Life Sciences Commons^™

Providing Experimental Evidence For Mouse Telomerase Reverse Transcriptase As A Novel And Unique Adult Neural Stem Cell Marker, Caroline Dean Curtis

Electronic Theses and Dissertations

With the prevalence of neurodegenerative pathologies in our society today it is imperative that we begin to look at novel approaches to the underlying problem of dying neurons that are not replaced. Adult neural stem cells exist naturally and could potentially be manipulated into targeted repair of damaged brains, given substantial research. The first step in this process is to find a way to specifically mark the earliest subset of these cells, the quiescent adult neural stem cells. Here we provide evidence for the existence of a novel and unique qANSC marker in mouse telomerase reverse transcriptase (mTERT). mTERT has …

Chemically Modified Monolayer Surfaces Influence Valvular Interstitial Cell Attachment And Differentiation For Heart Valve Tissue Engineering, Matthew N. Rush

Nanoscience and Microsystems ETDs

As a cell mediated-process, valvular heart disease (VHD) results in significant morbidity and mortality world-wide. In the US alone, valvular heart disease VHD is estimated to affect 2.5% of the population with a disproportionate impact on an increasing elderly populous. It is well understood that the primary driver for valvular calcification is the differentiation of valvular interstitial cells (VICs) into an osteoblastic-like phenotype. However, the factors leading to the onset of osteoblastic-like VICs (obVICs) and resulting calcification are not fully understood and a more complete characterization of VIC differentiation and phenotypic change is required before treatment of valve disease or …

Mob-Ndr Kinase Signaling Components Are Required For Epithelial Tube Formation In The Drosophila Follicular Epithelium, Juan Carlos Duhart

UNLV Theses, Dissertations, Professional Papers, and Capstones

A major goal of developmental biology is to understand how a single fertilized cell can give rise to the many functional tissues and organs, of specific sizes and shapes, that make up the adult body plan. Over the last 25 years, developmental geneticists have uncovered much concerning the cell-to-cell communication systems that are necessary to build complex tissues and organs. For example, throughout development, cells communicate with their neighbors using specialized signaling molecules. These signals are instructive and provide “signal-receiving” cells with information about space and time. That is, signal-receiving cells “learn” precisely where they are located, and, how far …

The Essential Roles Of The Chromatin Factor Gon4l In Heart Development, Terin Elise Budine

Arts & Sciences Electronic Theses and Dissertations

Heart development and the genetic pathways underlying it are highly conserved among vertebrates. During heart development, an embryo must induce mesoderm formation, pattern the mesoderm, specify cardiomyocytes, increase the population of cardiomyocytes through proliferation, and pattern the cardiac chambers. It is becoming increasingly clear that chromatin modifications help mediate the complex processes of heart development by providing spatiotemporal regulation of gene expression. My thesis work focuses on characterizing functions of the chromatin factor Gonad-4-like (Gon4l), encoded by the gene ugly duckling (udu), in zebrafish heart development. Previous works established a requirement for Gon4l in the formation of many mesoderm derivatives …

Genetic Basis Of Thermal Divergence In Saccharomyces Species, Xueying C. Li

Arts & Sciences Electronic Theses and Dissertations

The genetic architecture of phenotypic divergence is a central question in evolutionary biology. Genetic architecture is impacted by whether evolution occurs through accumulation of many small-effect or a few large-effect changes, the relative contribution of coding and cis-regulatory changes, and the prevalence of epistatic effects. Our empirical understanding of the genetic basis of evolutionary change remains incomplete, largely because reproductive barriers limit genetic analysis to those phenotypes that distinguish closely related species. In this dissertation, I use hybrid genetic analysis to examine the basis of thermal divergence between two post-zygotically isolated species, Saccharomyces cerevisiae and S. uvarum. S. cerevisiae is …

The Mitochondrial Calcium Uniporter Regulator 1 (Mcur1) Matrix Domain Is A Self-Associated Multimer Sensitive To Divalent Cations, Naveed Siddiqui

Electronic Thesis and Dissertation Repository

Mitochondria are primarily appreciated for the generation of adenosine triphosphate (ATP), a chemical store of energy required by all cells. These organelles, however, also play key roles in apoptosis, autophagy and shaping cytosolic calcium (Ca²⁺) signaling via Ca²⁺ uptake into the mitochondrial matrix. This Ca²⁺ uptake is mediated chiefly via the mitochondrial Ca²⁺ uniporter (MCU), an inner mitochondrial membrane protein that oligomerizes to form a Ca²⁺ selective pore. MCU is regulated by several protein binding partners, including the recently identified MCU regulator-1 (MCUR1). MCUR1 stabilizes a higher order MCU heterocomplex through interactions with MCU …

Regulation Of Canonical And Non-Canonical Hippo Pathway Components In Mitosis And Cancer, Seth Stauffer

Theses & Dissertations

The Hippo pathway is conserved regulator of organ size through control of proliferation, apoptosis, and stem-cell self-renewal. In addition to this important function, many of the canonical signaling members have also been shown to be regulated during mitosis. Importantly, Hippo pathway components are frequently dysregulated in cancers and have attracted attention as possible targets for improved cancer therapeutics. Further exploration of Hippo-YAP (yes-associated protein) signaling has revealed new regulators and effectors outside the canonical signaling network and has revealed a larger non-canonical network of signaling proteins in which canonical Hippo pathway components crosstalk with important cellular homeostasis and apoptosis signaling …

Delineation Of New Mechanisms Of Dna Double Strand Break Repair, Songli Zhu

Theses & Dissertations

DNA damage is frequently induced in cells by both endogenous and exogenous agents. DNA damage, particular double strand breaks (DSBs) may lead to genomic instability, and the progression of cancer, aging, neurodegeneration, and other human diseases. The cell employs two major DSB repair pathways, including homologous recombination (HR) and Non-homologous end joining (NHEJ), but the detailed mechanisms of DSB repair remain to be further revealed.

In the first part of this study, we characterized a plasmid-based assay to investigate NHEJ repair in Xenopus egg extracts. Our data argued for a preference for the precise repair by the NHEJ machinery and …

Intra- And Inter-Molecular Signaling In A Cardiac Connexin: Role Of Cytoplasmic Domain Dimerization And Phosphorylation, Andrew J. Trease

Theses & Dissertations

As critical mediators of cell-to-cell communication, gap junctions (GJs) are comprised of membrane channels that directly link the cytoplasm of adjacent coupled cells thereby allowing for the passage of ions, small metabolites, and secondary messengers. Each channel is formed by the apposition of two connexons from adjacent cells, each composed of six connexin (Cx) proteins. Each GJ channel functions to promote signal propagation and synchronization of cells and tissues in organs. Furthermore, GJs are essential for proper propagation of cardiac action potentials from one cell to the next, leading to the coordinated contraction and relaxation of heart muscle powering circulation. …

Towards An In Vitro Model Of Testing Osteoblast Cellular Function In Contact With Various Surfaces, Raheleh Miralami

Theses & Dissertations

Past studies have shown that the success of total joint replacements depends on the biocompatibility of orthopaedic materials, which can be improved by modifying the implant surface. However, the exact roles of these modifications and their effective mechanisms are poorly understood. The objective of this study was to develop and evaluate a model system to investigate the impact of nano-structured surfaces, produced by the ion beam-assisted deposition (IBAD) technique, on biomarkers of osteointegration using an in vitro model. The IBAD technique was employed to deposit zirconium oxide (ZrO₂), Titanium oxide (TiO₂), and Titanium (Ti) nano-films on …

19f-Mri To Monitor Therapeutic Cell Migration And Distribution, Corby Fink

Electronic Thesis and Dissertation Repository

A major hurdle hindering the advancement of cell-based cancer vaccines is the inability to non-invasively track in vivo therapeutic cell migration. Within cancer immunotherapy, antigen presenting cell-based (APC) immunotherapies can be dendritic cell-based (DC) and elicit de novo anti-tumor immune responses or formulated as mixed APC-based immunotherapy generated from peripheral blood mononuclear cells (PBMC). Mixed APC-based therapies contain monocytes and B cells as APC and can enhance a pre-existing anti-tumor immune response such that objective anti-tumor responses are observed. In either APC-based formulation, the quantity of anti-tumor APC reaching a secondary lymphoid organ post injection is predictive of the magnitude …

Frost450, A Novel Microtubule Targeting Agent: Resistance And Cross-Resistance In An Ovarian Cancer Cell Line, Jonathan Chancellor Fox

Undergraduate Honors Theses

Drug resistance remains problematic for chemotherapy treatments. As primary treatment strategies tend to lose potency against cancer, there is a need for treatment options that overcome drug resistance mechanisms. FROST450, a novel microtubule destabilizing agent, has proven anticancer effects that overcome taxane and multidrug resistance. However, to better understand the clinical potential of FROST450, in this study we examine the mechanisms and implications of acquired FROST450 resistance in an A2780 ovarian cancer cell line. Results indicate FROST450 resistance is associated with increased mitochondrial respiration and with changes in gene sequence or expression level of proteins associated with microtubules, and that …

Investigating Autophagy Dysfunction Induced By A Parkinson's Disease-Causing Mutation In Vps35, Abir Ashfakur Rahman

Boise State University Theses and Dissertations

Parkinson’s Disease (PD) is an idiopathic disorder with no known cure. With number of cases steadily rising around the world, it is imperative to turn to the underlying cellular and molecular mechanisms of the disease manifestation and neurodegeneration to craft novel modes of therapy. VPS35 is one of the few genes that have identified and definitively linked to familial PD. The particular mutation that has been associated is known to cause dysfunction of a key cellular process known as autophagy. This process is primarily responsible for clearance of unwanted, damaged or misfolded proteins, among other things. Our study reveals an …

Microenvironmental Forces Regulate Notch Signaling Through Integrins, Michael Allen Detweiler

Boise State University Theses and Dissertations

The extracellular microenvironment contributes significantly to a cell’s function and behavior. For instance, cell-cell interactions, cell-substrate interactions, and physical forces are all factors of the extracellular environment that can alter cellular behavior. Cells can receive these signals and forces through various membrane channels and receptors that transmit the signals from the extracellular to the intracellular space. Canonical Notch signaling is induced by ligand interactions with neighboring cells, but recent evidence has revealed that Notch signaling can occur through a variety of extracellular stimuli including hyperglycemia, hypoxia, multiple growth factors, fluid shear stress, and extracellular matrix (ECM) composition. Although Notch activation …

Impedance Sensing Of Cancer Cells Directly On Sensory Bioscaffolds Of Bioceramics Nanofibers, Hanan Alismail

Graduate Theses and Dissertations

Cancer cell research has been growing for decades. In the field of cancer pathology, there is an increasing and long-unmet need to develop a new technology for low-cost, rapid, sensitive, selective, label-free (i.e. direct), simple and reliable screening, diagnosis, and monitoring of live cancer and normal cells in same shape and size from the same anatomic region. For the first time on using an impedance signal, the breast cancer and normal cells have been thus screened, diagnosed and monitored on a smart bioscaffold of entangled nanowires of bioceramics titanate grown directly on the surface of implantable Ti-metal and characterized by …

Role Of P300 Zz Domain In Chromatin Association And Histone Acetylation, Yongming Xue

Dissertations & Theses (Open Access)

Transcription is strictly regulated by numerous factors including transcription coactivators. The p300 protein and its close paralogue CREB-binding protein (CREBBP, aka CBP) are well-known transcriptional coactivators that have intrinsic lysine acetyltransferase activity. The functions of p300/CBP largely rely on their capabilities to bind to chromatin and to acetylate the histone substrates. However, the molecular mechanisms underlying the regulation of these processes are not fully understood.

Through combination of various biochemical, biophysical and molecular approaches, we show that the ZZ-type zinc finger (ZZ) domain of p300 functions as a histone reader that specifically binds the N-terminal tail of histone H3. Crystal …

Prioritizing Chemical Constituents In Tobacco Products And Smoke To Predict Developmental Osteotoxicity In Human Embryonic Stem Cells, Joseph Madrid

Electronic Theses, Projects, and Dissertations

Though it is well known that tobacco related products can cause prenatal maldevelopment, very little is known on how tobacco products affect bone tissue as it develops in the embryo. Identifying which chemicals can induce the greatest harm to the prenatal skeletal system is an improbable task as there are over 7,000 chemicals in tobacco smoke alone. We hypothesized that the Toxicological Priority Index (ToxPI) program can be used to rank osteogenic cytotoxicity potential to aid in the assessment of what chemicals out of the thousands can cause osteogenic differentiation inhibition. ToxPI aggregates information from various assays and incorporates them …

Epidermal Growth Factor-Like Ligands Regulate Dimer Selection., Jamie S. Rush

Electronic Theses and Dissertations

There are thirteen known endogenous EGF-like ligands. We previously reported that Betacellulin (BTC) increases ligand-mediated corneal wound healing more than Epidermal Growth Factor (EGF) [Peterson et al. (2014) IOVS 55(5):2870-80], although the molecular reason for this is unknown. Despite being better at promoting wound healing via enhanced cell migration, BTC has reduced receptor affinity and weaker induction of EGFR phosphorylation. These data indicate that BTC’s response is not due to enhanced affinity or EGFR-kinase activity. Receptor phosphorylation and proximity ligation assays indicate that BTC treatment significantly increases ErbB3 phosphorylation and EGFR:ErbB3 heterodimers. BTC traffics EGFR at a faster rate than …

Engineering Yeast To Evaluate Human Proteins Involved In Selective Rna Packaging During Hiv Particle Production, Ryan M. Bitter

Master's Theses

Despite recent advances in antiretroviral therapy, nearly 37 million people continue to live with human immunodeficiency virus (HIV). Basic and applied research on the assembly of HIV could be enhanced by using a genetically tractable organism, such as yeast, rather than mammalian cells. While previous studies showed that expression of the HIV Gag polyprotein in Saccharomyces cerevisiae spheroplasts resulted in the production of virus-like particles (VLPs), many questions regarding the utility of yeast in HIV assembly remain uninvestigated. Here, we report use of S. cerevisiae for both the production of VLPs with selectively packaged RNA and to evaluate the human …

Molecular Dynamics Of P21 And Fluorescent Sphingomyelin In Keratinocytes Exposed To Uvb, Tyler Malcolm Fraser

Master's Theses

Non-melanoma skin cancer (NMSC) is the most common malignant tumor, representing more than a third of all malignant tumors combined and the incidence is increasing every year. Ultraviolet (UV) radiation from the sun is the most dominant factor contributing to tumor initiation and progression. The condition is most prevalent in populations with lighter skin and older age. Current pharmaceutical molecular research targets the inhibition of the Epidermal Growth Factor Receptor (EGFR), a receptor which is commonly over-expressed or dysregulated in skin malignancies. This study evaluates the content and location of the damage marker p21 within keratinocytes that were incubated in …

Determining The Molecular Mechanisms Of Pacs-1-Mediated Protein Sorting, Brennan S. Dirk

Electronic Thesis and Dissertation Repository

Membrane trafficking events are required to direct proteins to their precise subcellular locations. The cellular Phosphofurin Acidic Cluster Sorting protein – 1 (PACS-1) has emerged as a protein of interest in controlling the localization of a multitude of cellular and viral proteins. Specifically, PACS-1 is hijacked by type-1 Human Immunodeficiency Virus (HIV-1) to contribute to immune evasion in addition to regulating neuroendocrine hormone storage and release. To accomplish this, PACS-1 connects the cytoplasmic tail of cellular receptors to the heterotetrameric adaptor proteins (APs) to form a functional trafficking unit. Throughout this dissertation, I explored the role of PACS-1 and AP-1 …

Actinomycin D And Telmisartan Combination Therapy Targets Lung Cancer Stem Cells, Ryan Green

USF Tampa Graduate Theses and Dissertations

The failure of lung cancer treatments has been attributed partly to the development of drug resistance, however the underlying cellular and molecular mechanisms are poorly understood. It has been suggested that a very small group of specific cells within the heterogeneous tumors, cancer initiating stem cells (CSC), develop resistance to treatment, survive and later initiate the growth of new tumors. Due to their pivotal role in maintenance and relapse of tumors following the acquisition of drug resistance, we reasoned that novel drugs targeting cancer cells and CSC might provide the most effective treatments, if not a cure. To this end, …

Fatty Acid Amides And Their Biosynthetic Enzymes Found In Insect Model Systems, Ryan L. Anderson

USF Tampa Graduate Theses and Dissertations

A fatty acid amide is precisely as the name suggests: A fatty acid (CHn-COOH), in which the hydroxyl group of the carboxylic acid is displaced by an amine functional group from a biogenic amine (R-NH2), ultimately forming an amide bond. Furthermore, these fatty acid amides can be composed of a variety of different acyl chain lengths donated by the fatty acid and a myriad of different biogenic amines. Thus, these molecules can be subdivided in a number of different ways including the separation of short chain (acetyl to heptanoyl) and long chain (palmitoyl to arachidonoyl) and also based off the …

Investigating The Roles Of Fucosylation And Calcium Signaling In Melanoma Invasion, Tyler S. Keeley

USF Tampa Graduate Theses and Dissertations

Melanoma is the deadliest form of skin cancer. Prognosis for early stage melanoma patients is excellent, and surgery is often curative for these patients. However, once patients have presented with invasive disease, the average 5-year survival rate drops significantly from over 90% to between 10 and 15%. Several therapies have been developed to target a commonly mutated oncogene BRAF, or its downstream effectors. Unfortunately, while these treatments show robust initial response, most patients relapse within a year. Moreover, therapy-resistant tumors are often more invasive and metastatic. Therefore, it is important to investigate the molecular mechanisms underlying melanoma invasion and metastasis, …

Investigation Of Anemarrhena Asphodeloides And Its Constituent Timosaponin-Aiii As Novel, Naturally Derived Adjunctive Therapeutics For The Treatment Of Advanced Pancreatic Cancer, Catherine B. Marelia

USF Tampa Graduate Theses and Dissertations

Pancreatic adenocarcinoma (PDAC) remains one of the most lethal cancer types, with 92% of patients succumbing to the disease within five years. One of the hallmarks of this intractable disease is its nearly universal resistance to chemotherapeutic intervention. The deoxycytidine analog gemcitabine persists as the primary chemotherapy for advanced PDAC patients despite little improvement to its toxicity and tenuous efficacy over the past two decades. Plant-based drug discovery has a longstanding history and has led to some of the most significant drugs of the modern era, including some chemotherapeutic agents like paclitaxel and camptothecin. Our study analyzed the potential inhibitory …

Studies Of Dynein Anchoring Protein In Budding Yeast Saccharomyces Cerevisiae, Safia Omer

Doctoral Dissertations

Inheritance of equal genomic content is vital for cellular growth and survival. During cell division, the mitotic spindle is built to ensure equal chromosome segregation between dividing cells. Fidelity of segregation is achieved by the precise positioning of the mitotic spindle by force generators located at the cell cortex where they exert pulling on microtubule (MT). Cortical dynein, a minus end directed motor, generates pulling forces via microtubule end-on capture-shrinkage and lateral MT sliding mechanisms. In Saccharomyces cerevisiae, the dynein attachment molecule Num1 interacts with plasma membrane (PM), endoplasmic reticulum (ER), and mitochondria to facilitate spindle positioning across the …

Epigenetic Regulation Of Genomic Imprinting During Early Mammalian Embryonic Development, Chelsea Marcho

Doctoral Dissertations

Mammalian development involves remarkable changes, starting from a single-cell, totipotent zygote and ending with a developed organism comprised of diverse cells types with distinct morphologies, structures, and functions. Within three days of murine development, the two parental genomes merge into a single nucleus, begin zygotic gene expression, undergo epigenetic remodeling, and make the first lineage decisions. Diversity in cell-types is possible even though cells share the same genome. This diversity is achieved by the tight regulation of differential transcriptional programs. There are many ways these transcriptional programs can be initiated. Epigenetic alterations to the genome can drive transcriptional changes. Epigenetic …

Identifying Functional Components Of The Endoplasmic Reticulum Quality Control And Degradation Factor Edem1, Lydia Lamriben

Doctoral Dissertations

The ER Degradation-Enhancing Mannosidase-Like protein 1 (EDEM1) is a critical endoplasmic reticulum (ER) quality control factor involved in identifying and directing non-native proteins to the ER-associated protein degradation (ERAD) pathway. However, its recognition and binding properties have remained enigmatic since its discovery. Here we provide evidence for an additional redox-sensitive interaction between EDEM1 and Z/NHK that requires the presence of the single Cys on the α-1 antitrypsin ERAD clients. Moreover, this Cys-dependent interaction is necessary when the proteins are isolated under stringent detergent conditions, ones in which only strong covalent interactions can be sustained. This interaction is inherent to the …

Combination Regimens Using Dietary Components For The Chemoprevention Of Colorectal Cancer And Inflammation, Christina Dimarco-Crook

Doctoral Dissertations

ABSTRACT COMBINATION REGIMENS USING DIETARY COMPONENTS FOR THE CHEMOPREVENTION OF COLORECTAL CANCER AND INFLAMMATION September 2018 CHRISTINA DIMARCO-CROOK, B.S., IMMACULATA UNIVERSITY M.S., DREXEL UNIVERSITY Ph.D., UNIVERSITY OF MASSACHUSETTS AMHERST Directed by: Professor Hang Xiao Dietary components have been found to effectively modulate multiple deregulated signaling pathways associated with the initiation and progression of carcinogenesis and inflammation in cellular and animal models. However, clinical studies have shown mixed results when examining the efficacy of individual dietary components, perhaps suggestive of the synergism that exists between multiple components within a particular food and the diet as a whole. Additional research is needed …

Cd147 As A Potential Therapeutic Target In Glioblastoma Treatment, Beau Adams

All NMU Master's Theses

Glioblastoma (GBM) tumors are the most common and lethal form of cancer in the central nervous system (CNS). GBM tumors appear to contain a mixture of different cell types, which makes them difficult to treat. GBM cells exhibit altered morphology from normal cells on several different levels, which highlights different pathways to potentially target for therapeutic treatments. The human surface glycoprotein CD147, also known as basigin, is expressed at significantly higher levels in GBMs compared to non-neoplastic brain tissue. Furthermore, levels of CD147 expression correlate with brain tumor progression and show the highest expression in GBM. Here, we suppressed tumor …