Medicine and Health Sciences Commons^™

Targeting Epigenetic Mechanisms In Endometriosis, Sarah Elizabeth Brunty

Theses, Dissertations and Capstones

Endometriosis is a complex and elusive gynecological disease in which the inner lining of the uterus grows in locations outside of the uterus and forms lesions. It is known to affect 1 in 9 women of reproductive age worldwide. Symptoms of endometriosis include severe pain, heavy periods, and infertility. While multiple theories of origin exist, none fully encompass all aspects of the disease, although all theories agree that this is an inflammation-driven disease. Due to this, many researchers are turning towards epigenetics to explain the initiation and progression of endometriosis. However, what is causing these epigenetic changes is still a …

Endogenous Antioxidant Overexpression As An Adjuvant To Diet Or Exercise Intervention As Therapy To Counteract Obesity And Beneficially Shift The Gut Microbiome, Deborah Lynn Amos

Theses, Dissertations and Capstones

Oxidative stress plays a key role in metabolic syndrome which includes obesity and cardiometabolic diseases. It is implicated that oxygen-derived free radicals generated during the mitochondrial electron transport chain alter the function of specific biological components, thus activating obesogenic pathways such as glucose and lipid signaling. Research on this topic is of vital importance as obesity is a high-risk factor in the development and progression of severe, debilitating, life-threatening maladies such as cardiometabolic diseases, chronic inflammatory pathologies, and cancer. Furthermore, there is no universal effective therapy to combat the rising rates of obesity with over 1.9 billion (39%) adults classified …

Employing The Enzyme Cofactor Function Of Ascorbic Acid To Affect Oncogenic Pathways In Human Melanoma: Modulating Hypoxia Inducible Factor-1Α And Dna Demethylation To Reduce Malignant Potential, Adam Patrick Fischer

Theses, Dissertations and Capstones

Dioxygenase enzymes such as the HIF hydroxylases (PHD1-3, FIH) and the Ten-eleven translocation (TET1-3) enzymes regulate the activity of the hypoxia inducible factor-1_a (HIF1_a) transcription factor and the DNA methylation status of cells, respectively. Aberrant accumulation and activation of HIF-1_a can allow malignant cells to acquire attributes that promote progression, chemotherapy resistance, and survival, while aberrant hypermethylation of gene promoters can silence the expression of tumor suppressor genes essential to preventing tumorigenesis. Inadequate levels of intracellular ascorbic acid (AA), a necessary cofactor for optimal dioxygenase enzyme function, could potentiate these tumorigenic conditions. In fact, plasma levels …

Investigation Of Ultrasound Targeted Microbubbles As A Therapeutic Gene Delivery System For Prostate Cancer, Rounak Paramjeet Nande

Theses, Dissertations and Capstones

A major challenge for effective gene therapy is systemic delivery of viruses carrying therapeutic genes into affected tissue. The immunogenic nature of human adenoviruses (Ads) limits their use for intratumoral (IT) injection in gene therapy. Ads transfection is further hampered by the fluctuating presence of Coxsackie and Adenovirus Receptor (CAR) and integrins on the cells’ surface. To circumvent these limitations we developed a novel approach wherein Ads are encapsulated inside the shell of lyophilized, lipid-encapsulated, perfluorocarbon microbubbles (MBs)/ultrasound (US) contrast agents, which act as delivery vehicles for a sitespecific gene transfer system.

We performed infection studies with Ad.GFP (Green Fluorescent …

Chmp1 Negatively Regulates Epidermal Growth Factor Signaling In The Drosophila Wing, Meagan Elisabeth Valentine

Theses, Dissertations and Capstones

A critical step in cellular signaling through transmembrane receptors is the down-regulation of activated receptors through the multivesicular body (MVB) pathway to the lysosome. MVB generation is mediated by the highly conserved ESCRT (0, I, II, and III) protein complexes. Though the ESCRT-III complex provides the core function of the ESCRT machinery, it is the least characterized of the ESCRT complexes. The Chmp1 protein is an ESCRT-III component and a putative tumor suppressor that has been linked to pancreatic and renal cancers in humans. However, published data on Chmp1 activity are conflicting and its role during tissue development is not …

Role Of Iron In Ethanol Derived Hepatic Stress, Jesse A. Thornton

Theses, Dissertations and Capstones

Chronic alcohol abuse is the third leading cause of preventable death in the United States. Ethanol metabolism causes liver injury through alterations in hepatic metabolic state, redox status, and acetaldehyde adduct formations. Increased iron absorption is associated with chronic ethanol consumption and may play a role in ethanol induced oxidative stress. We tested the hypothesis that normal labile iron in the liver plays a role in ethanol related pathological stress, using C57/Bl6 mice pair-fed Lieber-DeCarli liquid ethanol diets for 11 and 22 weeks. Normal iron group mice received 55mg/kg iron as ferric citrate, whereas the low iron groups received 5mg/kg. …

Age-Associated Alterations In The F344xbn Rat Heart And The Efficacy Of Chronic Acetaminophen Ingestion As A Therapeutic Approach, Sunil K. Kakarla

Theses, Dissertations and Capstones

Despite advances in therapeutic and diagnostic strategies cardiovascular disease still remains the leading cause of death in older people. Recent studies suggest that age- related elevations in oxidative stress are associated with increases in cardiac apoptosis and compensatory hypertrophy of the remaining cardiomyocytes. Acetaminophen has been shown to exhibit cardioprotective effects following ischemia-reperfusion injury by acting as an antioxidant. Here, we investigate the molecular mechanisms underlying aging associated cardiac remodeling in male F344XBN rats and examine the efficacy of chronic acetaminophen ingestion as a therapeutic approach.

Aging in male F344XBN rats is characterized by a significant increase in heart size, …

Muscle Plasticity And Intramuscular Signaling In The Insulin-Resistant Obese Zucker Rat, Anjaiah Katta

Theses, Dissertations and Capstones

The ability to increase skeletal muscle mass may have important implications for the treatment of insulin resistance (IR) and diabetes [1-3]. Recent data suggest that IR muscle may adapt differently than normal muscle; however, molecular mechanism(s) responsible for this finding are not well understood [4]. Herein, we investigate the molecular mechanisms underlying the skeletal muscle remodeling in the IR Obese Zucker (OZ) rat.

The OZ rat is characterized by skeletal muscle insulin resistance, hyperglycemia, and hyperlipidemia. Compared to LZ rats, our data demonstrate that soleus muscle hypertrophy was significantly attenuated in the OZ rats after 3-weeks of muscle overload and …

Identification And Characterization Of Novel Sir3/Mecp2-Chromatin Interactions, Nicholas L. Adkins

Theses, Dissertations and Capstones

The eukaryotic genome is packaged into chromosomes that are made up of a highly organized and heavily regulated structure called chromatin. The proteins involved in the compaction of DNA into this condensed state are mostly understood at the level of the structure of the nucleosome. The higher order arrangement of chromatin and how it effects gene regulation is only partially understood and characterized. The compaction of nucleosomal arrays into 30-nm and higher structures are partially the responsibility of architectural, or structural, chromatin associated proteins. The following dissertation analyzes the individual chromatin contributions of two well studied architectural proteins, the yeast …

Age-Related Dgc Structure And Function In The F344/N X Bn Rat Heart, Sunil K. Kakarla

Theses, Dissertations and Capstones

Recent data has suggested that disruption of the dystrophin-glycoprotein complex (DGC) may be involved in mediating the progression of cardiac hypertrophy and failure. Here we examined the regulation of DGC proteins in the hearts of adult (6 months), aged (30 months), and very aged (36 months) F344/N X BN rats . Compared to adult animals, the content of α- and β-dystroglycan were 6.93 ± 5.16% and 58.36 ± 3.64% higher, respectively (P < 0.05) in very aged animals. Immunoblotting and immunhistochemical analysis suggested that aging appeared to diminish alpha-sarcoglycan, beta-sarcoglycan and delta-sarcoglycan content by 13.89 ± 3.1%,15.8 ± 2.8% and 18.63 ± 3.04%, respectively (P < 0.05). These alterations in the DGC occurred coincident with age- associated alterations in cytoplasmic anti-rat IgG immunoreactivity, TUNEL positive nuclei, alpha-fodrin cleavage, indices of caspase-3 activation and diminished AKT phosphoryation (Ser 308). Taken together, these data suggest that aging alters cardiac DGC structure and function.

Lean And Obese Zucker Rats Exhibit Different Patterns Of P70s6kinase Regulation In The Tibialis Anterior Muscle In Response To High Force Muscle Contraction, Anjaiah Katta

Theses, Dissertations and Capstones

Increased muscle loading results in phosphorylation of the 70 kDa ribosomal S6 kinase (p70s6k) signaling pathway and this event strongly correlates with degree of muscle adaptation following resistance exercise. Here, we compared the basal and contraction- induced phosphorylation of p70s6k, Akt and mTOR in tibialis anterior muscles of lean and obese Zucker rats. Immunoblotting demonstrated differences in level of basal p70S6k phosphorylation (Thr 389) in the normal and diabetic TA. HFES had an increase in p70S6k (Thr389) phosphorylation at 0-, 1- and 3-hr in lean TA and only at 3-hr in obese TA. mTOR (Ser 2448) Phosphorylation was elevated in …

The Genetic And Molecular Characterization Of The Polycystic Kidney Disease-Causing Mouse Gene Bicc1, Sarah J. Price

Theses, Dissertations and Capstones

Polycystic kidney disease (PKD) is one of the most common hereditary diseases and is characterized by progressive cyst formation, substantial renal enlargement, and frequently, progression to end-stage renal disease. One way to learn more about the etiology of this disease is to study mouse models that imitate the human situation. The juvenile congenital polycystic kidney disease (jcpk) gene on mouse Chromosome 10 has been found to cause a severe, early onset form of PKD when inherited in an autosomal recessive manner (Flaherty et al., 1995). Previous genetic studies mapped the jcpk locus to a 1 cM region on mouse Chromosome …

Cytoanalysis Of Pancreatic B-Cells: Using An Avian Model, Mammalian Tissue Culture And Implications Of Antisense Oligonucleotides Transfection, Ayman Salah-El-Deen Amer

Theses, Dissertations and Capstones

Calbindin-D_28k (CaBP_28K) is a vitamin D-dependent calcium-binding protein that may alter intracellular calcium ion levels, [Ca²⁺]i. This dissertation describes experiments done to gain an understanding of the potential role of CaBP_28k in pancreatic B-cells in control of insulin secretion. The localization of CaBP_28k and insulin in chicken pancreas are shown in Chapter 1. CaBP_28k expression was found to be highest in ventral and dorsal lobes and lowest in splenic lobe. Insulin concentrations were distributed similarly among these lobes. Confocal microscopic studies demonstrated colocalization of insulin and CaBP_28k in Bcells. These findings …

The Neural Connectivity Of The Torus Semicircularis In The Cave Dwelling Form Of Astyanax Fasciatus, Timothy B. Angel

Theses, Dissertations and Capstones

Afferent projections of the torus semicircularis were traced in the blind cave form of Astyanax fasciatus using HRP tract-tracing techniques, and the results were compared to those found in previous studies of closely related sighted teleosts.

Following unilateral injections of tracer to the nucleus centralis of the torus semicircularis, retrogradely labeled cells were found in two octaval nuclei of the medulla. The descending octavolateralis nucleus was labeled bilaterally and the anterior octaval nucleus was labeled primarily on the ipsilateral side. Both the descending and anterior octavolateralis nuclei are known to receive auditory input from CN VIII in sighted fish. In …

The Characterization Of Two Differentially Expressed Plasma Proteins In Obese Versus Lean Rats In Two Rodent Models Of Obesity, Roger D. Boggs

Theses, Dissertations and Capstones

Zucker fa and La/N faf rats are widely studied models of genetic obesity and its complications. These two rodent models of obesity were utilized to search for a circulating protein marker for obesity. Plasma samples from both of these models of obesity were collected and analyzed via SDS-PAGE analysis. Two proteins were found which demonstrate differential expression between lean and obese rats. Both proteins demonstrated increased expression in the obese rats compared to the lean. One differentially expressed protein migrated on SDS-PAGE gels at 116 KD while the second migrated at 22 KD compared to molecular weight markers. The 22 …