Matching Items (220)
152192-Thumbnail Image.png
Description
ABSTRACT Peptide microarrays may prove to be a powerful tool for proteomics research and clinical diagnosis applications. Fodor et al. and Maurer et al. have shown proof-of-concept methods of light- and electrochemically-directed peptide microarray fabrication on glass and semiconductor microchips respectively. In this work, peptide microarray fabrication based on the

ABSTRACT Peptide microarrays may prove to be a powerful tool for proteomics research and clinical diagnosis applications. Fodor et al. and Maurer et al. have shown proof-of-concept methods of light- and electrochemically-directed peptide microarray fabrication on glass and semiconductor microchips respectively. In this work, peptide microarray fabrication based on the abovementioned techniques were optimized. In addition, MALDI mass spectrometry based peptide synthesis characterization on semiconductor microchips was developed and novel applications of a CombiMatrix (CBMX) platform for electrochemically controlled synthesis were explored. We have investigated performance of 2-(2-nitrophenyl)propoxycarbonyl (NPPOC) derivatives as photo-labile protecting group. Specifically, influence of substituents on 4 and 5 positions of phenyl ring of NPPOC group on the rate of photolysis and the yield of the amine was investigated. The results indicated that substituents capable of forming a π-network with the nitro group enhanced the rate of photolysis and yield. Once such properly substituted NPPOC groups were used, the rate of photolysis/yield depended on the nature of protected amino group indicating that a different chemical step during the photo-cleavage process became the rate limiting step. We also focused on electrochemically-directed parallel synthesis of high-density peptide microarrays using the CBMX technology referred to above which uses electrochemically generated acids to perform patterned chemistry. Several issues related to peptide synthesis on the CBMX platform were studied and optimized, with emphasis placed on the reactions of electro-generated acids during the deprotection step of peptide synthesis. We have developed a MALDI mass spectrometry based method to determine the chemical composition of microarray synthesis, directly on the feature. This method utilizes non-diffusional chemical cleavage from the surface, thereby making the chemical characterization of high-density microarray features simple, accurate, and amenable to high-throughput. CBMX Corp. has developed a microarray reader which is based on electro-chemical detection of redox chemical species. Several parameters of the instrument were studied and optimized and novel redox applications of peptide microarrays on CBMX platform were also investigated using the instrument. These include (i) a search of metal binding catalytic peptides to reduce overpotential associated with water oxidation reaction and (ii) an immobilization of peptide microarrays using electro-polymerized polypyrrole.
ContributorsKumar, Pallav (Author) / Woodbury, Neal (Thesis advisor) / Allen, James (Committee member) / Johnston, Stephen (Committee member) / Arizona State University (Publisher)
Created2013
152556-Thumbnail Image.png
Description
Earth's topographic surface forms an interface across which the geodynamic and geomorphic engines interact. This interaction is best observed along crustal margins where topography is created by active faulting and sculpted by geomorphic processes. Crustal deformation manifests as earthquakes at centennial to millennial timescales. Given that nearly half of Earth's

Earth's topographic surface forms an interface across which the geodynamic and geomorphic engines interact. This interaction is best observed along crustal margins where topography is created by active faulting and sculpted by geomorphic processes. Crustal deformation manifests as earthquakes at centennial to millennial timescales. Given that nearly half of Earth's human population lives along active fault zones, a quantitative understanding of the mechanics of earthquakes and faulting is necessary to build accurate earthquake forecasts. My research relies on the quantitative documentation of the geomorphic expression of large earthquakes and the physical processes that control their spatiotemporal distributions. The first part of my research uses high-resolution topographic lidar data to quantitatively document the geomorphic expression of historic and prehistoric large earthquakes. Lidar data allow for enhanced visualization and reconstruction of structures and stratigraphy exposed by paleoseismic trenches. Lidar surveys of fault scarps formed by the 1992 Landers earthquake document the centimeter-scale erosional landforms developed by repeated winter storm-driven erosion. The second part of my research employs a quasi-static numerical earthquake simulator to explore the effects of fault roughness, friction, and structural complexities on earthquake-generated deformation. My experiments show that fault roughness plays a critical role in determining fault-to-fault rupture jumping probabilities. These results corroborate the accepted 3-5 km rupture jumping distance for smooth faults. However, my simulations show that the rupture jumping threshold distance is highly variable for rough faults due to heterogeneous elastic strain energies. Furthermore, fault roughness controls spatiotemporal variations in slip rates such that rough faults exhibit lower slip rates relative to their smooth counterparts. The central implication of these results lies in guiding the interpretation of paleoseismically derived slip rates that are used to form earthquake forecasts. The final part of my research evaluates a set of Earth science-themed lesson plans that I designed for elementary-level learning-disabled students. My findings show that a combination of concept delivery techniques is most effective for learning-disabled students and should incorporate interactive slide presentations, tactile manipulatives, teacher-assisted concept sketches, and student-led teaching to help learning-disabled students grasp Earth science concepts.
ContributorsHaddad, David Elias (Author) / Arrowsmith, Ramon (Thesis advisor) / Reynolds, Stephen (Committee member) / Semken, Steven (Committee member) / Shirzaei, Manoochehr (Committee member) / Whipple, Kelin (Committee member) / Zielke, Olaf (Committee member) / Arizona State University (Publisher)
Created2014
153110-Thumbnail Image.png
Description
The healthcare system in this country is currently unacceptable. New technologies may contribute to reducing cost and improving outcomes. Early diagnosis and treatment represents the least risky option for addressing this issue. Such a technology needs to be inexpensive, highly sensitive, highly specific, and amenable to adoption in a clinic.

The healthcare system in this country is currently unacceptable. New technologies may contribute to reducing cost and improving outcomes. Early diagnosis and treatment represents the least risky option for addressing this issue. Such a technology needs to be inexpensive, highly sensitive, highly specific, and amenable to adoption in a clinic. This thesis explores an immunodiagnostic technology based on highly scalable, non-natural sequence peptide microarrays designed to profile the humoral immune response and address the healthcare problem. The primary aim of this thesis is to explore the ability of these arrays to map continuous (linear) epitopes. I discovered that using a technique termed subsequence analysis where epitopes could be decisively mapped to an eliciting protein with high success rate. This led to the discovery of novel linear epitopes from Plasmodium falciparum (Malaria) and Treponema palladium (Syphilis), as well as validation of previously discovered epitopes in Dengue and monoclonal antibodies. Next, I developed and tested a classification scheme based on Support Vector Machines for development of a Dengue Fever diagnostic, achieving higher sensitivity and specificity than current FDA approved techniques. The software underlying this method is available for download under the BSD license. Following this, I developed a kinetic model for immunosignatures and tested it against existing data driven by previously unexplained phenomena. This model provides a framework and informs ways to optimize the platform for maximum stability and efficiency. I also explored the role of sequence composition in explaining an immunosignature binding profile, determining a strong role for charged residues that seems to have some predictive ability for disease. Finally, I developed a database, software and indexing strategy based on Apache Lucene for searching motif patterns (regular expressions) in large biological databases. These projects as a whole have advanced knowledge of how to approach high throughput immunodiagnostics and provide an example of how technology can be fused with biology in order to affect scientific and health outcomes.
ContributorsRicher, Joshua Amos (Author) / Johnston, Stephen A. (Thesis advisor) / Woodbury, Neal (Committee member) / Stafford, Phillip (Committee member) / Papandreou-Suppappola, Antonia (Committee member) / Arizona State University (Publisher)
Created2014
149856-Thumbnail Image.png
Description
Nucleosomes are the basic repetitive unit of eukaryotic chromatin and are responsible for packing DNA inside the nucleus of the cell. They consist of a complex of eight histone proteins (two copies of four proteins H2A, H2B, H3 and H4) around which 147 base pairs of DNA are wrapped

Nucleosomes are the basic repetitive unit of eukaryotic chromatin and are responsible for packing DNA inside the nucleus of the cell. They consist of a complex of eight histone proteins (two copies of four proteins H2A, H2B, H3 and H4) around which 147 base pairs of DNA are wrapped in ~1.67 superhelical turns. Although the nucleosomes are stable protein-DNA complexes, they undergo spontaneous conformational changes that occur in an asynchronous fashion. This conformational dynamics, defined by the "site-exposure" model, involves the DNA unwrapping from the protein core and exposing itself transiently before wrapping back. Physiologically, this allows regulatory proteins to bind to their target DNA sites during cellular processes like replication, DNA repair and transcription. Traditional biochemical assays have stablished the equilibrium constants for the accessibility to various sites along the length of the nucleosomal DNA, from its end to the middle of the dyad axis. Using fluorescence correlation spectroscopy (FCS), we have established the position dependent rewrapping rates for nucleosomes. We have also used Monte Carlo simulation methods to analyze the applicability of FRET fluctuation spectroscopy towards conformational dynamics, specifically motivated by nucleosome dynamics. Another important conformational change that is involved in cellular processes is the disassembly of nucleosome into its constituent particles. The exact pathway adopted by nucleosomes is still not clear. We used dual color fluorescence correlation spectroscopy to study the intermediates during nucleosome disassembly induced by changing ionic strength. Studying the nature of nucleosome conformational change and the kinetics is very important in understanding gene expression. The results from this thesis give a quantitative description to the basic unit of the chromatin.
ContributorsGurunathan, Kaushik (Author) / Levitus, Marcia (Thesis advisor) / Lindsay, Stuart (Committee member) / Woodbury, Neal (Committee member) / Yan, Hao (Committee member) / Arizona State University (Publisher)
Created2011
151229-Thumbnail Image.png
Description
It has been well established that mitochondria play a critical role in the pathology of Friedreich's Ataxia. This disease is believed to be caused by a deficiency of frataxin, which research suggests is responsible for iron sulfur cluster assembly. This incomplete assembly of iron sulfur clusters is believed to be

It has been well established that mitochondria play a critical role in the pathology of Friedreich's Ataxia. This disease is believed to be caused by a deficiency of frataxin, which research suggests is responsible for iron sulfur cluster assembly. This incomplete assembly of iron sulfur clusters is believed to be linked with dysfunctional complexes in the mitochondrial respiratory chain, increased oxidative stress, and potential cell death. Increased understanding of the pathophysiology of this disease has enabled the development of various therapeutic strategies aimed at restoring mitochondrial respiration. This thesis contains an analysis of the biological activity of several classes of antioxidants against oxidative stress induced by diethyl maleate in Friedreich's Ataxia lymphocytes and CEM leukemia cells. Analogues of vitamin E α-tocopherol have been shown to protect cells under oxidative stress. However, these same analogues show various levels of inhibition towards the electron transport chain complex I. Bicyclic pyridinols containing a ten carbon substituent provided favorable cytoprotection. N-hydroxy-4-pyridone compounds were observed to provide little protection. Similarly, analogues of CoQ10 in the form of pyridinol and pyrimidinol compounds also preserved cell viability at low concentrations.
ContributorsJaruvangsanti, Jennifer (Author) / Hecht, Sidney (Thesis advisor) / Woodbury, Neal (Committee member) / Skibo, Edward (Committee member) / Arizona State University (Publisher)
Created2012
156131-Thumbnail Image.png
Description
Though DNA nanostructures (DNs) have become interesting subjects of drug delivery, in vivo imaging and biosensor research, however, for real biological applications, they should be ‘long circulating’ in blood. One of the crucial requirements for DN stability is high salt concentration (like ~5–20 mM Mg2+) that is unavailable in a

Though DNA nanostructures (DNs) have become interesting subjects of drug delivery, in vivo imaging and biosensor research, however, for real biological applications, they should be ‘long circulating’ in blood. One of the crucial requirements for DN stability is high salt concentration (like ~5–20 mM Mg2+) that is unavailable in a cell culture medium or in blood. Hence DNs denature promptly when injected into living systems. Another important factor is the presence of nucleases that cause fast degradation of unprotected DNs. The third factor is ‘opsonization’ which is the immune process by which phagocytes target foreign particles introduced into the bloodstream. The primary aim of this thesis is to design strategies that can improve the in vivo stability of DNs, thus improving their pharmacodynamics and biodistribution.

Several strategies were investigated to address the three previously mentioned limitations. The first attempt was to study the effect length and conformation of polyethylene glycol (PEG) on DN stability. DNs were also coated with PEG-lipid and human serum albumin (HSA) and their stealth efficiencies were compared. The findings reveal that both PEGylation and albumin coating enhance low salt stability, increase resistance towards nuclease action and reduce uptake of DNs by macrophages. Any protective coating around a DN increases its hydrodynamic radius, which is a crucial parameter influencing their clearance. Keeping this in mind, intrinsically stable DNs that can survive low salt concentration without any polymer coating were built. Several DNA compaction agents and DNA binders were screened to stabilize DNs in low magnesium conditions. Among them arginine, lysine, bis-lysine and hexamine cobalt showed the potential to enhance DN stability.

This thesis also presents a sensitive assay, the Proximity Ligation Assay (PLA), for the estimation of DN stability with time. It requires very simple modifications on the DNs and it can yield precise results from a very small amount of sample. The applicability of PLA was successfully tested on several DNs ranging from a simple wireframe tetrahedron to a 3D origami and the protocol to collect in vivo samples, isolate the DNs and measure their stability was developed.
ContributorsBanerjee, Saswata (Author) / Yan, Hao (Thesis advisor) / Angell, Austen (Committee member) / Woodbury, Neal (Committee member) / Liu, Yan (Committee member) / Arizona State University (Publisher)
Created2018
156594-Thumbnail Image.png
Description
Aquifers host the largest accessible freshwater resource in the world. However, groundwater reserves are declining in many places. Often coincident with drought, high extraction rates and inadequate replenishment result in groundwater overdraft and permanent land subsidence. Land subsidence is the cause of aquifer storage capacity reduction, altered topographic gradients which

Aquifers host the largest accessible freshwater resource in the world. However, groundwater reserves are declining in many places. Often coincident with drought, high extraction rates and inadequate replenishment result in groundwater overdraft and permanent land subsidence. Land subsidence is the cause of aquifer storage capacity reduction, altered topographic gradients which can exacerbate floods, and differential displacement that can lead to earth fissures and infrastructure damage. Improving understanding of the sources and mechanisms driving aquifer deformation is important for resource management planning and hazard mitigation.

Poroelastic theory describes the coupling of differential stress, strain, and pore pressure, which are modulated by material properties. To model these relationships, displacement time series are estimated via satellite interferometry and hydraulic head levels from observation wells provide an in-situ dataset. In combination, the deconstruction and isolation of selected time-frequency components allow for estimating aquifer parameters, including the elastic and inelastic storage coefficients, compaction time constants, and vertical hydraulic conductivity. Together these parameters describe the storage response of an aquifer system to changes in hydraulic head and surface elevation. Understanding aquifer parameters is useful for the ongoing management of groundwater resources.

Case studies in Phoenix and Tucson, Arizona, focus on land subsidence from groundwater withdrawal as well as distinct responses to artificial recharge efforts. In Christchurch, New Zealand, possible changes to aquifer properties due to earthquakes are investigated. In Houston, Texas, flood severity during Hurricane Harvey is linked to subsidence, which modifies base flood elevations and topographic gradients.
ContributorsMiller, Megan Marie (Author) / Shirzaei, Manoochehr (Thesis advisor) / Reynolds, Stephen (Committee member) / Tyburczy, James (Committee member) / Semken, Steven (Committee member) / Werth, Susanna (Committee member) / Arizona State University (Publisher)
Created2018
156515-Thumbnail Image.png
Description
The fundamental photophysics of fluorescent probes must be understood when the probes are used in biological applications. The photophysics of BODIPY dyes inside polymeric micelles and rhodamine dyes covalently linked to proteins were studied. Hydrophobic boron-dipyrromethene (BODIPY) dyes were noncovalently encapsulated inside polymeric micelles. Absorbance and fluorescence measurements were employed

The fundamental photophysics of fluorescent probes must be understood when the probes are used in biological applications. The photophysics of BODIPY dyes inside polymeric micelles and rhodamine dyes covalently linked to proteins were studied. Hydrophobic boron-dipyrromethene (BODIPY) dyes were noncovalently encapsulated inside polymeric micelles. Absorbance and fluorescence measurements were employed to study the photophysics of these BODIPY dyes in the micellar environments. Amphiphilic polymers with a hydrophobic character and low Critical Micelle Concentration (CMC) protected BODIPYS from the aqueous environment. Moderate dye loading conditions did not result in ground-state dimerization, and only fluorescence lifetimes and brightnesses were affected. However, amphiphilic polymers with a hydrophilic character and high CMC did not protect the BODIPYS from the aqueous environment with concomitant ground-state dimerization and quenching of the fluorescence intensity, lifetime, and brightnesses even at low dye loading conditions. At the doubly-labeled interfaces of Escherichia coli (E. coli) DNA processivity β clamps, the interchromophric interactions of four rhodamine dyes were studied: tetramethylrhodamine (TMR), TMR C6, Alexa Fluor 488, and Alexa Fluor 546. Absorbance and fluorescence measurements were performed on doubly-labeled β clamps with singly-labeled β clamps and free dyes as controls. The absorbance measurements revealed that both TMR and TMR C6 readily formed H-dimers (static quenching) at the doubly-labeled interfaces of the β clamps. However, the TMR with a longer linker (TMR C6) also displayed a degree of dynamic quenching. For Alexa Fluor 546 and Alexa Fluor 488, there were no clear signs of dimerization in the absorbance scans. However, the fluorescence properties (fluorescence intensity, lifetime, and anisotropy) of the Alexa Fluor dyes significantly changed when three methodologies were employed to disrupt the doubly-labeled interfaces: 1) the addition of sodium dodecyl sulfate (SDS) detergent to denature the proteins, 2) the addition of clamp loader (γ complex) to open one of the two interfaces, and 3) the use of subunit exchange to decrease the number of dyes per interface. These fluorescence measurements indicated that for the Alexa Fluor dyes, other interchromophoric interactions were present such as dynamic quenching and homo-Förster Resonance Energy Transfer (homo-FRET).
ContributorsDonaphon, Bryan Matthew (Author) / Levitus, Marcia (Thesis advisor) / Van Horn, Wade (Committee member) / Woodbury, Neal (Committee member) / Arizona State University (Publisher)
Created2018
156312-Thumbnail Image.png
Description
Glycans are monosaccharide-based heteropolymers that are found covalently attached to many different proteins and lipids and are ubiquitously displayed on the exterior surfaces of cells. Serum glycan composition and structure are well known to be altered in many different types of cancer. In fact, glycans represent a promising but only

Glycans are monosaccharide-based heteropolymers that are found covalently attached to many different proteins and lipids and are ubiquitously displayed on the exterior surfaces of cells. Serum glycan composition and structure are well known to be altered in many different types of cancer. In fact, glycans represent a promising but only marginally accessed source of cancer markers. The approach used in this dissertation, which is referred to as “glycan node analysis”, is a molecularly bottom-up approach to plasma/serum (P/S) glycomics based on glycan linkage analysis that captures features such as α2-6 sialylation, β1-6 branching, and core fucosylation as single analytical signals.

The diagnostic utility of this approach as applied to lung cancer patients across all stages as well as prostate, serous ovarian, and pancreatic cancer patients compared to certifiably healthy individuals, nominally healthy individuals and/or risk-matched controls is reported. Markers for terminal fucosylation, α2-6 sialylation, β1-4 branching, β1-6 branching and outer-arm fucosylation were most able to differentiate cases from controls. These markers behaved in a stage-dependent manner in lung cancer as well as other types of cancer. Using a Cox proportional hazards regression model, the ability of these markers to predict progression and survival in lung cancer patients was assessed. In addition, the potential mechanistic role of aberrant P/S glycans in cancer progression is discussed.

Plasma samples from former bladder cancer patients with currently no evidence of disease (NED), non-muscle invasive bladder cancer (NMIBC), and muscle invasive bladder cancer (MIBC) along with certifiably healthy controls were analyzed. Markers for α2-6 sialylation, β1-4 branching, β1-6 branching, and outer-arm fucosylation were able to separate current and former (NED) cases from controls; but NED, NMIBC, and MIBC were not distinguished from one another. Markers for α2-6 sialylation and β1-6 branching were able to predict recurrence from the NED state using a Cox proportional hazards regression model adjusted for age, gender, and time from cancer. These two glycan features were found to be correlated to the concentration of C-reactive protein, a known prognostic marker for bladder cancer, further strengthening the link between inflammation and abnormal plasma protein glycosylation.
ContributorsRoshdiferdosi, Shadi (Author) / Borges, Chad R (Thesis advisor) / Woodbury, Neal (Committee member) / Hayes, Mark (Committee member) / Arizona State University (Publisher)
Created2018
156778-Thumbnail Image.png
Description
The dynamic Earth involves feedbacks between the solid crust and both natural and anthropogenic fluid flows. Fluid-rock interactions drive many Earth phenomena, including volcanic unrest, seismic activities, and hydrological responses. Mitigating the hazards associated with these activities requires fundamental understanding of the underlying physical processes. Therefore, geophysical monitoring in combination

The dynamic Earth involves feedbacks between the solid crust and both natural and anthropogenic fluid flows. Fluid-rock interactions drive many Earth phenomena, including volcanic unrest, seismic activities, and hydrological responses. Mitigating the hazards associated with these activities requires fundamental understanding of the underlying physical processes. Therefore, geophysical monitoring in combination with modeling provides valuable tools, suitable for hazard mitigation and risk management efforts. Magmatic activities and induced seismicity linked to fluid injection are two natural and anthropogenic processes discussed in this dissertation.

Successful forecasting of the timing, style, and intensity of a volcanic eruption is made possible by improved understanding of the volcano life cycle as well as building quantitative models incorporating the processes that govern rock melting, melt ascending, magma storage, eruption initiation, and interaction between magma and surrounding host rocks at different spatial extent and time scale. One key part of such models is the shallow magma chamber, which is generally directly linked to volcano’s eruptive behaviors. However, its actual shape, size, and temporal evolution are often not entirely known. To address this issue, I use space-based geodetic data with high spatiotemporal resolution to measure surface deformation at Kilauea volcano. The obtained maps of InSAR (Interferometric Synthetic Aperture Radar) deformation time series are exploited with two novel modeling schemes to investigate Kilauea’s shallow magmatic system. Both models can explain the same observation, leading to a new compartment model of magma chamber. Such models significantly advance the understanding of the physical processes associated with Kilauea’s summit plumbing system with potential applications for volcanoes around the world.

The unprecedented increase in the number of earthquakes in the Central and Eastern United States since 2008 is attributed to massive deep subsurface injection of saltwater. The elevated chance of moderate-large damaging earthquakes stemming from increased seismicity rate causes broad societal concerns among industry, regulators, and the public. Thus, quantifying the time-dependent seismic hazard associated with the fluid injection is of great importance. To this end, I investigate the large-scale seismic, hydrogeologic, and injection data in northern Texas for period of 2007-2015 and in northern-central Oklahoma for period of 1995-2017. An effective induced earthquake forecasting model is developed, considering a complex relationship between injection operations and consequent seismicity. I find that the timing and magnitude of regional induced earthquakes are fully controlled by the process of fluid diffusion in a poroelastic medium and thus can be successfully forecasted. The obtained time-dependent seismic hazard model is spatiotemporally heterogeneous and decreasing injection rates does not immediately reduce the probability of an earthquake. The presented framework can be used for operational induced earthquake forecasting. Information about the associated fundamental processes, inducing conditions, and probabilistic seismic hazards has broad benefits to the society.
ContributorsZhai, Guang (Author) / Shirzaei, Manoochehr (Thesis advisor) / Garnero, Edward (Committee member) / Clarke, Amanda (Committee member) / Tyburczy, James (Committee member) / Li, Mingming (Committee member) / Arizona State University (Publisher)
Created2018