Life Sciences Commons^™

Mitochondrial Dna Analysis By Denaturing High-Performance Liquid Chromatography For The Characterization And Separation Of Mixtures In Forensic Samples, Richard Kristinsson

Electronic Theses and Dissertations

A mixture of different mtDNA molecules in a single sample is a significant obstacle to the successful use of standard methods of mtDNA analysis (i.e., dideoxy dye-terminator sequencing). Forensic analysts often encounter either naturally occurring mixtures (e.g., heteroplasmy) or situational mixtures typically arising from a combination of body fluids from separate individuals. The ability to accurately resolve and interpret these types of samples in a timely and cost efficient manner would substantially increase the power of mtDNA analysis and potentially provide valuable investigative information by allowing its use in cases where the current approach is limited or fails. Therefore, this …

Sequence Detection And Comparative Analysis Of The Hv1 And Hv2 Control Regions Of Human Mitochondrial Dna By Denaturing High-Performance Liquid Chromatography, Sarah E. Lewis

Electronic Theses and Dissertations

The objective of this research was the evaluation and forensic validation of Denaturing High-Performance Liquid Chromatography (DHPLC) as a sequencingindependent means of detecting the presence of sequence differences in pair-wise mixtures of non-concordant amplicons of human mitochondrial DNA (mtDNA). The reproducibility and efficacy of DHPLC results, including amplification reproducibility, injection reproducibility, and column-to-column reproducibility were measured, showing negligible assay-to-assay variability. In addition, cross-contamination on the DHPLC columns demonstrated very low level DNA carryover between a high-abundance sample and subsequent zero-volume injections.

The accuracy with which DHPLC technology can be used to screen both evidence and control samples in the context …

Does Sabp2 Exist As A Dimer?, Mir Ashad Hossain

Electronic Theses and Dissertations

Salicylic acid binding protein 2 (SABP2) is one of the key enzymes in salicylic acid-dependent plant defense pathway. SABP2 is a 29 kDa protein present in extremely low abundance in plants and it catalyzes the conversion of signaling molecule methyl salicylate into salicylic acid. Although it has been shown that 6x His-tagged SABP2 over expressed in E. coli is a homodimer, its exact conformation in planta is still unknown. Therefore, we proposed to determine if SABP2 exist as a dimer and/or monomer under natural condition. To verify the exact conformation of native SABP2 protein in plant, SABP2 was purified from …

Camp And Calcium Interactions In Endocrine Cells, Colette Crane

Electronic Theses and Dissertations

Inhibitors of cAMP phosphodiestereses (PDEs) promoted an increase in intracellular [cAMP]_c and an influx of Ca²⁺, in both MMQ pituitary cells and α-Pancreatic cells. The specific targets for PDEs and the domains of PDEs are still unknown. Pituitary MMQ cells were used as a model to test modulation of calcium influx through the L-type calcium channel with PDE inhibitors. cAMP analogs were used to determine cAMP molecular targets. 8-cpt-cAMP initiated an Epac mediated pathway and had strong effects on calcium influx in the cell; suggesting, a relatively strong cAMP target. Experiments of α-Pancreatic cells were done in …

Alternative Scheduling In The Middle School: Considering Circadian Rhythms., James Edward Carter

Electronic Theses and Dissertations

The passage of No Child Left Behind has increased the level of accountability for all educators. There are many factors that affect student achievement. One factor that may be overlooked is the schedule configuration of schools. Addressing student needs through scheduling options may assist school systems and students in performing at the level they are being held accountable.

The population for this study was students from a rural East Tennessee middle school with a population of approximately 700 students. The low socioeconomic students represent 68% of the school total enrollment while 18% of the students have an individual education plan …

A Predictive Model Which Uses Descriptors Of Rna Secondary Structures Derived From Graph Theory., Alissa Ann Rockney

Electronic Theses and Dissertations

The secondary structures of ribonucleic acid (RNA) have been successfully modeled with graph-theoretic structures. Often, simple graphs are used to represent secondary RNA structures; however, in this research, a multigraph representation of RNA is used, in which vertices represent stems and edges represent the internal motifs. Any type of RNA secondary structure may be represented by a graph in this manner. We define novel graphical invariants to quantify the multigraphs and obtain characteristic descriptors of the secondary structures. These descriptors are used to train an artificial neural network (ANN) to recognize the characteristics of secondary RNA structure. Using the ANN, …

Flow Cytometric Analysis Of Crayfish Hemocytes., Sarah Kathryn Allen

Electronic Theses and Dissertations

Crayfish exhibit innate immune responses via hemocytes and their products. There are 3 hemocyte populations: hyaline cells, granular cells, and semigranular cells. Hemocytes from laboratory housed, untreated crayfish (normal crayfish) have been quantified on the basis of cell type, cell size, and cell granularity using Flow Cytometry. These data present the first overall picture of normal hemocytes from Red Swamp Crayfish with regard to cell type, cell size, and cell granularity and will serve as a baseline for all future studies in our lab. Experiments using crayfish injected with Pseudomonas aeruginosa, Staphylococcus aureus, or crayfish saline alone showed …

Requirement Of Ssdelseed-Motif Of Escherichia Coli F1FO Atp Synthase In Antimicrobial Peptide Binding., Junior Kom Tayou

Electronic Theses and Dissertations

F₁F_O ATP synthase is a membrane bound enzyme capable of synthesizing and hydrolyzing ATP. Lately, α-helical cationic peptides such as melittin and melittin related peptide (MRP) were shown to inhibit E. coli ATP synthase. The proposed but unconfirmed site of inhibition is βDELSEED-motif formed by the residues 380-386, located at the interface of α/β subunit of ATP synthase. This project was a mutagenic analysis of βDELSEED-motif residues to understand the binding mechanism and mode of action of peptide inhibitors. The study addressed 2 main questions: Are the antibacterial/anticancer effects of these peptides related to their inhibitory action …

Spatial Ecology Of The Spotted Salamander Ambystoma Maculatum During The Nonbreeding Season., Dale Patrick Ledford

Electronic Theses and Dissertations

Spotted Salamanders, Ambystoma maculatum, are a widely distributed pond-breeding amphibian that spends an estimated 95% of its life in woodlands surrounding breeding ponds. Their terrestrial ecology remains poorly understood. Few studies have tracked the movement patterns of Spotted Salamanders, and they have given limited information on fine scale patterns. Studies of distribution around vernal pool environments have brought increased attention to their terrestrial ecology. To this end I tracked a sample of Passive Integrated Transponder (PIT) tagged animals with a Radio Frequency Identification Device (RFID). Quadrat-variance methods and a Poisson distribution were used to analyze spatial distribution patterns over …

A Study Of Habitat Selection And Fluctuating Asymmetry Of Amybstoma Tigrinum At Henderson Island Wildlife Management Area In Jefferson County Tn., Christopher Scott Ogle

Electronic Theses and Dissertations

Studies were conducted on a population of tiger salamanders, Ambystoma tigrinum, at Henderson Island Wildlife Management Area in Jefferson County, TN. Tests were conducted to locate the nonbreeding habitat of the salamanders and to detect any difference in fluctuating asymmetry (FA) between larval populations in a large, permanent pond and an ephemeral wetland. Drift fences were installed with pitfall traps at selected locations around each pond to determine nonbreeding habitat use by adults. Most adult salamanders were found using a blackberry (Rubus sp.) dominated old-field, a grassy field, and a shrub-grass mix field, which were all adjacent …

Molecular Modulation Of A-Subunit Visit-Dg Sequence Residue Asp-350 In The Catalytic Sites Of Escherichia Coli Atp Synthase., Sneha R. Jonnalagadda

Electronic Theses and Dissertations

ATP Synthase is the fundamental means of cellular energy production in animals, plants, and almost all microorganisms. In order to understand the mechanism of ATP catalysis, critical amino acid residues involved in Pi binding have to be identified. The αVISIT-DG sequence at the interface of α/β subunits that contains residues from 345-351 is highly conserved and αAsp-350 has been chosen because of its negative charge side chain and its close proximity (~2.8 Å) to the known phosphate binding residue αArg-376. The mutant's αD350R, αD350Q, αD350A, αR376A/D, and αG351R/A/D were generated by site directed mutagenesis and several biochemical assays were performed …

Novel Copper Loaded Core-Shell Silica Nanoparticles With Improved Copper Bioavailability Synthesis, Characterization And Study Of Antibacterial Properties, Pavithra Maniprasad

Electronic Theses and Dissertations

A novel core-shell silica based antimicrobial nanoparticle was synthesized. The Stöber silica shell has been engineered to accommodate copper. Synthesis of the core-shell Cu-silica nanoparticle (C-S CuSiNP) involves preparation of base-hydrolyzed Stöber silica “seed” particles first, followed by the acid-catalyzed seeded growth of the Cu-silica shell layer around the core. Scanning electron microscopy and transmission electron microscopy showed monodispersed, spherical shaped nanoparticles with smooth surface morphology. Characterization of particle size distribution in solution by the Dynamic Light Scattering (DLS) technique was fairly consistent with the electron microscopy results. Loading of Cu to nanoparticles was confirmed by the SEM-Energy Dispersive X-Ray …

A Solid Phase Assay For Topoisomerase I Interfacial Poisons And Catalytic Inhibitors, Vidusha Cyril

Electronic Theses and Dissertations

We report a mechanism based screening technique to rapidly identify eukaryotic topoisomerase I targeting agents. The method is based on genetic tagging of topoisomerase I to immobilize the enzyme on a solid surface in a microtiter well format. DNA is added to the wells and retained DNA is detected by Picogreen fluorescence. Compounds that result in an increase in Picogreen staining represent potential topoisomerase interfacial poisons while those that reduce fluorescence report catalytic inhibitors; therefore, the solid phase assay represents a „bimodal‟ readout that reveals mechanisms of action. The method has been demonstrated to work with known interfacial poisons and …

Biophysical Characterization Of The Membrane Binding Domain Of The Pro-Apoptotic Protein Bax, Pranav Garg

Electronic Theses and Dissertations

The BCL-2 family of proteins tightly regulates the delicate balance between life and death. The pore forming Bax is a pro-apoptotic member belonging to this protein family. At the onset of apoptosis, monomeric cytoplasmic Bax translocates to the outer mitochondrial membrane, forms oligomeric pores thereby letting mitochondrial cytochrome c enter the cytosol and initiate the apoptotic cascade. The C-terminal "helix 9" is thought to mediate the membrane binding of BAX. A 20-amino acid peptide corresponding to Bax C-terminus (VTIFVAGVLTASLTIWKKMG) and two mutants where the two lysines are replaced with Glu (charge reversal mutant, EE) or Leu (charge neutralization mutant, LL) …

Memory Access Patterns For Cellular Automata Using Gpgpus, James Michael Balasalle

Electronic Theses and Dissertations

Today's graphical processing units have hundreds of individual processing cores that can be used for general purpose computation of mathematical and scientific problems. Due to their hardware architecture, these devices are especially effective when solving problems that exhibit a high degree of spatial locality. Cellular automata use small, local neighborhoods to determine successive states of individual elements and therefore, provide an excellent opportunity for the application of general purpose GPU computing. However, the GPU presents a challenging environment because it lacks many of the features of traditional CPUs, such as automatic, on-chip caching of data. To fully realize the potential …

Plasma Membrane Dynamics In The Drosophila Embryo, Amelia E. Zommer

Electronic Theses and Dissertations

Convergent extension is a highly conserved process among mammals, in which the tissue narrows in one axis, and extends across another. Tissue elongation is directed by the regulation of cell interface behaviors, which guides cell intercalation and rosette formation. Rosette formation occurs through the contraction of vertically oriented cell interfaces, and the subsequent elongation of new horizontal interfaces. It has been shown that actomyosin-generated tension functions to direct rosette formation. In this thesis, I have tested the function of regulators of F-actin networks, as well as endocytic and exocytic mechanisms, to identify new components that control interface behaviors and cell …

Micrornas 9a, 9b, 9c And 315 Regulate Expression Of A Reporter For The Neuronal Microtubule-Associated Protein Futsch/Map1b, Leslie M. Rozeboom

Electronic Theses and Dissertations

Fragile X syndrome (FXS) is the most common form of inherited mental retardation in humans. FXS is caused by loss of the Fragile X Mental Retardation Protein (FMRP), an important regulator of neuronal mRNA translation. Patients with FXS display cognitive deficits including memory problems. Protein synthesis-dependent long-term changes in synaptic plasticity are involved in the establishment and maintenance of long-term memory. One prevalent theory of FXS pathology predicts that FMRP is required to negatively regulate the translation of important mRNAs at the synapse. We are investigating microRNAs (miRNAs) as a potential regulator of synaptic FMRP-regulated mRNAs that have previously been …

Genetic Polymorphisms And The Treatment Outcome In Standard-Risk Pediatric Acute Lymphoblastic Leukemia, Qi Wei

Electronic Theses and Dissertations

Acute lymphoblastic leukemia (ALL) is the most common childhood cancer and accounts for 30 - 35% of all cancers in children. Significant improvement in the treatment of pediatric ALL has been achieved in recent years. Only 50 years ago, the disease was uniformly fatal with an Overall Survival (OS) rate < 5%. Modern-day, multi-drug chemotherapy is associated with an overall survival rate over 80%. Standard-risk ALL comprises the majority of ALL with an overall survival approaching 90%. Despite this success, children who relapse from this disease accounts for the majority of cancer-related deaths in children. The backbone of treatment protocols have incorporated somatic but not host genetic features in the treatment regimens. The current study examined 12 genetic polymorphisms affecting the pharmacodynamics of antileukemic drugs in an attempt to identify biologic markers related to the risk of disease relapse.

In the current research program, 125 standard-risk ALL patients who were treated at The Children's Hospital were enrolled in a retrospective study. Statistical analysis was performed to evaluate the association between genetic polymorphisms and risk of disease relapse in this study cohort. The GSTM1 null genotype was associated with a …

Functional Activation Of Cartilaginous Fish Melanocortin Receptors, Christina Lee Reinick

Electronic Theses and Dissertations

The evolution of the melanocortin receptor (MCR) gene family has been dictated by two genome duplication events (2R hypothesis). The gnathostomes are thus predicted to possess a minimum of four MCR genes. Previous studies on cartilaginous fish have shown evidence for the presence of only three MCRs. The purpose of this thesis is to functionally express the MC5R from the spiny dogfish (Squalus acanthias) and the putative MC2R and MC3R from the elephant shark (Callorhinchus milii); these receptors have not yet been fully characterized. In this study, SacMC5R was able to be expressed in CHO cells …

Hepatitis C Virus-Induced Hepatocellular Carcinoma: Cancer Stem Cell And Gene Therapy, Heba Samir Esmaeil Allam

Electronic Theses and Dissertations

Hepatitis C virus (HCV) is an important human pathogen that causes chronic hepatitis cirrhosis, steatosis and hepatocellular carcinoma (HCC) worldwide. Although current FDA-approved drugs are partially effective in the treatment of HCV, methods for cure and vaccine against the virus are still awaited. My thesis work presented here is focused on finding mechanism (s) of HCV-induced hepatocarcinogenesis and devising novel means to combat the HCV infection.

Herein, we have shown that long-term HCV expression results in the acquisition of cancer stem-like cells (CSCs) traits in liver derived cell lines. These traits include enhanced expression of putative stem cell markers DCAMKL-1, …

The Sheddase Activity Of Adam10/Adam17 On Cxcl16 Increases Proliferation And Survival Of Colorectal Cancer Cells, Tamu C. Talton

Electronic Theses and Dissertations

CXCL16 is an interferon-inducible chemokine of the CXC-subfamily and functions as an adhesion molecule, when membrane bound, and a chemoattractant when soluble. Upregulation of cell associated CXCL16 (cCXCL16) in colorectal cancer is associated with increased tumor infiltrating lymphocytes and good prognosis. ADAM10 and ADAM17 are metalloproteinases responsible for cleaving CXCL16, releasing soluble CXCL16 (sCXCL16) and contributing to proliferation and migration of mesangial cells, in kidney inflammatory disease. We hypothesize that cCXCL16 is a substrate for ADAM10 and ADAM17 cleavage in colorectal cancer, releasing sCXCL16, which mediates cell proliferation. To this end, we first identified CXCL16 in the human colon carcinoma …

Acute Synaptic Activity Causes Differential Mirna Expression In The Drosophila Melanogaster Larval Central Nervous System, Robert Ian Sand

Electronic Theses and Dissertations

The primary goal of this thesis was to determine if spaced synaptic stimulation induced the differential expression of microRNAs (miRNAs) in the Drosophila melanogaster central nervous system (CNS). Prior to attaining this goal, we needed to identify and validate a spaced stimulation paradigm that could induce the formation of new synaptic growth at a model synapse, the larval neuromuscular junction (NMJ). Both Channelrhodopsin- and high potassium-based stimulation paradigms adapted from (Ataman, et al. 2008) were tested. Once validation of these paradigms was complete, we sought to characterize the miRNA expression profile of the larval CNS by miRNA array. Following attainment …

Adaptive Responses Of Branchial Morphology To Hypoxia In The Neotropical Electric Fish Genus Brachyhypopomus, Leilani B. Pathak

Electronic Theses and Dissertations

Many tropical aquatic environments worldwide are characterized by intermittent or prolonged hypoxia (low dissolved oxygen). Nevertheless, many tropical freshwater fishes are able to inhabit these challenging environments via a range of morphological, physiological and behavioral adaptations. Brachyhypopomus is a diverse genus of weakly electric fishes represented by 28 known species distributed from Panama to Argentina. 17 species are restricted to permanently normoxic habitats (blackwater rivers and terra firme streams), eight species are restricted to seasonally or perennially hypoxic habitats (whitewater floodplains of large tropical rivers or permanent swampy habitats), and three species are eurytopic (occur in both seasonally hypoxic and …

Mechanisms Of Alpha]-Synuclein-Induced Neurodegenertaion In Parkinson's Disease And Stroke, Cherine Belal

Electronic Theses and Dissertations

Parkinson’s disease (PD) is a debilitating neurodegenerative disorder affecting one million Americans. Despite its social and economic impact, the pathological cascades that lead to neuron dysfunction and degeneration in PD are poorly understood. Endoplasmic reticulum (ER) stress has been implicated as an initiator or contributing factor in neurodegenerative diseases including PD. The ER is an organelle central to protein folding and intracellular Ca2+ homeostasis. Perturbations of these functions result in ER stress and upregulation of ER stress proteins, of which some have been implicated in counteracting ER stress-induced cell death. The mechanisms that lead to ER stress and how ER …

Identification Of Fabrics Likely To Collect And Disperse Fel D 1, Mary Janice Jones

Electronic Theses and Dissertations

Individuals sensitive to domestic cat allergen Fel d 1 experience a variety of symptoms including eye irritation, respiratory irritation, asthma, and severe respiratory distress. Fel d 1 is a protein produced in the saliva and on the skin of domestic cats. Previous studies have demonstrated that Fel d 1 adheres to clothing, upholstery, and human hair and has been found in non-cat environments in levels high enough to cause allergic reactions in sensitive individuals. In a general sense, two very different approaches have been adopted to study Fel d 1. One area of the literature focuses on the molecular biology …

Effects Of Block Predator Management On Duck And Pheasant Nest Success In Eastern South Dakota, Nicholas R. Docken

Electronic Theses and Dissertations

Nest success is the biggest limiting factor governing waterfowl production in the Prairie Pothole Region which supports greater than 50% of North America’s breeding duck population. Depredation by predators accounts for 80% or more of nest losses each year (Klett 1988). Township size block predator management (BPM) has been effective at increasing duck nest success in North Dakota and Canada, but no BPM work has been done in South Dakota. There has also been no research on the effects of BPM on pheasants. The objectives of this study were to evaluate the effectiveness of increasing duck and pheasant nest success …

Characterization Of Cis-Regulatory Elements Controlling Repo Transcription In Drosophila Melanogaster, Robert Johnson

Electronic Theses and Dissertations

The glial cells missing (gcm) gene has been identified as a "master regulator" of glial cell fate in the fruit fly Drosophila . However, the gcm gene is also expressed in and required for the development of larval macrophages and tendon cells, and lamina neurons in the adult CNS. Thus, the Gcm protein activates the transcription of different sets of genes in different developmental contexts. How the Gcm protein regulates these different outcomes is not known. My long-term goal is to identify proteins that collaborate with Gcm to promote the transcriptional activation of Gcm target genes specifically in glial cells, …

In Vitro And In Vivo Pharmacodynamic Characterization Of The Novel Plasma Kallikrein Inhibitor Pf-04886847, Dhaval Sanjeev Kolte

Electronic Theses and Dissertations

Plasma kallikrein plays an important role in the pathogenesis of inflammation and thrombosis. Kallikrein cleaves high molecular weight kininogen (HK) to liberate the potent pro-inflammatory peptide bradykinin (BK). BK upon activation of its constitutive B2 receptors on endothelial cells leads to an increase in intracellular Ca2+ level and subsequent production of nitric oxide (NO) and prostacyclin (PGI2), ultimately leading to vasodilation, hypotension, increased vascular permeability and vascular leakage. Further, kallikrein activates the complement system and stimulates neutrophil chemotaxis, aggregation and elastase release. Furthermore, kallikrein mediates the conversion of factor XII (FXII) to activated factor XII (FXIIa) thereby potentiating the intrinsic …

Invasion Of Hurricane-Disturbed Scrub Communities On Barrier Islands In Mississippi: A Battle Between Native And Non-Native Ecosystem Engineers, Christine Annette Bertz

Electronic Theses and Dissertations

Some communities may resist invasion because they are dominated by ecosystem engineers. If these dominant species are removed by some catastrophic disturbance, they can be replaced by invasive species that will dramatically alter the ecosystem in a way that may prevent its recovery. I tested several hypotheses related to this idea within barrier island scrub communities dominated by Florida rosemary (Ceratiola ericoides Michx.). Hurricanes represent a major source of disturbance to coastal Florida rosemary communities, and individual rosemary shrubs in three populations recently impacted by major hurricanes were not protected from the storm surge at higher elevation. Unlike inland populations …

Effects Of Tornado Damage, Prescribed Fire, And Salvage Logging On Oak (Quercus Spp.) Saplings In Upland Oak Forests In Northern Mississippi, Jeffery B. Cannon

Electronic Theses and Dissertations

After European colonization and extensive logging folloby long periods of fire suppression, oak dominated forests, woodlands, and savannahs are being replaced by an unprecedented forest ecosystem. In Mississippi, these new forest systems are dominated in the mid- and understory by mesophytic species such as red maple (Acer rubrum L.) and sweetgum (Liquidambar styraciflua L.). Partial thinning of trees folloby prescribed fire can regenerate oaks in managed timber stands. A severe tornado occurred on a monitored oak stand in northern Mississippi. Of the damaged plots, some were treated with either prescribed fire or salvage logging or were left alone. I examined …