Cell and Developmental Biology Commons^™

Adenoviral-Vectored Epigraph Vaccine Elicits Robust, Durable, And Protective Immunity Against H3 Influenza A Virus In Swine, Erika M. Petro-Turnquist, Matthew J. Pekarek, Nicholas Jeanjaquet, Cedric Wooledge, David J. Steffen, Hiep Vu, Eric A. Weaver

Nebraska Center for Virology: Faculty Publications

Current methods of vaccination against swine Influenza A Virus (IAV-S) in pigs are infrequently updated, induce strain-specific responses, and have a limited duration of protection. Here, we characterize the onset and duration of adaptive immune responses after vaccination with an adenoviral-vectored Epigraph vaccine. In this longitudinal study we observed robust and durable antibody responses that remained above protective titers six months after vaccination. We further identified stable levels of antigen-specific T cell responses that remained detectable in the absence of antigen stimulation. Antibody isotyping revealed robust class switching from IgM to IgG induced by Epigraph vaccination, while the commercial comparator …

Anillin And Dunk: The Conserved And Non-Conserved Players Regulating Myosin Dynamics In Drosophila Cleavage, Jiayang Chen

Dartmouth College Ph.D Dissertations

Cleavage is a common process during early embryogenesis. Similar to conventional cytokinesis, Drosophila cellularization, a unique form of cleavage, requires precise regulation of non-muscle myosin II ("myosin") at cleavage furrows, where myosin forms an interconnected basal array before reorganizing into individual cytokinetic rings. However, how myosin is regulated through both conserved and cellularization-specific mechanisms during this special form of cytokinesis remains unclear.

In my first project, I investigated the molecular mechanisms by which the cellularization-specific zygotic gene dunk regulates myosin at the basal array. I identified anillin (Scraps in Drosophila), a conserved scaffolding protein in cytokinesis, as the primary …

Quantitative And Qualitative Analysis Of Mutation In Pam-1 Of Model Organism Caenorhabditis Elegans., Jessica Stein, Jessica Stein

Honors College Theses

The pam-1 gene in the model roundworm Caenorhabditis elegans governs meiotic exit and establishment of cellular polarity in the single-celled C. elegans embryo. Mutation of the pam-1 gene results in reduced fertility and fecundity in adult C. elegans and disrupts the anatomy of the germinal gonad. The aim of this study is to qualitatively and quantitatively define the changes in the germline cells associated with mutations to the pam-1 gene. Specifically, we examined the stages of germ cell development within the gonads of adult worms, both wild-type and pam-1 compromised, and identified the changes in the length of the mitotic, …

Evaluation Of Nutrient Digestibility Of Weaned Calves From Early And Late Shedding Dams, Jennifer Keele

Theses and Dissertations

Researchers have investigated several factors that could alter fetal growth, including nutrient restriction (Valiente et al., 2021), hair shedding (Gray et al., 2011), and extreme hot and cold temperatures (Davidson et al., 2022). Hot temperatures and increased humidity percentages in the southeast United States caused researchers to investigate the hair coats of Angus cattle in the commercial production setting. An improvement in fiber digestibility and calf birth and weaning weights has been observed in Angus dams that shed 50% of the winter hair coat by May (Gray et al., 2011; Burnett et al., 2021). Our objective of this experiment was …

A Review Of The Causes Of Optic Nerve Atrophy And Potential Ways Of Axon Regeneration In The Optic Nerve, Cole Wyatt

Honors College Theses

The optic nerve connects the eye to the lateral geniculate nucleus in the brain and is a key component in the human visual pathway. The optic nerve is made up of the axons of approximately 770,000 to 1.2 million specialized neurons called retinal ganglion cells. Like any other organ and tissue in the body, the optic nerve is susceptible to damage or injury. However, unlike some other organs and tissues, once damaged, the optic nerve cannot regenerate. Individuals with damaged optic nerves are said to have Optic Nerve Atrophy and, as of now, there are no cures or treatment options …

Understanding The Regulatory Context And Mechanism Of Hlh-25 Mediated Transcriptional Repression, Dagmawi Mamo

Masters Theses, 2020-current

The HES family of transcription factors are primarily repressors of transcription that play critical roles in mammalian embryogenesis by regulating cell differentiation and proliferation. Proper functioning of HES proteins is crucial in humans, as mutations or misexpression of HES1 is associated with lung, pancreas, colon, and other cancers. HES1 may mediate short-range or long-range repression through DNA-binding and or protein-protein interactions with other factors. However, prior genetic studies have not fully elucidated how the context-specific binding of HES1, or its C. elegans ortholog HLH-25, promotes the repression of transcription. Characterization of the cis-regulatory context and mechanism of HLH-25 mediated repression …

Microglia Refine Early Multisensory Circuits In The Lateral Cortex Of The Inferior Colliculus In A Complement-Dependent Manner, Shaida Hamidi

Masters Theses, 2020-current

Microglia cells (MGCs), the resident macrophage of the brain, play a pivotal role in synaptic pruning of newly established circuits during early critical periods. Classical complement cascade signaling enables MGCs to identify unnecessary or underutilized contacts and specifically target them for removal and subsequent degradation. Such recognition and engulfment behaviors require receptor-ligand interactions, whereby “eat me” tags (complement component 3, C3) bind their cognate receptor (complement receptor 3, CR3) expressed on MGCs. Compromised complement signaling in certain systems results in under-pruning or unrefined map configurations, and is thought perhaps to underlie certain autism-related behavioral phenotypes (e.g. sensory hypersensitivities, decreased social …

The G-Signaling Protein Rcp Controls The Polarized Basement Membrane Deposition In Epithelial Cells, Rebecca M. Brnot

Honors Capstones

Epithelial tissues are the most common type of tissue in the human body, forming the outer layer of the skin and most organs. They are composed of epithelial cells and rely heavily on their cellular architecture, which is organized by an apical-basal polarity, for their function. One critical component for the establishment and maintenance of the epithelial cell architecture is the proper placement of the basement membrane. The basement membrane is a specialized sheet within the extracellular matrix that lines the basal side of epithelial cells, and there is a biological pathway that is responsible for the proper placement of …

An In-Depth Study Of Border Cell Migration Within Drosophila Melanogaster, Leif R. Verace

Honors Capstones

Cell migration plays a key role in many biological processes including embryonic development, tissue morphogenesis, and the metastasis of cancer cells; thus, understanding how this behavior works is highly important. To study cell migration, we use the process of border cell (BC) migration in Drosophila melanogaster, which sees a cluster of cells migrate through a multicellular structure known as the egg chamber found in the Drosophila ovary. Here, I will describe my results in studying the role of a particular kinesin-like gene, CG14535, in BC migration. Through analysis of a mutant for this gene (labelled KC53), RNAi …

Growth Plate Cartilage: Understanding The Contribution Of Adhesion To Column Formation And Matrix Structure, Sydney E. Greer

Theses & Dissertations

Throughout fetal and adolescent development, bone growth is regulated by fine-tuned and controlled maturation of chondrocytes through a cartilaginous template called the growth plate. Bone growth rate is controlled through cell enlargement and extracellular matrix deposition, while the polarized arrangement of proliferative chondrocytes into columns aligned with the long axis of the bone potentiate growth. Chondrocytes are surrounded by a complex three-dimensional arrangement of matrix molecules, all of which are secreted by chondrocytes and assembled/remodeled to support the biological functions of the cell. Adhesion receptors found on the cell membrane of chondrocytes are crucial to the organization of matrix proteins …

Development Of Combination Therapy Strategies To Treat Cancer Using Dihydroorotate Dehydrogenase Inhibitors, Nicholas Mullen

Theses & Dissertations

Cancer cells require supraphysiologic levels of nucleotide triphosphates to fuel their malignant behaviors, including uncontrolled proliferation, immune evasion, metastasis, and therapy resistance. This requirement is met by hyperactive flux through the de novo pyrimidine and de novo purine pathways. Dihydroorotate dehydrogenase (DHODH) is an essential enzyme in the de novo pyrimidine pathway that can be targeted by FDA approved and experimental drug compounds. A robust body of preclinical evidence, encompassing hundreds of independent studies over several decades, demonstrates impressive activity of DHODH inhibitors in animal models of cancer. Paradoxically, however, all clinical trials to date have failed to demonstrate efficacy …

Targeting The Cxcr1/2 Axis In Pancreatic Ductal Adenocarcinoma, Caitlin Molczyk

Theses & Dissertations

Pancreatic ductal adenocarcinoma (PDAC) is notoriously challenging to diagnose and treat. This tumor has an aggressive phenotype and benign clinical features until severe disease dissemination, which makes it too late to eradicate. Although it is only the 10th most common cancer in the United States, it is the fourth leading cause of cancer-related deaths. The treatment options are complicated further due to tumor heterogeneity, meaning not all tumor cells are in the same state phenotypically or epigenetically. They also express cytokines, chemokines, and receptors differently. Here we explore the usage of CXCR1 and CXCR2 and their ligands in PDAC tumor …

The Involvement Of Ubiquitin In Med13 Cyclin C Degradation Following Cellular Stress, Ayesha Gurnani, Brittany Friedson, Katrina Cooper

Rowan-Virtua Research Day

The Cdk8 Kinase Module is a dissociable regulator of cellular stress response genes, with degradation of its components Med13 and cyclin C eventually determining cell fate decisions such as engaging cell survival or cell death mechanisms. We aimed to explore the roles of ubiquitin in degradation of the Cdk8 Kinase Module following nitrogen starvation, with respect to the potential involvement of deubiquitinating enzyme Doa4, lysine linkage at position K63, and E2 ubiquitin conjugating enzymes Ubc4 and Ubc5. We utilized Western blot analysis to observe nitrogen starvation-induced degradation of Med13-HA in wild-type, doa4 mutant, and K63R yeast strains; degradation of cyclin …

Swallowing Disrupts Tongue-Jaw Coordination During Chewing In A Rat Model Of Parkinson's Disease, Meejan Palhang, N. Charles, Francois Gould

Rowan-Virtua Research Day

The primary motor symptoms of Parkinson’s disease, including bradykinesia, rigidity, and tremor, are associated with difficulties regulating transitions between motor behaviors due to basal ganglia dysfunction. Chewing and swallowing, which are disordered in most patients with Parkinson’s disease, are two complex motor behaviors which overlap in time and share some neuromuscular components. The objective of this study is to identify how Parkinson’s disease affects the coordination of chewing and swallowing. We hypothesize that as a result of impaired regulation of shift between motor patterns, chewing cycles that occur with a swallow will be more affected that chewing cycles occurring in …

Orthologs Of The C. Elegans Heterochronic Genes Have Divergent Functions In C. Briggsae, Maria Ivanova, Eric G. Moss

Rowan-Virtua Research Day

The heterochronic genes of C. elegans comprise the best-studied pathway controlling the timing of tissue and organ formation in an animal. To begin to understand the evolution of this pathway, the significance of each factor, and the relationships among the components, we characterized 11 C. briggsae orthologs of C. elegans heterochronic genes. Using CRISPR/Cas9, we made a variety of alleles and found that several mutant phenotypes differ in significant ways from those of C. elegans. Although most orthologs displayed defects in developmental timing, those phenotypes could differ in which stages they controlled, the penetrance and expressivity of the phenotypes, or …

Extravasated Brain-Reactive Autoantibodies Perturb Neuronal Surface Protein Expression In Alzheimer's Pathology, Wardah Bajwa, Mary Kosciuk, Randel L. Swanson, Anuradha Krishnan, Venkat Venkataraman, Robert Nagele, Nimish Acharya

Rowan-Virtua Research Day

Background: Increased blood-brain barrier (BBB) permeability is reported in both the neuropathological and in vivo studies in both Alzheimer’s Disease (AD) and age matched cognitively normal, no cognitive impairment (NCI), subjects. Impaired BBB allows various vascular components such as immunoglobulin G (IgG) to extravasate into the brain and specifically bind to various neuronal surface proteins (NSP), also known as brain reactive autoantibodies (BrABs). This interaction is predicted to further enhance deposition of amyloid plaques.

Hypothesis: Interaction between extravasated BrABs and its cognate NSPs lower the expression of that NSPs in AD patients.

Methods: We selected Western blotting technique to study …

Parkin Ubiquitination Of Kindlin-2 Enables Mitochondria-Associated Metastasis Suppression, Minjeong Yeon, Irene Bertolini, Ekta Agarwal, Jagadish C Ghosh, Hsin-Yao Tang, David W. Speicher, Frederick Keeney, Khalid Sossey-Alaoui, Elzbieta Pluskota, Katarzyna Bialkowska, Edward F. Plow, Lucia R. Languino, Emmanuel Skordalakes, M Cecilia Caino, Dario C. Altieri

Department of Pharmacology, Physiology, and Cancer Biology Faculty Papers

Mitochondria are signaling organelles implicated in cancer, but the mechanisms are elusive. Here, we show that Parkin, an E3 ubiquitination (Ub) ligase altered in Parkinson's disease, forms a complex with the regulator of cell motility, Kindlin-2 (K2), at mitochondria of tumor cells. In turn, Parkin ubiquitinates Lys581 and Lys582 using Lys48 linkages, resulting in proteasomal degradation of K2 and shortened half-life from ∼5 h to ∼1.5 h. Loss of K2 inhibits focal adhesion turnover and β1 integrin activation, impairs membrane lamellipodia size and frequency, and inhibits mitochondrial dynamics, altogether suppressing tumor cell-extracellular matrix interactions, migration, and invasion. Conversely, Parkin does …

Does Vdac2 Have A Bh3 Domain?, Lillian Ferkany

Honors Theses

Mitochondrial outer membrane permeabilization (MOMP) by Bax oligomerization triggers apoptosis. BCl-2 family proteins, classified as BH3 only proteins, pro-survival proteins, or pro-apoptotic proteins, control apoptosis partly through their agonist or antagonistic effects on Bax, which are mediated by their conserved BH3 domains. All BH3 domains form an alpha helix containing 5-7 conserved hydrophobic residues, designated H0-H5, and one conserved aspartic acid that drive interaction with Bax and other ‘multi-domain’ BCl-2 members. BH3 agonists induce Bax oligomerization, while BH3 antagonists sequester Bax to prevent MOMP. We discovered that voltage dependent anion channels (VDACs) in the MOM contain a putative BH3-like domain …

Immunomodulatory Effects Of Resolvin D2 In A Model Of Infection, Prem Yugandhar Kadiyam Sundarasivarao

Graduate School of Biomedical Sciences Theses and Dissertations

Dysregulated hyperinflammatory host immune response to underlying bacterial infections is a characteristic of sepsis. In sepsis, bacteria often trigger abnormal hyperinflammatory responses which can cause multiple organ failure and if sustained can lead to an immunosuppressive phase where the host is susceptible to secondary infections caused by opportunistic bacteria like Pseudomonas aeruginosa (P. aeruginosa). In our studies, we used a 2-hit model of cecal ligation and puncture (CLP) followed by P. aeruginosa secondary lung infection to investigate cellular and molecular mechanisms in the beneficial action of resolvin D2 (RvD2). Resolvins of the D-series are a group of fatty acids known …

Macromolecular Crowding On The Formation Of Amyloid Fiber Hydrogel, Rachael Marie Crews

Theses and Dissertations

The aggregation of proteins to form fibrillar deposits is a hallmark of numerous medical conditions, including Alzheimer’s Disease (AD). Protein monomers aggregate to form amyloid fibers, followed by these fibers further aggregating to form larger structures such as fiber bundles and hydrogels. Previous studies into protein aggregation were mostly done under dilute solutions that poorly reflect the crowded nature of the tissue environment. Can a liquid solution truthfully represent what is going on in a solid tissue? Utilizing lysozyme as a model protein and two different polysaccharides as crowding agents, dextran and hyaluronic acid (HA), the effects of macromolecular crowding …

Investigating The Phenotypic Effects Of Ring1- And Yy1-Binding Protein (Rybp) In Glioblastoma Multiforme, Ronald W. Bucher

Graduate Theses

Glioblastoma multiforme (GBM) is an aggressive form of brain cancer that has horrendous survival outcomes with the use of current therapies. Further study into its molecular mechanisms will inform development of new, more effective treatments. The Polycomb protein RING1- and YY1- Binding Protein (RYBP) has emerged as an important gene in multiple cancers. In complex with other Polycomb proteins, RYBP acts to repress regions of chromatin, though it also performs other functions independent of these complexes. RYBP has a tumor suppressive role in various cancers, but may act as an oncogene in others, demonstrating its context-specific effects. The role of …

Crispr-Based Genome Editing Of Caenorhabditis Elegans And Cultured Human Cells, Katie Marie Toperzer

Theses and Dissertations

The fields of genetics and molecular biology have grown exponentially in the knowledge and ability to edit DNA sequences. The discovery of DNA structure in the 1950s and enzymes that are able to make site-specific cuts in the 1970s allowed scientists to harness growth in the field of genetic engineering and develop realistic gene therapies for diseases. In my thesis, I will utilize cutting-edge technology to be the first person at Florida Tech to genetically engineer two different model systems. Using CRISPR, I will study the molecular mechanisms of fertilization by introducing a genetically encoded reporter in C. elegans. Additionally, …

A Review Of The Neural Basis Underlying The Acoustic Startle Response With A Focus On Recent Developments In Mammals, Alice Zheng, Susanne Schmid

Anatomy and Cell Biology Publications

The startle response consists of whole-body muscle contractions, eye-blink, accelerated heart rate, and freezing in response to a strong, sudden stimulus. It is evolutionarily preserved and can be observed in any animal that can perceive sensory signals, indicating the important protective function of startle. Startle response measurements and its alterations have become a valuable tool for exploring sensorimotor processes and sensory gating, especially in the context of pathologies of psychiatric disorders. The last reviews on the neural substrates underlying acoustic startle were published around 20 years ago. Advancements in methods and techniques have since allowed new insights into acoustic startle …

The Effects Of Salt And Temperature On Three Methanogen Species: Implications For Mars, Katy Dunlap

Biological Sciences Undergraduate Honors Theses

The question of whether life has ever existed on Mars – either in the past or currently – has been pursued for decades. This debate has been prompted by a variety of discoveries regarding similarities between Mars and Earth and more recently concerns over human extinction. Mars, like Earth, once had large amounts of water, a key ingredient for life. H₂O exists on Mars in various forms now, and it is theorized that there is liquid water beneath the surface as well. Mars also contains salt and is very cold at its surface, pointing researchers towards the idea …

Investigating The Role Of The Sumo-Activating Enzyme, Uba2, In Infection-Related Morphogenesis Of Magnaporthe Oryzae, Rachel Taylor

Biological Sciences Undergraduate Honors Theses

The plant pathogenic fungus Magnaporthe oryzae forms a specialized pressure-generating infection cell called an appressorium on the leaf surface, which it uses to mechanically rupture the otherwise impenetrable cuticle, enabling colonization of the underlying tissue and the establishment of blast disease. Appressorium differentiation by M. oryzae is cell cycle-regulated and requires programmed cell death of a three-celled propagative spore, as well as extensive remodeling of the microtubule, actin, and septin cytoskeletons. Recent studies have proposed the importance of sumoylation – the enzymatic conjugation of a small ubiquitin-like modifier to target proteins, for various aspects of infection-related development by M. oryzae, …

Measuring Energy Demands Of Dynamic Remodeling Events During Embryogenesis In Live Drosophila Embryos, Grayson Young

Biological Sciences Undergraduate Honors Theses

Embryonic development is a complex process requiring dynamic cell movements to create complex tissue structures. We assume that these embryonic dynamic remodeling events are highly energy intensive and that this energy expenditure is mediated by mitochondria. However, this has been difficult to demonstrate in live embryos. Convergent extension of the Drosophila neuroectoderm is a promising system to study the bioenergetics of tissue remodeling because it changes from a static to a dynamically remodeling tissue with temporal precision. We hypothesize that the switch from static to dynamic development at the onset of convergent extension will be accompanied by an increase in …

Effects Of Sow Vocalization And Scent On Complete Blood Cell Count During Early Weaning Period, Mason Paladino

Biological Sciences Undergraduate Honors Theses

Piglets are deprived of their sows’ touch, sound, and smell after weaning which results in biological stress. The present study investigated continuing exposure of sow vocalization and scent during the early weaning period on complete blood cell count. A total of 12 litters, 8 piglets (4 male and 4 female) per litter were selected at weaning and assigned to 1 of 4 experimental treatments: control, O (olfactory group with scent and no vocal stimulation), V (non-olfactory with vocal stimulation), VO (vocal stimulation and olfactory stimulation) as a 2x2 factorial arrangement. Vocal stimulation was emulated by an audio recording of the …

Studying The Phosphorylation Of Isocitrate Dehydrogenase In Humans, Hannah Smith

Chemistry & Biochemistry Undergraduate Honors Theses

Isocitrate dehydrogenase is an important enzyme in the citric acid cycle where it catalyzes the oxidative decarboxylation of isocitrate to alpha-ketoglutarate. While there are three isoforms of isocitrate dehydrogenase (IDH1, IDH2, and IDH3), this research will focus on IDH1. The phosphorylation of isocitrate dehydrogenase is a process that has been linked to the formation of both luminal-like and basal-like breast cancer. Despite these correlations, the mechanisms that cause breast cancer development are unknown. To examine this, an enzyme activity assay for each phosphorylation variant and crystallization were conducted. The results of these indicate that phosphorylation at each site (IDH1-T77, IDH1-S188, …

Investigation Of The Role Of Dax-1 In The Hpa Axis In Human Adrenal Cells, Katerina N. Fargas

Undergraduate Honors Theses

DAX-1 is a nuclear hormone orphan receptor that plays a key role in the development of reproductive tissues and steroid hormone production. The hypothalamic-pituitary-adrenal axis is the pathway for steroid hormone production, primarily glucocorticoids, mineralocorticoids, and adrenal androgens, in the human body. The mechanisms of DAX-1 in hormone production in the adrenal cortex of the HPA axis are not completely understood and, therefore, were the focus of this honors thesis research project. Due to the high level of DAX-1 expression, SW13 adrenal carcinoma cells were conducted for these experiments. We investigated whether glucocorticoids, specifically the synthetic glucocorticoid dexamethasone, had any …

Review Of Biomedical Applications Of Cardiovascular Tissue Engineering, Natalie M. Howard

Honors College Theses

Tissue engineering can be defined as processes that aim to generate three-dimensional functional tissues in vitrothat have been favorably altered according to the structural, biochemical, electrophysiological, and biomechanical properties of the desired tissue before implantation into the human body. In relation to cardiac tissues, these properties would include the ability to conduct action potentials, withstand systolic pressure, permit sufficient O₂ and CO₂penetration, sufficient vascularization to supply nutrients for cellular activity, surface topology that enables cellular communication, and more. As heart diseases and instances of myocardial infarction continue to rise worldwide, there is an increasing need for …