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Description
There is a critical need for the development of clean and efficient energy sources. Hydrogen is being explored as a viable alternative to fuels in current use, many of which have limited availability and detrimental byproducts. Biological photo-production of H2 could provide a potential energy source directly manufactured from water and sunlight. As a part of the photosynthetic electron transport chain (PETC) of the green algae Chlamydomonas reinhardtii, water is split via Photosystem II (PSII) and the electrons flow through a series of electron transfer cofactors in cytochrome b6f, plastocyanin and Photosystem I (PSI). The terminal electron acceptor of PSI is ferredoxin, from which electrons may be used to reduce NADP+ for metabolic purposes. Concomitant production of a H+ gradient allows production of energy for the cell. Under certain conditions and using the endogenous hydrogenase, excess protons and electrons from ferredoxin may be converted to molecular hydrogen. In this work it is demonstrated both that certain mutations near the quinone electron transfer cofactor in PSI can speed up electron transfer through the PETC, and also that a native [FeFe]-hydrogenase can be expressed in the C. reinhardtii chloroplast. Taken together, these research findings form the foundation for the design of a PSI-hydrogenase fusion for the direct and continuous photo-production of hydrogen in vivo.
ContributorsReifschneider, Kiera (Author) / Redding, Kevin (Thesis advisor) / Fromme, Petra (Committee member) / Jones, Anne (Committee member) / Arizona State University (Publisher)
Created2013
Description
Specificity and affinity towards a given ligand/epitope limit target-specific delivery. Companies can spend between $500 million to $2 billion attempting to discover a new drug or therapy; a significant portion of this expense funds high-throughput screening to find the most successful target-specific compound available. A more recent addition to discovering highly specific targets is the application of phage display utilizing single chain variable fragment antibodies (scFv). The aim of this research was to employ phage display to identify pathologies related to traumatic brain injury (TBI), particularly astrogliosis. A unique biopanning method against viable astrocyte cultures activated with TGF-β achieved this aim. Four scFv clones of interest showed varying relative affinities toward astrocytes. One of those four showed the ability to identify reactive astroctyes over basal astrocytes through max signal readings, while another showed a statistical significance in max signal reading toward basal astrocytes. Future studies will include further affinity characterization assays. This work contributes to the development of targeting therapeutics and diagnostics for TBI.
ContributorsMarsh, William (Author) / Stabenfeldt, Sarah (Thesis advisor) / Caplan, Michael (Committee member) / Sierks, Michael (Committee member) / Arizona State University (Publisher)
Created2013
Description
The ribosome is a ribozyme and central to the biosynthesis of proteins in all organisms. It has a strong bias against non-alpha-L-amino acids, such as alpha-D-amino acids and beta-amino acids. Additionally, the ribosome is only able to incorporate one amino acid in response to one codon. It has been demonstrated that reengineering of the peptidyltransferase center (PTC) of the ribosome enabled the incorporation of both alpha-D-amino acids and beta-amino acids into full length protein. Described in Chapter 2 are five modified ribosomes having modifications in the peptidyltrasnferase center in the 23S rRNA. These modified ribosomes successfully incorporated five different beta-amino acids (2.1 - 2.5) into E. coli dihydrofolate reductase (DHFR). The second project (Chapter 3) focused on the study of the modified ribosomes facilitating the incorporation of the dipeptide glycylphenylalanine (3.25) and fluorescent dipeptidomimetic 3.26 into DHFR. These ribosomes also had modifications in the peptidyltransferase center in the 23S rRNA of the 50S ribosomal subunit. The modified DHFRs having beta-amino acids 2.3 and 2.5, dipeptide glycylphenylalanine (3.25) and dipeptidomimetic 3.26 were successfully characterized by the MALDI-MS analysis of the peptide fragments produced by "in-gel" trypsin digestion of the modified proteins. The fluorescent spectra of the dipeptidomimetic 3.26 and modified DHFR having fluorescent dipeptidomimetic 3.26 were also measured. The type I and II DNA topoisomerases have been firmly established as effective molecular targets for many antitumor drugs. A "classical" topoisomerase I or II poison acts by misaligning the free hydroxyl group of the sugar moiety of DNA and preventing the reverse transesterfication reaction to religate DNA. There have been only two classes of compounds, saintopin and topopyrones, reported as dual topoisomerase I and II poisons. Chapter 4 describes the synthesis and biological evaluation of topopyrones. Compound 4.10, employed at 20 µM, was as efficient as 0.5 uM camptothecin, a potent topoisomerase I poison, in stabilizing the covalent binary complex (~30%). When compared with a known topoisomerase II poison, etoposide (at 0.5 uM), topopyorone 4.10 produced similar levels of stabilized DNA-enzyme binary complex (~34%) at 5 uM concentration.
ContributorsMaini, Rumit (Author) / Hecht, Sidney M. (Thesis advisor) / Gould, Ian (Committee member) / Yan, Hao (Committee member) / Arizona State University (Publisher)
Created2013
Description
The biological and chemical diversity of protein structure and function can be greatly expanded by position-specific incorporation of non-natural amino acids bearing a variety of functional groups. Non-cognate amino acids can be incorporated into proteins at specific sites by using orthogonal aminoacyl-tRNA synthetase/tRNA pairs in conjunction with nonsense, rare, or 4-bp codons. There has been considerable progress in developing new types of amino acids, in identifying novel methods of tRNA aminoacylation, and in expanding the genetic code to direct their position. Chemical aminoacylation of tRNAs is accomplished by acylation and ligation of a dinucleotide (pdCpA) to the 3'-terminus of truncated tRNA. This strategy allows the incorporation of a wide range of natural and unnatural amino acids into pre-determined sites, thereby facilitating the study of structure-function relationships in proteins and allowing the investigation of their biological, biochemical and biophysical properties. Described in Chapter 1 is the current methodology for synthesizing aminoacylated suppressor tRNAs. Aminoacylated suppressor tRNACUAs are typically prepared by linking pre-aminoacylated dinucleotides (aminoacyl-pdCpAs) to 74 nucleotide (nt) truncated tRNAs (tRNA-COH) via a T4 RNA ligase mediated reaction. Alternatively, there is another route outlined in Chapter 1 that utilizes a different pre-aminoacylated dinucleotide, AppA. This dinucleotide has been shown to be a suitable substrate for T4 RNA ligase mediated coupling with abbreviated tRNA-COHs for production of 76 nt aminoacyl-tRNACUAs. The synthesized suppressor tRNAs have been shown to participate in protein synthesis in vitro, in an S30 (E. coli) coupled transcription-translation system in which there is a UAG codon in the mRNA at the position corresponding to Val10. Chapter 2 describes the synthesis of two non-proteinogenic amino acids, L-thiothreonine and L-allo-thiothreonine, and their incorporation into predetermined positions of a catalytically competent dihydrofolate reductase (DHFR) analogue lacking cysteine. Here, the elaborated proteins were site-specifically derivitized with a fluorophore at the thiothreonine residue. The synthesis and incorporation of phosphorotyrosine derivatives into DHFR is illustrated in Chapter 3. Three different phosphorylated tyrosine derivatives were prepared: bis-nitrobenzylphosphoro-L-tyrosine, nitrobenzylphosphoro-L-tyrosine, and phosphoro-L-tyrosine. Their ability to participate in a protein synthesis system was also evaluated.
ContributorsNangreave, Ryan Christopher (Author) / Hecht, Sidney M. (Thesis advisor) / Yan, Hao (Committee member) / Gould, Ian (Committee member) / Arizona State University (Publisher)
Created2013
Description
This study sought to analyze the messages being conveyed through the discourse utilized in presenting the public face of The Arizona Early Childhood Development and Health Board, popularly known as First Things First (FTF) and to reveal how the different discourses and ideologies within FTF have been in the past and currently are "contending and struggling for dominance (Wodak, 2007)." FTF is located within the policy realm of Early Childhood Education and Care (ECEC). The people and the system have been very influential in guiding the course and policies set forth in Arizona since the citizen initiative, Proposition 203, passed in 2006, which allowed for the creation of the Early Childhood Development and Health Board. Lakoff's techniques for analyzing frames of discourse were utilized in conjunction with critical discourse analysis in order to tease out frames of reference, shifts in both discourse and frames, specific modes of messaging, and consistencies and inconsistencies within the public face presented by FTF.
ContributorsMiller, Lisa (Author) / Swadener, Elizabeth B (Thesis advisor) / Nakagawa, Kathy (Committee member) / Romero, Mary (Committee member) / Arizona State University (Publisher)
Created2013
Description
As digital media practices become readily available in today's classrooms, literacy and literacy instruction are changing in profound ways (Alvermann, 2010). Professional organizations emphasize the importance of integrating new literacies (New London Group, 1996) practices into language-arts instruction (IRA, 2009; NCTE, 2005). As a result, teachers search for effective ways to incorporate the new literacies in an effort to engage students. Therefore, this study was designed to investigate the potential of digital storytelling as participatory media for writing instruction. This case study was conducted during the fall semester of 2012 in one first-grade classroom and one second-grade classroom in the Southwestern United States. The study addressed ten interrelated research questions relating to how primary-grade students performed in relation to the Common Core writing standards, how they were motivated, how they formed a meta- language to talk about their writing, how they developed identities as writers, and how they were influenced by their teachers' philosophies and instructional approaches. Twenty-two first-grade students and 24 second-grade students used the MovieMaker software to create digital stories of personal narratives. Data included field notes, interviews with teachers and students, teacher journals, my own journal, artifacts of teachers' lesson plans, photographs, students' writing samples, and their digital stories. Qualitative data were analyzed by thematic analysis (Patton, 1990) and discourse analysis (Gee, 2011). Writing samples were scored by rubrics based on the Common Core State Standards. The study demonstrated how digital storytelling can be used to; (a) guide teachers in implementing new literacies in primary grades; (b) illustrate digital storytelling as writing; (c) develop students' meta-language to talk about writing; (d) impact students' perceptions as writers; (e) meet Common Core State Standards for writing; (f) improve students' skills as writers; (g) build students' identities as writers; (h) impact academic writing; (i) engage students in the writing process; and (j) illustrate the differences in writing competencies between first- and second-grade students. The study provides suggestions for teachers interested in incorporating digital storytelling in primary-grade classrooms.
ContributorsFoley, Leslie M (Author) / Guzzetti, Barbara J. (Thesis advisor) / Hayes, Elisabeth R. (Committee member) / Gee, James P (Committee member) / Arizona State University (Publisher)
Created2013
Description
The purpose of this study was to investigate critical literacy practices in two prehistoric exhibits in a natural history museum. Bourdieu's habitus and Bakhtin's dialogism served as theoretical frames to collect and analyze data. Data were collected and triangulated using field notes, interview transcriptions, archives, and other data sources to critically scrutinize textual meaning and participant responses. Spradley's (1979) domain analysis was used to sort and categorize data in the early stage. Glaser and Strauss's (1967) constant comparative method was used to code data. My major findings were that museum texts within this context represent embedded beliefs and values that were interwoven with curators` habitus, tastes and capital, as well as institutional policies. The texts in the two Hohokam exhibits endorse a certain viewpoint of learning. Teachers and the public were not aware of the communicative role that the museum played in the society. In addition, museum literacy/ies were still practiced in a fundamental way as current practices in the classroom, which may not support the development of critical literacy. In conclusion, the very goal for critical museum literacy is to help students and teachers develop intellectual strategies to read the word and the world in informal learning environments.
ContributorsLiang, Sheau-yann (Author) / Mccarty, Teresa (Thesis advisor) / Marsh, Josephine (Committee member) / Blumenfeld-Jones, Donald (Committee member) / Welsh, Peter (Committee member) / Arizona State University (Publisher)
Created2013
Description
Researchers have postulated that math academic achievement increases student success in college (Lee, 2012; Silverman & Seidman, 2011; Vigdor, 2013), yet 80% of universities and 98% of community colleges require many of their first-year students to be placed in remedial courses (Bettinger & Long, 2009). Many high school graduates are entering college ill prepared for the rigors of higher education, lacking understanding of basic and important principles (ACT, 2012). The desire to increase academic achievement is a wide held aspiration in education and the idea of adapting instruction to individuals is one approach to accomplish this goal (Lalley & Gentile, 2009a). Frequently, adaptive learning environments rely on a mastery learning approach, it is thought that when students are afforded the opportunity to master the material, deeper and more meaningful learning is likely to occur. Researchers generally agree that the learning environment, the teaching approach, and the students' attributes are all important to understanding the conditions that promote academic achievement (Bandura, 1977; Bloom, 1968; Guskey, 2010; Cassen, Feinstein & Graham, 2008; Changeiywo, Wambugu & Wachanga, 2011; Lee, 2012; Schunk, 1991; Van Dinther, Dochy & Segers, 2011). The present study investigated the role of college students' affective attributes and skills, such as academic competence and academic resilience, in an adaptive mastery-based learning environment on their academic performance, while enrolled in a remedial mathematics course. The results showed that the combined influence of students' affective attributes and academic resilience had a statistically significant effect on students' academic performance. Further, the mastery-based learning environment also had a significant effect on their academic competence and academic performance.
ContributorsFoshee, Cecile Mary (Author) / Atkinson, Robert K (Thesis advisor) / Elliott, Stephen N. (Committee member) / Horan, John (Committee member) / Arizona State University (Publisher)
Created2013
Description
Alzheimer's Disease (AD) is the sixth leading cause of death in the United States and the most common form of dementia. Its cause remains unknown, but it is known to involve two hallmark pathologies: Amyloid Beta plaques and neurofibrillary tangles (NFTs). Several proteins have been implicated in the formation of neurofibrillary tangles, including Tau and S100B. S100B is a dimeric protein that is typically found bound to Ca(II) or Zn(II). These experiments relate to the involvement of S100B in Alzheimer's Disease-related processes and the results suggest that future research of S100B is warranted. Zn(II)-S100B was found to increase the rate at which tau assembled into paired helical filaments, as well as affect the rate at which tubulin polymerized into microtubules and the morphology of SH-SY5Y neuroblastoma cells after 72 hours of incubation. Zn(II)-S100B also increased the firing rate of hippocampal neurons after 36 hours of incubation. Together, these results suggest several possibilities: Zn(II)-S100B may be a key part of the formation of paired helical filaments (PHFs) that subsequently form NFTs. Zn(II)-S100B may also be competing with tau to bind tubulin, which could lead to an instability of microtubules and subsequent cell death. This finding aligns with the neurodegeneration that is commonly seen in AD and which could be a result of this microtubule instability. Ultimately, these results suggest that S100B is likely involved in several AD-related processes, and if the goal is to find an efficient and effective therapeutic target for AD, the relationship between S100B, particularly Zn(II)-S100B, and tau needs to be further studied.
ContributorsNaegele, Hayley (Author) / Mcgregor, Wade C (Thesis advisor) / Baluch, Debra (Committee member) / Francisco, Wilson (Committee member) / Arizona State University (Publisher)
Created2013
Description
The purpose of professional development is to enhance educator practices so that students may achieve at high levels. Too often, professional development tends to be too broad, general, or unrelated to problems of practice that teachers face in their own classrooms. This action research project builds upon the scholarly research that recognizes the need for professional development to be sustained, connected to teachers' own contexts, focused on specific subject matter, collaborative, and reflective. The goal of this action research study was to facilitate a culture of continuous improvement in teaching and learning by utilizing a model of professional development that challenges teachers to question their practices, utilize research to support their instruction, design an inquiry project that supports a change in practice, and examine changes in student growth. Results suggest that although teachers recognize the complexities that surround professional development, they found that this professional development model focused on their needs as professionals, was sustained over time, and was supported by a variety of professional influences. As a result of the model implemented, teachers reported shifts in their instructional practices and student growth related to personal inquiry projects.
ContributorsHudak, Michele (Author) / Roe, Mary (Thesis advisor) / Weber, Catherine (Committee member) / Chalex, Nancy (Committee member) / Arizona State University (Publisher)
Created2013