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A Comparison Of Pm-Nato3’S Influence On Neural Progenitors And Mature Dopamine Neurons, Mary E. West

Masters Theses

This thesis presents significant findings regarding the role of PM-Nato3 in its interaction with developing neurons in the context of Parkinson's disease (PD) and regenerative medicine. We investigated the effects of PM-Nato3 on dopamine (DA) neurogenesis under different culture conditions, both in vitro and in vivo. In the standard dopaminergic culture condition, PM-Nato3 potentially increased the speed of DA neuron production but did not significantly increase the yield of DA neurons. In a minimal culture condition, there was no notable difference between the control and PM-Nato3 conditions, suggesting minimal impact on DA neurogenesis. In vivo studies using a mouse model …

Migratory Material: Epigenetics & Weaving At The Us-Mexico Border, Valerie Navarrete

Masters Theses

Discourse often sutures the body shut, disallowing representations of identity to outgrow sociopolitical interests. This issue may originate from borders, but also from the unnamable pathology that generational colonial trauma transmits to the mind, body, and environment. Without a direct form of translatability, this thesis proposes a new materialism that deviates from any object-oriented ontology. Untethered and intra-active, epigenetics and weaving represent objects that transform typical ways of knowing and seeing. Their sensitivity to the environment, in addition to their mobility across generations of time, broaden the spatiotemporal loci of the body and its embodiment. Proposing new materials that expand …

The Effects Of Deleting Lrb E3 Ligase Region 2 On Red Light Responses In Arabidopsis Thaliana, Sabina Liskey

Masters Theses

The ability of plants to absorb light through photoreceptors is essential for successful plant development. Phytochromes are important plant photoreceptors that detect and absorb red and far-red light and must function properly for plant survival. The Light Response BTB proteins (LRB) play an important role in a plant’s response to red light by degrading phytochrome B (PhyB) via the ubiquitin proteasome system (UPS). Although there has been a successful investigation into understanding the function of the LRBs, the function of a few regions within LRB proteins remains unknown, including a portion located on the N-terminal end of the protein, termed …

Microbial Experience Increases Cytotoxicity Of Tumor-Infiltrating Cd8+ T Cells And Controls Tumor Growth, Nicholas Aaron Bunda

Masters Theses

Cancer immunotherapy research is traditionally conducted with specific pathogen-free (SPF) mice, which most accurately mimic the immune system of a human newborn. This makes translational research challenging, as this mouse model is not an accurate reflection of the adult patients who ultimately receive newly developed treatments. It is necessary to further develop a mouse model that bridges this gap and increases translatability of current cancer immunotherapy research.

By cohousing specific pathogen-free mice with regular pet store (PS) mice, we have generated a cohoused (CoH) mouse that reflects the microbial experience of an adult human immune system. We investigated antigen experience, …

The Effect Of Bovine Endometrial Cell Conditioned Media On In Vitro Produced Bovine Embryo Development And Gene Expression, Mary A. Oliver

Masters Theses

In vitro produced (IVP) bovine embryos provide enhanced genetic value and a means for studying early embryonic development. However, IVP embryos are developmentally compromised and are lower quality compared to in vivo derived (IVD) embryos. Consequently, after transfer pregnancy rates from IVP embryos are consistently lower, as they exhibit the highest amounts of pregnancy failure during the early embryonic phase (prior to day 28). Progesterone (P4) has been shown to influence bovine endometrial epithelial (EPI) and stromal fibroblast (SF) cells to secrete developmentally important molecules that promote early embryo development and conceptus elongation. However, IVP embryos are not exposed to …

Retinal Regeneration In A Genetic Mouse Model Of Glaucoma, Joshua Paris

Masters Theses

Glaucoma is a degenerative retinal disease characterized by progressive loss of retinal ganglion cells (RGCs). Previous studies have shown that application of a specific α7 nicotinic acetylcholine receptor agonist, PNU-282987 (PNU), onto the murine retina induces neurogenesis of numerous retinal cell types, including RGCs. The aim of this study is to characterize the short-term and long-term effects of PNU in a glaucoma model. The effects of PNU were analyzed in a DBA/2J mouse model that auto-induces a glaucoma-like condition in adulthood. These mice manifest an elevated intraocular pressure (IOP) starting at 6 months, followed by loss of RGCs. To assess …

The Investigation Of Group 1 Late Embryogenesis Abundant Protein 6 And Its Role In Arabidopsis Thaliana Desiccation Tolerance, Joann Eise Thomson

Masters Theses

Like most seeds, Arabidopsis thaliana seed development is divided into three major phases: morphogenesis, storage deposition, and maturation drying. During maturation drying the seed becomes desiccation tolerant (DT) at some point and the seed rapidly loses water until its moisture level is approximately 10% on a fresh weight basis. The ability of the seeds to withstand this extreme water loss is hypothesized to be due, in part, to the accumulation of late embryogenesis abundant (LEA) proteins. In A. thaliana, there are 51 LEA proteins which are divided into seven groups based on amino acid sequence similarities. Group 1 LEA …

Putative Flagellar Channel-Lining Residues And Their Role In Filament Synthesis, Morphology And Stability, Troy Burtchett

Masters Theses

Bacterial flagella are comprised of ~20,000 repeating subunits called flagellin and are synthesized by transporting unfolded flagellin monomers through the central channel of the flagellar filament to the distal end where they are folded and incorporated into the growing structure. The transport mechanism is predicted to be facilitated by a hydrophilic coating of amino acid side chains on the inside wall of the central channel that repels the mostly hydrophobic surface of the unfolded flagellin monomer. The lack of interaction between the flagellin monomer and the central channel wall is thought to allow for the passive diffusion of protein monomers. …

Evaluation Of Autoreactivity In Vh4-34-Containing Antibodies From Human B-1 Vs B-2 Cells, Michelle E. Ray

Masters Theses

B-1 cells are a unique population of lymphocytes, with distinct phenotypic and functional characteristics in comparison to conventional B-2 cells. B-1 cells are thought to be more closely related to the innate immune system than the adaptive, and B-1 cells constitutively secrete antibody that is recognized as broadly reactive with low levels of autoreactivity. The antibody heavy chain segment, VH4-34, is overutilized by the human B-1 cell population. VH4-34 antibodies are associated with autoreactivity, although not all VH4-34 antibodies are autoreactive. The primary objective of this study was to determine whether autoreactive forms of VH4-34 antibodies are localized only to …

Design, Synthesis, And Characterization Of Chemical Tools To Study Peroxisomal Import, Jhalak N. Timilsena

Masters Theses

Peroxisomes are dynamic and interconnected single lipid membrane bound organelles found in the eukaryotic cells which are involved in various biochemical processes including the b-oxidation of very long chain and branched chain fatty acids, metabolism of reactive oxygen and nitrogen species, and reduction of hydrogen peroxide among others. These organelles are known to host numerous proteins and enzymes depending on the cellular environment. All of the proteins needed in the peroxisomes are encoded in the nucleus and synthesized in the cytosol which are then transported to the peroxisomes with the help of a sophisticated protein-transport machinery. Pex5 is one of …

Seasonal Variation In Mitochondrial Bioenergetics Of The Bluegill Sunfish, Lepomis Macrochirus, From A Shallow Midwest River, Derick Isaac Lamptey

Masters Theses

As average global temperature increase, the frequency and magnitude of extreme temperatures in shallow aquatic ecosystems are more ubiquitous. In order to understand how these changing thermal regimes affect aquatic ectotherms, it is essential to develop studies evaluating the response of ectotherms to seasonal fluctuating thermal regimes. Previous studies on fluctuating temperature regimes have reported an increased physiological stress leading to morphological, behavioral and biochemical adaptations. From the latter, the adaptive capacity and seasonal performance associated with optimal function of the oxidative phosphorylation system (OXPHOS) are key for species persistence. However, studies on this matter are scarce. This study explores …

Investigation Of Glutamate Carboxypeptidase Ii Roles In The Folate Cycle Dependent Reproduction And Development Of Caenorhabditis Elegans, Jessica M. Derham

Masters Theses

Glutamate Carboxypeptidase II (GCPII) is a transmembrane, zinc metallopeptidase that is expressed in a wide range of organisms, including roundworms, mice, and humans. In humans, GCPII is primarily expressed in the prostate, kidneys, small intestine, and central nervous system. Within the small intestine, the expected function of GCPII is to aid in the absorption of dietary folate from the intestinal lumen. GCPII cleaves excess glutamates from folate to yield monoglutamated folate which is then readily transported into the enterocyte. Folate can then be used through the one carbon metabolic cycle for the synthesis of nucleotides, conversion of homocysteine to methionine, …

Reprograming Neuronal Cells By Overexpression Of Fibroblast-Specific Transcription Factors, Abdulmohsen Alanazi

Masters Theses

In mammals, a complex system of regulatory signals distinguishes tissues, structures and functions. Combinations of transcription factors and co-factors regulate activation and repression of genes that result in cellular differentiation. Whole genome arrays allow the monitoring of genomic expression in specific tissues. Fibroblast microarray studies have shown candidate genes that may be involved in fibroblast identification, including genes that express transcription factors Prrx1, Snai2 and Twist1. A previous study showed that the Prrx1 and Snai2 could reactivate a fibroblast phenotype in hybrid cells that had lost fibroblast identity. Furthermore, overexpression of these factors in liver-derived cells strongly repressed liver gene …

Therapeutic Dna: Delivery And As A Delivery Vehicle, Natalie J. Holl

Masters Theses

"A review of gene delivery methods and gene editing methods, as well as original research utilizing DNA as a delivery vehicle is presented in the following thesis. Thousands of diseases have been linked to genes. Gene therapy, either delivering therapeutic genes or editing DNA bases, has arisen as a treatment option with the potential to cure diseases, rather than just ease symptoms. Genes and editing tools need to be delivered to cells for these therapies to be effective and many techniques have been developed to address the issue of delivery. Nonviral and viral methods have been used to deliver nucleic …

Reprogramming Of Rat Hepatoma Cells With Fibroblast-Specific Protein Twist1, Kezban Ucar Cifci

Masters Theses

In mammalian development, a complex system comprised of regulatory signals causes tissue distinction with unique structures as well as functions. Gene expression controls these functions through specific combinations of transcription factors and cofactors that influence cell differentiation by both activation and repression of genes.

Whole genome microarray studies of fibroblasts have identified candidate genes that can serve as master regulators of fibroblast identity. A previous study showed that Prrx1 and Snai2 play important roles in activating expression of fibroblast identity, and Snai2 overexpression in hepatoma cells (Fg14) activated expression of fibroblast specific genes. Moreover, Snai2 overexpression resulted in repression of …

Xylan Metabolism By Caulobacter Crescentus, Viet Bui

Masters Theses

Bacteria present unique opportunities to explore the molecular mechanisms of nutrient acquisition processes at single cell level under various experimental conditions. Caulobacter crescentus is a gram-negative oligotrophic environmental bacterium with adaptive physiological responses, such as slow growth or complete arrest of cell cycle, presence of sessile and motile progenies with the sessile cells growing stalk to enhance nutrient uptake for growth in low-nutrient conditions. Details of plant polysaccharide-derived carbon compound, such as cellobiose, maltose, and xylose utilization by C. crescentus can be found but molecular details or mechanisms of polysaccharide utilization, such as xylan, has not been reported. Database analyses …

14-3-3 Protein Rad24 And Its Effect On Mid1 Localization And Contractile Acto- Myosin Ring Assembly During Cytokinesis In S. Pombe, Anissa P. Grawe

Masters Theses

In animal and yeast cells, cell division (cytokinesis) is facilitated by the formation of a contractile acto-myosin ring (CAR). Proper CAR formation and constriction is heavily reliant on the temporal regulation, phosphorylation, and localization of key proteins. In the fission yeast S. pombe, Mid1 is an important dimeric CAR scaffolding protein that connects the contractile apparatus to the plasma membrane at the right place and time during cytokinesis. Mid1 is confined to both the nucleus and protein assemblies called nodes during interphase, and transitions to the cell cortex at mitotic entry as nodes mature and coalesce into the CAR. Rad24 …

Evaluation Of The Phylogenetic Relationships Of The Marine Rivulariaceae., Jose Alberto Martinez Yerena

Masters Theses

Historically, the Rivulariaceae family has grouped all heteropolar, tapered and heterocitous Cyanobacteria, however recent description of multiple cyanobacterial populations with this morphotype using 16S rRNA phylogenies have reported that the populations with this morphotype are polyphyletic, and many new and old tapered and heteropolar genera (Cyanomargarita, Gloeothrichia, Rohoaltiella, etc.) have been classified and reclassified to different families than the Rivulariaceae. This is also the case of the main genera of the family (Rivularia and Calothrix), which have been repeatedly reported as polyphyletic, and whose separation to different families has been called for. In this project I carried out the evaluation …

The Role Of Prrx1 And Snai2 As Master Regulators Of Fibroblast Identity, Huda A. Alzahrani

Masters Theses

Mammalian development involves a complex system of regulatory signals and reactions, leading to the creation of distinct tissues with specific structures and functions controlled by various gene expression profiles. Tissue-specific gene expression is by controlled specific combinations of transcription factors and cofactors that dictate cell differentiation via activation and repression of genes in mammalian cells.

Whole genome microarray analyses of fibroblasts have revealed candidate genes that may serve as master regulators of fibroblast identity, such as Prrx1 and Snai2, that activate downstream genes such as Bmp3, Twist, Shox2, Cfos, Slug, Sema3A, Spp1, and Col1a1. A previous study showed that Prrx1 …

Defining The Immune Response And Cellular Pathologies In Multiple System Atrophy, Annamarie Valenti

Masters Theses

Multiple System Atrophy (MSA) is a neurodegenerative disease characterized by the accumulation of misfolded α-synuclein (α-Syn) in oligodendrocytes of the central nervous system (CNS). A previous study in our lab used Affymetrix DNA microarray analysis to show the downregulation of various mitochondrial related genes, as well as the upregulation of genes involved in inflammatory/immune responses in MSA brain tissue. Therefore, it is hypothesized that an increased presence of immune proteins exists in the MSA brain accompanied by glial cell pathology including decreases in cell metabolism and cell viability. This study uses immunohistochemistry (IHC) to show the increased presence of CD68 …

The Role Of Calcium In Regulation Of Glial Cell Line Derived Neurotrophic Factor By Skeletal Muscle Cells, Alicia M. Boynton

Masters Theses

Glial cell line-derived neurotrophic factor (GDNF) is an important signaling molecule for the somatic motor nervous system. GDNF protein is produced and secreted by skeletal muscle cells and helps maintain motor neuron innervation at the neuromuscular junction. Treatment with exogenous GDNF prevents denervation which is characteristic of aging and neurodegenerative disease. The therapeutic potential of GDNF cannot be fully explored without understanding the mechanisms by which GDNF protein production is regulated. The primary objective of this study was to determine the role of calcium in regulating GDNF protein expression by skeletal muscle cells. Skeletal muscle cells (C2C12) were grown in …

The Role Of Creg1 As A Master Regulator Of Liver Function, Abdulrahman Siran Aldaghmi

Masters Theses

The liver is known as the chemical factory of the body because it performs a wide range of biochemical functions required for life. Since the liver has such an important role in regulation of normal physiological processes, liver diseases cause a high rate of morbidity and mortality. Therefore, understanding the mechanisms of liver development will shed light on the causes of liver disease. In this study, a cell line model that utilizes rat hepatoma cells (Fg14) and hepatoma variant cells (H11) was used to identify master regulators of liver gene expression. Whole genome expression studies identified the gene CREG1 (Cellular …

Metabolic And Morphologic Shifts In Neuro2a Cells Cultured In Galactose Medium, Leah Welker

Masters Theses

It has been observed that highly-proliferating cells, such as cancer cells, rely mainly on glycolysis for ATP production, regardless of presence of oxygen. This effect, however, can be reversed by changing the main energy substrate in the medium from glucose to galactose. The oxidation of galactose in glycolysis yields less net ATP, presumably forcing the cell into OXPHOS. This has been established in many cell lines, including HeLA, HepG2, and skeletal muscle cells. As of yet, this has not been reproduced in neuronal cells. Using Neuro2a, a murine neuroblastoma cell line, this study exposes neuronal cells to galactose medium, and …

Investigating Ph Banding Kinetics Of Chara Corallina In Alternating Light Conditions With Rectangular Pulse Voltammetry, Matthew C. Zandee

Masters Theses

Chara corallina is an important biological model due to its large internodal cells, the simple shape, and the wide range of techniques that can be used to study it. The alternating acidic and basic banding pattern of Chara corallina upon illumination has been studied and well described over the past several decades. However, much of this complex mechanism is not fully understood. Few studies have shown how the acidic and basic regions have responded in real time as lighting conditions change. Utilizing rectangular pulse voltammetry (RPV) and pH-sensitive carbon microelectrodes along the cell wall allows for a real time profile …

Impact Of Transcription Factors On Fibroblast Specific Gene Silencing In Somatic Cell Hybrids, Tania Ray

Masters Theses

As totipotent cells differentiate to specific cell types in multicellular organisms, certain sets of genes are turned off (gene silencing/gene extinction) while others are turned on (gene activation). Cell type specific gene silencing and gene activation is the basis of cell differentiation. Somatic cell hybrids which are produced by fusing different types of somatic cells from two different tissues have been used as a valuable resource to discern the phenomenon of lineage- specific gene extinction. Identification of regulatory factors that mediate gene extinction in hybrids are key to understanding regulatory mechanisms that govern cell differentiation in a multicellular organism. Transcription …

Expression Of Artemia Lea Proteins In Drosophila Melanogaster Cells Using Multicistronic Vector Constructs, Kazi Nazrul Islam

Masters Theses

In nature several organisms exhibit anhydrobiosis, the outstanding feature to survive in extreme desiccation by entering into a state of dormancy known as diapause. The cyst of the brine shrimp Artemia franciscana shows anhydrobiosis by entering into a diapause phase. Previous studies showed a correlation between anhydrobiosis and expression of highly hydrophilic polypeptides termed late embryogenesis abundant (LEA) proteins. However, the precise molecular mechanisms of LEA proteins are still unknown. The presence of multiple LEA proteins in Artemia suggests that some of them might work together. Here, I aimed to express different combination of two LEA proteins from Artemia franciscana …

One Fish, Two Fish, Red Fish, Blue Fish: The Analysis Of The Specter Cell Cycle Mutant In Zebrafish, Tetiana Petrachkova

Masters Theses

Cell division is controlled by genes that regulate the cell cycle. Here we show that the zebrafish specter (spr) mutant is mutation in the cyclin B1 gene, a gene necessary for the G2 to M transition of the cell cycle.

The spr mutation mapped to the cyclin B1 gene. Sequencing showed a transition (C139→T) that caused a nonsense mutation in exon 2 of the cyclin B1 gene. In situ hybridization of cyclin B1 revealed that the mRNA is absent in the mutant embryo by gastrulation. We found that the earliest visible mutant phenotype was a darkening of the head caused …

Bridging In Vi Tro & In S Il Ico Techniques: Uncovering The Structural Characteristics And Light-Dependent Modifications Of The Light-Response Btb Proteins In Arabidopsis Thaliana, Nicholas Reitz

Masters Theses

The conjugation of the NEDD8 protein has been found to cause a wide range of changes to the overall function of a protein. The addition of NEDD8 to a conserved lysine on Cul proteins has been found to provide Cul with increased flexibility, while the transcription factor protein p53 was prevented from binding to DNA after NEDD8 conjugation. Using Arabidopsis thaliana, we investigated the potential conjugation of NEDD8 to the AtLRB1 and AtLRB2 proteins, which are both members of Cul3-RING E3 Ubiquitin Ligase complexes, using both in vitro and in silico techniques. The AtLRBs are negative regulators of the red …

The Role Of Creg1 And Pitx1 As Master Regulators Of Liver Function, Anit Shah

Masters Theses

Development in mammals requires a complex array of positive and negative regulatory signals and responses. The liver is a major organ that has been studied extensively to understand underlying genetic processes responsible for specification, establishment and maintenance of tissue identity. Hepatoma and hepatoma variant cell lines have been used as a model to understand genetic networks responsible for liver function. Whole genome analysis of these cell types has identified candidate genes that might serve as master regulators of liver identity. In this study, the role of two candidate genes - cellular repressor of E1A stimulated gene (CREG1) and paired-liked homeodomain-1 …

The Effects Of Solutes In The Cryopreservation Of Adherent Neuroblastoma (Neuro-2a) Cells, Trisha L. Bailey

Masters Theses

A simple method to cryopreserve adherent monolayers of neuronal cells is currently not available, but the development of this technique could facilitate numerous applications in the field of biomedical engineering, cell line development, and drug screening. However, complex tissues of some exceptional animals survive freezing in nature. These animals are known to accumulate several small molecular weight solutes prior to freezing. Following a similar strategy, we investigated the effects of osmolytes such as trehalose, proline, and sucrose as additives to the traditional cryoprotectant dimethyl sulfoxide (Me2SO) in modulating the cryopreservation outcome of mouse neuroblastoma (Neuro-2a) cells. Neuro-2a cells adhered to …