regulatory policies are recommended.
ContributorsFlowers, Taylor Nicole (Author) / Gaughan, Monica (Thesis director) / Shin, Hoon Cheol (Committee member) / School of International Letters and Cultures (Contributor) / School of Human Evolution and Social Change (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05
Description
Throughout history, terrorism has had major effects on tourists and tourism destinations that are targeted. When terrorists execute an attack in order to communicate a message, resulting impacts go far beyond lives lost. Included in these impacts is the fear that terror attacks leave on tourists, sometimes creating such an impression that tourists change travel plans as a result. Although there are many studies on destination image, risk perception, and decision-making, very few focus specifically on the millennial generation. Because millennials are changing the travel and tourism scene - as they now make up the largest percentage of the population \u2014 it is important for stakeholders in the tourism industry and DMOs to understand this shift and the implications that come with these changes. This study provides a qualitative analysis of millennials' views, attitudes, and beliefs regarding terror impacted tourist destinations. Specifically, it will explore how millennials develop their destination images, if millennials view international travel to be risky, and if millennials are willing to travel to terror impacted tourist destinations. Using focus group methodology, this study gathered data from students from a number of disciplines, genders, and travel experience, in order to understand the themes listed above. Analysis of the findings and implications for officials and DMOs follow, as well as a conclusion including limitations and recommendations for future studies.
ContributorsBacon, Laura Elizabeth (Author) / Buzinde, Christine (Thesis director) / Nyaupane, Gyan (Committee member) / Department of Management and Entrepreneurship (Contributor) / W. P. Carey School of Business (Contributor) / College of Public Service and Community Solutions (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05
Description
Genetic manipulation of human cell lines has widespread applications in biomedical research ranging from disease modeling to therapeutic development. Human cells are generally difficult to genetically engineer, but exogenous nucleic acids can be expressed by viral, chemical, or nonchemical means. Chemical transfections are simpler in practice than both viral and nonchemical delivery of genetic material, but often suffer from cytotoxicity and low efficiency. Novel aminoglycoside antibiotic-derived lipopolymers have been synthesized to mediate transgene delivery to human cells. These polymers are comprised of either paromomycin or apramycin crosslinked with glycerol diglycidylether and derivatized with stearoyl chloride in varying molar ratios. In this work, three previously identified target lipopolymers were screened against a library of human embryonic and induced pluripotent stem cell lines. Cells were transfected with a plasmid encoding green fluorescent protein (GFP) and expression was quantified with flow cytometry 48 hours after transfection. Transfection efficiency was evaluated between three distinct lipopolymers and four lipopolymer:DNA mass ratios. GFP expression was compared to that of cells transfected with commercially available chemical gene delivery reagent controls\u2014JetPEI, Lipofectamine, and Fugene\u2014at their recommended reagent:DNA ratios. Improved transgene expression in stem cell lines allows for improved research methods. Human stem cell-derived neurons that have been genetically manipulated to express phenotypic characteristics of aging can be utilized to model neurodegenerative diseases, elucidating information about these diseases that would be inaccessible in unmanipulated tissue.
ContributorsMehta, Frea (Author) / Brafman, David (Thesis director) / Rege, Kaushal (Committee member) / Chemical Engineering Program (Contributor) / School of Life Sciences (Contributor) / Barrett, The Honors College (Contributor)
Created2017-05
Description
This study examines the The Disney College Program, a semester-long paid internship hosted by the Walt Disney Company employing more than 10,000 students each year. With over 120,000 alumni in the past 10 years, this program offers students housing and community building opportunities within the "Living" component, college credit courses within the "Learning" component, and on-the-job experience at Disneyland Resort and Walt Disney World theme parks through the "Earning" component. Specifically, the research focuses on Walt Disney World in Orlando, Florida. The researcher conducted a 39-question online survey prompting 1,749 responses from Disney College Program alumni to help answer the following research questions: (1) Who are Disney College Program Cast Members, (2) What is their level of satisfaction with the program, and (3) Are they influencers? This study uses theoretical elements (e.g. levels of adoption, influencers and brand loyalty) to describe influence and psychological effects to describe satisfaction (e.g. indoctrination, human motivation and Stockholm Syndrome). With the findings showing discrepancies between the ratings of "Living," "Learning," and "Earning" and the average overall rating, some questions arise about the program's tendencies to form tightly cohesive groups approaching elements of Stockholm Syndrome and cult-like ethos. Focusing on the 1,490 of 1,749 respondents from Walt Disney World in the past 10 years, the study concludes that Walt Disney World College Program alumni are not influencers nor advocates, but rather evangelists (i.e., zealous advocate) and loyalists.
ContributorsMcgowan, Sloane Shea (Author) / Matera, Fran (Thesis director) / Archer, Ciara (Committee member) / School of Community Resources and Development (Contributor) / Walter Cronkite School of Journalism and Mass Communication (Contributor) / College of Public Service and Community Solutions (Contributor) / Barrett, The Honors College (Contributor)
Created2018-05