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Description
Coccidioidomycosis, also known as Valley Fever, is a disease caused by the dimorphic soil-dwelling fungus, Coccidioides sp. Coccidioidomycosis is difficult to diagnose because symptoms are similar to community-acquired pneumonia. Current diagnostic tests rely on antibody responses, but immune responses can be delayed and aberrant, resulting in false negative diagnoses. Unlike serology, detection of coccidioidal proteins or other fungal components in blood could distinguish valley fever from other pulmonary infections and provide a definitive diagnosis. Using mass spectrometry (LC-MS/MS) we examined the plasma peptidome from patients with serologically confirmed coccidioidomycosis. Mass spectra were searched using the protein database from the Coccidioides species, generated and annotated by the Broad Institute. 15 of 20 patients with serologically confirmed coccidioidomycosis demonstrated the presence of a peptide in plasma, "PGLDSKSLACTFSQV" (PGLD). The peptide is derived from an open reading frame from a "conserved hypothetical protein" annotated with 2 exons, and to date, found only in the C. posadasii strain Silviera RMSCC 3488 genomic sequence. In this thesis work, cDNA sequence analysis from polyadenylated RNA confirms the peptide sequence and genomic location of the peptide, but does not indicate that the intron in the gene prediction of C. posadasii strain Silviera RMSCC 3488 is present. A monoclonal antibody generated against the peptide bound to a 16kDa protein in T27K coccidioidal lysate. Detecting components of the fungus plasma could be a useful diagnostic tool, especially when serology does not provide a definitive diagnosis.
ContributorsDuffy, Stacy Leigh (Author) / Lake, Douglas (Thesis advisor) / Magee, Dewey Mitch (Committee member) / Antwi, Kwasi (Committee member) / Arizona State University (Publisher)
Created2013
Description
Since Darwin popularized the evolution theory in 1895, it has been completed and studied through the years. Starting in 1990s, evolution at molecular level has been used to discover functional molecules while studying the origin of functional molecules in nature by mimicing the natural selection process in laboratory. Along this line, my Ph.D. dissertation focuses on the in vitro selection of two important biomolecules, deoxynucleotide acid (DNA) and protein with binding properties. Chapter two focuses on in vitro selection of DNA. Aptamers are single-stranded nucleic acids that generated from a random pool and fold into stable three-dimensional structures with ligand binding sites that are complementary in shape and charge to a desired target. While aptamers have been selected to bind a wide range of targets, it is generally thought that these molecules are incapable of discriminating strongly alkaline proteins due to the attractive forces that govern oppositely charged polymers. By employing negative selection step to eliminate aptamers that bind with off-target through charge unselectively, an aptamer that binds with histone H4 protein with high specificity (>100 fold)was generated. Chapter four focuses on another functional molecule: protein. It is long believed that complex molecules with different function originated from simple progenitor proteins, but very little is known about this process. By employing a previously selected protein that binds and catalyzes ATP, which is the first and only protein that was evolved completely from random pool and has a unique α/β-fold protein scaffold, I fused random library to the C-terminus of this protein and evolved a multi-domain protein with decent properties. Also, in chapter 3, a unique bivalent molecule was generated by conjugating peptides that bind different sites on the protein with nucleic acids. By using the ligand interactions by nucleotide conjugates technique, off-the shelf peptide was transferred into high affinity protein capture reagents that mimic the recognition properties of natural antibodies. The designer synthetic antibody amplifies the binding affinity of the individual peptides by ∼1000-fold to bind Grb2 with a Kd of 2 nM, and functions with high selectivity in conventional pull-down assays from HeLa cell lysates.
ContributorsJiang, Bing (Author) / Chaput, John C (Thesis advisor) / Chen, Julian (Committee member) / Liu, Yan (Committee member) / Arizona State University (Publisher)
Created2013
Description
Unauthorized immigrants account for approximately one fourth of all immigrants in the United States, yet they dominate public perceptions and are at the heart of a policy impasse. Caught in the middle are the children of these immigrants--youth who are coming of age and living in the shadows; they are an invisible cohort. An estimated 5.5 million children and adolescents are growing up with unauthorized immigrant parents, and are experiencing multiple, and yet unrecognized developmental consequences of their families' existence in the shadow of the law. Although these youth are American in spirit and voice, they are, nonetheless, members of families that are "illegal" in the eyes of the law. Many children have been exiled to México; these are the children living in the shadows of Mexican diaspora, Los Retornos. This phenomenological study developed a conceptual framework to examine the effects in which being an exiled United States citizen living in Morelia, Michoacán, affected these many children and adolescents. Bourdieu's (1977) theoretical framework is used in this study and is based on his social, cultural capital concept; the assumption is that Los Retornos carry valuable sociocultural, bilingual and monoliterate capital that is endangered, unrecognized, replaceable, and not used to the best interest of students in schools. This study made use of this framework to answer the following questions: 1. How do Retorno families (nuclear and extended) develop the self-efficacy needed to preserve the social and cultural capital they bring with them to Michoacán? 2. How are communities and identity forms imagined and created in the context of this new migration shift? 3. How are Los Retornos responding to the involuntary shift (N=7) from the U.S to Michoacán? 4. How are teachers adjusting their classroom practices and curriculum to meet the academic needs of Los Retornos? The purpose of this qualitative phenomenological study is to improve understanding of Los Retornos. This phenomenological case study is focused on identifying experiences Los Retornos encounter in their schools and family lives through their personal migration experience to illuminate how best to help them preserve the social and cultural, capital they bring with them. The findings from this study may assist educators and policy makers in developing interventions and policies that meet the needs of this cohort.
ContributorsQuezada Sanders, Irene Genevieve (Author) / Ovando, Carlos J. (Thesis advisor) / Mccarty, Teresa L. (Committee member) / De Los Santos Jr., Alfredo G. (Committee member) / Arizona State University (Publisher)
Created2013
Description
Speciation is the fundamental process that has generated the vast diversity of life on earth. The hallmark of speciation is the evolution of barriers to gene flow. These barriers may reduce gene flow either by keeping incipient species from hybridizing at all (pre-zygotic), or by reducing the fitness of hybrids (post-zygotic). To understand the genetic architecture of these barriers and how they evolve, I studied a genus of wasps that exhibits barriers to gene flow that act both pre- and post-zygotically. Nasonia is a genus of four species of parasitoid wasps that can be hybridized in the laboratory. When two of these species, N. vitripennis and N. giraulti are mated, their offspring suffer, depending on the generation and cross examined, up to 80% mortality during larval development due to incompatible genic interactions between their nuclear and mitochondrial genomes. These species also exhibit pre-zygotic isolation, meaning they are more likely to mate with their own species when given the choice. I examined these two species and their hybrids to determine the genetic and physiological bases of both speciation mechanisms and to understand the evolutionary forces leading to them. I present results that indicate that the oxidative phosphorylation (OXPHOS) pathway, an essential pathway that is responsible for mitochondrial energy generation, is impaired in hybrids of these two species. These results indicate that this impairment is due to the unique evolutionary dynamics of the combined nuclear and mitochondrial origin of this pathway. I also present results showing that, as larvae, these hybrids experience retarded growth linked to the previously observed mortality and I explore possible physiological mechanisms for this. Finally, I show that the pre-mating isolation is due to a change in a single pheromone component in N. vitripennis males, that this change is under simple genetic control, and that it evolved neutrally before being co-opted as a species recognition signal. These results are an important addition to our overall understanding of the mechanisms of speciation and showcase Nasonia as an emerging model for the study of the genetics of speciation.
ContributorsGibson, Joshua D (Author) / Gadau, Jürgen (Thesis advisor) / Harrison, Jon (Committee member) / Pratt, Stephen (Committee member) / Verrelli, Brian (Committee member) / Willis, Wayne (Committee member) / Arizona State University (Publisher)
Created2013
Description
Examining the elements of the hidden curriculum in theatre education allows theatre educators the opportunity to reflect on their own pedagogy and its effects on the learner. The hidden curriculum refers to the unspoken or implicit values, norms, and beliefs that are transmitted through tacit messages. When the hidden curriculum remains veiled, the impact on the learner's education and socialization process can perpetuate gender, race, and class inequalities. In order to understand how the hidden curriculum manifests itself in theatre classrooms, we have to look at schools as "agents of legitimation, organized to produce and reproduce the dominant categories, values, and social relationships necessary for the maintenance of the larger society" (Giroux, 1983, p. 72). This qualitative study examined the hidden curriculum in theatre at the secondary level and looked at theatre teachers' pedagogy in reproducing elements of the hidden curriculum. Interviews, naturalistic observation, and a researcher reflective journal were employed in the data collection process to better understand: a) the elements of hidden curriculum that appear in theatre education at the secondary level, b) how the pedagogical practices of theatre teachers support societal structures, and c) how the hidden curriculum in theatre reinforces gender, race, and social class distinctions. Data were then coded and analyzed to find emergent themes. Multiple theoretical perspectives serve as a conceptual framework for understanding the hidden curriculum, and provide a neglected perspective of the hidden curriculum in theatre education. The theatre classroom provides a unique space to view hidden curriculum and can be viewed as a unique agent of social change. Themes related to the first research question emerged as: a) privileges for older students, b) school rules, c) respect for authority, d) acceptance of repetitive tasks, and c) punctuality. Themes related to the second research question emerged as: a) practices, b) procedures, c) rules, d) relationships, and e) structures. Finally, themes related to the third question emerged as: a) reinforcement of social inequality, b) perpetuation of class structure, and c) acceptance of social destiny. The discussion looks at the functions of theatre pedagogy in the reproduction of class, inequality, and institutionalized cultural norms.
ContributorsHines, Angela R (Author) / Saldana, Johnny (Thesis advisor) / Malewski, Erik (Committee member) / Fischman, Gustavo (Committee member) / Arizona State University (Publisher)
Created2013
Description
The principle of Darwinian evolution has been applied in the laboratory to nucleic acid molecules since 1990, and led to the emergence of in vitro evolution technique. The methodology of in vitro evolution surveys a large number of different molecules simultaneously for a pre-defined chemical property, and enrich for molecules with the particular property. DNA and RNA sequences with versatile functions have been identified by in vitro selection experiments, but many basic questions remain to be answered about how these molecules achieve their functions. This dissertation first focuses on addressing a fundamental question regarding the molecular recognition properties of in vitro selected DNA sequences, namely whether negatively charged DNA sequences can be evolved to bind alkaline proteins with high specificity. We showed that DNA binders could be made, through carefully designed stringent in vitro selection, to discriminate different alkaline proteins. The focus of this dissertation is then shifted to in vitro evolution of an artificial genetic polymer called threose nucleic acid (TNA). TNA has been considered a potential RNA progenitor during early evolution of life on Earth. However, further experimental evidence to support TNA as a primordial genetic material is lacking. In this dissertation we demonstrated the capacity of TNA to form stable tertiary structure with specific ligand binding property, which suggests a possible role of TNA as a pre-RNA genetic polymer. Additionally, we discussed the challenges in in vitro evolution for TNA enzymes and developed the necessary methodology for future TNA enzyme evolution.
ContributorsYu, Hanyang (Author) / Chaput, John C (Thesis advisor) / Chen, Julian (Committee member) / Yan, Hao (Committee member) / Arizona State University (Publisher)
Created2013
Description
Teacher attrition and the migration between schools and districts can have a negative impact on quality of education and teacher performance. Novice teachers leave the profession because they are overwhelmed by the workload and responsibilities of the job. In a previous action research cycle, I found that novice teachers' perceptions of isolation and lack of opportunities to share experiences had a negative effect on teacher perceptions of efficacy. This action research project examines the effect of leveraging social media and professional learning communities to provide opportunities for a group of novice teachers to share experiences and seek advice. By addressing the challenges that novice teachers face and providing solutions for common problems, it is the hope of this researcher that highly effective teachers will remain in the classroom. The results of the study indicate that the combined use of Twitter and YouTube in collaboration with professional learning communities will improve teacher perceptions of efficacy. Teachers who participated in the social media based professional learning communities are also more likely to remain in the classroom.
ContributorsBostick, Bradley Alan (Author) / Zambo, Ronald (Thesis advisor) / Heck, Thomas (Committee member) / Isai, Shelley (Committee member) / Arizona State University (Publisher)
Created2013
Description
Modified and artificial water sources can be used as a management tool for game and non-game wildlife species. State, federal, and private agencies allocate significant resources to install and maintain artificial water sources (AWS) annually. Capture mark recapture methods were used to sample small mammal communities in the vicinity of five AWS and five paired control sites (treatments) in the surrounding Sonoran desert from October 2011 to May 2012. I measured plant species richness, density, and percent cover in the spring of 2012. A Multi-response Permutation Procedure was used to identify differences in small mammal community abundance, biomass, and species richness by season and treatment. I used Principle Component Analysis to reduce 11 habitat characteristics to five habitat factors. I related rodent occurrence to habitat characteristics using multiple and logistic regression. A total of 370 individual mammals representing three genera and eight species of rodents were captured across 4800 trap nights. Desert pocket mouse (Chaetodipus penicillatus) was the most common species in both seasons and treatments. Whereas rodent community abundance, biomass, and richness were similar between seasons, community variables of AWS were greater than CS. Rodent diversity was similar between treatments. Desert pocket mouse abundance and biomass were twice as high at AWS when compared to controls. Biomass of white-throated woodrat (Neotoma albigula) was five times greater at AWS. Habitat characteristics were similar between treatments. Neither presence of water nor distance to water explained substantial habitat variation. Occurrence of rodent species was associated with habitat characteristics. Desert rodent communities are adapted for arid environments (i.e. Heteromyids) and are not dependent on "free water". Higher abundances of desert pocket mouse at AWS were most likely related to increased disturbance and debris and not the presence of water. The results of this study and previous studies suggest that more investigation is needed and that short term studies may not be able to detect interactions (if any) between AWS and desert small mammal communities.
ContributorsSwitalski, Aaron (Author) / Bateman, Heather L (Thesis advisor) / Miller, William (Committee member) / Alford, Eddie (Committee member) / Arizona State University (Publisher)
Created2013
Description
Cyanovirin-N (CV-N) is a naturally occurring lectin originally isolated from the cyanobacteria Nostoc ellipsosporum. This 11 kDa lectin is 101 amino acids long with two binding sites, one at each end of the protein. CV-N specifically binds to terminal Manα1-2Manα motifs on the branched, high mannose Man9 and Man8 glycosylations found on enveloped viruses including Ebola, Influenza, and HIV. wt-CVN has micromolar binding to soluble Manα1-2Manα and also inhibits HIV entry at low nanomolar concentrations. CV-N's high affinity and specificity for Manα1-2Manα makes it an excellent lectin to study for its glycan-specific properties. The long-term aim of this project is to make a variety of mutant CV-Ns to specifically bind other glycan targets. Such a set of lectins may be used as screening reagents to identify biomarkers and other glycan motifs of interest. As proof of concept, a T7 phage display library was constructed using P51G-m4-CVN genes mutated at positions 41, 44, 52, 53, 56, 74, and 76 in binding Domain B. Five CV-N mutants were selected from the library and expressed in BL21(DE3) E. coli. Two of the mutants, SSDGLQQ-P51Gm4-CVN and AAGRLSK-P51Gm4-CVN, were sufficiently stable for characterization and were examined by CD, Tm, ELISA, and glycan array. Both proteins have CD minima at approximately 213 nm, indicating largely β-sheet structure, and have Tm values greater than 40°C. ELISA against gp120 and RNase B demonstrate both proteins' ability to bind high mannose glycans. To more specifically determine the binding specificity of each protein, AAGRLSK-P51Gm4-CVN, SSDGLQQ-P51Gm4-CVN, wt-CVN, and P51G-m4-CVN were sent to the Consortium for Functional Glycomics (CFG) for glycan array analysis. AAGRLSK-P51Gm4-CVN, wt-CVN, and P51G-m4-CVN, have identical specificities for high mannose glycans containing terminal Manα1-2Manα. SSDGLQQ-P51Gm4-CVN binds to terminal GlcNAcα1-4Gal motifs and a subgroup of high mannose glycans bound by P51G-m4-CVN. SSDGLQQ-wt-CVN was produced to restore anti-HIV activity and has a high nanomolar EC50 value compared to wt-CVN's low nanomolar activity. Overall, these experiments show that CV-N Domain B can be mutated and retain specificity identical to wt-CVN or acquire new glycan specificities. This first generation information can be used to produce glycan-specific lectins for a variety of applications.
ContributorsRuben, Melissa (Author) / Ghirlanda, Giovanna (Thesis advisor) / Allen, James (Committee member) / Wachter, Rebekka (Committee member) / Arizona State University (Publisher)
Created2013
Description
The purpose of this study was to examine the attitudes and opinions of Navajo students toward the Navajo language and culture programs within the schools they were attending. Although in the final year of the No Child Left Behind, a majority of the 265 schools on and near the Navajo reservation have not been making Adequate Yearly Progress, a concern for the parents, teachers, administrators, school board members, and the Navajo Nation. The study entailed conducting a survey at five schools; three of which were not meeting the requirements of the No Child Left Behind. The purpose of the survey instrument (27 questions) administered to the students at the five schools was to examine their attitudes and opinions as to participating in Navajo language and culture programs, to determine if the programs assisted them in their academic achievements, and to examine whether these programs actually made a difference for schools in their Adequate Yearly Progress requirement Approximately 87% of 99 Navajo students, 55 boys and 58 girls, ages 9 through 14, Grades 3 through 8, who lived off the reservation in Flagstaff, Arizona and Gallup, New Mexico, and took the survey knew and spoke Navajo, but less fluently and not to a great extent. However, the students endorsed learning Navajo and strongly agreed that the Navajo language and culture should be part of the curriculum. Historically there have been schools such as the Rock Point Community School, Rough Rock Demonstration School, Borrego Pass Community School, and Ramah Community School that have been successful in their implementation of bilingual programs. The question presently facing Navajo educators is what type of programs would be successful within the context of the No Child Left Behind federal legislation. Can there be replications of successful Navajo language and culture programs into schools that are not making Adequate Yearly Progress?
ContributorsTsosie, David J (Author) / Spencer, Dee A. (Thesis advisor) / Appleton, Nicholas A. (Committee member) / Koerperich, Robbie (Committee member) / Arizona State University (Publisher)
Created2013