Life Sciences Commons^™

Characterization Of Radiotolerance In Potato And Development Of A Gamma Radiation Phytosensor., Robert Graham Sears

Doctoral Dissertations

As humans pursue space travel and nuclear energy, the risk of harm from ionizing radiation increases. On Earth or in space, plants are essential to our personal and environmental health. Plants serve as sentinels, bioremediators and food sources in areas of high ionizing radiation, therefore it is essential to understand how ionizing radiation affects plant biology. This work aimed to understand plant responses to ionizing radiation in the potato chassis and apply that knowledge to generate novel phenotypes for nuclear energy and space applications. The first gamma radiation phytosensor was developed for monitoring at standoff distances greater than three meters. …

Cell-Free Metabolic Engineering Strategies For Accelerated Biomanufacturing, Jaime Lorenzo N. Dinglasan

Doctoral Dissertations

Biomanufacturing propels the bioeconomy. Accelerating bioeconomic growth thus requires the expedited development of biomanufacturing processes that can expand the current bioproduct portfolio. Lysate-based cell-free systems provide unique advantages for simplified metabolic pathway construction. Their central metabolic pathways and transcriptional-translational (TX-TL) machineries are free from genome regulation and are amenable to direct manipulation, enabling the streamlined construction of biomanufacturing processes. While their utility as prototyping platforms for accelerating cellular metabolic engineering has been demonstrated, the potential to rapidly build commercial “cell-free factories” capable of sophisticated bioconversion has not been fully realized. Lysates with high-yield pathways are projected to enable commercialized cell-free …

What I Talk About When I Talk About Integration Of Single-Cell Data, Yang Xu

Doctoral Dissertations

Over the past decade, single-cell technologies evolved from profiling hundreds of cells to millions of cells, and emerged from a single modality of data to cover multiple views at single-cell resolution, including genome, epigenome, transcriptome, and so on. With advance of these single-cell technologies, the booming of multimodal single-cell data creates a valuable resource for us to understand cellular heterogeneity and molecular mechanism at a comprehensive level. However, the large-scale multimodal single-cell data also presents a huge computational challenge for insightful integrative analysis. Here, I will lay out problems in data integration that single-cell research community is interested in and …

A Rapid And Ultra-Sensitive Biosensing Platform Based On Tunable Dielectrophoresis For Robust Poc Applications, Yu Jiang

Doctoral Dissertations

With the ongoing pandemic, there have been increasing concerns recently regarding major public health issues such as abuse of organophosphorus compounds, pathogenic bacterial infections, and biosecurity in agricultural production. Biosensors have long been considered a kernel technology for next-generation diagnostic solutions to improve food safety and public health. Significant amounts of effort have been devoted to inventing novel sensing mechanisms, modifying their designs, improving their performance, and extending their application scopes. However, the reliability and selectivity of most biosensors still have much to be desired, which holds back the development and commercialization of biosensors, especially for on-site and point-of-care (POC) …

Interactions Between Soft Nanoparticles And Mammalian Cells, Mitchell Raith

Doctoral Dissertations

Nanoparticles have been of interest to the pharmaceutical industry since the 1980s. The first FDA approved nanoparticle-based therapies included liposomal anesthesia agents. Since then, the amount of FDA-approved nanoparticle therapies remains low. This is because nanoparticle-patient interactions can be very complex and are not well understood. Complicating factors also include increasing obesity rates among the patient population and many small animal pre-clinical trials are completed with healthy, lean animals. The biochemical differences between lean and obese patients prevents early studies from accurately predicting nanoparticle clinical behaviors. Many nanoparticles fail in trails. In this thesis, I aimed to uncover how nanoparticles …

Improving The Biocompatibility Of The Bio-Inorganic Interface For Enhanced Photosystem I-Based Biophotovoltaic Device Performance, Alexandra H. Teodor

Doctoral Dissertations

The world’s energy demands are projected to increase by nearly 50% by the year 2040, and consumption of carbon-based fuels continues to release greenhouse gases such as carbon dioxide and methane into the atmosphere. This has been causally linked with climate change and increased extreme weather events, which has been further linked to adverse health outcomes and negative effects on biodiversity, food security, and increased disease transmission. Clearly, there is a need for a sustainable, carbon-free, and cost-effective method of energy production to meet growing energy production demands. The sun irradiates Earth’s surface annually with ~80,000 terawatts (TW), making solar …

Alteration Of Potato Plastids As Platforms For Biotechnology And Synthetic Biology, Alexander C. Pfotenhauer

Doctoral Dissertations

Plastids represent a unique opportunity for plant biotechnology and synthetic biology. Their fundamental uses for photosynthesis, starch storage, and other processes can be exploited to benefit food production and other industries. In this work, we attempt to modulate these plastids in order to produce potato plants with beneficial characteristics for further use in synthetic biology. The first chapter of this dissertation involves genome-editing of the FtsZ1 gene, which is involved in plastid division. Plants were generated with large plastids and starch granules, and similar methods could be used to produce these plants without foreign DNA integration. The second chapter involves …

Engineering Modularity Of Ester Biosynthesis Across Biological Scales, Hyeongmin Seo

Doctoral Dissertations

Metabolic engineering and synthetic biology enable controlled manipulation of whole-cell biocatalysts to produce valuable chemicals from renewable feedstocks in a rapid and efficient manner, helping reduce our reliance on the conventional petroleum-based chemical synthesis. However, strain engineering process is costly and time-consuming that developing economically competitive bioprocess at industrial scale is still challenging. To accelerate the strain engineering process, modular cell engineering has been proposed as an innovative approach that harnesses modularity of metabolism for designing microbial cell factories. It is important to understand biological modularity and to develop design principles for effective implementation of modular cell engineering. In this …

Overcoming Dna Restriction-Modification Systems To Enable Transformation And Genetic Engineering In Non-Model Organisms, Lauren Ann Riley

Doctoral Dissertations

Many organisms naturally possess complex physiological traits that are of interest for biotechnology research. These non-model organisms are not routinely used though, due to a lack of foundational knowledge and genetic tools. The development of genetic tools is limited in part by the inability to efficiently transform DNA into these organisms. One of the major barriers to efficient DNA transformation is the native DNA Restriction-Modification systems within bacteria. DNA Restriction-Modification systems act as an immune system to cut heterologous DNA methylated differently than the host. To overcome these systems, DNA needs to be methylated in the same manner as the …

Needles In A Haystack Of Protein Diversity: Interrogation Of Complex Biological Samples Through Specialized Strategies In Bottom-Up Proteomics Uncover Peptides Of Interest For Diverse Applications, Manuel I. Villalobos Solis

Doctoral Dissertations

Peptide identification is at the core of bottom-up proteomics measurements. However, even with state-of the-art mass spectrometric instrumentation, peptide level information is still lost or missing in these types of experiments. Reasons behind missing peptide identifications in bottom-up proteomics include variable peptide ionization efficiencies, ion suppression effects, as well as the occurrence of chimeric spectra that can lower the efficacy of database search strategies. Peptides derived from naturally abundant proteins in a biological system also have better chances of being identified in comparison to the ones produced from less abundant proteins, at least in regular discovery-based proteomics experiments. This dissertation …

Bioinformatic And Experimental Approaches For Deeper Metaproteomic Characterization Of Complex Environmental Samples, Ramsunder Mahadevan Iyer

Doctoral Dissertations

The coupling of high performance multi-dimensional liquid chromatography and tandem mass spectrometry for characterization of microbial proteins from complex environmental samples has paved the way for a new era in scientific discovery. The field of metaproteomics, which is the study of protein suite of all the organisms in a biological system, has taken a tremendous leap with the introduction of high-throughput proteomics. However, with corresponding increase in sample complexity, novel challenges have been raised with respect to efficient peptide separation via chromatography and bioinformatic analysis of the resulting high throughput data. In this dissertation, various aspects of metaproteomic characterization, including …

Development Of Ms-Based Proteomics Approaches To Examine Metabolic Pathways And Protein:Protein Interactions In Microbial Systems, Chen Qian

Doctoral Dissertations

The proteome is perhaps the most functional operating machinery for almost all biological processes, serving as the bridge to link the genome and phenotypes. The proteome undergoes dynamic changes in terms of the abundance or interactions, responding to the environmental stimuli. Understanding this dynamic of protein alterations is the key to delineate critical biological mechanisms. Mass-spectrometry-based proteomics is a powerful tool to systematically monitor the heterogeneous alterations of the proteome, including the changes of abundance, modifications and interactions. In this dissertation, a research project was built upon current proteomics approaches to solve the issues regarding to the sample preparation and …

Managing Exoelectrogenic Microbial Community Development Through Bioprocess Control For Conversion Of Biomass-Derived Streams, Alex James Lewis

Doctoral Dissertations

Bioelectrochemical systems are an emerging technology capable of utilizing aqueous waste streams generated during biomass conversion of lignocellulosic feedstocks to produce valuable co-products and thus, have potential to be integrated into biorefineries. In a microbial electrolysis cell, organic compounds are converted to electrons, protons, and CO₂ by fermentative and exoelectrogenic bacteria in the anode compartment. By having the ability to extract electrons from waste streams, these systems can treat water while also producing hydrogen, and thus can improve the efficiency of biomass to fuel production by minimizing external hydrogen requirement and enabling water recycle. The overall goal of this …

Bioremediation Of Chlorinated Ethenes: Ph Effects, Novel Dechlorinators And Decision-Making Tools, Yi Yang

Doctoral Dissertations

Chlorinated solvents have been widely used in different areas of modern society. Usage of these chlorinated solvents was not necessarily accompanied with proper handling and disposal of these hazardous compounds, which caused a variety of environmental problems and continues to affect human health. Remediation of chlorinated ethenes contaminated sites has high priority for state regulators and site owners. Among the available treatment technologies, bioremediation shows great promise as a cost-effective corrective strategy for a variety of environmental pollutants. Prerequisites are that the microbiology involved in contaminant degradation and geochemical factors, such as pH, are understood, so that bioremediation technologies can …

Electron Transport To Photosystem I By Soluble Carriers: Evolution Of The Interacting Pair, Khoa Dang Nguyen

Doctoral Dissertations

Oxygenic photosynthesis is driven via sequential action of Photosystem II (PSII) and (PSI) reaction centers via the Z-­‐scheme. Both of these pigment– membrane protein complexes are found in cyanobacteria, algae, and plants. PSI, unlike PSII, is remarkably stable and does not undergo limiting photo-­‐damage. This stability, as well as other fundamental structural differences, makes PSI the most attractive reaction centers for applied photosynthetic applications. These applied applications exploit the efficient light harvesting and high quantum yield of PSI where the isolated PSI particles are redeployed providing electrons directly as a photocurrent or, via a coupled catalyst to yield H2. Here, …

Modification Of Carbohydrate Active Enzymes In Switchgrass (Panicum Virgatum L.) To Improve Saccharification And Biomass Yields For Biofuels, Jonathan Duran Willis

Doctoral Dissertations

The natural recalcitrance of plant cell walls is a major commercial hurdle for plant biomass to be converted into a viable energy source as alternative to fossil fuels. To circumvent this hurdle manipulation of carbohydrate enzymes active in the cellulose and hemicellulose portions of the plant cell wall can be utilized to improve feedstocks. Production of cellulolytic enzymes by plants have been evaluated for reducing the cost associated with lignocellulosic biofuels. Plants have successfully served as bioreactors producing bacterial and fungal glycosyl hydrolases, which have altered plant growth to improve saccharification. A bioprospecting opportunity lies with the utilization of insect …

Nanocomposite Adhesive Of English Ivy (Hedera Helix): Bioproduction, Nanoparticle Isolation, And Molecular Analysis, Jason Neil Burris

Doctoral Dissertations

English ivy naturally produces organic nanoparticles from its adventitious root hairs, and possesses characteristics that may allow them to replace metal-based nanoparticles in common applications, such as sunscreen. At the onset of the research, it was hypothesized that a physical mechanism of attachment, similar to the gecko footpad, was used to generate the adhesive force for attachment; however, through the results obtained from recent work, it is clear that a biochemical mechanism is involved in the generation of the strength of adhesion. Therefore, the goal of this research was to provide a better understanding of the genetic basis of nanoparticle …

Biochemistry And Evolution Of The Phytohormone-Methylating Sabath Methyltransferase In Plants, Minta Chaiprasongsuk

Doctoral Dissertations

Known members of Phytohormone-methylating compounds are plant synthesis compounds that serve as attractants of other living organisms beneficial to the plants or as defense against other biotic as well as abiotic agents. To increase their fitness and survival in a stressful environment plants produce distinct sets of phytohormone-methylating compounds. Plant genomes can encode the necessary enzymes to acquire the ability to make new specialized compounds during evolution. This dissertation aims to investigate the biochemical and biological functions and evolution of SABATH genes in different lineages of plants. Black cottonwood, Brachypodium and Norway spruce genome were used as the model for …

Engineering Photosystem I Complexes For Use In Bio-Hybrid Dye-Sensitized Solar Cells, Richard Franklin Simmerman

Doctoral Dissertations

Increasing global population, growing per capita energy needs, diminishing fossil fuels, and climate change collectively will require new, innovative, and sustainable alternatives to meet the world’s growing energy needs. One of the most promising yet simple approaches are dye-sensitized solar cells (DSSCs). However, conventional DSSCs use semi-conductor anodes sensitized with complex synthetic organometallic dyes. Most dyes utilize ruthenium complexes to absorb photons, which upon excitation, inject electrons into the anode, while holes migrate to the cathode via liquid electrolyte. However, these dyes are expensive, difficult to make, and resource-limited. This dissertation focuses on replacing synthetic dyes with the naturally occurring, …

Genetic Modification Of Switchgrass (Panicum Virgatum L.) For Improvement Of Plant Architecture, Biomass Productivity And Sugar Release Efficiency For Biofuel, Wegi Aberra Wuddineh

Doctoral Dissertations

Switchgrass (Panicum virgatum L.) is a leading candidate bioenergy crop for sustainable biofuel production. To ensure its economic viability, tremendous improvements in switchgrass biomass productivity and recalcitrance to enzymatic saccharification are needed. Genetic manipulation of lignin biosynthesis by targeting transcriptional regulators of higher level domains of lignin biosynthesis and other complex traits could alter several bioenergy-desirable traits at once. A three-pronged approach was made in the dissertation research to target one plant growth regulator and transcription factors to alter plant architecture and cell wall biosynthesis.

Gibberellin (GA) catabolic enzymes, GA 2-oxidases (GA2oxs), were utilized to alternatively modify the lignin …

Glyr3 Regulation In Clostridium Thermocellum, Jinlyung Choi

Doctoral Dissertations

Bio-ethanol from cellulosic biomass is a promising candidate as a liquid transportation fuel because of its high-energy content and the abundance of cellulose. Consolidated bioprocessing (CBP) helps to reduce the traditional 2-step process of bio-ethanol production into single-step to improve cost efficiency. A bacterium, Clostridium thermocellum, has a multi-enzyme complex for hydrolyzing cellulase, called the Cellulosome that enables the organism to have high rates of cellulose utilization. However, the ethanol yield of C. thermocellum needs to be improved in order to make consolidated bioprocessing with C. thermocellum commercially viable. It is essential to understand the regulation of carbohydrate-degrading enzyme …

Applications And Improvements In The Molecular Modeling Of Protein And Ligand Interactions, Jason Bret Harris

Doctoral Dissertations

Understanding protein and ligand interactions is fundamental to treat disease and avoid toxicity in biological organisms. Molecular modeling is a helpful but imperfect tool used in computer-aided toxicology and drug discovery. In this work, molecular docking and structural informatics have been integrated with other modeling methods and physical experiments to better understand and improve predictions for protein and ligand interactions. Results presented as part of this research include:

1.) an application of single-protein docking for an intermediate state structure, specifically, modeling an intermediate state structure of alpha-1-antitrypsin and using the resulting model to virtually screen for chemical inhibitors that can …

Toward Direct Biosynthesis Of Drop-In Ready Biofuels In Plants: Rapid Screening And Functional Genomic Characterization Of Plant-Derived Advanced Biofuels And Implications For Coproduction In Lignocellulosic Feedstocks, Blake Lee Joyce

Doctoral Dissertations

Advanced biofuels that are “drop-in” ready, completely fungible with petroleum fuels, and require minimal infrastructure to process a finished fuel could provide transportation fuels in rural or developing areas. Five oils extracted from Pittosporum resiniferum, Copaifera reticulata, and surrogate oils for Cymbopogon flexuosus, C. martinii, and Dictamnus albus in B20 blends were sent for ASTM International biodiesel testing and run in homogenous charge combustion ignition engines to determine combustion properties and emissions. All oils tested lowered cloud point. Oils derived from Copaifera reticulata also lowered indicated specific fuel consumption and had emissions similar to the ultra-low sulfur diesel control. Characterization …

Femtosecond Laser Patterned Templates And Imprinted Polymer Structures, Deepak Rajput

Doctoral Dissertations

Femtosecond laser machining is a direct-write lithography technique by which user-defined patterns are efficiently and rapidly generated at the surface or within the bulk of transparent materials. When femtosecond laser machining is performed with tightly focused amplified pulses in single-pulse mode, transparent substrates like fused silica can be surface patterned with high aspect ratio (>10:1) and deep (>10 μm) nanoholes. The main objective behind this dissertation is to develop single-pulse amplified femtosecond laser machining into a novel technique for the production of fused silica templates with user-defined patterns made of high aspect ratio nanoholes. The size of the …

Characterization Of Field Evolved Resistance To Transgenic Cry1fa Maize In Spodoptera Frugiperda (J. E. Smith), Siva Rama Krishna Jakka

Doctoral Dissertations

Transgenic Bt crops expressing Cry and Vip toxins from Bacillus thuringiensis (Bt) have been increasingly planted to manage insect pest damage on agricultural crops. The high adoption of Bt-based insecticidal technologies suggests an increase selection pressure for the evolution of resistance in insect populations. So far, nine insect species have developed field evolved resistance to Bt crops, yet the mechanisms involved in field evolved resistance are unknown. In the present study, the resistance mechanism in field evolved resistance to maize producing Cry1Fa in Spodoptera frugiperda collected in fields from Puerto Rico was characterized. High levels of resistance to …

Optimization Of Bacterial Bioluminescence (Lux) Expression And Development Of Autonomous Lux-Based Reporters In Human Cell Lines, Tingting Xu

Doctoral Dissertations

Despite its extensive use as a prokaryotic bioreporter, only recently has the bacterial bioluminescence (lux) system been adapted to express at a functional level in the mammalian cellular background. While this novel strategy has the potential to contribute to the fields of high throughput screening and non-invasive in vivo imaging due to its autonomous substrate production, it is still constrained because its signal intensity is lower than that of other bioluminescent reporters. This work demonstrates the development of strategies that optimize human cell lux-based bioluminescence to overcome this detriment for advancement towards a fully functional lux reporter …

Engineering Transcriptional Control And Synthetic Gene Circuits In Cell Free Systems, Sukanya Iyer

Doctoral Dissertations

Engineering gene networks offers an opportunity to harness biological function for biotechnological and biomedical applications. In contrast to cell-based systems, cell free extracts offer a flexible and well-characterized context in which to implement predictable gene circuits. Critical to these efforts is the availability of a library of ligand sensitive gene regulatory systems. Here, I describe efforts to develop molecular tools to control gene expression and implement a negative feedback circuit in E.coli cell extracts. First, a strategy to regulate T7 RNA polymerase using DNA aptamers is detailed. I test the hypothesis that a DNA aptamer, when placed near the transcription …

Synthesis, Characterization And Self-Assembly Of Amphiphilic Block Copolymers, Xiaojun Wang

Doctoral Dissertations

This dissertation presents a review on state-of-the-art research of well-defined charged block copolymers, including synthesis, characterization, bulk morphology and self-assembly in aqueous solution of amphiphilic block polyelectrolytes. In Chapter 1, as a general introduction, experimental observations and theoretical calculations devoted towards understanding morphological behavior in charged block copolymer systems are reviewed along with some of the new emerging research directions. Further investigation of charged systems is urged in order to fully understand their morphological behavior and to directly target structures for the tremendous potential in technological applications. Following this background, in Chapters 2, 3, 4 and 5 are presented the …

Measuring Single Nucleotide Polymorphism In The Vegetable Pathogen Phytophthora Capsici, Daniel Jara Gobena

Doctoral Dissertations

Phytophthora capsici is a eukaryotic plant pathogen that causes root, fruit, and foliar disease on a variety of important vegetables including pepper, tomato, eggplant, snap and lima beans, and essentially all cucurbits. At some locations, populations utilize sexual and asexual reproduction, whereas at others, populations appear to be entirely asexual. The differing population structure has important implications for developing control strategies. Our objective was to develop SNP markers to characterize natural populations and laboratory crosses. In chapter two, a novel technique to assay SNP genotypes based on high-resolution DNA melting analysis is presented. In chapter three, the genetic diversity for …

Comparing The Midgut Regenerative Responses In Bacillus Thuringiensis-Susceptible And Resistant Heliothis Virescens Larvae, Anais Severiana Castagnola

Doctoral Dissertations

The crystal (Cry) toxins from Bacillus thuringiensis (Bt) display high specificity and toxicity against relevant insect pests and the use of Bt-based products continues to contribute to insect pest management. To protect this investment, further its potential, and investigate possible unintended effects, various research questions have been proposed. One issue related to Bt usage is the evolution of pest resistance to Bt toxins. The midgut epithelium is targeted by Cry toxins killing enterocytes, facilitating invasion of the hemocoel, leading to septicemia and mortality. While resistance may emerge from alterations to these steps, most research efforts have been focused on reduced …