Digital Commons Network^™

Novel Constitutively Active Point Mutations In The Nh2 Domain Of Cxcr2 Capture The Receptor In Different Activation States, Giljun Park

Doctoral Dissertations

Chemokines are structurally and functionally related 8-10 kDa proteins defined by four conserved cysteine residues. They consist of a superfamily of proinflammatory mediators that promote the recruitment of various kinds of leukocytes and other cell types through binding to their respective chemokine receptor, a member of the GPCR family. Abnormal control of this system results in various diseases including tumorigenesis and cancer metastasis. Deregulation can occur when constitutively active mutant (CAM) chemokine receptors are locked in the “on” position. This can lead to cellular transformation/tumorigenesis. A viral CAM receptor, ORF74, that can cause tumors in humans, also has homology to …

Biomechanical Risk Factors For Knee Osteoarthritis In Young Adults: The Influence Of Obesity And Gait Instruction, Julia Ann Freedman

Doctoral Dissertations

With increasing rates of obesity, research has begun to focus of co-morbidities of obesity such as osteoarthritis. The majority of existing research has focused on older adults as the group most likely to suffer from osteoarthritis. The purpose of this study was to determine if overweight and obese young adults exhibit biomechanical risk factors for knee osteoarthritis, and to determine if young adults with biomechanical risk factors of osteoarthritis can modify these with instruction. This purpose was divided into two separate studies.

Study 1: Thirty adults between 18-35 years old were recruited into three groups according to body mass index: …

Characterization Of The Extracellular Proteome Of A Natural Microbial Community With An Integrated Mass Spectrometric / Bioinformatic Approach, Brian Keith Erickson

Doctoral Dissertations

Proteomics comprises the identification and characterization of the complete suite of expressed proteins in a given cell, organism or community. The coupling of high performance liquid chromatography (LC) with high throughput mass spectrometry (MS) has provided the foundation for current proteomic progression. The transition from proteomic analysis of a single cultivated microbe to that of natural microbial assemblages has required significant advancement in technology and has provided greater biological understanding of microbial community diversity and function.

To enhance the capabilities of a mass spectrometric based proteomic analysis, an integrated approach combining bioinformatics with analytical preparations and experimental data collection was …

Human Decomposition Ecology At The University Of Tennessee Anthropology Research Facility, Franklin Edward Damann

Doctoral Dissertations

The University of Tennessee Anthropology Research Facility (ARF) is well known for its unique history as a site of human decomposition research in a natural environment. It has been integral to our understanding of the processes of human decomposition. Over the last 30 years 1,089 bodies have decomposed at this 1.28 acre facility, producing a density of 850 corpses per acre of land. This project evaluated the abiotic and biotic characteristics of the soil exposed to various levels of human decomposition in order to determine the effect on the physicochemical properties and the indigenous bacterial communities.

Specifically, 75 soil samples …

Mechanisms Associated With The Chemotherapeutic Effects Of Zyflamend, A Multi-Herbal Extract, On Advanced Prostate Cancer, E-Chu Huang

Doctoral Dissertations

Advanced prostate cancer (PrC) is the second leading cause of death from cancer in US males. Advanced PrC cells are initially androgen-sensitive and thus androgen ablation therapy causes tumors to undergo regression and fall into a remission phase where residual cells remain dormant while androgen levels remain very low. Unfortunately, this phase usually lasts 3 to 5 years prior to tumor relapse, where the tumor cells re-grow in the absence of androgens. This form of the disease is aggressive and invariably fatal. In this study, we investigated the effects of a combination of herbal extracts on various stages of PrC, …

Sublethal Toxicity Of Microcystis And Microcystin-Lr In Fish, Emily Dawn Rogers

Doctoral Dissertations

The occurrence of blooms of toxic cyanobacteria in freshwater environments is a global ecological and public health concern. Species of Microcystis are of particular importance because blooms occur in many freshwater environments throughout the world and microcystin toxin concentrations can exceed World Health Organization advisory levels. While microcystin has been associated with fish kills, sublethal effects of chronic exposure at environmentally relevant concentrations are relatively unknown. The objective of this research was to evaluate toxicity of microcystin and Microcystis in fish during all life history stages. We evaluated global gene expression response in larval zebrafish (Danio rerio), and …

Phylogenetic Analysis Of The Nearctic Blepharicera Macquart (Diptera: Blephariceridae) With An Emphasis On The Eastern Blepharicera Tenuipes Group Hogue, Amanda Jane Jacobson

Doctoral Dissertations

The eastern Nearctic fauna of Blepharicera Macquart (Diptera: Blephariceridae) is revised to include twenty-three species, six of which are new to science. Descriptions of the larvae, pupae, and adults of B. amnicula n. sp., B. conifera n. sp., B. crista n. sp., B. enoristera n. sp., B. hillabee n. sp., and B. opistera n. sp. are presented. Keys to instar IV larvae, pupae, and adults of all eastern Blepharicera (except B. caudata Courtney) are provided. Phylogenetic studies were conducted to determine the relationships between eastern and western Nearctic Blepharicera and among species within these groups. Larvae, pupae, and adults were …

Novel Delivery Systems Of Nisin To Enhance Long-Term Efficacy Against Foodborne Pathogen Listeria Monocytogenes, Dan Xiao

Doctoral Dissertations

Novel intervention strategies such as food grade antimicrobials are used to enhance food safety. Nisin is a widely used naturally occurring antimicrobial effective against many pathogenic and spoilage microorganisms. However, its antimicrobial efficacy in food matrices is much reduced because of interactions with food components. Novel delivery systems may overcome this problem by protecting nisin in capsules and releasing it in a controlled manner. The overall goal of this research was to develop delivery systems of nisin to improve its long-term antimicrobial effectiveness. The first objective was to develop a low-cost method to extract nisin from a commercial preparation containing …

Diagnosis And Management Of Horses With Equine Metabolic Syndrome (Ems), Kelly Ann Chameroy

Doctoral Dissertations

In horses, a painful and often debilitating disease known as laminitis can result in impaired function and, in severe cases, euthanasia. Equine Metabolic Syndrome (EMS) is a syndrome in horses that results in development of laminitis and is characterized by the presence of general and/or regional adiposity (“cresty neck”), aberrations in blood lipid concentrations, insulin resistance (IR) and/ or hyperinsulinemia. Therapies have focused on improving the state of obesity and insulin resistance with the goal of diminishing the likelihood of laminitis development. A definitive cause for laminitis has not been established, but hyperinsulinemia and IR are likely candidates as experimental …

A Comparison Of Commonly Used Accelerometer Based Activity Monitors In Controlled And Free-Living Environment, Yuri Feito

Doctoral Dissertations

This dissertation was designed to determine the effects of body mass index (BMI) and walking speed on activity monitor outputs. A secondary purpose was to compare the activity monitors’ performance in a free-living environment. In the first experiment, 71 participants wore three waist-mounted activity monitors (Actical, ActiGraph, and NL-2000) and an ankle-mounted device (StepWatch 3) while walking on a treadmill (40, 67 and 94 m/min). The tilt angle of each device was measured. The Actical recorded 26% higher activity counts (P < 0.01) in obese persons with a tilt <10 degrees, compared to normal weight persons. The ActiGraph was unaffected by BMI or tilt angle. In the second experiment, the steps recorded by the devices were compared to actual steps. Speed had the greatest influence on the accuracy these devices. At 40 m/min, the ActiGraph was the least accurate device for normal weight (38%), overweight (46%) and obese (48%) individuals. The Actical, NL-2000 and StepWatch averaged 65%, 73% and 99% of steps taken, respectively. Lastly, several generations of the ActiGraph (7164, GT1M, and GT3X), and other research grade activity monitors (Actical; ActivPAL; and Digi-Walker) were compared to a criterion measure of steps. Fifty-six participants performed treadmill walking (40, 54, 67, 80 and 94 m/min) and wore the devices for 24-hours under free-living conditions. BMI did not affect step count accuracy during treadmill walking. The StepWatch, PAL, and the AG7164 were the most accurate across all speeds; the other devices were only accurate at the faster speeds. In the free-living environment, all devices recorded about 75% of StepWatch-determined steps, except the AG7164 (99%). Based on these findings, we conclude that BMI does not affect the output of these activity monitors. However, waist-borne activity monitors are highly susceptible to under-counting steps at walking speeds below 67 m/min, or stepping rates below 100 steps/min. An activity monitor worn on the ankle is less susceptible to these speed effects and provides the greatest accuracy for step counting.

Alternative Splicing In Human Colorectal Cancer, Jae Hoon Bahn

Doctoral Dissertations

Most human genes undergo alternative splicing, and many abnormal splicing processes are associated with human diseases. However, the molecular relationship between alternative splicing and tumorigenesis is not well understood. Here, we identified novel Krüppel-like factor 4 (KLF4) splicing variants produced by exon skipping in human cancer cell lines as well as colon tumor tissues. To elucidate the mechanism involved in KLF4 alternative splicing, we developed KLF4 minigene system and found that RNA binding motif protein 5 (RBM5) plays an important role in KLF4 splicing, as assessed by gain and loss of functional studies. Several anti-tumorigenic compounds were also tested for …

The Role Of The Suppressor Of Hairy-Wing Insulator Protein In Chromatin Organization And Expression Of Transposable Elements In Drosophila Melanogaster, Heather Anne Wallace

Doctoral Dissertations

ABSTRACT Chromatin insulators are required for proper temporal and spatial expression of genes in metazoans. Insulators are thought to play an important role in the regulation of gene expression through the formation of higher-order chromatin structures. One of the best characterized insulators is the Drosophila gypsy insulator, which is located in the gypsy retrovirus. Several proteins are required for gypsy insulator function, including Su(Hw), Mod(mdg4), and CP190. In addition to the gypsy insulator, these proteins are located throughout the genome at sites which are thought to correspond to endogenous insulators. Analysis of the distribution of insulator proteins across a region …

“Development Of Genetic And Genomic Predictors Of Fertility In Argentinean Holstein Cattle.”, Fernando Alfonso Di Croce

Doctoral Dissertations

The overall aim of the studies described herein was to evaluate genetic variation in cattle fertility traits for development of genetic and genomic predictors in breeding strategies. Results from these experiments suggest that improvements in fertility through genetic selection are a possible approach to increase reproductive efficiency. Experiment 1 evaluated the development of genetic parameters associated with multiple ovulation and embryo transfer schemes in an attempt to assist producers in identifying animals with greater genetic merit for these protocols. This study confirmed that genetic selection of donors or sires appears to be a potential approach to improve efficiency of MOET …

Analysis Of Polymorphism In Human Cytomegalovirus (Hcmv) Chemokine, Vcxcl-1, And Its Role In Cellular Activation, Jinho Heo

Doctoral Dissertations

The human cytomegalovirus (HCMV) viral chemokine gene, UL146, shows a high degree of variability in clinical isolates. The UL146-produced viral chemokine, vCXCL-1, has homology to CXC chemokines and is predicted to be an immune modulator that may contribute to the pathogenesis of HCMV infections. In the analysis of clinical isolates from congenitally infected infants, we found 11 distinct vCXCL-1 clades. Although the four cysteine residues that create two disulfide bonds providing the essential structure for CXC chemokines,are conserved, the N-loop region, which is important for receptor binding and activation, was hypervariable. One clade also contained a modified glutamic acid-leucine-arginine (ELR) …

Identifying Mechanisms Associated With Innate Immunity In Cows Genetically Susceptible To Mastitis, Alexandra Alida Elliott

Doctoral Dissertations

Mastitis, or mammary gland inflammation, causes the greatest loss in profit for dairy producers. Mastitis susceptibility differs among cows due to environmental, physiological, and genetic factors. Prior research identified a genetic marker in a chemokine receptor, CXCR1, associated with mastitis susceptibility and decreased neutrophil migration. Current research seeks to identify reasons behind mastitis susceptibility by validating this model through in vivo challenge with Streptococcus uberis and studying specific mechanisms causing impaired neutrophil migration. Holstein cows with GG (n=19), GC (n=28), and CC (n=20) genotypes at CXCR1+777 were challenged intramammarily with S. uberis strain UT888. After challenge 68% of quarters from …

Mechanisms Of Antimicrobial Peptide Resistance In Campylobacter, Ky Van Hoang

Doctoral Dissertations

Campylobacter is the major bacterial cause of human gastroenteritis in the United States and other developed countries. Poultry are considered a main source of human Campylobacter infections. Thus, reduction of Campylobacter load in poultry is significant in food safety and public health. However, no effective measure is commercially available to prevent Campylobacter colonization in poultry to date. Antimicrobial peptides (AMPs) are short and bactericidal peptides widely present in intestine to limit bacterial infections. Recently, AMPs have been increasingly recognized as a novel class of antibiotics (peptide antibiotics) to control foodborne pathogens. Notably, several potent anti-Campylobacter bacteriocins, a group of AMPs …

Steps Toward Butternut (Juglans Cinerea L.) Restoration, Sunshine L. Brosi

Doctoral Dissertations

Butternut (Juglans cinerea L.), a lesser-known relative of black walnut (Juglans nigra L.), is a native tree species beneficial for wildlife, valuable for timber, and part of the great diversity of species in the eastern forests of North America. Populations of butternut are being devastated by butternut canker disease, caused by the fungus Sirococcus clavigignenti-juglandacearum (V.M.G. Nair, Kostichka, & Kuntz), which is thought to be introduced to North America. The disease causes multiple branch and stem cankers that eventually girdle trees. Small population sizes, lack of sprouting, and shade intolerance exacerbates the disease and results in permanent losses of butternut …

Conservation Genetics And The Ctenosaura Palearis Clade, Stesha Ann Pasachnik

Doctoral Dissertations

We are now in the midst of a mass extinction crisis. The top threats to biodiversity include habitat destruction, pollution, over-harvesting, and invasive species. The field of conservation genetics seeks to understand these threats and devise management to preserve taxa with the ability to cope with environmental change. Preserving genetic variation and the processes in which variation is created and maintained is vital to long-term conservation goals. Limited conservation resources are cause for the prioritization of taxa and areas. Nine basic methods of prioritization have been developed. Though there are differences in these methods, and thus in the resulting target …

Characterizing The Role Of Dna Repair Proteins In Telomere Length Regulation And Maintenance: Fanconi Anemia Complementation Group C Protein And 8-Oxoguanine Dna Glycosylase, David Beomjin Rhee

Doctoral Dissertations

Telomeres are the chromosome end structures consisting of telomere-associated proteins and short tandem repeat sequences, TTAGGG, in humans and mice. Telomeres prevent chromosome termini from being recognized as broken DNA ends. The structural integrity of DNA including telomeres is constantly threatened by a variety of DNA damaging agents on a daily basis. To counteract the constant threats from DNA damage, organisms have developed a number of DNA repair pathways to ensure that the integrity of genome remains intact. A number of DNA repair proteins localize to telomeres and contribute to telomere maintenance; however, it is still unclear as to what …

Translational Regulation In Arabidopsis Thaliana: Genetic And Functional Characterization Of Eukaryotic Initiation Factor 3, Bijoyita Roy

Doctoral Dissertations

Molecular functions of eukaryotic initiation factor 3 (eIF3) in translation are manifold, encompassing events from initiation complex assembly to translation termination. The contribution of the individual subunits of eIF3 to its multiple activities is quite unclear. It has been hypothesized that several of its 13 subunits contribute to mRNA specific regulation. Prior research had established that the h subunit of eIF3 in Arabidopsis was required for translation of specific mRNAs as well as for organ formation and meristem development. This study aims towards understanding the functions of individual subunits of eIF3 in the context of plant development and to further …

An Analysis Of Global Gene Expression Resulting From Exposure To Energetic Materials, Vernon L. Mcintosh Jr.

Doctoral Dissertations

AN ANALYSIS OF GLOBAL GENE EXPRESSION RESULTING FROM EXPOSURE TO ENERGETIC MATERIALS

A Dissertation

Presented for the Doctor of Philosophy Degree

University of Tennessee, Knoxville

VERNON LASHAWN MCINTOSH JR.

August 2010

Dedication

This dissertation is dedicated to my family. My mother and father Debra and Vernon McIntosh instilled in me the respect for academic excellence and the drive maximize my potential. Early on, my younger brother Kyle started showing signs of a shared interest in biology thus my desire to be a positive role model for him kept me motivated. Last but certainly not least, my loving wife and best …

Investigating Cold Hardiness And Management Practices Of Warm-Season Putting Green Species In The Transition Zone, John M. Kauffman

Doctoral Dissertations

Warm-season turf species are becoming increasingly popular for putting green use in the transition zone. Ultradwarf bermudagrass (Cynodon dactylon (L.)  C. transvaalensis Burtt-Davy) is the prevalent warm-season putting green species, but seashore paspalum (Paspalum vaginatum Swartz) and ‘Diamond’ zoysiagrass [Zoysia matrella (L.) Merr.] may also be grown in the transition zone. Warm-season species are susceptible to winter injury and may require different management regimes than cool-season species. Therefore, the objectives of this research were to assess the impacts of various management practices on warm-season putting green species and characterize the physiological basis for differences in freeze tolerance of various …

Spatiotemporal Dynamics In A Lower Montane Tropical Rainforest, Robert Michael Lawton

Doctoral Dissertations

Disturbance in a forest’s canopy, whether caused by treefall, limbfall, landslide, or fire determines not only the distribution of well-lit patches at any given time, but also the ways in which the forest changes over time. In this dissertation, I use a 25 year record of treefall gap formation find a novel and highly patterned process of forest disturbance and regeneration, providing a local mechanism by examining the factors that influence the likelihood of treefall. I then develop a stochastic cellular automaton for disturbance and regeneration based on the analysis of this long term data set and illustrate the potential …

The Effect Of Body Mass Index On Pedometer Accuracy In A Free-Living Environment, Brian Matthew Tyo

Doctoral Dissertations

The purpose of this dissertation was to determine if the New Lifestyles NL-2000 (NL) and the Digi-Walker SW-200 (DW), waist-mounted devices, yield similar daily step counts as compared to the StepWatch 3 (SW), an ankle-mounted device, worn by adults and children in the free-living environment.

For the first study, fifty-six adults (32.7 + 14.5 y) wore the devices for seven consecutive days. There were 20 normal weight, 18 overweight, and 18 obese participants. The NL and DW undercounted (pedometer error) similarly in the normal weight and overweight groups (-15.4% to -18.2%, respectively). However, the DW undercounted more than the NL …

Fibrinolytic Proteins And Brain-Derived Neurotrophic Factor Modulation Of Suprachiasmatic Nucleus Circadian Clock, Xiang Mou

Doctoral Dissertations

Mammalian circadian rhythms are controlled by a clock located in the suprachiasmatic nucleus (SCN). The mechanisms through which light phase-shifts the SCN circadian clock are similar to those underlying memory formation and long-term potentiation (LTP). Several secreted proteins, including tissue-type plasminogen activator (tPA), plasminogen, and brain-derived neurotrophic factor (BDNF), have been implicated in this process. These same proteins are important for photic phase-shifts of the SCN circadian clock. Early night glutamate application to SCN containing brain slices resets the circadian clock. Our experiments find that the endogenous tPA inhibitor, plasminogen activator inhibitor 1(PAI-1), blocked these shifts in slices from wildtype …

5’-Proximal Cis-Acting Rna Signals For Coronavirus Genome Replication, Bo-Jhih Guan

Doctoral Dissertations

RNA sequences and higher-order structures in the 5’ and 3’ untranslated regions (UTRs) of positive-strand RNA viruses are known to function as cis-acting elements for translation, replication, and transcription. In coronaviruses, these are best characterized in the group 2a bovine coronavirus (BCoV) and mouse hepatitis virus (MHV), yet their precise mechanistic features are largely undefined. Here, we use a reverse genetics system in MHV to exploit the ~30% nt sequence divergence between BCoV and MHV to establish structure/function relationships of 5’ UTR cis-replication elements. It had been previously shown that a precise replacement of the 391-nt MHV 3’ UTR with …

Methicillin Resistance In Staphylococcus Pseudintermedius, Chad Christopher Black

Doctoral Dissertations

Staphylococcus pseudintermedius affecting dogs is analogous to S. aureus on humans, acting as both normal flora and opportunistic pathogen. Methicillin resistance in S. pseudintermedius is recent, with the first documented occurrence of an isolate bearing the methicillin resistance gene, mecA, in 1999. This gene encodes penicillin binding protein 2a, which renders all beta-lactam drugs ineffective and functions as a “gateway” antibiotic resistance determinant. In the presence of ineffective antibiotics, opportunities for mutational events and acquisition of mobile genetic elements increase as microbial densities increase, often leading to multi-drug resistance. Methicillin-resistant S. pseudintermedius (MRSP) infections have become increasingly common. For …

Linking Community Ecology And Biogeography: The Role Of Biotic Interactions And Abiotic Gradients In Shaping The Structure Of Ant Communities., Jean-Philippe Lessard

Doctoral Dissertations

Understanding what drives variation in species diversity in space and time and limits coexistence in local communities is a main focus of community ecology and biogeography. My doctoral work aims to document patterns of ant diversity and explore the possible ecological mechanisms leading to these patterns. Elucidating the processes by which communities assemble and species coexist might help explain spatial variation in species diversity. Using a combination of manipulative experiments, broad-scale surveys, behavioral assays and phylogenetic analyses, I examine which ecological processes account for the number of species coexisting in ant communities. Ants are found in most terrestrial habitats, where …

The Maximum Clique Problem: Algorithms, Applications, And Implementations, John David Eblen

Doctoral Dissertations

Computationally hard problems are routinely encountered during the course of solving practical problems. This is commonly dealt with by settling for less than optimal solutions, through the use of heuristics or approximation algorithms. This dissertation examines the alternate possibility of solving such problems exactly, through a detailed study of one particular problem, the maximum clique problem. It discusses algorithms, implementations, and the application of maximum clique results to real-world problems. First, the theoretical roots of the algorithmic method employed are discussed. Then a practical approach is described, which separates out important algorithmic decisions so that the algorithm can be easily …

Studies On The Role Of Vitronectin And Plasminogen-Activator Inhibitor-1 Complexes Beyond Inhibiting Proteases: Binding To The Extracellular Matrix, Cell Interactions And Pathogenesis, Sumit Goswami

Doctoral Dissertations

Plasminogen activator inhibitor-1 (PAI-1), a member of the serine protease inhibitor (serpin) superfamily of proteins, circulates in blood in a complex with vitronectin (VN). These two proteins are also found localized together in the extracellular matrix in many different pathophysiological conditions. Both of these proteins are involved with a number of physiologically important processes. Though PAI-1 is a well-known inhibitor of serine proteases, more emphasis is now geared towards its protease independent functions. VN, on the other hand, is a binding protein that exists in the circulation in a preferred monomeric conformation. However, in the extracellular matrix, VN exists as …