Substrate Trafficking Within The Type Vii Secretion Systems Of Pathogenic Mycobacteria, Zachary A. Williamson

Theses and Dissertations--Molecular and Cellular Biochemistry

Tuberculosis (TB), primarily caused by infection of Mycobacterium tuberculosis (Mtb) in the lungs, is the deadliest infectious bacterial disease killing 1.5 million people annually. A major determinant of virulence is active secretion through three specialized type VII secretion (ESX) systems; ESX-1, ESX-3, and ESX-5. A large group of substrates exported by the ESX systems is the PE (Proline-Glutamine) and PPE (Proline-Proline-Glutamate) families of proteins, which are highly expanded in the pathogenic species of Mycobacteria and encompass over 7% of Mtb’s genome coding capacity. PE and PPE proteins interact together to form PE-PPE heterodimers, and are secreted through …

Reversible Glucan Phosphorylation In The Red Alga, Cyanidioschyzon Merolae, Corey Owen Brizzee

Theses and Dissertations--Molecular and Cellular Biochemistry

Starch and glycogen are an essential component for the majority of species and have been developed to maintain homeostasis in response to environmental changes. Water-soluble glycogen is an excellent source of quick, short-term energy in response to energy demands. In contrast, plants and algae have developed the macromolecule starch that is elegantly suitable for their dependence on external circumstances. Semi-crystalline starch is water-insoluble and inaccessible to most amylolytic enzymes, thus plants and algae have developed a coordinated system so that these enzymes can gain access to the denser starch energy cache. Starch-like semi-crystalline polysaccharides are also found in red algae, …

Characterization Of Spontaneous Preeclampsia In The African Green Monkey (Chlorocebus Aethiops Sabaeus), Chelsea Christina Weaver

Theses and Dissertations--Biology

Hypertensive pregnancy disorders are a major contribution to maternal and neonatal mortality worldwide. Two of these disorders, preeclampsia and chronic hypertension in pregnancy, affect up to 10% of all pregnancies. These hypertensive disorders of pregnancy are associated with long-term, postnatal risk factors for both mother and offspring. Despite numerous recent advances in preeclampsia research, the underlying mechanisms are still not understood. This could be due to lack of a spontaneous animal model. This dissertation presents the African Green Monkey (AGM; Chlorocebus aethiops sabaeus) as the first known spontaneous animal model of preeclampsia and a highly translational model of chronic …

Molecular Mechanisms Regulating Optic Fissure Fusion During Zebrafish Eye Development, Megan Weaver

Theses and Dissertations--Biology

Vertebrate retinal development requires timely and precise fusion of the optic fissure. Failure of this event leads to congenital vision impairment in the form of coloboma. Recent studies have suggested hyaloid vasculature to be involved in OF fusion. In order to examine this link, we analyzed optic fissure fusion and hyaloid vasculogenesis in the zebrafish pax2a noi mutant line. We first determined that pax2a^-/- embryos fail to accumulate F-actin in the optic fissure prior to basement membrane (BM) degradation. Furthermore, using 3D and live imaging we observed reduced OF hyaloid vascularization in pax2a^-/- embryos. When examining the connection …

Spontaneous Postpartum Hypertension In The African Green Monkey, Patrick Rivera

Theses and Dissertations--Biology

Postpartum hypertension (PPHT) is a hypertensive disorder of the puerperium that occurs in women at a rate between 0.8-28% although the exact incidence is unknown due primarily to its transient presentation during a time of reduced medical supervision. The etiology of PPHT is currently unknown with no present experimental animal model. We present the African green monkey (Chlorocebus aethiops sabaeus; AGM) as a potentially translational NHP model of PPHT in humans. AGMs were identified as PPHT using Doppler sphygmomanometry and American Heart Association standards of hypertension for systolic blood pressure (systolic blood pressure > 140 mmHg). Disease characteristics were …

The Social Behavior Of Pine Sawflies In The Genus Neodiprion, John W. Terbot Ii

Theses and Dissertations--Biology

Group living is found across the animal kingdom ranging from temporary mating aggregations to complex, eusocial lifestyles. A particularly common form of group living found among insects are larval or nymphal herds. This lifestyle consists of immature insects living together and results in several proposed costs and benefits. Benefits of this lifestyle include improved ability to regulate a group’s microenvironment, more efficient use of their host, and the ability to engage in collective predator defenses. Offsetting these benefits are costs resulting from living in close proximity to conspecifics which include increased competition, greater visibility to predators, and heightened disease risks. …

Leveraging Transcriptomic Approaches To Identify Differences In Genetic Programming Driving Two Distinct Wound Healing Mechanisms, Regeneration And Fibrosis, In Acomys And Mus, Shishir K. Biswas

Theses and Dissertations--Biology

Why can some animals and others cannot? This fundamental question has fueled scientists studying regeneration for hundreds of years since early observations in crayfish, salamanders and many other organisms. While most contemporary work in regeneration is done in a handful of species including salamanders, zebrafish and flatforms, these organisms lack a closely-related, non-regenerating sister species from which unique genetic differences can be identified. Additionally, while much has been learned from these organisms, they do not share fundamental biological traits with mammals (endothermy, metabolism and immune system) which limits the ability to translate this research for clinical medicine. To this end, …

Cellular Bioenergetics Regulates Cell Proliferation During Mammalian Regeneration, Sandeep Saxena

Theses and Dissertations--Biology

Mammalian system consists of stress-sensing molecules that regulates their cellular response against damage, injury and oncogenic stress. During vertebrate regeneration, cells responding to injury re-enter the cell cycle and proliferate to form new tissue. Cell cycle re-entry or arrest is at least partly regulated by cellular senescence which negatively impacts the proliferative pool of cells during regeneration. What remains unclear is whether cells in regenerating systems possess an increased propensity to proliferate and are refractory to signals that induce senescence. My thesis work has focused on how fibroblasts from the ear pinna differentially regulate healing in highly regenerative mammals (e.g., …

Pre-Clinical Advancements In Biomarkers, Tools, And Therapeutics For A Metabolic Neurodegenerative Disease, Zoë Simmons

Theses and Dissertations--Molecular and Cellular Biochemistry

Glycogen is the storage form of glucose and a highly important substrate for cellular metabolism. Characterization of the enzymes and mechanisms of glycogen metabolism began over 70 years ago and over the last 20 years, a previously unknown protein called laforin has emerged as an important contributor to glycogen metabolism homeostasis. Multiple labs demonstrated that laforin is a glycogen phosphatase and mutations in the gene encoding laforin cause the formation of aberrant glycogen-like aggregates called Lafora bodies (LBs). LBs are cytoplasmic, water-insoluble aggregates that drive neurodegeneration and early death in Lafora disease (LD) patients. The direct relationship between mutated laforin, …

Toward Intelligent Welding By Building Its Digital Twin, Qiyue Wang

Theses and Dissertations--Electrical and Computer Engineering

To meet the increasing requirements for production on individualization, efficiency and quality, traditional manufacturing processes are evolving to smart manufacturing with the support from the information technology advancements including cyber-physical systems (CPS), Internet of Things (IoT), big industrial data, and artificial intelligence (AI). The pre-requirement for integrating with these advanced information technologies is to digitalize manufacturing processes such that they can be analyzed, controlled, and interacted with other digitalized components. Digital twin is developed as a general framework to do that by building the digital replicas for the physical entities. This work takes welding manufacturing as the case study to …

Machine Learning Approach For Vigilance State Classification In Mice, Anik Muhury

Theses and Dissertations--Electrical and Computer Engineering

Sleep has a significant impact on cognitive abilities such as memory, reaction time, productivity, and creative thinking; however, there are many aspects of this important activity that are not clearly understood. Over the last century, researchers have developed technology and animal models to assist in the study of sleep. Manual sleep scoring is time consuming, reduces productivity, and is impacted by human scorer subjectivity. On the other hand, automatic sleep stage categorization can enhance consistency and reliability, aiding professionals in identifying sleep related health problems.

In recent times various studies reported significant achievements for automatic vigilance detection and overcome the …

Accelerometer-Based Vigilance State Classification In Dairy Cows, Evan King

Theses and Dissertations--Electrical and Computer Engineering

Globally, dairy farming is a $700 billion industry, with more than 9 million dairy cows in the United States alone. Depriving cows of required activities such as sleep has been shown to negatively impact reproductive efficiency, decrease the volume of milk produced, and increase the risk of culling. Overcrowded herds can decrease individual animal health, demanding the need for automatic behavior detection that would provide insight into their state of health.

Using electroencephalography (EEG), electrooculography (EOG), and electromyography (EMG) to characterize the phases of sleep is a technique which has been used for decades. While these techniques are considered the …

Home Point Of Care Inr Testing In Left Ventricular Assist Device Patients: A Clinical Evaluation, Emily Corsentino

DNP Projects

Purpose: The purpose of this project was to evaluate the complications and adverse events in left ventricular assist device (LVAD) patients when comparing those using home point of care (POCT) international normalized ratio (INR) testing versus lab testing. Additionally, the time in therapeutic window for both these groups was studied. A cost benefit analysis was performed. All this data was compiled into a provider education evaluated by a pre and post-survey design.

Methods: The project is a single-center study that will occur at the University of Kentucky Chandler Medical Center. It is a cross-sectional, pre/post-test design. Data will be collected …

Positive Reflective Journaling As A Tool For Managing Stress And Promoting A Healthy Work Environment In Nursing Academia, Roxanne Gall

DNP Projects

Abstract

Background: Teaching and nursing are consistently cited as the two occupations that work under the highest levels of occupational stress. Positive Reflective Journaling may be a tool that helps to manage stress and influence a healthy work environment in academic nursing settings.

Purpose: The purpose of this study was to investigate whether Positive Reflective Journaling could be used as an adjunct to reduce stress and support a healthy working environment in

nursing academia.

Methodology: Eighty-six faculty and staff from a University based School of Nursing were invited to participate in the study. A positive reflective journaling exercise was evaluated …

Optical Seed Sorter-Based Selection Lowers Deoxynivalenol Accumulation In Soft Red Winter Wheat, William Jesse Carmack

Theses and Dissertations--Plant and Soil Sciences

Fusarium head blight (FHB) of wheat (Triticum aestivum L.) results in discolored grain contaminated with deoxynivalenol (DON). DON accumulation, an indicator of FHB resistance, can be quantified and used as the basis for direct phenotypic selection, but testing is expensive. The primary objective of this study was to evaluate an optical seed sorter as an alternative to DON testing for FHB resistance breeding. Three hundred F4 derived soft red winter wheat (SRWW) breeding lines were grown in an inoculated FHB nursery over several years in Lexington, KY. Grain from each breeding line was sorted using an optical seed sorter calibrated …

Incorporation Of Summer Annual Mixtures Into Grazing Systems In Kentucky, Kelly Marie Mercier

Theses and Dissertations--Plant and Soil Sciences

Utilizing summer annual grass-legume forage mixtures has the potential to improve forage yield and nutritive characteristics, and/or animal performance during times when cool-season pasture growth is limited by high temperatures. Legumes can utilize atmospheric nitrogen, which can increase crude protein and forage digestibility in mixtures. As nitrogen application generally improves both the yield and nutritive characteristics of summer annual forages, but can have a negative effect on legume competitiveness, nitrogen fertilizer recommendations for legume-containing summer annual mixtures are not well established.

Two experiments were conducted to determine the feasibility of utilizing summer annual mixtures in Kentucky, USA. The first experiment …

Genomic Selection Strategies To Predict Grain Yield And Disease Resistance Traits In A Wheat Breeding Program, Virginia Laura Verges

Theses and Dissertations--Plant and Soil Sciences

Genomic selection (GS) is a form of marker-assisted selection (MAS) that simultaneously estimates all locus, haplotype or marker effects across the entire genome to calculate genomic estimated breeding values (GEBVs). Since its inception, it has had the attention of breeders keen on finding tools to accelerate genetic gain and reduce phenotyping costs in the breeding program. A first objective of this study was to evaluate strategies to design the training population (TP) and validating population (VP) to estimate GEBVs for grain yield and agronomic traits for wheat breeding lines. Our results demonstrate that, despite the small family size, an approach …

Topographic Gradients In Bulk Soil Properties In Central Kentucky Karst Sinkholes, Anne Kathryn Mcgraw

Theses and Dissertations--Plant and Soil Sciences

The Bluegrass physiographic region of Kentucky is underlain by the late Ordovician Lexington Limestone formation. This area has many identifiable karst features, including sinkholes. Karst sinkholes and associated soils coevolve, which may influence the distribution of bulk soil properties and storage of carbon across karst landscapes. Two sinkholes on the University of Kentucky’s C. Oran Little Animal Research Center in Woodford County, central Kentucky, were selected for analysis. We described and sampled nine pedons at the dominant landscape positions (e.g., summit, shoulder, backslope, footslope, and toeslope) within the two sinkholes; we characterized the physical, chemical, and mineralogical soil properties across …

Optimization Of Gelatin-Based Cellular Coating Of Msc For Myocardial Infarction Therapy, Kara Amelle Davis

Theses and Dissertations--Chemical and Materials Engineering

Cardiovascular disease remains the number one threat to American lives. During an acute myocardial infarction (AMI), blood flow is blocked and results in the formation of scar tissue. As the body’s immune system responds, inflammatory signaling causes an increase in both scar tissue size and the patient’s risk for further chronic heart failure. In order to reduce the risk of continued heart disease inflammatory signaling must be reduced. Stem cell therapies have the ability to alter the immune system’s pro-inflammatory signal. However, stem cell retention is limited due to blood flow shear. Gelatin methacrylate (GelMA) based coatings have been shown …

Assessment Of A Decentralized Solution For Waste Plastic Management In Developing Regions, Chandniben Ashishkumar Joshi

Theses and Dissertations--Chemical and Materials Engineering

Rapid population growth, urbanization and availability of pre-packaged consumer goods have led to increased generation and consumption of plastic – so much so that it has become ubiquitous in the environment. An affordable, durable, and lightweight material of construction, plastic is used in innumerable products in every country on earth. However, this explosion of consumption coupled with the material’s significantly low degradability have led to serious plastic accumulation challenges, which are now an imminent threat to terrestrial and marine species globally. These challenges are especially acute in developing countries, where capital and infrastructure constraints, poor governmental regulation and lack of …

Electrochemical Performance Of Biomass-Derived Activated Carbon Supercapacitor Under Compression, Yulin Zhang

Theses and Dissertations--Chemical and Materials Engineering

Supercapacitors are important energy storage device of high-power density, fast charge/discharge rate, environmental friendliness and long service life. Currently, flexible supercapacitors have attracted widespread interest in academia and industry. Supercapacitors under the action of external force will inevitably cause structural changes, performance degradation and even loss of energy storage and supply functionalities. As the most abundant renewable resource so far, biomass is an important energy source for human survival and development. Most electrode materials for supercapacitors are porous activated carbons, which can be derived from abundant renewable biomass, contributing to the sustainable development of society and environmental protection. This thesis …

Clpxp Degradation System In Escherichia Coli, A Study Of Its Energy Sources And Its Applications In Managing The Expression Levels Of Targeted Membrane And Soluble Proteins, Thilini Abeywansha

Theses and Dissertations--Chemistry

ClpXP is an Escherichia coli protease that carries out energy-dependent intracellular proteolysis. In recent years, this system has been widely studied due to its importance as protein regulatory machinery and a virulence factor. Protein substrates of ClpXP contain degrons with a specific protein sequence. SsrA tag is one of the five degrons known to designate proteins for ClpXP degradation. SsrA is an 11 amino acid peptide added to the C-terminus of nascent polypeptide chains translated from aberrant messenger RNAs lacking stop codons via a process called trans-translation.

ClpXP was known to targets only cytosolic proteins with degrons until recently, …

Rational Design Of Small Molecule Disruptors Of Protein-Protein Interactions: Pd-1/Pd-L1; C-Myc, Arid4b, Samuel Ofori

Theses and Dissertations--Chemistry

Protein-protein interactions (PPIs) are vital to many biological processes, including gene expression, and immune reactions to pathogens. There are approximately 650,000 PPIs in humans with pertinent physiological functions. Aberrant expression of PPIs lead to improper function and contribute to a plethora of disease conditions including cancer. Thus, PPIs represent an enormous target space for drug discovery and chemical probes. Direct targeting of clinically relevant PPIs with small molecules remains an unmet medical need. Development of small-molecule inhibitors of PPIs is a challenging enterprise and in most cases considered undruggable due to large protein surfaces, lack of deep binding pockets and …

Investigation Of Multidrug Efflux Transporter Acrb In Escherichia Coli: Assembly, Degradation And Dynamics, Prasangi Irosha Rajapaksha

Theses and Dissertations--Chemistry

The Resistant Nodulation Division (RND) super family member, tripartite AcrA-AcrB-TolC efflux pump, is a major contributor in conferring multidrug-resistance in Escherichia coli. The structure of the pump complex, and drug translocation by functional rotation mechanism have been widely studied. Despite of all these data, the dynamics of the assembly process of the pump and AcrB during functional rotation in the process of drug efflux remains poorly understood. My thesis focuses on understanding the pump assembly process, dynamics of AcrB in functional rotation mechanism, and also investigate the mechanism of degradation of AcrB facilitated by a C-terminal ssrA tag.

In the …

Developing Synthetic Strategies For Multifaceted Applications Of Stable Gold-Based Complexes, Randall Tyler Mertens

Theses and Dissertations--Chemistry

Development of stable gold-based complexes has been a rapidly advancing field due to the popularity of gold complexes, particularly for use in biomedical research and catalytic transformations. Given that auranofin, a gold(I) complex with FDA approval for the treatment of rheumatoid arthritis is used in the clinic, the development of stable gold-based molecules of clinical relevance is urgently needed. Herein are reported, synthetic strategies used for the development of new classes of gold(I) and gold(III) complexes for advancement in mitochondrial modulation for use as chemotherapeutics as well as application to gold catalysis due to the unique geometry of complexes presented …

Mass Spectrometry Method Development For The Discovery And Characterization Of Secondary Metabolites, Zachary Kelley

Theses and Dissertations--Chemistry

Secondary metabolites are organic compounds produced by an organism for reasons other than growth and development. In plants, secondary metabolites generally act as defense agents produced to deter predators and inhibit other competitive species. For humans, these compounds can often have a beneficial effect and are pursued and utilized as natural pharmaceuticals. The development of sensitive, high-throughput analytical screening methods for plant derived metabolites is crucial for natural pharmaceutical product discovery and plant metabolomic profiling. Here, metabolomic profiling methods were developed using a microfluidic capillary zone electrophoresis device and evaluated against traditional separation approaches. An alkaloid screening assay was constructed …

Flavin Modification And Redox Tuning In The Bifurcating Electron Transfer Flavoprotein From Rhodopseudomonas Palustris: Two Arginines With Different Roles, Nishya Mohamed-Raseek

Theses and Dissertations--Chemistry

Electron bifurcation is considered as a third fundamental mode of energy conservation mechanism in addition to two well-known mechanisms, substrate level phosphorylation and Oxidative phosphorylation, in electron bifurcation endergonic and exergonic redox reactions are coupled. The newly discovered flavin based electron bifurcation in electron transfer flavoproteins (ETFs) helps to reduce low potential ferredoxin, which provides electrons to drive biologically demanding reactions such as atmospheric dinitrogen fixation in diazotroph and methane production in methanogens.

Current research demonstrates the capacity for electron bifurcation in the Rhodopseudomonas palustris ETF (RpalETF) system. RpalETF contains two chemically identical but functionally different FADs: …

The Effects Of Escalated Cocaine Intake On Decision-Making Dynamics, Mcallister Stephens

Theses and Dissertations--Psychology

Cocaine Use Disorder (CUD) is characterized partly by the use of cocaine at the expense of other alternatives, in other words, it is a decision-making pathology (Kalivas & Volkow, 2005). Concurrent choice tasks assess decision-making in a dynamic scenario that more closely resembles real life. Value-based decision-making is an important facet of understanding the addictive properties of drugs of abuse. In order to compare two value-based theories of addiction (habit theory and relative value theory), a concurrent choice task was run in tandem with an escalation procedure. First, animals were trained on a choice task until stable, then trained on …

Physiological Stress Responding During Negative Memory Recollection And The Habitual Use Of Psychological Coping Strategies, Kyle P. Rawn

Theses and Dissertations--Psychology

Coping represents an important process for stress: how effectively one copes dictates how a person experiences stress. However, research has not yet examined how a person’s long-term coping habits are related to their physiological stress responding during a particular situation. The current study addresses this through examination of skin conductance level (SCL) trajectories and coping tendencies (i.e. habitual use of coping strategies) during an autobiographical interview. 167 college students completed questionnaires assessing their demographics and their frequency of use for fourteen coping strategies. SCL was collected while participants recalled and discussed a negative family memory from their childhood. Multilevel modeling …

Distractors As A Tool To Increase “Self-Control” In Pigeons (Columba Livia), Peyton Mueller

Theses and Dissertations--Psychology

In the successive delay-discounting task, all trials start with a stimulus to which a response results in a small amount of reinforcement (smaller-sooner). If no response is made, the stimulus changes and a response results in a larger reinforcer (larger-later). The purpose of this study was to examine the use of a non-reinforced distractor (a stimulus to which responding has no programmed consequence) as a method of increasing the proportion of larger-later (LL) choices in a successive delay-discounting task. Earlier research studying the use of distractors may have inadvertently associated the distractor with reinforcement. Four experiments were conducted and each …