Digital Commons Network^™

Targeting Autophagy In Multiple Myeloma, Yun Dai

Theses and Dissertations

Apoptosis (Type I) and autophagy (Type II) represent two major forms of programmed cell death. Numerous anticancer agents employed in standard chemotherapy or novel targeted therapy induce both apoptosis and autophagy. Of note, a cytoprotective autophagic response often counteracts apoptosis triggered by such agents, potentially contributing to drug-resistance. Mechanistically, autophagy and apoptosis share molecular regulatory mechanisms primarily governed by the Bcl-2 family proteins. However, since autophagy acts as the double-edge sword in cancer, whether autophagy should be inhibited or activated in cancer treatment remains the subject of debate. Here we report a) a novel autophagy-targeted strategy that targeting the adaptor …

Immunotherapy Of Cancer: Reprogramming Tumor/Immune Cellular Crosstalk To Improve Anti-Tumor Efficacy, Kyle K. Payne

Theses and Dissertations

Immunotherapy of cancer has been shown to be promising in prolonging patient survival. However, complete elimination of cancer and life-long relapse-free survival remain to be major challenge for anti-cancer therapeutics. We have previously reported that ex vivo reprogramming of tumor-sensitized immune cells by bryostatin 1/ionomycin (B/I) and the gamma-chain (γ-c) cytokines IL-2, IL-7, and IL-15 resulted in the generation of memory T cells as well as CD25+ NKT cells and CD25+ NK cells. Adoptive cellular therapy (ACT) utilizing these reprogrammed immune cells protected FVBN202 mice from tumor challenge, and overcame the suppressive functions of myeloid-derived suppressor cells (MDSCs). We then …

Mast Cells In Kainate Receptor Knockout Mice, Andrea J. Elkovich

Theses and Dissertations

Kainate receptor knockout mice have unique differences within their immune system. They exhibit an attenuated T_H2 branch, while maintaining a robust T_H1 response. Specifically, blocking the formation of functional kainate receptors affects mast cells and their related pathologies. While they seem to develop and activate normally in vivo and in vitro, KAR KO mast cells release more inflammatory mediators upon degranulation. These mice experience severe anaphylactic shock due to two compounding abnormalities. First, KAR KO mast cells release significantly more histamine in vivo upon IgE-mediated activation. Second, the animals over-respond to exogenous histamine with drastic …

Characterization Of Transfer Of The Mobile Genomic Island Encoding Methicillin Resistance Among Staphylococci, Melissa D. Ray

Theses and Dissertations

The gene encoding methicillin resistance in Staphylococcus aureus (MRSA) is carried in the chromosome on a large genomic island called SCCmec and is always inserted at the att site within orfX. SCCmec has been designated a mobile genetic element but a mechanism by which it moves among different strains and species of staphylococci has never been demonstrated. This work shows that bacteriophage 80α is capable of transducing SCCmec into a recipient cell, after which it can integrate into the bacterial chromosome via homologous recombination. More importantly, this work characterizes a conjugative mechanism of SCCmec transfer. …

Mirnas As Biomarkers For Prostate Cancer Progression, Gene C. Clark

Theses and Dissertations

Prostate cancer is one of the most challenging global medical issues today. In 2011, prostate cancer was the most diagnosed malignancy in the United States, making up 29% of new cancer cases. In that year it was the second leading cause of cancer related deaths among men in the USA and the second most common cause of cancer related death overall from the EU. The prostate remains, however, an under studied organ, making insights into the anatomy and biology of prostate cancer difficult to achieve. After 30 years, PSA screening of men of the appropriate age is still the first …

Automating The Process Of Fabricating Uniform-Sized Cell Spheroids For Three-Dimensional Bioprinting, Ganesh Sosale

Theses and Dissertations

Although researchers have been able to print small, simple, and avascular tissues, they have been unsuccessful in creating large, complex and vascularized organs. Printing large and complex three-dimensional tissues or organs involves utilizing a large quantity of cellular spheroids and layer-by-layer addition of spheroids. In this study, an in-house cell spheroid fabrication system was developed to produce cell spheroids with human liver cells (hepG2), human endothelial cells (hEC), human neural stem cells (hNSC), and induced pluripotent stem cells (iPSC). It offers the ability of fabricating uniform-sized spheroids repeatedly, which is essential when large and complex structures need to be produced. …

Multi-Platform Arabinoxylan Scaffolds As Potential Wound Dressing Materials, Donald C. Aduba Jr

Theses and Dissertations

Biopolymers are becoming more attractive as advanced wound dressings because of their naturally derived origin, abundance, low cost and high compatibility with the wound environment. Arabinoxylan (AX) is a class of polysaccharide polymers derived from cereal grains that are primarily used in food products and cosmetic additives. Its application as a wound dressing material has yet to be realized. In this two-pronged project, arabinoxylan ferulate (AXF) was fabricated into electrospun fibers and gel foams to be evaluated as platforms for wound dressing materials. In the first study, AXF was electrospun with varying amounts of gelatin. In the second study, AXF …

Dispersal: A Multidisciplinary Investigation Of Plant Life, Alexandra E. Arzt

Theses and Dissertations

Using plants as a basis for exploring the interstices between the human and nonhuman, this thesis investigates ideas of awareness, intelligence, deep time, animism, and the fluctuating human perception of the agency of Nature. It outlines environmental art practices since the 1950s involving vegetal life. In addition, the paper provides a critical analysis of plant perception of Jakob von Uexküll’s work and theories of vital materialism and “critical plant studies” while noting recent studies in plant neurobiology. In my work, plants become active participants via their movement, seeding, and smell. This study takes the form of imitation, purposeful symbiosis, anthropomorphism, …

Emergent Non-Consumptive Predator Effects Alter Habitat Colonization By Dipteran Prey, Ethan G. Staats

Theses and Dissertations

When ovipositing, prey organisms avoid habitat patches containing predator cues because predators consume, and negatively affect the fitness of their prey. Richness of predator species often enhances the strength of consumptive predator effects, but little is known about how multiple predators combined affect prey non-consumptively. We quantified dipteran colonization in aquatic mesocosms in response to varied predator richness. Multiple predator species combined reduced oviposition by Culex mosquitoes, chironomid midges, and the general colonizing dipteran community more than predicted by the effects of the independent predator species. Previous research which quantifies effects of multiple predators on prey as prey abundance, but …

Prothonotary Warbler Nestling Diet And Growth In Response To Variation In Aquatic And Terrestrial Food Availability, Jenna C. Dodson

Theses and Dissertations

Food supply has been suggested as the main determinant of reproductive success in birds. Riparian species can take advantage of seasonal pulses of both terrestrial and aquatic prey, though aquatic resources are often overlooked in studies of diet and reproductive ecology. This study investigates the flux of both aquatic (mayfly) and terrestrial (caterpillar) prey resources and nestling diet of the Prothonotary Warbler throughout the breeding season in two eastern Virginia sites. One site had significantly higher aquatic prey (mayfly) availability. Nestling diet was generally reflective of prey availability, and nestlings grew faster at the site with high aquatic prey availability. …

Spatio-Temporal Modeling Of Anatomic Motion For Radiation Therapy, Elizabeth Zachariah

Theses and Dissertations

In radiation therapy, it is imperative to deliver high doses of radiation to the tumor while reducing radiation to the healthy tissue. Respiratory motion is the most significant source of errors during treatment. Therefore, it is essential to accurately model respiratory motion for precise and effective radiation delivery. Many approaches exist to account for respiratory motion, such as controlled breath hold and respiratory gating, and they have been relatively successful. They still present many drawbacks. Thus, research has been expanded to tumor tracking.

The overall goal of 4D-CT is to predict tumor motion in real time, and this work attempts …

The Role Of Srsf3 In Control Of Alternative Splicing Of Cpeb2 In Triple Negative Breast Cancer, Brian P. Griffin

Theses and Dissertations

In the presented study, we identified that SRSF3 controls the alternative splicing of CPEB2 and consequently promotes a metastatic phenotype in triple negative breast cancer (TNBC). TNBC causes thousands of deaths annually, frequently due to a lack of effective treatments and a high rate of metastasis in patients. Alternative splicing has been found to be dysregulated in numerous cancers, while splicing factors such as SRSF3 are variably expressed. In this study we performed a siRNA panel to screen potential splicing factors, then used specific siRNA to study the effect of its knockdown on cellular function. These results showed that SRSF3 …

Generation And Delivery Of Charged Aerosols To Infant Airways, Landon T. Holbrook

Theses and Dissertations

The administration of pharmaceutical aerosols to infants on mechanical ventilation needs to be improved by increasing the efficiency of delivery devices and creating better ways of evaluating potential therapies. Aerosolized medicines such as surfactants have been administered to ventilated infants with mixed results, but studies have shown improvement in respiratory function with a much lower dose than with liquid instillation through an endotracheal tube (ETT). An aerosolized medicine must be transported through the ventilation tubing and deposit in the lungs to have the desired therapeutic response.

This work has taken a systematic approach to (i) develop new devices for the …

Effects Of Brain-Derived Neurotrophic Factor And Its Signaling Pathway On Sensory Neuronal Activation During Colitis, Fiza Hashmi

Theses and Dissertations

Visceral hypersensitivity is the heightened response to sensory stimuli. Visceral sensations are transmitted through primary afferent neurons in the dorsal root ganglion (DRG) and the sensitization of the neural pathway leads to modification in spinal ascending and descending neurons. The aim of this investigation is to determine the effects of brain-derived neurotrophic factor (BDNF) and its signaling pathway on sensory neuronal activation during colitis. In order to evaluate this, levels of calcitonin-gene related peptide (CGRP), a neuropeptide marker for nociceptive transmission, and phosphorylated cAMP-response element binding protein (pCREB), a molecular switch in neuronal plasticity, were studied in response to BDNF …

Xlf-Dependent Nonhomologous End Joining Of Complex Dna Double-Strand Breaks With Proximal Thymine Glycol And Screening For Xrcc4-Xlf Interaction Inhibitors, Mohammed Al Mohaini

Theses and Dissertations

DNA double-strand breaks induced by ionizing radiation are often accompanied by ancillary oxidative base damage that may prevent or delay their repair. In order to better define the features that make some DSBs repair-resistant, XLF-dependent nonhomologous end joining of blunt-ended DSB substrates having the oxidatively modified nonplanar base thymine glycol (Tg) at the first (Tg1) , second (Tg2), third (Tg3) or fifth (Tg5) positions from one 3’ terminus was examined in human whole-cell extracts. Tg at the third position had little effect on end-joining even when present on both ends of the break. However, Tg as the terminal or penultimate …

Domination Numbers Of Semi-Strong Products Of Graphs, Stephen R. Cheney

Theses and Dissertations

This thesis examines the domination number of the semi-strong product of two graphs G and H where both G and H are simple and connected graphs. The product has an edge set that is the union of the edge set of the direct product of G and H together with the cardinality of V(H), copies of G. Unlike the other more common products (Cartesian, direct and strong), the semi-strong product is neither commutative nor associative.

The semi-strong product is not supermultiplicative, so it does not satisfy a Vizing like conjecture. It is also not submultiplicative so it shares these two …

Expression And Pharmacological Modulation Of Pain-Depressed Behavior In Rats, Michael D. Leitl

Theses and Dissertations

Pain is often associated with depression of behavior and mood, and relief of pain-related depression is a common goal of treatment. This goal of this dissertation was to conduct preclinical research experiments designed to address a set of three inter-related aims that examine the expression, mechanisms and treatment of pain-related depression of Intracranial Self-Stimulation (ICSS) in rats. First, studies evaluated the hypothesis that acute acid-induced depression of ICSS was mediated by a kappa opioid receptor mediated decrease in mesolimbic dopamine release in the nucleus accumbens. Results support a role for depressed mesolimbic dopamine release in pain-related depression of ICSS; however, …

Depletion Of The Chromatin Remodeler Chd4 Sensitizes Aml Blasts To Genotoxic Agents And Reduces Tumor Formation, Justin Sperlazza

Theses and Dissertations

Chromodomain Helicase DNA-Binding Protein 4 (CHD4) is an ATPase that alters the phasing of nucleosomes on DNA and has recently been implicated in DNA double stranded break (DSB) repair. Here, we show that depletion of CHD4 in Acute Myeloid Leukemia (AML) blasts induces a global relaxation of chromatin that renders cells more susceptible to DSB formation, while concurrently impeding their repair. Furthermore, CHD4 depletion renders AML blasts more sensitive both in vitro and in vivo to genotoxic agents used in clinical therapy: daunorubicin (DNR) and cytarabine (ara-C). Sensitization to DNR and ara-C is mediated in part by activation of the …

Pemetrexed, A Modulator Of Amp-Activated Kinase Signaling And An Inhibitor Of Wild Type And Mutant P53, Stuti Agarwal

Theses and Dissertations

New drug discoveries and new approaches towards diagnosis and treatment have improved cancer therapeutics remarkably. One of the most influential and effective discoveries in the field of cancer therapeutics was antimetabolites, such as the antifolates. The interest in antifolates increased as some of the antifolates showed responses in cancers, such as mesothelioma, leukemia, and breast cancers. When pemetrexed (PTX) was discovered, our laboratory had established that the primary mechanism of action of pemetrexed is to inhibit thymidylate 22 synthase (TS) (E. Taylor et al., 1992). Preclinical studies have shown that PTX has a broad range of antitumor activity in human …

Investigating The Role Of The Nucleosome Remodeling Factor Nurf As A Regulator Of Gene Expression, Aiman S. Alhazmi

Theses and Dissertations

The nucleosome remodeling factor (NURF) is an evolutionary conserved ATP-dependent chromatin remodeling factor. It was first isolated from Drosophila as a complex with enzymatic activity that once recruited to nucleosome, it slides the nucleosome to provide accessibility for transcription factors. Since then, numerous works from animal models and cell lines show the role of NURF as a regulator of gene expression. NURF interacts with H3K4me3 and sequence specific transcription factors that recruit the complex to promoter regions. Whether this is the only mechanism by which NURF regulates gene expression is not known. However, other ATP-dependent chromatin remodeling complexes are known …

Effects Of Hiv-1 Viral Protein Tat On The Viability And Function Of Oligodendroglial Cells, Shiping Zou

Theses and Dissertations

Myelin pallor is frequently reported in HIV patients, and can occur in the CNS prior to other evidence of disease process. Our exploratory studies showed that oligodendrocytes (OLs) are direct targets of HIV-1 Tat (transactivator of transcription). Tat induces a dose-dependent increase of intracellular Ca²⁺ level ([Ca²⁺]_i) in cultured murine OLs, which can be attenuated by ionotropic glutamate receptor (iGluR) antagonists MK801 and CNQX. The Tat-induced [Ca²⁺]_i increase leads to increased death in immature (O4⁺, MBP^-), but not mature (O4⁺, MBP⁺) OLs, over 96 …

Consequences Of Vine Infestation: Linking Abiotic Influences And Biotic Interactions To Successional And Structural Changes In Coastal Communities, Spencer N. Bissett

Theses and Dissertations

Located at the interfaces of terrestrial and marine environments, coastal habitats are inherently vulnerable to the effects of global change. Barrier island systems in particular serve not only as protective buffers against storm events, but also as sentinel ecosystems for observation of the impacts of sea level rise, and of increasing storm frequency and intensity. In the mid-Atlantic region, shrub thickets of Morella species compose the dominant forest community. The often monospecific nature of these plant community assemblages is advantageous to ecological studies and cross-scale applications; the relatively low diversity facilitates transitions between scales. My objective was to investigate the …

Effects Of Constant Vs. Fluctuating Temperatures On Performance And Life History Of The Herbivorous Pest Lymantria Dispar (Lepidoptera: Eribidae), Brendan E. Sostak

Theses and Dissertations

The role of temperature variation in organismal performance is understudied, but is critically important for understanding the response of biodiversity to climate change. To address this issue in herbivorous insects, I studied the direct and interactive effects of thermal regime (constant vs. fluctuating temperatures) and nutrition (dietary nitrogen) on gypsy moth (Lymantria dispar) performance under laboratory conditions. Predictions for differences between constant and fluctuating thermal conditions were derived from Jensen’s inequality, and artificial diets of differing nutritional quality were made by modifying nitrogen (casein) content. Larvae were reared in the laboratory under four temperature regimes (22°C constant, 22°C …

Avatar, Cyborg, Icevorg: Simulacra’S Scion, Guido E. Alvarez

Theses and Dissertations

I propose a theoretical framework that describes how avatars incorporate media as an inherent part of their nature and find a hosting body in cyborgs to navigate and spawn in media. I propose the birth of a new scion that combines avatar, medium and cyborg into a conceptual being that I call “ICEVORG.”

The ICEVORG expands beyond representation into the actual physical world by means of media transgression—more specifically, by the use of the Strange Loop also known as Metalepsis ICEVORG find an effective soil to thrive and interrogate our ideas of reality by means of iteration, expansion, fragmentation and …

Expanding Eco-Visualization: Sculpting Corn Production, Jennifer E. Figg

Theses and Dissertations

This dissertation expands upon the definition of eco-visualization artwork. EV was originally defined in 2006 by Tiffany Holmes as a way to display the real time consumption statistics of key environmental resources for the goal of promoting ecological literacy. I assert that the final forms of EV artworks are not necessarily dependent on technology, and can differ in terms of media used, in that they can be sculptural, video-based, or static two-dimensional forms that communicate interpreted environmental information. There are two main categories of EV: one that is predominantly screen-based and another that employs a variety of modes of representation …

The Role Of Α3Β4* Subtype Of Nicotinic Acetylcholine Receptors In Reversing Opioid-Induced Constipation In Mice, Aravind Gade

Theses and Dissertations

Opioids are excellent pain relievers. A major side-effect of chronic opioid treatment is constipation whereas withdrawal following chronic exposure leads to diarrhea and increased gastrointestinal motility. These effects of chronic opioids are mediated by μ-opioid receptors expressed on enteric neurons. Previous studies have shown that chronic opioids enhance sensitivity to nicotine in the gastrointestinal tract. This suggested that prokinetic effects of nicotine mediated through the activation of nicotinic acetylcholine receptors (nAChRs) may be useful in reversing opioid-induced constipation. The goal of this dissertation was to investigate the nAChR subtype expressed on enteric neurons and their role in reversing opioid-induced constipation. …

Psychoactive Synthetic Cathinones (Or 'Bath Salts'): Investigation Of Mechanisms Of Action, Farhana Sakloth

Theses and Dissertations

Synthetic cathinones represent threatening and high abuse-potential designer drugs. These are analogs of cathinone (the b-keto analog of amphetamine (AMPH)) a naturally occurring stimulant found in the plant Catha Edulis. Methcathinone (MCAT) was the first synthetic analog of cathinone to be identified in 1987 by Glennon and co-workers and it exerted its action primarily through the dopamine transporter (DAT). Most central stimulants exert their action via monomaine transporters by causing either the release (e.g. cathinone analogs such as MCAT) or by preventing the reuptake (e.g. cocaine) of the neurotransmitter dopamine (DA) thus increasing the extracellular synaptosomal concentration of this …

Interaction Between Atm Kinase And P53 In Determining Glioma Radiosensitivity, Syed F. Ahmad

Theses and Dissertations

Glioblastoma multiforme (GBM) is the most common primary brain tumor. Studies have shown that targeting the DNA damage response can sensitize cancer cells to DNA damaging agents. Ataxia telangiectasia mutated (ATM) is involved in signaling DNA double strand breaks. Our group has previously shown that ATM inhibitors (ATMi) sensitize GBM cells and tumors to ionizing radiation. This effect is greater when the tumor suppressor p53 is mutated.

The goals of this work include validation of a new ATM inhibitor, AZ32, and elucidation of how ATMi and p53 status interact to promote cell death after radiation. We propose that ATMi and …

Light Controlled Drug Activation And Release, Jonathon Sheldon

Theses and Dissertations

Cancer constitutes a terrible burden on modern society. In the United States there are an estimated 1,658,370 new cancer diagnoses resulting in 589,430 deaths in 2015 alone.^[1] An estimated 41,170 of these cases will be diagnosed right here in Virginia. With new cancer patients comes the expanding demand for new treatments. As we all know, many modern chemotherapeutics cause adverse reactions to patients. This is because the toxic nature of these therapies often affects normal tissue alongside the tumors that are infesting the body. Therefore, researching novel ways to make chemotherapeutics selective for cancer, while leaving healthy tissue unscathed, …

Normalizing Tumor Vasculature Using Sepiapterin To Increase Radiosensitivity, Ninu Bruno

Theses and Dissertations

Our studies examine the role of nitric oxide synthase (NOS) in tumor vasculature. NOS is “uncoupled” in tumor cells, resulting in peroxynitrite (ONOO^-) formation in lieu of nitric oxide (NO). NO signaling is critical for vascular function, thus uncoupling of eNOS in endothelial cells may partly explain the poor vasculature found within tumors. NOS can be “recoupled” through Sepiapterin (SP) treatment of tumor cells. We examined whether SP could normalize tumor vasculature, promoting radiosensitivity. Optoacoustic tomography of flank xenografts and spontaneous tumor models demonstrate that SP significantly enhances percent oxyhemoglobin in tumors. Immunohistochemical analysis of SP-treated tumors showed …