Life Sciences Commons^™

Role Of Atp-Binding Cassette (Abc) Transporters As A Protector Of Testis Stem Cell Niche From Chemotherapy Drugs In Drosophila Melanogaster, Swagata Maity

Master's Theses

In chemotherapy treatments most tumor cells are destroyed by drugs, but some tumor cells are drug resistant, and these are known as cancer stem cells (CSCs). Normal adult stem cells are also chemo resistant. Therefore, investigation of normal stem cells can be useful to provide further knowledge of CSCs. One of the primary ways that CSCs achieve drug resistance is by drug efflux through transmembrane pumps called ATP-binding cassette (ABC) transporters. Many studies on ABC transporters have been performed in vitro using cultured cell lines, but very few studies have been performed on normal stem cell populations in vivo. Drosophila …

The Role Of B Cell Activation State And Sex In Aryl Hydrocarbon Receptor Mediated Induction Of Chemokine Receptor 9 And Alpha4beta7 Expression In Vitro, Logan Bauerle

Master's Theses

Defense of mucosal tissues from microbial infection and allergy is reliant on continual production of antibodies. The aryl hydrocarbon receptor (AhR) is known to regulate B cell development and is associated with suppression of systemic humoral immunity. Recent attention has been paid to the role of the AhR in altering expression of cell adhesion molecules (CAMs). B cells express CAMs and chemokine receptors to migrate around the body for localized secretion of antibodies. AhR agonists promote B cell migration to the small intestine through upregulation of chemokine receptor 9 (CCR9) and integrin α₄β₇. Both the AhR …

Species Delimitation Of Slimy Salamanders, Plethodon Kisatchie And Plethodon Mississippi, Across The Lower Mississippi River, Brock Hunter Stevenson

Master's Theses

Species are fundamental units of biodiversity yet delimiting species can be challenging. Slimy Salamanders of the Plethodon glutinosus species complex are a classic example of cryptic species for which species boundaries and relationships have proved difficult to determine. Once thought to be a single species ranging across the eastern United States, protein analysis revealed high genetic divergences among geographically distinct groups of populations, leading to 16 species being recognized within the group. Two of these species, the Louisiana Slimy Salamander (Plethodon kisatchie) and the Mississippi Slimy Salamander (Plethodon mississippi), are closely related but occur on opposite sides of the Mississippi …

Biomarkers In Borrelia Burgdorferi Infected Triple-Negative Breast Cancer Cells, Sambuddha Paul

Master's Theses

Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer with limited treatment options and a poor prognosis. Although some studies have attempted to identify new biomarkers and therapies, only a few have shown promise in clinical trials. In the context of cancer development, infectious agents rank as the third major risk factor, contributing to approximately 15-20% of cancer cases, according to the American Cancer Society. Hence, the identification of biomarkers associated with infection-originated cancer is crucial for enhancing cancer prevention, early detection, diagnosis, and treatment. Borrelia burgdorferi, the causative agent of Lyme disease, has been linked to TNBC …

Development Of Wiskott-Aldrich Syndrome Knock Out Protocol For Drug Substance Assay Development, Julia C. Hanna

Master's Theses

Wiskott-Aldrich Syndrome (WAS) is a rare X-linked primary immunodeficiency affecting approximately 1 in 100,000 live XY births in North America and is caused by a mutation to the WAS gene which is expressed across hematopoietic lineages. The WAS protein (WASp) plays a role in regulating actin polymerization. On a cellular level, there are a variety of effects of a lack of WASp or expression of a dysfunctional WASp protein for patients including issues with migration, adhesion, chemotactic response, phagocytosis, activation, and proliferation across different cell types in addition to reduced platelet size and output. This can lead to several systematic …

Rattus Norvegicus As A Biological Detector Of Clandestine Remains And The Use Of Ultrasonic Vocalizations As A Locating Mechanism, Gabrielle M. Johnston

Master's Theses

In investigations, locating missing persons and clandestine remains are imperative. One way that first responder and police agencies can search for the remains is by using cadaver dogs as biological detectors. Cadaver dogs are typically used due to their olfactory sensitivity and ability to detect low concentrations of volatile organic compounds produced by biological remains. Cadaver dogs are typically chosen for their stamina, agility, and olfactory sensitivity. However, what is not taken into account often is the size of the animal and the expense of maintaining and training the animal. Cadaver dogs are typically large breeds that cannot fit in …

Trace Dna Detection Using Diamond Dye: A Recovery Technique To Yield More Dna, Leah Davis

Master's Theses

This study aspires to find a new screening approach to trace DNA recovery techniques to yield a higher quantity of trace DNA from larger items of evidence. It takes the path of visualizing trace DNA on items of evidence with potential DNA so analysts can swab a more localized area rather than attempting to recover trace DNA through the general swabbing technique currently used for trace DNA recovery. The first and second parts consisted of observing trace DNA interaction with Diamond Dye on porous and non-porous surfaces.

The third part involved applying the Diamond Dye solution by spraying it onto …

How To Build A Cortex: Coordinated Assembly Of Cortical Septins And Actomyosin In The Leader Bleb, Asia Marie Paguntalan

Master's Theses

In confined spaces, migrating cells can undergo mesenchymal-to-amoeboid transitions by altering their cortical dynamics and adhesion with the environment. Septins frequently associate with cortical actin and non-muscle myosin (NMII), but the functional nature of these interactions remains unclear. Upon non-adhesive confinement and NMII enrichment, fibroblasts can switch to a fast, leader bleb-based mode of motility, characterized by the absence of adhesions and stress fibers and formation of a single, elongated leader bleb. During this transition, cortical actin remodeling and polarized NMII contractility drive leader bleb stabilization by generating long-range cortical flows, in coordination with changes in septin localization and assembly …

The Role Of Med12 In Adipogenesis, Caroline Helen Rinderle

Master's Theses

In order for any function to occur within a cell, transcription factors must be able to interact with genes. When this occurs, genes are expressed, and ultimately, proteins are translated and perform the specific function that needs to be done within the cell. In order for this to occur, genes must interact with transcription machinery. The Mediator complex recruits transcription factors to genes in order to promote cell-type specific gene expression. The Mediator complex is a multi-protein complex consisting of four modules: head, middle, tail, and kinase. The kinase module is known to dissociate from the rest of the complex …

The Role Of Med31 And Med12 In Directing Adipogenesis Of Human Adult-Derived Mesenchymal Stem Cells, Joseph Straub

Master's Theses

Selective gene expression is crucial in maintaining the self-renewing and multipotent properties of stem cells. Mediator is a large, evolutionarily conserved, multisubunit protein complex that modulates gene expression by relaying signals from cell type-specific transcription factors to RNA polymerase II. In humans, this complex consists of 30 subunits arranged in four modules: head, middle, tail, and kinase. In our introduction, we show the state of the field of Mediator study with a focus on the critical kinase module. In the following chapters, we used siRNA knockdowns to investigate the roles of the highly-conserved core subunit MED31 and the kinase module …

Effects Of Bacterial Metabolites In Conditioned Media On Colon Cancer Proliferation, Hunter Panier

Master's Theses

Cancer is the second leading cause of death in the United States with colorectal cancer (CRC) being the third most common type. Available treatments include a combination of surgery and chemotherapy but have debilitating side effects. These also have limited effectiveness in some cases, creating the need for additional treatment options, or supplementary treatments to increase their effectiveness. This is leading scientists to consider the microbiome to fix this shortcoming. Current research is focusing on the microbiome and its interactions with certain diseases, which could lead to pro- or prebiotic therapies. This work aims to establish specific bacterial species can …

Using Zebrafish As A Model System For Studying The Autism Risk Gene Adnp In Early Embryonic Development, William Theune

Master's Theses

Autism Spectrum Disorders (ASD) are a group of debilitating neurodevelopmental disorders, estimated to affect 1 in 54 children. Despite the identification of many autism-risk genes, the underlying mechanisms for ASD remain unclear. One of the most common forms of ASD is associated with de novo mutations in the ADNP gene (Activity-Dependent Neuroprotector Homeobox; accounting for 0.17% of ASD individuals). Termed ADNP syndrome, this disorder is characterized by intellectual disability, facial dysmorphia, and is comorbid with multiple organ system deficits. We set out to use zebrafish as a model organism to gain mechanistic insights into ADNP function. Zebrafish have the advantage …

Characterization Of Alcohol-Mediated Promotion Of Breast Cancer Stem Cells, Mckenna Below

Master's Theses

Breast cancer is the leading cause of cancer-related death in women globally, and one in eight women will be diagnosed with breast cancer at some point in her life. Alcohol consumption has been linked to increase breast cancer risk and increased risk of tumor recurrence. Understanding the molecular mechanisms of modifiable lifestyle factors such as alcohol consumption will help to lower these risks and design more effective therapeutic approaches. Estrogen Receptor positive (ER+) breast cancer comprises approximately 70% of all breast cancers, and these tumors are effectively targeted by endocrine therapy in the form of estrogen deprivation with aromatase inhibitors, …

Transgenerational Plasticity Causes Differences In Uv-Tolerance Of Intertidal And Subtidal Populations Of The Purple Sea Urchin, Strongylocentrotus Purpuratus, Yareli Alvarez, Nikki L. Adams

Master's Theses

Planktonic larvae of marine organisms are increasingly being exposed and required to respond to a changing physical environment. Adult sea urchins occupy both intertidal and subtidal waters and broadcast spawn gametes into the water column to contend with variable physical conditions. To answer how populations of invertebrates residing at different depths adequately prepare their offspring to cope with different levels of ultraviolet radiation (UVR), we collected adult purple sea urchins, Strongylocentrotus purpuratus, from four sites (two intertidal and two subtidal (~15 m deep)) on the central coast of CA to compare UV tolerance in offspring. Our measurements of UVA …

Characterizing Erad And Antioxidant Response In Ixodes Scapularis Ise6 Cells During Borrelia Infection, Latoyia Downs

Master's Theses

Ticks are hematophagous ectoparasites that transmit a multitude of diseases to humans. Borrelia burgdorferi (BB) and Borrelia miyamotoi (BM) are both tick-borne pathogens that cause disease in humans and are transmitted by the black-legged tick (Ixodes scapularis). A byproduct of blood digestion generates reactive oxygen species that are toxic and cause oxidative stress which promotes cellular damage and dysfunction. The endoplasmic reticulum (ER) is especially affected by oxidative stress, resulting in a buildup of improperly folded proteins in the ER lumen called ER stress. To prevent cellular damage, the tick utilizes an antioxidant system to neutralize ROS and mechanisms …

Investigating The Role Of The 5’Utr Region Of Tmev In Viral Myocarditis, Maryam Alqahtani

Master's Theses

Viral myocarditis is described as the inflammation of the heart muscle, and it is a common condition that can affect the heart. It is estimated that myocarditis causes 1.5 million deaths annually. In mice, Theiler’s encephalomyelitis virus (TMEV) is a common cause of myocarditis. This non-attenuated virus can be used in a cell culture model system to study viral replication and persistence. These processes have been linked to the severity of the heart damage, which in most clinical scenarios, can lead to dilated cardiomyopathy. The 5’ untranslated region (5‘UTR) found in the viral genome of TMEV is well known to …

Investigating The Relationship Between Hormone Receptors Dax-1 And Estrogen Receptor Within Prostate Cancer Cell Lines, Meghana G. Vijayraghavan

Master's Theses

Prostate cancer (PCa) is estimated to be the second leading cause of cancer-related deaths among men in the United States in 2019. While most prostate tumors rely on androgens for growth signaling, there are subsets of tumors that become androgen-resistant, therefore resisting conventional androgen-depravation therapy. Consequently, research has focused on targeting different hormone receptors for novel therapeutics. One such receptor is Dosage-sensitive sex reversal, adrenal hypoplasia critical region, on chromosome X, gene 1 (DAX-1). DAX-1 is known to interact with other hormone receptors, such as androgen receptor (AR) and estrogen receptors (ERs) to suppress the proliferative effects …

The Democratization And Development Of Cell-Free Protein Synthesis, Max Z. Levine

Master's Theses

Cell-free protein synthesis (CFPS) using crude lysates has developed into a robust platform technology over the last 60 years to express numerous types of recombinant proteins. The open-nature, elimination of reliance on cell viability, and focus of all energy towards production of the protein of interest represent substantial advantages of CFPS over in vivo protein expression methods. CFPS has provided new opportunities across a series of research fields that include metabolic engineering, therapeutic and vaccine development, education, biosensors, and many more. In recent years, optimizations of CFPS have even allowed the platform to reach the industrial level of protein production. …

Development Of A Prolyl Endopeptidase Expression System In Lactobacillus Reuteri To Reduce The Clinical Manifestation Of Celiac Disease, Kara Lynn Jew

Master's Theses

Celiac Disease (CD) is an autoimmune disorder that emerges due to the ingestion of gluten, a protein found in a variety of common grains such as wheat, rye, and barley. Approximately 1 in 100 individuals in the US suffer from CD, making it the most commonly diagnosed gastrointestinal disorder (Ciclitira et. al., 2005). These proline-rich gluten peptides are resistant to proteolysis and accumulate in the duodenum of the small intestine. Once in the duodenum, these peptides illicit an autoimmune response resulting in villous atrophy. Current treatment for CD requires a rigorous adherence to a gluten-free diet. Nevertheless, gluten-containing grains are …

Dna And Protein Sequence Analysis Of Neuronal Markers Neuronal Nuclei (Neun) And Doublecortin (Dcx) In The Northern Pacific Rattlesnake (Crotalus Oreganus) And Western Fence Lizard (Sceloporus Occidentalis)., Brett M. Vassar

Master's Theses

Neuronal Nuclei (NeuN) and Doublecortin (DCX) are neuron specific proteins that are used in histological studies of brain structure in a variety of vertebrate taxa.Antibodies against NeuN (anti-NeuN) bind to the Fox-3 protein, an RNA binding protein common in mature neurons. Anti-DCX labels a microtubule-associated protein expressed in actively dividing neural progenitor cells and migrating neurons. The DCX gene encodes a protein that is well conserved across mammalian, avian, and a few reptilian species, therefore anti-DCX staining has been used successfully across a range of vertebrate taxa. Successful neuronal staining using anti-NeuN has been demonstrated in mammals, birds, and the …

Attenuated Responses To Inflammatory Cytokines In Mouse Embryonic Stem Cells: Biological Implications And The Molecular Basis, Bohan Chen

Master's Theses

Embryonic stem cells (ESCs) have attracted intense interest due to their great potential for regenerative medicine. However, their immune property is an overlooked but a significant issue that needs to be thoroughly investigated not only to resolve the concern for therapeutic applications but also for further understanding the early stage of organismal development. Recent studies demonstrated that ESCs are deficient in innate immune responses to viral/bacterial infections and inflammatory cytokines. Inflammatory conditions generally inhibit cell proliferation, which could be detrimental to ESCs, since cell proliferation is their dedicated task during early embryogenesis. Thus, I hypothesize that the attenuated innate immunity …

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 …

Evaluation Of Endothelial Cell Responses To Elevated Glucose, Gabriella Sugerman

Master's Theses

Developing a tissue-engineered Blood Vessel Mimic (BVM) to represent diabetic macrovascular disease could expedite design of new vascular devices specifically tailored to diabetic patients. In contribution toward this model, this thesis assessed Human Umbilical Vein Endothelial Cell (HUVEC) responses to high glucose conditions. Interleukin 6 (IL-6) and Cluster of Differentiation 36 (CD36) were selected to signify oxidative stress activity, a hallmark of diabetic macrovascular disease. Next, activity of potential reference genes B2M, HPRT1, and ACTB was assessed. All genes were found to exceed acceptable variability, so the E-ΔC T method of data analysis was selected. Next, cellular responses to high …

Zinc Sunscreens Affect Development Of Strongylocentrotus Purpuratus Embryos, Brittany E. Cunningham, Nikki L. Adams

Master's Theses

The growing popularity of physical sunscreens will also lead to an increased release of the ingredients from zinc oxide (ZnO) sunscreens into marine environments. Though zinc (Zn) is a necessary micronutrient in the ocean, greater than natural Zn concentrations are being released into marine environments by use of sunscreens. The extent of the consequences of the addition of Zn to the ocean are not fully understood. We investigated effects of materials released by zinc oxide (ZnO) sunscreens on the development of California purple sea urchin, Strongylocentrotus purpuratus. Embryos developed in various concentrations of Zn, the sources of which included …

Investigating Autophagy, Extracellular Vesicles, And Glycobiology, Benjamin Cook

Master's Theses

Autophagy is a cellular process of degradation which degrades cellular materials in acidic organelles called lysosomes. Impairment of lysosomal degradation can lead to lysosomal storage disorders such as Parkinson's Disease. In Parkinson's Disease neurotoxic α-synuclein can spread from cell-to-cell via extracellular vesicles. We investigated the changes in extra vesicular glycans upon perturbation of autophagy using a class of glycan binding molecules termed lectins.

We found that upon induction of autophagy causes EV glycans to increase while others remained consistent. Additionally, we found that specific glycans are increased more than others. Conversely, when lysosomal degradation was impaired, we observed a decrease …

Proteomic Analysis Of The Crustacean Molting Gland (Y-Organ) Over The Course Of The Molt Cycle, Talia B. Head

Master's Theses

Molting in crustaceans is a highly complex physiological process involving negative regulation by two paired endocrine glands, the X-organ/sinus gland complex (XO/SG) and the Y-organ (YO). The XO/SG complex is responsible for making molt-inhibiting hormone (MIH) which negatively regulates synthesis of the molting hormones, ecdysteroids, by the YO. Analysis of gene expression in the XOs and YOs has led to the development of a proposed molecular signaling pathway which regulates ecdysteroidogenesis and subsequent molting in crustaceans. In this study, changes in protein abundance in the YO were characterized over the course of a molt cycle (intermolt, early premolt, mid premolt, …

Effect Of Oxygen-Limiting Tidal Conditions On Muscle Metabolism And Structure In The Giant Acorn Barnacle, Balanus Nubilus, Katie O. Grady

Master's Theses

Crustacean muscle fibers are some of the largest cells in the animal kingdom, with fiber diameters in the giant acorn barnacle (Balanus nubilus) exceeding 3 mm. Sessile animals with extreme muscle sizes and that live in the hypoxia-inducing intertidal zone – like B. nubilus – represent ideal models for probing the effects of oxygen limitation on muscle cells. We investigated changes in metabolism and structure of B. nubilus muscle in response to: normoxic immersion, anoxic immersion, or air emersion, for acute (6h) or chronic (6h exposures twice daily for 2wks) time periods. Following exposure, we immediately measured hemolymph …

Reconstitution Of Munc18-Dependent Mast Cell Degranulation, Matthew Grant Arnold

Master's Theses

Mast cells are specialized secretory leukocytes that play diverse roles in the body, mediated by release of histamine and other pharmacologically active substances. Although offering essential protection in innate and adaptive immunity, mast cells are also essential to the progression of inflammatory diseases, including allergy and asthma, (Theoharides et al., 2012). Exocytosis of pro-inflammatory mast cell mediators in response to otherwise innocuous antigens relies on evolutionarily-conserved membrane fusion machinery. These proteins, called SNAREs are regulated by the Sec1/Munc18 (SM) protein family (Axle Lorentz, Baumann, Vitte, & Blank, 2012). Mast cells express three mammalian Munc18 isoforms (a, b, and c), which …

Regulation Of Gonad Morphogenesis In Drosophila Melanogaster By Broad Complex, Tramtrack And Bric À Brac Transcription Factors, Diane Silva

Master's Theses

During embryogenesis, primordial germ cells (PGCs) and somatic gonadal precursor cells (SGPs) migrate and coalesce to form the early gonad. A failure of SGPs and PGCs to form a gonad with the proper architecture not only affects germ cell development, but also can result in infertility. We have characterized the phenotypes of two genes, longitudinals lacking (lola) and ribbon (rib), required for the coalescence and compaction of the embryonic gonad in Drosophila melanogaster. Both rib and lola are expressed in the SGPs and to a lesser extent in the PGCs of the developing gonad, and genetic interaction analysis suggests these …