Medicine and Health Sciences Commons^™

Regulation Of The Mouse And Human Β-Globin Genes By Krüppel Like Transcription Factors Klf1 And Klf2, Yousef N. Alhashem

Theses and Dissertations

Krüppel-like factors KLF1 and KLF2 are closely related transcription factors with three zinc finger domains in their carboxy-termini. KLF1 (erythroid Krüppel-like factor, or EKLF) plays essential roles in embryonic and adult erythropoiesis. KLF2 is a positive regulator of the mouse and human embryonic β- globin genes. KLF1 and KLF2 have overlapping roles in embryonic erythropoiesis, as demonstrated using single and double knockout (KO) mouse models. Ablation of the KLF1 or KLF2 gene causes embryonic lethality, and double KO embryos are more anemic and die sooner than either single KO. We have shown that KLF1 and KLF2 positively regulate the human …

Association Of Genetic Variation, Gene Expression, And Protein Abundance Within The Natriuretic Peptide Pathway, Bipin Sunkara

Honors College Theses

Background: The natriuretic peptide (NP) system is a critical physiologic pathway in heart failure, but there is marked individual variability in its functioning which may be genetic in origin. We investigated how genetic variations in NP pathway genes correlate with gene expression and protein abundance.

Methods: DNA, RNA, and tissue from human kidney (n=77) were obtained. Kidney was chosen as target tissue because each of the four candidate genes and proteins (natriuretic peptide receptor 1 (NPR1), NPR2, NPR3 and membrane metallo-endopeptidase (MME)) are expressed there. DNA samples were genotyped for 120 single nucleotide polymorphisms (SNP) in these four genes using …

Peptide-Mediated Targeting Of Angiogenesis For Molecular Imaging And Treatment Of Cancer, Choi-Fong Cho

Electronic Thesis and Dissertation Repository

The development of screening approaches to identify novel affinity ligands has paved the way for a new generation of molecular targeted nanomedicines. To identify novel targeting ligands, several studies have demonstrated the advantages in screening one-bead-one-compound (OBOC) libraries. Conventional methods typically bias the display of the target protein to ligands during the screening process. We have developed an unbiased multiplex ‘beads on a bead’ strategy to isolate, characterize, and validate high affinity ligands from OBOC libraries. In addition, due to the advantages associated with screening OBOC libraries directly against living cells, we sought to combine cell-based screen methods with automated …

The Role Of Aldhhicd44+ Cells In Breast Cancer Metastasis And Therapy Resistance, Alysha K. Croker

Electronic Thesis and Dissertation Repository

Breast cancer is a leading cause of death in women, due primarily to ineffective treatment of metastatic disease. Despite being a lethal process, metastasis is surprisingly inefficient, with less than 0.5% of cancer cells able to successfully navigate the metastatic cascade, indicating that only a small proportion of cancer cells possess the necessary characteristics to establish metastases. Cancer stem cells (CSCs) have recently been identified in leukemia and solid tumors; however, the role of CSCs in breast cancer metastasis and therapy resistance remains poorly understood. Sub-populations of cells demonstrating stem-cell like characteristics (high expression of CSC markers and/or high ALDH …

The Role Of Ca2+/Calmodulin Dependent Protein Kinase Ii Alpha In Group 1 Metabotropic Glutamate Receptor Endocytosis And Signalling, Stephanie C. Kulhawy

Electronic Thesis and Dissertation Repository

Group 1 metabotropic glutamate receptors (mGluR1 and mGluR5) are G-protein coupled receptors (GPCRs) activated by glutamate. mGluR1/5 couples to Gα_q/11 and releases Ca²⁺ from the endoplasmic reticulum. Ca²⁺/calmodulin-dependent protein kinase II alpha (CaMKIIα) can be activated by Gα_q/11-mediated Ca²⁺ release through binding of Ca²⁺/calmodulin. Results from a proteomic screen identified CaMKII as a novel mGluR-interacting protein. Therefore, we hypothesized that CaMKIIα associates with group 1 mGluRs and this association alters mGluR1/5 signalling and internalization. Firstly, we demonstrated the novel association between CaMKIIα and mGluR1/5 by co-immunoprecipitation of transiently transfected proteins in …

Immunohistochemical Studies Of Expression And Correlation Of Osteopontin, Cd44, And Integrin Αvβ3 In Selected Benign And Malignant Salivary Gland Tumours, Tommy Fok

Electronic Thesis and Dissertation Repository

Osteopontin (OPN) is a phosphorylated acidic glycoprotein that has been shown to play a vital role in tumourigenesis and metastases. The objective of this study was to determine the expression of OPN and its receptors (CD44 and integrin α_vβ₃) in several salivary gland cancers, and assess its clinical correlations.

Immunohistochemical techniques were used to semi-quantify the level of OPN expression and its receptors in normal salivary glands and different types of salivary gland tumour.

Our results show that in tumours of salivary gland origin, higher levels of OPN were detected in tumours which were malignant compared …

Roles Of Pgc-1Α/Ppars Pathway In Regulating Insulin Sensitivity In Mouse Skeletal Muscle Cells Under Prolonged Hypoxia, Yunyan Zhang

Electronic Thesis and Dissertation Repository

Using the C₂C₁₂ mouse myoblast cell line, I investigated how prolonged hypoxia affected components of the insulin signalling and FAO/PGC-1α/PPARs pathways, as they might impact insulin sensitivity in skeletal muscle cells.

Hypoxia resulted in lower p-Akt (Thr 308) and higher total cellular GLUT4 protein levels after 7 days of differentiation. This coincided with higher triglyceride content and alterations of the FAO/PGC-1α/PPARs components, both of which could contribute to the changes observed in the components of the insulin signalling pathway. Specifically, cells differentiating in 1% O₂ had lower SIRT1, PPAR- α, FATP4 and MCAD mRNA; accompanied by …

The Role Of Non-Canonical Wnt/Planar Cell Polarity Signalling In Breast Cancer Progression, Connor D. Macmillan

Electronic Thesis and Dissertation Repository

The role of planar cell polarity (PCP) signalling in breast cancer is unclear, as evidence shows that activation of this pathway with WNT5A can either promote or inhibit progression. Using the 21T cells, which represent distinct stages of breast cancer progression when grown in a mouse xenograft model (21PT, atypical ductal hyperplasia; 21NT ductal carcinoma in situ; 21MT-1, invasive mammary carcinoma), I hypothesized that WNT5A will promote progression only when VANGL1, another component of PCP signalling, is elevated.

Each 21T cell line showed distinct stage-specific morphologies when grown in a Matrigel “on-top” assay. WNT5A overexpression promoted progression to an invasive …

From Linkage To Gwas: A Multifaceted Exploration Of The Genetic Risk For Alcohol Dependence, Amy Adkins

Theses and Dissertations

Family, twin and adoption studies consistently suggest that genetic factors strongly influence the risk for alcohol dependence (AD). Although the literature supports the role of genetics in AD, identification of specific genes contributing to the etiology of AD has proven difficult. These difficulties are due in part to the complex set of risk factors contributing to the development of AD. These risk factors include comorbidities with other clinical diagnoses and behavioral phenotypes (e.g., major depression), physiological differences that contribute to the differences between people in their level of response to ethanol (e.g., initial sensitivity) and finally the large number of …

Gender Does Not Influence The Relationship Between Posterior Cruciate Ligament Design And Patient-Reported Outcomes In Patients Receiving Primary Unilateral Total Knee Arthroplasty, Ryan Milan

Electronic Thesis and Dissertation Repository

The effect of the interaction between gender and posterior cruciate ligament (PCL) prosthesis design on patient-reported outcomes is an understudied area of research. We evaluated 1613 patients, from the Ontario Joint Replacement Registry (2001-2006), who underwent primary total knee replacement. This study investigated the impact of the gender-PCL design interaction on Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) change scores and patient satisfaction, by performing linear regression analysis, using full-adjusted models that also included the gender-PCL prosthesis design interaction variable. PCL prosthesis design did not affect WOMAC change scores or satisfaction (p>0.05). Moreover, Gender did not influence either …

Regulation Of Nk Cell-Mediated Tubular Epithelial Cell Death And Kidney Ischemia-Reperfusion Injury By The Nkr-P1b Receptor And Clr-B, James Yip

Electronic Thesis and Dissertation Repository

Renal ischemia-reperfusion injury (IRI) occurs following reduced renal blood flow and is a major cause of acute injury in both native and transplanted kidneys. We have previously demonstrated that NK cells can mediate tubular cell death and kidney IRI. Natural killer receptor-protein 1B (NKR-P1B) has been shown to interact with C-type lectin-related protein B (Clr-b), resulting in the suppression of NK cell-mediated cytotoxicity.

Clr-b mRNA and protein expression in the kidney were up-regulated after renal IRI. Similar upregulation of Clr-b expression was seen in cytokine-challenged primary-cultured tubular epithelial cells (TEC). Furthermore, NK cytotoxicity assays demonstrated enhanced necrotic death in TEC …

A Beginning Education In The Anatomy And Function Of The Human Visual Apparatus For Grades 2-4, Evan Andrews

Honors Theses

The human visual apparatus is a remarkable feat of evolution that is nearly a complete mystery to many in the general population. Introducing such a challenging topic is difficult, but, if done in an enjoyable and interactive way to a young audience, the eye can become exciting instead of confusing. The primary objective of this project was to create a short lesson for elementary school students that gave an understanding of the eye's basic anatomy and functions. A fast-paced, interactive format was decided on that included units on the major eye components; color vision and how it can be affected; …

The Adverse Effects Periodontal Disease Has On Women, Jennifer Long

Honors Theses

The Adverse Effects Periodontal Diseases Has on Women Periodontal diseases are challenging to define given the number and variety of different oral diseases. This review sought to answer the question: How does a periodontal disease impact a woman throughout the life span? Specifically, this document discussed how, because of the cycling hormones and the effects those hormones have on the different bacteria, women's periodontal diseases differ from men's. Diseases of the mouth were first discovered during the Egyptian period and the information and knowledge about the different diseases advanced as time progressed. From here, non-surgical and surgical treatments were discussed …

Diffuse Traumatic Axonal Injury Within The Visual System: Implications For Visual Pathway Reorganization, Jiaqiong Wang

Theses and Dissertations

Traumatic brain injury is a major health problem with much of its morbidity associated with traumatic axonal injury (TAI). To date, significant insight has been gained into the initiating pathogenesis of TAI. However, the specific anterograde and retrograde sequelae of TAI are poorly understood because the diffuse nature of TAI complicates data analysis. To overcome this limitation, we subjected transgenic mice expressing yellow fluorescent protein (YFP) within the visual system to central fluid percussion injury, and consistently generated diffuse TAI within the optic nerve that could easily be followed in the organized YFP positive fibers. We demonstrated progressive axonal swelling, …

Hematopoietic Stem Cell Threshold Sensing Controls Regulatory Pathways Facilitating Clinically Relevant Ex Vivo Expansion For Stem Cell Transplantation, Andrew D. Lasiter

Theses and Dissertations (ETD)

The ex vivo expansion of hematopoietic stem cells (HSCs) for transplantation has threefold importance: 1.) First, because of the rarity of stem cells there often isn’t a sufficient supply obtainable from common sources for larger children and adults. 2.) Secondly, patient morbidity and time to hematopoietic reconstitution following myeloablative preconditioning is improved by administering a larger pool of HSCs. 3.) Lastly, gene therapies for hematological diseases still require a robust supply of HSCs to offset varying degrees of inefficiency in vector mediated transfection protocols. These reasons, and others, have been an impetus for many discoveries made within four primary subdivisions …

Characterization Of Beryllium As A Novel Agent To Study Cell Cycle Arrest And Cellular Senescence, Priyatham Gorjala

UNLV Theses, Dissertations, Professional Papers, and Capstones

Cancer cells evade senescence, apoptosis, and other constraints on proliferation, often via mutation of the p53 tumor suppressor gene (TP53). Normal human lung fibroblasts have been shown to enter premature senescence upon exposure to beryllium. In these cells, BeSO₄ stabilizes p53 protein, increases p21 gene expression, induces senescence-associated β-galactosidase activity and causes cell proliferation arrest. In the present study, we have investigated whether BeSO₄ is able to induce similar effects in cancer cells that have wildtype p53. We have demonstrated that beryllium salt at low concentration can induce molecular changes in the p53 signaling pathway leading to cell …

Credentialing Success In Respiratory Therapy Education: Revisiting Bourdieu's Concepts Of Field And Capital, Karen Lightbody Shaw

UNLV Theses, Dissertations, Professional Papers, and Capstones

The field of Respiratory Therapy (RT) is expected to experience a workforce shortfall over the next decade. The numbers of both program applicants and graduates have declined in recent years, necessitating strategies to improve board exam pass rates for future graduates. In response to the pending employment crisis, the Commission on Accreditation for Respiratory Care has published Programmatic Outcomes Data detailing individual program statistics. A theoretical framework adapted from Pierre Bourdieu's Concepts of Field and Capital was proposed to explain a possible re-stratification of RT programs. It states, in part, that a modification of position-takings within the RT educational field …

Evaluation Of Polymer Gel Dosimeters For Measurements Of Dose And Let In Proton Beams, Kevin M. Vredevoogd

Dissertations & Theses (Open Access)

This project assessed the effectiveness of polymer gel dosimeters as tools for measuring the dose deposited by and LET of a proton beam. A total of three BANG® dosimeter formulations were evaluated: BANG®-3-Pro-2 BANGkits™ for dose measurement and two BANG®-3 variants, the LET-Baseline and LET-Meter dosimeters, for LET measurement. All dosimeters were read out using an OCT scanner. The basic characteristics of the BANGkits™ were assessed in a series of photon and electron irradiations. The dose-response relationship was found to be sigmoidal with a threshold for response of approximately 15 cGy. The active region of the dosimeter, the volume in …

Development Of A Beam-Specific Planning Target Volume And A Robust Plan Analysis Tool For Proton Therapy, Peter Park

Dissertations & Theses (Open Access)

Proton therapy is growing increasingly popular due to its superior dose characteristics compared to conventional photon therapy. Protons travel a finite range in the patient body and stop, thereby delivering no dose beyond their range. However, because the range of a proton beam is heavily dependent on the tissue density along its beam path, uncertainties in patient setup position and inherent range calculation can degrade thedose distribution significantly. Despite these challenges that are unique to proton therapy, current management of the uncertainties during treatment planning of proton therapy has been similar to that of conventional photon therapy. The goal of …

Scopolamine-Induced Dry Eye Model In Female Rats, Ryan Alderson

Masters Theses

The objective of this study will be to establish, validate, and characterize a dry eye model in female rats induced by continuous systemic administration of scopolamine via Alzet® osmotic pumps for 28 days. This study will use female rats as a model for dry eye and will be implanted with osmotic pumps that will continuously distribute scopolamine or saline systemically over 28 days. Scopolamine will be used to induce the dry eye condition and saline will be used as a negative control. The clinical signs of dry eye will be evaluated by tear volume measurements with sterilized phenol-red thread (FCI …

The Role Of Bip Nucleotide Exchange Factor Sil1 In Immunoglobulin Biosynthesis, Tyler Sanford

Theses and Dissertations (ETD)

BiP is an essential endoplasmic reticulum resident molecular chaperone of the HSP70 family that binds exposed hydrophobic regions of unfolded proteins. Substrates bound by BiP are protected from dangerous non-specific interaction with other unfolded proteins via their aggregation prone exposed hydrophobic regions, which are normally buried in the native state, and act as BiP binding targets. For these substrates to mature into the native state, BiP must be released. Like all HSP70 family members, cycles of BiP binding and release are nucleotide dependent and facilitated by two genres of cochaperones, HSP40s, and nucleotide exchange factors (NEFs). HSP40s function in stimulating …

Metabotropic Glutamate Receptor 5 In The Medial Prefrontal Cortex: Role In Cocaine Sensitization And Addiction, Kristin Marie Timmer

Theses and Dissertations (ETD)

Cocaine sensitization is associated with cocaine-induced hyperexcitability of pyramidal projection neurons within the medial prefrontal cortex (mPFC). Such hyperexcitability presumably results in increased glutamatergic input to reward-affiliated brain regions such as the ventral tegemental area (VTA) and nucleus accumbens (NAc), consequently facilitating drugseeking behavior. Metabotropic glutamate receptor 5 (mGluR5) has been implicated in cocaine addiction and demonstrated to increase neuronal excitability, therefore, the aim of the present study was to investigate the effect of intra-mPFC mGluR5 manipulation on behavioral and neurochemical sensitization and drug-seeking. Bilateral cannulae were implanted into the mPFC of male Sprague-Dawley rats and mGluR5 antagonist MTEP (15 …

N200 And P300 Evoked By Stimuli Straddling Category Boundary In Lexical Context, Ji Young Lee

Theses and Dissertations (ETD)

Background and Objectives: Event-related potentials (ERPs) like N200 and P300 have been reported to reflect the categorical perception of speech. The purpose of the present study is to explore whether these ERP components reflect the influence of lexical context on categorical perception. Findings may provide evidence for bottom-up or top-down processing of speech. Methods: On a seven step series of the /bi/-/pi/ continuum, a two-forced choice labeling test was administered in two conditions: /bi/ context (e.g. bee sting) and /pi/ context (e.g. pea soup). From the labeling results, Stimulus 1which is a prototypical /bi/ was selected for standard stimuli, and …

Mrp4 Is A Crucial Regulator Of Testosterone Biosynthesis, Jessica Ann Morgan

Theses and Dissertations (ETD)

The physiological role of multidrug resistance protein 4 (Mrp4) in the testes is unknown. It was discovered that Mrp4 is expressed primarily in mouse and human Leydig cells; however, there is no current evidence that Mrp4 regulates testosterone biosynthesis. The role of Mrp4 was investigated in Leydig cells where testosterone production is regulated by cAMP, an intracellular second messenger formed when the luteinizing hormone (LH) receptor (Lhr) is activated. As Mrp4 regulates cAMP, we compared testosterone levels in our Mrp4 WT and KO mice. Prepubertal KO mice had significantly reduced testicular testosterone, impaired gametogenesis, and disrupted cAMP homeostasis, resulting in …

Anti-Apoptotic Mcl-1 Localizes To The Mitochondrial Matrix And Couples Mitochondrial Fusion To Respiration, Rhonda Perciavalle

Theses and Dissertations (ETD)

MCL-1, an anti-apoptotic BCL-2 family member that is essential for the survival of multiple cell lineages, is also among the most highly amplified genes in cancer. Although MCL-1 is known to oppose cell death, precisely how it functions to promote survival of normal and malignant cells is poorly understood. Here, I report that different forms of MCL-1 reside in distinct mitochondrial locations and exhibit separable functions. On the outer mitochondrial membrane, a MCL-1 isoform acts like other anti-apoptotic BCL-2 molecules to antagonize apoptosis, whereas an amino-terminally truncated isoform of MCL-1 that is imported into the mitochondrial matrix is necessary to …

Post-Transcriptional Regulation Of Mammalian Gene Expression In Non-Coding Region Of Target Rna, Jing Lin

Dissertations & Theses (Open Access)

Tumor Suppressor Candidate 2 (TUSC2) is a novel tumor suppressor gene located in the human chromosome 3p21.3 region. TUSC2 mRNA transcripts could be detected on Northern blots in both normal lung and some lung cancer cell lines, but no endogenous TUSC2 protein could be detected in a majority of lung cancer cell lines. Mechanisms regulating TUSC2 protein expression and its inactivation in primary lung cancer cells are largely unknown. We investigated the role of the 5’- and 3’-untranslated regions (UTRs) of the TUSC2 gene in the regulation of TUSC2 protein expression. We found that two small upstream open-reading frames (uORFs) …

Role Of Tyrosyl-Dna Phosphodiesterase (Tdp 1) On Repair Of 3′-Phosphoglycolate (3′- Pg) Terminated Dna Double-Strand Breaks (Dsbs) And In Response To Oxidative Stress, Tong Zhou

Theses and Dissertations

DNA DSBs are most toxic to cells because they can lead to genomic rearrangements and even cell death. Most DSBs induced by ionizing radiation or radiomimetic drugs such as calicheamicin and bleomycin, bear 3′-phosphate or 3′- PG moieties that must be removed to allow subsequent gap filling and ligation. DSBs can be repaired by two main pathways: the homologous recombination (HR) pathway and the non-homologous end-joining (NHEJ) pathway, NHEJ is the primary repair pathway in mammalian cells. While HR repairs single strand breaks (SSBs) or DSBs accurately by using an undamaged copy of the sequence mostly at late S phase …

Investigating A Model For Fetal Alcohol Damage In Caenorhabditis Elegans, Lindsay Kondo

Theses and Dissertations

Alcohol use and abuse has many harmful effects, especially to children exposed prenatally, including fetal alcohol spectrum disorders (FASDs). The disabilities due to fetal alcohol exposure continue throughout life and cause major financial burdens to society. The molecular mechanisms underlying FASDs are not well understood. We have taken a genetic approach to characterize ethanol’s effect on changing a discrete cell fate decision during embryogenesis in the nematode, Caenorhabditis elegans (C. elegans). Our preliminary data suggest that ethanol can affect the development of AWC neurons, a pair of olfactory neurons in C. elegans. We suggest that lipids can protect AWC neurons …

The Role Of P62 In Osteoclastogenesis And Paget’S Disease Of Bone, Tamer Hadi

Theses and Dissertations

Paget’s disease (PDB) is the second most common metabolic bone disease after osteoporosis, affecting up to 3% of adults over age 55. It is characterized by focal lesions of bone resorbed by hyperactive osteoclasts coupled with rapid formation of highly disorganized, low quality bone formed by osteoblasts. Such lesions cause skeletal deformity, fractures, and other symptoms that significantly decrease quality of life. In 2001, mutations in the SQSTM1/p62 gene were found in a subset of Paget’s patients. The work summarized in this dissertation sought to answer two broad questions: what is the function of p62 in normal bone homeostasis and …

The Role Of Magl Inhibition In Nicotine Withdrawal And Reward, Pretal Muldoon

Theses and Dissertations

ROLE OF MAGL INHIBITION IN NICOTINE WITHDRAWAL AND REWARD. A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at Virginia Commonwealth University. by Pretal Ishvarlal Patel Muldoon Director: M. Imad Damaj, PhD Professor, Department of Pharmacology and Toxicology Tobacco use is one of the leading causes of preventable deaths worldwide. Nicotine, the main psychoactive component of tobacco, sustains and initiates tobacco addiction. Cessation of nicotine induces a dependence withdrawal syndrome. Recent in vivo studies indicate that the endocannabinoid (EC) system modulates both nicotine reward and withdrawal. The purpose of this proposal is to …