Life Sciences Commons^™

Genetic Engineering Of Black Cherry (Prunus Serotina) For Reproductive Sterility And Insect Pest Resistance, Ying Wang

Open Access Dissertations

Black cherry (Prunus serotina Ehrh.) is one of the most valuable hardwoods for high- end cabinetry, furniture, architectural millwork, paneling, and veneer. However, the damage caused by cambial-mining insect pests triggers gummosis in black cherry, a non-specific defense response in which resinous gum is deposited at the site of injury. The gum defects dramatically decrease the yield of high-quality black cherry lumber, and the value can be reduced by as much as 90%. The goal of this project was to optimize the transformation and in vitro rooting system, and to develop transgenic black cherry for reproductive sterility and insect pest …

Gas-Phase Covalent And Non-Covalent Ion/Ion Chemistry Of Biological Macromolecules, John Robert Stutzman

Open Access Dissertations

Gas-phase ion/ion chemistry involves the interaction of oppositely charged ions inside of the mass spectrometer. During this gas-phase chemistry, particle transfer (i.e., proton and electron) or synthesis can occur at rapid reaction rates. Particle transfer represents a mature area of ion/ion chemistry, while selective covalent modification represents a fairly new area of gas-phase chemistry. Gas-phase covalent chemistry is based on traditional solution phase organic chemistry.

The work demonstrated in this dissertation greatly involves gas-phase covalent and non-covalent Schiff base chemistry on peptide and protein ions. The reagent dianion, 4-formyl 1,3-benzene disulfonic acid, has been used to covalently modify unprotonated primary …

Measuring And Modeling The Response Characteristics Of The Environmental Phosphate Transducer In Escherichia Coli, Chetan Sood

Open Access Dissertations

The PhoR/PhoB two-component system in Escherichia coli is a biological transducer that senses the limitation of environmental inorganic orthophosphate, the bacteria's preferred source of the essential nutrient phosphate, and transmits that information to the interior of the cell initiating a response that mitigates phosphate starvation. In the first part of this study, we present and apply a fluorescence microscopy technique to measure, in vivo, the dynamic response characteristics of the transducer with single-cell resolution. We report that the transience in the PhoR/PhoB TCS response is consistent with the transducer having a threshold sensitivity to the concentration of environmental phosphate, …

Estimation Of Variation For High-Throughput Molecular Biological Experiments With Small Sample Size, Danni Yu

Open Access Dissertations

Motivation: In the quantification of molecular components, a large variation can affect and even potentially mislead the biological conclusions. Meanwhile, the high-throughput experiments often involve a small number of samples due to the limitation of cost and time. In such cases, the stochastic information may dominate the outcome of an experiment because there may not be enough samples to present the true biological information. It is challenging to distinguish the changes in phenotype from the stochastic variation.

Methods: Since the biological molecules have been quantified with different technologies, different statistical methods are required. Focusing on three types of important high-throughput …

Genes With Physiological Roles In Callipyge Muscle Hypertrophy, Hui Yu

Open Access Dissertations

Callipyge sheep is an excellent model to study genes regulation in muscle growth since the up-regulation of DLK1 and/or RTL1 results in extreme postnatal muscle hypertrophy in loin and hindquarters. DLK1 and/or RTL1 are the primary inducers of muscle hypertrophy due to the inheritance model for the callipyge phenotype. The overall aim of this dissertation is to study the physiological pathways responding to the up-regulation of DLK1 and / or RTL1 in the hypertrophied muscles. Microarray analysis of gene expression identified 375 genes that were differentially expressed in callipyge semimembranosus. Twenty-five transcripts were further verified by quantitative PCR in two …

Functional Characterization Of Metallothioneins In Arabidopsis And Barley, Nimnara Yookongkaew

Open Access Dissertations

Metalothioneins (MTs) are metal binding proteins that can bind metals such as Cu, Zn, and Cd but the functions of MTs in plants are largely unknown. To understand the function of MTs in Arabidopsis, T-DNA insertion mutants lacking 4 MT genes including MT1a, MT2a, MT2b and MT3 were developed. The quadruple mutant (mt1a/mt2a/mt2b/mt3) showed no visible phenotype under standard growth conditions. However, it accumulated higher Cu in leaves compared to wild type but lower Cu in seeds. Further analysis showed that triple mutants lacking both MT1a and MT2b, which are highly expressed in vascular tissues, had similar …

The City In Mind: Environmental Literacy And Adaptation In Nineteenth-Century British Literature, Adam Edward Watkins

Open Access Dissertations

This dissertation argues that a new paradigm of selfhood emerged in nineteenth-century British literature, one that recognized the individual will and environmental influence not as antithetical but as dialectical forces in the formation of the self. The concept of an externally negotiated subject challenges both the inward and socially determined conceptions of self that have dominated the relevant criticism. Informed by empiricist, associationist, and evolutionary theories of the mind, the portrayals of subject-formation in this study highlight the radical changes occurring in the human environment in nineteenth-century, which catalyzed the conception of a malleable yet self-forming subject. Along with the …

Statistical Models For Gene And Transcripts Quantification And Identification Using Rna-Seq Technology, Han Wu

Open Access Dissertations

RNA-Seq has emerged as a powerful technique for transcriptome study. As much as the improved sensitivity and coverage, RNA-Seq also brings challenges for data analysis. The massive amount of sequence reads data, excessive variability, uncertainties, and bias and noises stemming from multiple sources all make the analysis of RAN-Seq data difficult. Despite much progress, RNA-Seq data analysis still has much room for improvement, especially on the quantification of gene and transcript expression levels. The quantification of gene expression level is a direct inference problem, whereas the quantification of the transcript expression level is an indirect problem, because the label of …

Biophysical Studies Of Cholesterol In Unsaturated Phospholipid Model Membranes, Justin Adam Williams

Open Access Dissertations

Cellular membranes contain a staggering diversity of lipids. The lipids are heterogeneously distributed to create regions, or domains, whose physical properties differ from the bulk membrane and play an essential role in modulating the function of resident proteins. Many basic questions pertaining to the formation of these lateral assemblies remain. This research employs model membranes of well-defined composition to focus on the potential role of polyunsaturated fatty acids (PUFAs) and their interaction with cholesterol (chol) in restructuring the membrane environment. Omega-3 (n-3) PUFAs are the main bioactive components of fish oil, whose consumption alleviates a variety of health problems by …

Effects Of Dip-Coated Films On The Properties Of Implantable Intracortical Microelectrodes, Salah Sommakia

Open Access Dissertations

The successful clinical use of implantable intracortical microelectrodes (ICMs) to treat certain types of deafness, blindness, and paralysis is limited by a reactive tissue response (RTR) of the brain. This RTR culminates in the formation of a tight glial scar and a loss of neuronal density around implanted ICMs, and is accompanied by a decrease in signal to noise ratio and an increase in impedance. While no comprehensive mechanistic understanding of the underlying biology is currently agreed upon in the field, a general consensus exists around a highly volatile acute RTR phase. During this acute phase, the electrical properties of …

Effects Of Amur Honeysuckle (Lonicera Maackii [Rupr.] Herder) Invasion And Removal On Native Vegetation And White-Footed Mice (Peromyscus Leucopus) In Mixed-Hardwood Forests Of Indiana, Joshua Michael Shields

Open Access Dissertations

The threat of non-native invasive species continues to compromise the ecological and economic integrity of our natural resources. Numerous investigators have documented the negative effects of invasive species on native biota. However, much work is still needed with regard to how invasive species spread in space and time, factors contributing to impacts on native biota within invaded ecosystems, and resultant effects of removing invasive species. In terms of invasion patterns, few studies have documented local patterns and rates of woody plant invasions, and even less is known about changes in spatial patterning and factors influencing structural characteristics (diameter and height) …

Development Of Isotags For Nmr Based Metabolite Profiling And Applications, Fariba Tayyari

Open Access Dissertations

NMR spectroscopy is a powerful analytical tool for both qualitative and quantitative metabolite profiling analysis. However, accurate quantitative analysis of biological systems especially using one dimensional NMR has been challenging due to signal overlap. In contrast, the enhanced resolution and sensitivity offered by chemoselective isotope tags have enabled new and enhanced methods for detecting hundreds of quantifiable metabolites in biofluids using NMR spectroscopy or mass spectrometry. In this thesis we show improved sensitivity and resolution of NMR experiments imparted by 15N and 13C isotope tagging which enables the accurate analysis of plasma metabolites. To date, isotope tagging has been used …

Synthesis Of Novel Isoprenoid Diphosphate Analogs As Chemical Tools To Investigate Protein Geranylgeranylation, Kayla Jo Temple

Open Access Dissertations

Many proteins require prenylation in order to be biologically functional. Some such proteins include the small Ras and Rho GTPase superfamilies, nuclear lamins A and B, and the kinesin motor proteins CENP-E and F. Prenyltransferase (PTase) inhibition is currently being explored as a possible treatment not only for cancer but for a wide variety of other diseases.

Clinical studies revealed that the effectiveness of farnesyltransferase inhibitors (FTIs) to treat Ras-dependent tumors is determined by which isoform of Ras is overactive. Unfortunately the majority of Ras-dependent tumors have a mutation in either the N- or K-Ras isoforms; both of these isoforms …

Characterization Of Caxck31, A Bacterial Calcium/Proton Antiporter, Marc Robert Ridilla

Open Access Dissertations

To better understand a class of transporters known as Calcium/Cation Antiporters (CaCAs), the bacterial calcium/proton antiporter CAXCK31 was purified and characterized. New methods were developed for its heterologous overexpression and purification. These methods help to define stress responses to toxic membrane overproduction in E. coli and may be broadly applicable to studies of membrane proteins. The results from a variety of biochemical and biophysical experiments demonstrated that CAXCK31 exists as a dimer in the membrane and can be purified in the dimeric state. The methods used include chemical cross-linking, FRET, and SEC-MALS. In addition, various transport properties of CAXCK31, including …

Quantifying Crop Yield, Bioenergy Production And Greenhouse Gas Emissions From Cropland And Marginal Land Using A Model-Data Fusion Approach, Zhangcai Qin

Open Access Dissertations

Bioenergy is becoming increasingly attractive to many countries, but has sparked an intensive debate regarding energy, economy, society and environment. Biofuels provide alternative energy to conventional fossil fuels. However, the environmental impact of producing and using biofuel is a major concern to our society. This study is dedicated to quantifying and evaluating biofuel production and potential climate change mitigation due to potential large-scale bioenergy expansion in the conterminous United States, using model-data fusion approaches.

Biofuel made from conventional (e.g., maize (Zea mays L.)) and cellulosic crops (e.g., switchgrass (Panicum virgatum L.) and Miscanthus (Miscanthus × giganteus)) …

Welfare Impacts Of False Codling Moth Threatening California Oranges, Azza Mohamed Kamal Ahmed Mohamed

Open Access Dissertations

Welfare impacts of alternative pest management strategies of False Codling Moth (FCM) threatening California's oranges are examined. Different economic agents along the supply chain of fresh oranges and orange products in the United States are considered, including consumers, retailers, wholesalers, and growers. A partial equilibrium dynamic framework that accounts for supply response from the other US orange producing states is developed. Data for supply shocks (orange yield losses and control costs) are obtained from the Animal and Plant Health Inspection Service of the United States Department of Agriculture (APHIS).

FCM is not presently in the United States. If introduced to …

Invasive Silver Carp (Hypophthalmichthys Molitrix) Protein Hydrolysates- A Potential Source Of Natural Antioxidant, Sravanthi Priya Malaypally

Open Access Dissertations

Invasive silver carp (Hypophthalmichthys molitrix), continue to spread over the Mississippi River causing a great concern for the river ecosystem due to their impact on native fish species. To minimize the negative effects of silver carp, many strategies were implemented including using it for animal feed, as fertilizers or simply discarding them into waste. However, these fish are high in protein content, making them excellent starting material for protein-derived by-products. One alternative is to recover and modify the protein in the fish into bioactive food ingredient. In the functional/ nutraceutical food industry, there is high demand for antioxidant agents. In …

Structural Studies On The Rubella Virus Capsid Protein And Its Organization In The Virion, Vidya Mangala Prasad

Open Access Dissertations

Rubella virus is a leading cause of birth defects due to infectious agents. When contracted during pregnancy, rubella infection leads to severe damage in fetuses. Despite its medical importance, very little is known about the structure of the pleomorphic rubella virus as compared to its alphavirus relatives. The rubella capsid protein is a critical structural component of virions as well as a key factor in virus-host interactions. Three crystal structures of the structural domain of the rubella capsid protein have been described here. The polypeptide fold of the capsid protomer has not been observed previously. The capsid protein structure, along …

Notchless Interacts With Multiple Signaling Pathways During Mouse Peri-Implantation Development, Chiao-Ling Lo

Open Access Dissertations

During peri-implantation, the embryo transitions from a suspension environment in the fallopian tubes to an adherent system within the uterus. Successful transition requires maternal and fetal signaling cascades that establish maternal-fetal boundaries. Failure is common, as ~ 25% of all human pregnancies terminate during these steps. A large-scale mutation study in mice produced two mutants (l11Jus1 and l11Jus4) that are excellent models of this transition. l11Jus1 and l11Jus4 contain missense mutations in the Notchless homolog 1 (Drosophila) (Nle1) gene. NLE1 is thought to signal via the canonical NOTCH pathway in vertebrates. Although in invertebrates and lower vertebrates, NOTCH signaling directs …

Three Essays On The Interaction Between Global Trade And Greenhouse Gas Mitigation Agreements, Zeynep Burcu Irfanoglu

Open Access Dissertations

The role of trade sanctions in enforcing greenhouse gas mitigation agreements is the fundamental theme of the three essays comprising this dissertation. All three essays employ a multi-sector, multi-region computable general equilibrium model, GTAP-AEZ-GHG, documented in Golub et al. (2009) to investigate how the United States can use trade sanctions as an enforcement mechanism in a global greenhouse gas (GHG) mitigation agreement. The focus is placed on inducing China to comply with the global agreement.

The first essay contributes to the body of knowledge on global GHG emissions mitigation agreements by investigating the range of emission taxes that can be …

Development Of Tyrosine Kinase Peptide Biosensors And Methods For Detection, Andrew Michael Lipchik

Open Access Dissertations

New methods to monitor tyrosine kinase activity are critical for studying kinases in cell biology, drug discovery and the clinic. Peptide-based biosensors for detection of kinase activity utilitize a kinase specific artificial peptide substrate, which can report intercellular kinase activity through the incorporation of phosphate.

An artificial Syk substrate peptide was developed and incorporated with other functional modules to produce a Syk biosensor. These modules included a biotin-tag for affinity capture, a photo-cleavable amino acid to allow release of the substrate from the delivery module and the cell penetrating peptides TAT. A live cell kinase assay utilizing this biosensor was …

Hybrid Opto-Electrokinetic Technique For Micro/Nanomanipulation: Towards Application Of A Novel Non-Invasive Manipulation Technique In Microbiological Assay, Jae-Sung Kwon

Open Access Dissertations

This dissertation explores various physical mechanisms of the Rapid Electrokinetic Patterning (REP) technique suggested for rapid and precise on-chip manipulation of colloids and fluids, and bio-compatibility of the technique for biological applications. REP is a hybrid opto-electrokinetic technique that is driven by the simultaneous application of an AC electric field and a heating source. It can not only effectively transport and manipulate a fluid but also concentrate and pattern particles suspended in the fluid through the combined effect of an electrohydrodynamic flow, electrostatic colloidal interactions and an electrothermal microfluidic flow. These capabilities make REP a promising tool which can provide …

Establishing The Role Of The Pancreatic Transcription Factor Mist1 In Xbp1-Mediated Maintenance Of Pancreatic Acinar Cell Homeostasis, David Alan Hess

Open Access Dissertations

Pancreatic acinar cells (PACs) continuously produce more protein than any other cell type in the human body. As a result, PACs and other specialized secretory cells have a constant demand placed on their protein synthetic and packaging machinery. When demand for secreted products exceeds the capacity of the cell's basal protein production facilities, dangerous accumulations of misfolded proteins can build up, resulting in a condition known as ER stress. To ameliorate this stress, secretory cells activate a coordinated, three-part compensatory network collectively known as the unfolded protein response (UPR) to both expand the capacity of the ER and directly assist …

Generation And Statistical Modeling Of Active Protein Chimeras: A Sequence Based Approach, Nicholas Fico

Open Access Dissertations

Generation of active protein chimeras is a valuable tool to probe the functional space of proteins. Statistical modeling is the next logical step, allowing us to build a model of gene fragment replaceability between species. In this thesis I begin to develop the statistical tools that are needed to systematically describe combinatorial protein libraries. I present three sets of diverse chimeric protein libraries developed using sequence information. The statistical model of the human N-Ras and human K-Ras-4B genes reveal a set previously unidetifed surface residues on the N-Ras G-Domain that may be involved in cellular localization. Statistical modeling of a …

Ozonation Systems As A Non-Chemical Alternative For Stored Grain Protection, Carlos A. Campabadal

Open Access Dissertations

The use of ozone as a non-chemical alternative in stored grain protection was studied by conducting scale-up demonstrations using a fixed bed ozonation system and developing a semi-continuous counterflow and a continuous flow ozonation treatment system. The objectives of this research were to determine the efficacy of ozonation to control insect pests without affecting end-use quality; to prove the concept of the semi-continuous counterflow ozonation system to ozonate grain at a faster rate and quantify its effect on mold growth reduction; to evaluate the efficacy of a modified screw conveyor for pest control by treating grain in a continuous-flow ozonation …

Observation-Based Algorithm Development For Subsurface Hydrology In Northern Temperate Wetlands, Chun-Mei Chiu

Open Access Dissertations

This study investigates wetland subsurface hydrology, as well as biogeochemistry - which is strongly influenced by water and temperature dynamics - as these interactions are expected to be highly significant, yet remain poorly represented in current ecosystem and climate models.

Northern wetlands have received widespread public attention due to steadily increasing summer mean global temperatures, extreme precipitation events and higher rates of natural greenhouse gas emissions, as well as the significant impacts on them due to human activities. The goal of my graduate research has been to improve quantification of the role of subsurface hydrology in northern wetlands by using …

An Analysis Of The Impact Of Storage Temperature, Moisture Content & Duration Upon The Chemical Components & Bioprocessing Of Lignocellulosic Biomass, Arun Athmanathan

Open Access Dissertations

The successful utilization of lignocellulosic biomass as a feedstock for fuels and chemicals necessitates storage for 2-6 months. It is correspondingly important to understand the impact of storage parameters - moisture concentration, temperature and duration - on biomass quality.

As aerobic storage is the most viable large-scale solution, aerobic storage experiments were carried out with three projected bioenergy feedstocks - sweet sorghum (Sorghum bicolor) bagasse, corn (Zea mays) stover and switchgrass (Panicum virgatum). Stored samples of each were examined for dry matter loss and composition change to develop a material balance around carbohydrates and lignin.

A mean dry matter loss …

Neuroprotection By Chitosan And Chitosan Nanoparticles, Bojun Chen

Open Access Dissertations

In the U.S., about 200,000 people are currently living with spinal cord injury (SCI). An estimated of 50%-70% of all SCI cases occurs in the range of ages between 15-35 years old. The destructive neurotrauma results in the majority of adult disability, even after patients suffering with SCI survived from the acute death. There are two stages involved in the progression of SCI, the primary stage and the secondary stage. The primary stage is mainly the mechanical damage to the central nervous system. The rapid collapse of the integrity of cell membrane and tissue is often one of the initial …

Understanding Osmoregulation And Immunity In Heteromyid Rodents Through Insights Gained From Rna-Seq, Nicholas John Marra

Open Access Dissertations

Understanding the genetic basis of adaptation is one of the central questions of evolutionary genetics. In the face of selective pressure, individuals with adaptive phenotypes survive and pass on their beneficial alleles that conferred this fitness advantage. Over sufficient time scales and under strong selection, differences accumulate between taxa in the form of physical and/or behavioral traits that have been shaped differently. By looking at the transcriptomes of related taxa we can identify sequence and regulatory differences that represent molecular signatures of these different selection regimes. When we study these differences between taxa that have evolved under opposing selective forces …

Fate Of 17Α-Estradiol, 17Β-Estradiol, And Estrone In Agricultural Soils And Sediments, Michael L. Mashtare Jr

Open Access Dissertations

The shift to concentrated animal production facilities and increasing rural-urban migration has increased the localized land application of nearly 1 billion tons of manure and biosolids annually. Although these applications provide nutrients and contribute to soil tilth, they also serve as a source for an estimated 49 tons of the natural manure-borne estrogens, 17α-estradiol (17α-E2), 17β-estradiol (17β-E2), and estrone (E1). While these estrogens are critical to endocrine systems, the low concentrations observed in the environment can disrupt endocrine function in non-target organisms, e.g., altering secondary sex characteristics which can lead to changes in wildlife communities.

Research presented here focuses on …