Digital Commons Network^™

Engineering Proteins With Unique Characteristics For Diagnostics And Biosensors, Smita Joel

University of Kentucky Doctoral Dissertations

Proteins possess a broad range of structural and functional properties and, therefore, can be employed in a variety of biomedical applications. While a good number of protein-based biosensing systems and biosensors for target analytes have been developed, the search for versatile, highly sensitive and selective sensors with long term stability able to provide fast detection of target analytes continues to be a challenge. To that end, we now report the design and development of modified proteins with tailored characteristics and their further utilization in the development of biosensing systems.

We take advantage of binding proteins that undergo a change in …

Effects Of Corticosterone And Ethanol Co-Exposure On Hippocampal Toxicity: Potential Role For The Nmda Nr2b Subunit, Tracy Renee Butler

University of Kentucky Doctoral Dissertations

Chronic ethanol (EtOH) exposure produces neuroadaptations within the NMDA receptor system and alterations in HPA axis functioning that contribute to neurodegeneration during ethanol withdrawal (EWD). Chronic EtOH exposure and EWD, as well as corticosteroids, also promote increased synthesis and release of polyamines, which allosterically potentiate NMDA receptor open-channel time at the NR2B subunit. The current studies investigated effects of 10 day EtOH and corticosterone (CORT) co-exposure on toxicity during EWD in rat organotypic hippocampal slice cultures, and alterations in function and/or density of the NR2B subunit of the NMDA receptor that may mediate CORT-potentiation of toxicity during EWD. We hypothesized …

The Role Of The Nr4a Orphan Nuclear Receptor Nor1 In Vascular Cells And Atherosclerosis, Yue Zhao

University of Kentucky Doctoral Dissertations

The neuron-derived orphan receptor 1 (NOR1) belongs to the NR4A nuclear receptor subfamily. As an immediate early response gene, NOR1 is rapidly induced by a broad spectrum of physiological and pathological signals. Functional studies demonstrate NOR1 as a constitutively active ligand-independent nuclear receptor whose transcriptional activity is dependent on both expression level and posttranslational modifications. To date, an increasing number of studies have demonstrated a pivotal role of NOR1 in the transcriptional control of metabolism and the development of cardiovascular diseases.

In this dissertation, we demonstrate NOR1 expression in endothelial cells and sub-endothelial cells of human atherosclerotic lesions. In response …

Gene Expression Regulators Lin-11 And Let-711, In Modulating The Rate Of Aging And Lifespan, In C. Elegans., Tseten Yeshi Jamling

University of Kentucky Doctoral Dissertations

lin-11 and let-711 are early-developmental gene expression regulators with no previously known roles in aging regulation. Yet, they show strong aging-correlated expression profiles (Lund, Tedesco et al. 2002). lin-11 is strongly upregulated in very old worm populations, and let-711 is progressively downregulated in aging worm populations. Microarray studies were performed to identify their genome-wide targets, which were then subjected to further lifespan and genetic analysis to investigate their role in C. elegans aging.

The results indicate that the target pools of both lin-11 and let-711 are enriched for aging genes, since a significant number of tested genes increased lifespan. This …

The Ecology Of Disturbances And Global Change In The Montane Grasslands Of The Nilgiris, South India, Madhusudan P. Srinivasan

University of Kentucky Doctoral Dissertations

Biodiversity rich regions worldwide face threats from various global change agents. This research quantifies environmental influences on vegetation, and the impacts of exotic woody plant invasion and anthropogenic nitrogen (N) deposition in a global biodiversity hotspot. The study was conducted in the montane grasslands of the Nilgiris, Western Ghats, and outlines potential management options for this region. Specifically, I examined (1) the role of environmental factors in influencing native plant distribution and ecosystem properties, (2) the status and impact of exotic shrub (Scotch broom, henceforth broom) invasion, (3) the role of disturbances in the success of broom, (4) the role …

Evaluation And Enhancement Of Seed Lot Quality In Eastern Gamagrass [Tripsacum Dactyloides (L.) L.], Cynthia Hensley Finneseth

University of Kentucky Doctoral Dissertations

Eastern gamagrass [Tripsacum dactyloides (L.) L.] is a warm-season, perennial grass which is native to large areas across North America. Cultivars, selections and ecotypes suitable for erosion control, wildlife planting, ornamental, forage and biofuel applications are commercially available. Plantings are commonly established from seed; however, seed quality and dormancy are barriers to establishment. The objective here was to investigate parameters that contribute to inconsistent performance of this seed kind to subsequently improve seed lot quality.

Forty-two seed lots were used, with seed fill and germination potential ranging from 78- 100% and 11-91%, respectively. Actual germination and dormant seed ranged …

Analysis Of The Crmp Gene In Drosophila: Determining The Regulatory Role Of Crmp In Signaling And Behavior, Deanna Hardt Morris

University of Kentucky Doctoral Dissertations

The mammalian genome encodes five collapsin response mediator protein (CRMP) isoforms. Cell culture studies have shown that the CRMPs mediate growth cone dynamics and neuron polarity through associations with a variety of signal transduction components and cytoskeletal elements. CRMP is also a member of a protein family including the presumably ancestral dihydropyrimidinase (DHP) protein that catalyzes the second step in pyrimidine degradation. In Drosophila, CRMP and DHP proteins are produced by alternatively spliced transcripts of the CRMP gene. The alternative protein forms have a 91% sequence identity, but unique expression patterns. CRMP is found exclusively in neuronal tissues and …

Environmental Effects On Behavior And Physiology In Crayfish, Sonya M. Bierbower

University of Kentucky Doctoral Dissertations

Despite dramatic morphological differences between animals from different taxa, several important features in organization and sensory system processing are similar across animals. Because of this similarity, a number of different organisms including mammals, insects, and decapod crustaceans serve as valuable model systems for understanding general principles of environmental effects. This research examines intrinsic and extrinsic factors by behaviorally and physiologically means to identify the impact of environmental conditions on two distinct crayfish species- Procambarus clarkii (surface) and Orconectes australis packardi (cave).

The research identified behavioral and physiological responses in these two morphological and genetically distinct species. The studies also examined …

Substrate And Regulation Of Mitochondrial Μ-Calpain, Aashish Joshi

University of Kentucky Doctoral Dissertations

μ -Calpain is localized to the mitochondrial intermembrane space. Apoptosisinducing factor (AIF), which executes caspase-independent cell death, is also localized to the mitochondrial intermembrane space. Following processing at the N-terminus, AIF becomes truncated (tAIF) and is released from mitochondria. The protease responsible for AIF processing has not been established. The same submitochondrial localization of mitochondrial μ-calpain and AIF gives support to the hypothesis that mitochondrial μ-calpain may be responsible for processing AIF. Atractyloside-induced tAIF release in rat liver mitochondria was inhibited by cysteine protease inhibitor MDL28170, but not by calpain inhibitors PD150606 or calpastatin. Moreover, μ-calpain immunoreactivity was difficult to …

Physiological Genomics Of Spinal Cord And Limb Regeneration In A Salamander, The Mexican Axolotl, James Robert Monaghan

University of Kentucky Doctoral Dissertations

Salamanders have a remarkable ability to regenerate complex body parts including the limb, tail, and central nervous system. Although salamander regeneration has been studied for several hundred years, molecular-level studies have been limited to a relatively few important transcription factors and signaling molecules that are highly conserved among animals. Physiological genomic approaches were used here to investigate spinal cord and limb regeneration. Chapter 2 reports that hundreds of gene expression changes were identified during spinal cord regeneration, showing that a diverse injury response is activated in concert with extracellular matrix remodeling mechanisms during the early acute phase of natural spinal …

Transcriptional And Morphological Changes During Thyroxine-Induced Metamorphosis Of The Mexican Axolotl And Axolotl-Tiger Salamander Hybrids, Robert Bryce Page

University of Kentucky Doctoral Dissertations

For nearly a century, amphibian metamorphosis has served as an important model of how thyroid hormones regulate vertebrate development. Consequently metamorphosis has been studied in a number of ways including: morphologically, developmentally, ecologically, and from an endocrine perspective. Over the last two decades, much has been learned about the molecular basis of anuran (frog) metamorphosis. However, very little is known about the molecular underpinnings of urodele (salamander) metamorphosis. Using the axolotl and axolotl hybrids as models, I present some of the first studies on the gene expression changes that occur during urodele metamorphosis. In Chapter 1, the motivation for the …

Functional Characterization Of Upd3 In Drosophila Development, Liqun Wang

University of Kentucky Doctoral Dissertations

The JAK/STAT pathway is a non-receptor tyrosine kinase signaling pathway that is well conserved and highly re-utilized in many mammalian and Drosophila developmental processes. Compared to dozens of ligands and receptors in mammalian JAK/STAT, Drosophila JAK/STAT pathway is simpler with one receptor and three ligands, Upd, Upd2 and Upd3, which have similar amino acid sequences. Previous literature shows that upd and upd2 exhibit the same dynamic striped expression pattern in embryos and have semi-redundant functions during embryogenesis. Do Upd and Upd3 also have redundant functions? To answer this question, the functions of Upd3 in Drosophila development were investigated in this …

Comparative Biology Of Seed Dormancy-Break And Germination In Convolvulaceae (Asterids, Solanales), Kariyawasam Marthinna Gamage Gehan Jayasuriya

University of Kentucky Doctoral Dissertations

The biology of seed dormancy and germination of 46 species representing 11 of the 12 tribes in Convolvulaceae were compared in laboratory (mostly), field and greenhouse experiments. Seeds were tested for kind of dormancy and storage behavior; artificial or simulated natural treatments were applied to break physical dormancy (PY); the initial route of water entry (“water gap”) into seeds was identified; the morphoanatomy of the water gap was compared in seeds of 17 species; ontogenetical differences between water gap and seed coat away from the hilum were described in Ipomoea lacunosa seeds; cycling of sensitivity to dormancy break was elucidated …

The Influence Of Ca2+ Regulation In Synaptic Facilitation Of Motor Nerve Terminals In Crayfish And Drosophila As Well As In The Physiological Regulation Of Larval Drosophila Heart, Mohati Desai-Shah

University of Kentucky Doctoral Dissertations

Intracellular Ca²⁺ ions are highly regulated in animal cells for them to function normally. Since the tight regulation of [Ca²⁺]i is so ubiquitous among cells, it is not surprising that altered function in [Ca²⁺]i regulation is associated with a myriad of disease states in humans. This is particularly evident in pacing myocytes and nerve terminals related to synaptic transmission. A common thread through this dissertation is on the role of three regulators proteins that are common to many cell types. These are the plasmalemmal Na⁺/Ca²⁺ exchanger (NCX), the Ca²⁺-ATPase (PMCA) …

Ambystoma: Perspectives On Adaptation And The Evolution Of Vertebrate Genomes, Jeramiah James Smith

University of Kentucky Doctoral Dissertations

Tiger salamanders, and especially the Mexican axolotl (Ambystoma mexicanum), are important model organisms in biological research. This dissertation describes new genomic resources and scientific results that greatly extend the utility of tiger salamanders. With respect to new resources, this dissertation describes the development of expressed sequence tags and assembled contigs, a comparative genome map, a web-portal that makes genomic information freely available to the scientific community, and a computer program that compares structure features of organism genomes. With respect to new scientific results, this dissertation describes a quantitative trait locus that is associated with ecologically and evolutionarily relevant …