Medicine and Health Sciences Commons^™

Interrogation Of The Molecular Mechanisms That Finetune Fcεri Signaling And Mast Cell Outcomes, Derek A. Rinaldi

Biomedical Sciences ETDs

FcεRI is the ITAM receptor primarily responsible for the allergic response and immune hypersensitivity diseases such as asthma. Regulation of the strength of both FcεRI signaling, and the resulting outcomes, is accomplished via a complex array of molecular mechanisms whose interactions remain unclear. Here, I explored how the biophysical properties of antigens, crosstalk between FcεRI and integrin signaling and Arf6-mediated control of trafficking and exocytosis tune mast cell outcomes. I first found significant differences in FcεRI aggregation, phosphorylation and granule secretion after crosslinking with the multivalent antigen DNP-BSA or the small, trivalent antigen DF3. I also revealed a bi-directional, Syk-mediated …

Environmental Ozone Exposure, Gestational Health, And The Lung-Placental Axis, Russell Hunter

Biomedical Sciences ETDs

Adverse gestational outcomes of pregnancy are a globally occurring threat to fetomaternal health, with limited clinical prognostic and intervention strategies aside from premature labor induction. Epidemiological data shows strong correlations between gestational exposure to air pollution and the incidence of adverse gestational outcomes. The research presented here was designed to evaluate potential murine models of gestational exposure, as well as quantify maternal cardiovascular damage therein. These studies employ radiotelemetry, echocardiography, single cell RNA-sequencing, and untargeted lipidomic analyses in efforts to study these effects. Results implicate cardiovascular responses to mid-gestational ozone exposure as evidenced by decreased maternal cardiac output and increased …

Synthetic Biology Enabled Modification Of Bone Cells And Impact On Bone Mineral Resorption, Dulce C. Hayes

Biomedical Sciences ETDs

Development of a cell culture based bone model can contribute to biomedical research by producing a reproducible model system that mimics a bone remodeling disorder and reduces the use of animals in research studies. Synthetic biology was used to engineer osteoblast cell lines to alter a key protein in the RANKL pathway in efforts to control the bone remodeling process. Osteoblast cell lines, K7M2 and MC3T3-E1 Subclone 4 were engineered to express osteoprotegerin (OPG) and inhibit OPG expression, respectively. The upregulation of OPG expression was confirmed with Real Time PCR and Quantikine® Immunoassay. To identify the functionality of the engineered …

Investigation Of Orthohantavirus Genetics In Rodent Reservoirs And Clinical Samples, Samuel M Goodfellow

Biomedical Sciences ETDs

Orthohantaviruses are negative-sense, single stranded RNA viruses with trisegmented genomes that can cause severe disease in humans and are carried by several host reservoirs throughout the world. In the United States, Sin Nombre orthohantavirus (SNV) is the primary cause of hantavirus cardiopulmonary syndrome (HCPS) with a fatality rate of ~36% and the highest cases occuring in the southwest region. The primary SNV host reservoir is thought to be the western deer mouse, Peromyscus maniculatus, however it has been shown that other rodents can carry different orthohantaviruses. We designed a pan-orthohantavirus detection tool to survey several small mammal populations throughout New …

Determining The Existence And Regulation Of Microlipophagy In Primary Brown And White Adipocytes, Sara C. Arenas De Leon

Biomedical Sciences ETDs

Lipids are one of life’s four main macromolecules and provide essential functions to cells.

The degradation of lipid droplets and mobilization of lipids occurs through lipolysis. Emerging evidence demonstrates evidence of a selective form of autophagy in lipolysis. The process of microlipophagy has only recently been described, and many features of its regulation are still poorly understood. Our recent study showed that inhibiting mammalian target of rapamycin complex 1 (mTORC1) pathway by depletion of Raptor in adipocytes led to an influx of lysosomes and accumulation of lipid droplets within lysosome. Here, we expand on these previous findings. We were able …

Effects Of Acute And Chronic Gut Injury On Secretory Lineage Differentiation, Roger Atanga

Biomedical Sciences ETDs

Acute and chronic injury to the gut has several detrimental consequences. However, dysregulation of secretory lineage cells is a common observation. To model acute gut injury, we use non-fissile uranium bearing dust (UBD), an environmental toxicant prevalent in communities adjacent abandoned uranium mines, majority of which are located in the Southwestern United States. Repeated exposure to environmental toxins and heavy metals has been obscurely linked to intestinal inflammation, increased susceptibility to pathogen-induced diseases, and higher incidences of colorectal cancer. However, there is no molecular evidence that non-fissile uranium can cause damage to the intestinal epithelia. Human colonoids exposed to UBD …

Determining The Existence And Regulation Of Microlipophagy In Primary Brown And White Adipocytes, Sara C. Arenas De Leon

Biomedical Sciences ETDs

Lipids are one of life’s four main macromolecules and provide essential functions to cells, as they serve as building blocks of cellular membranes, mediate cell signaling, and are a critical energy source. Within cells, esterified fatty acids aggregate into lipid droplets in the form of triglycerides, consisting of a neutral lipid core surrounded by a phospholipid monolayer decorated with various proteins.

The degradation of lipid droplets and mobilization of lipids occurs through lipolysis, a process that hydrolyzes a molecule of triglyceride into a glycerol and three fatty acids. Emerging evidence demonstrates the involvement of a selective form of autophagy in …

Neuroinflammatory And Metabolomic Temporal Dynamics From Inhaled Wildfire Smoke With Attenuation Via Pharmacological Intervention, David Scieszka, David Scieszka

Biomedical Sciences ETDs

Wildfire smoke is a global issue affecting nearly all humans on the surface of this Earth. Smoke dispersion from wildfires can span continents and cross oceans with amazing ease. Furthermore, total acres of land burned per year in the United States has roughly doubled since 1980, meaning this issue will persist for an indeterminate amount of time. Exposure to wildfire smoke is hazardous and promotes aging-associated events in the cardiopulmonary systems. Yet long- and short-term effects on the brain remain relatively unknown. From these, it is critically important to understand the neurological impacts from wildfire smoke exposure; to compare these …

Effect Of Ligase I Deficiency On Replication Fork And Chromatin, Seema Khattri Bhandari

Biomedical Sciences ETDs

Half of all the eukaryotic nuclear genome is synthesized as Okazaki fragments on the lagging strand at the replication fork. Okazaki fragment synthesis begins with repeated production of short RNA-DNA primers synthesized by the DNA Polymerase α (Pol α)-primase complex. The primer terminus is recognized by Replication Factor C (RFC) which then loads the sliding clamp protein, proliferating cell nuclear antigen (PCNA), resulting in the recruitment of DNA Polymerase δ (Pol δ) which carries out the gap filling DNA synthesis displacing the 5’ end of the preceding Okazaki fragment which is cleaved by Flap Endonuclease 1 (FEN-1), and ultimately the …

Survival Percentage And Outgrowth Collection Kit (Spock) Development And Usage To Analyze Targets Of The Transcription Factor Gcn4 For Their Impact On Chronological Lifespan In The Model Organism S. Cerevisiae, Daniel Paul Felker

Biomedical Sciences ETDs

Plate reader-based methods for high-throughput measurement of growth rate, cellular survival, and chronological lifespan are a compelling addition to the already powerful toolbox of budding yeast Saccharomyces cerevisiae genetics. These methods have overcome many of the limits of traditional yeast biology techniques, but also present a new bottleneck at the point of data-analysis. Herein, we describe SPOCK (Survival Percentage and Outgrowth Collection Kit), an R-based package for the analysis of data created by high-throughput plate reader based methods. This package allows for the determination of chronological lifespan, cellular growth rate, and survival in an efficient, robust, and reproducible fashion. To …

Electroconvulsive Therapy Safety Biomarkers, Jeremy Duvall Miller

Biomedical Sciences ETDs

Objective: The two studies presented utilize neuroimaging data to evaluate potential therapeutic and safety biomarkers in depressed patients receiving Electroconvulsive Therapy (ECT).

Method: The first study enrolled 41 depressed subjects receiving ECT from two independent sites and evaluated gamma aminobutyric acid (GABA+) in the anterior cingulate cortex with proton-spectroscopy contrasted with healthy controls and therapeutic and cognitive outcomes. The second study investigated 17 subjects receiving ECT and evaluated ictal theta power during ECT to electric dose measured by E-field modeling, therapeutic and cognitive outcomes.

Results: No significant difference was seen in GABA+ in depressed subjects receiving ECT compared to healthy …

Prenatal Alcohol Exposure Alters The Expression Of Mir-150-5p And Its Vascular Targets In Brain Microvascular Endothelial Cells And Disrupts The Structure And Integrity Of The Developing Brain Vasculature, Gabriela Perales

Biomedical Sciences ETDs

Prenatal alcohol exposure (PAE) can result in fetal alcohol spectrum disorders (FASD) causing developmental disabilities and neurocognitive deficits. The effects of PAE on neuronal-specific alterations during brain development have been studied extensively but the contribution of the brain vascular system is limited. Recent studies have determined that there are alterations to the cortical microvasculature during PAE, posing new questions about the role of the vasculature and the development of PAE-mediated neurocognitive deficits. Even less studied are the molecular mechanisms that contribute to changes in the vasculature during PAE. As microRNAs (miRNAs) are master post-transcriptional regulators capable of regulating many mRNA …

Comparative Genomics Methods And Applications, Emily N. Alden

Biomedical Sciences ETDs

Virtually all fields of biology have benefited from the advancements in comparative genomics technologies, specifically in the study of evolution. In this dissertation I develop and use comparative genomic technologies to investigate the novel SARS-CoV-2 virus, assembly the first genome of the black lace domestic angelfish and identify germline genetic variants associated with altered breast cancer-specific survival. Our genome tiling array for the novel coronavirus presents a rapid and cost-effective method to sequence the entire viral genome and can be used to track the rapid evolution of viral variants in the population. The domestic angelfish is a member of the …

Exploring The Interaction Of Amphiphilic Mycobacterial Lipoarabinomannan With Lipoproteins: Implications For Blood-Based Diagnosis, Shailja Jakhar

Biomedical Sciences ETDs

Currently, over one-third of the global population is infected with tuberculosis (TB), one of the world’s most deadly infectious diseases. The current gold standard for TB diagnosis is microbiological culture, which can take up to 6-8 weeks, which is not conducive to timely treatment. There are no reliable diagnostics for TB. Disease manifestation and outcome of diagnostic tests for TB are influenced by co-morbidities such as HIV co-infection, further complicating diagnostics development. Thus, there is a need for reliable diagnostics for TB. This is especially true in pediatric populations where the differential disease presentation further complicates outcomes.

This dissertation describes …

Role Of The Acid Sensing Ion Channel 1a In The Systemic Circulation, Selina M. Garcia

Biomedical Sciences ETDs

Acid-sensing ion channel 1a (ASIC1a) is a non-selective cation channel that is expressed in vascular smooth muscle cells. However, little is known about the contribution of ASIC1a to vascular reactivity and cardiovascular homeostasis. We have previously shown that ASIC1a contributes to pulmonary vascular smooth muscle cell (VSMC) Ca²⁺ entry and vasoconstriction as well as chronic hypoxia-induced pulmonary hypertension. However, a role of ASIC1a in the systemic circulation is unclear. Previous reports have shown that ASIC1a is expressed in femoral and cerebral arteries, however, these studies suggest that ASIC1a contributes to vascular reactivity through neuronal regulation rather than vascular. Therefore, …

The Effects Of Glycosylation On Immunogenicity And Replication Of Emerging Viral Pathogens, Elizabeth C. Clarke

Biomedical Sciences ETDs

For filoviruses, a family of hemorrhagic fever viruses, the main antigen for vaccines in development is the glycosylated glycoprotein GP1,2. How the glycosylation of GP1,2 differs when the protein is produced in different cell lines is unclear. We showed significant differences in the types of glycans conferred onto filovirus GP1,2s in produced in mammalian-origin and insect-origin cell lines. Mammalian-derived filovirus GP1,2s have diverse N-linked glycans and extensive O-linked glycans, whereas insect-derived filovirus GP1,2s have N-linked glycans that are primarily paucimannose type and have no observable O-linked glycans. We then showed that differences in glycosylation impacted how immunogenic the proteins were …

The Role Of Hud In Alternative Splicing And Alternative Polyadenylation In The Brain, Rebecca M. Sena

Biomedical Sciences ETDs

RNA binding proteins (RBPs) regulate several processes in the cell, including alternative splicing and alternative polyadenylation. Hu proteins, a class of ELAV-like RBPs, are crucial for proper development and maintenance of the nervous system. Several Hu proteins, including HuD, have been shown to regulate alternative splicing and alternative polyadenylation of neuronal transcripts. However, previous studies have relied on molecular techniques to analyze individual transcripts, which do not provide a global overview of the transcriptome. The purpose of this study was to develop a bioinformatics pipeline to analyze splicing and polyadenylation using RNA-sequencing data. Once the pipeline was developed, it was …

Developing A Virus-Like Particle (Vlp)-Based Vaccine For Neisseria Gonorrhoeae, Ashvini Vaidya

Biomedical Sciences ETDs

Neisseria gonorrhoeae (Ng) is a sexually transmitted bacterium that infects human reproductive mucosal surfaces. Due to the risk of complications in untreated cases and evolving antibiotic resistance, a prophylactic vaccine is critical for preventing infection. There are four Ng outer-membrane proteins that contribute to Ng survival and resistance: PorB, BamA, TbpA, and MtrE. Each protein contains surface-exposed loops that are targets of bactericidal antibodies. Vaccines were developed by displaying Ng epitopes on the surface of bacteriophage virus-like particles (VLP). VLPs are immunogenic, multivalent, self-assembling complexes composed of viral structural proteins. Antigenic peptide sequences can be displayed on the surface …

Suicide, Heroin Use And Protective Resiliency Factors Amongst Transgender Adolescents In New Mexico, Richard Gadomski

Biomedical Sciences ETDs

Transgender people experience significant mental health inequities. The goal of this study was to evaluate disparities related to suicide attempt in the past 12 months and ever heroin use in cisgender versus transgender adolescents, and identify specific resiliency factors associated with a decrease in the prevalence of suicide attempt and heroin use in transgender adolescents. This study is a cross-sectional, secondary analysis of the 2017 New Mexico Youth Risk and Resiliency Survey among high school students. The odds of suicide attempt were 5.5-fold greater, and the odds of lifetime heroin use were nearly 14-fold greater, for transgender compared to cisgender …

Virus-Like Particle-Based Vaccines Induce Antibody-Mediated Neutralization Of Staphylococcus Aureus Virulence, Jason Alexander Joyner

Biomedical Sciences ETDs

Staphylococcus aureus is a pathogenic bacterium that inflicts a large amount of human suffering using an arsenal of virulence factors to cause minor to life-threatening infections. Both the accessory gene regulator (agr) operon, a virulence regulatory pathway that controls expression of over 200 virulence factors, and alpha-hemolysin (Hla), a key pore-forming toxin, play a significant role in infection severity. Given the clinical trial failures of S. aureus vaccine candidates and the prevalence of antibiotic-resistant strains, novel and effective treatment options are urgently needed. Chapter 2 describes a novel virus-like particle (VLP) -based vaccine inducing agr-I-inhibiting antibodies …

Sexual Dimorphism In Host Innate Response To Staphylococcus Aureus: Mechanisms Driving Sex Differences In Immune Cell Function And Host-Directed Interventions, Srijana Pokhrel

Biomedical Sciences ETDs

With the emerging antibiotic resistance, Staphylococcus aureus, primary causative agent of skin and soft tissue infections (SSTIs) as well as other life-threatening conditions has become a global health concern. While factors contributing to host defense against S. aureus SSTIs has been well studied, impact of sex as a variable has not been reported. In Chapter 2, we uncover the sex-specific differences in host response to S. aureus SSTIs and the contribution of female sex hormone, estrogen (E2) in providing protection in females. In Chapter 3, we identify mechanisms of innate differences in neutrophils bactericidal efficacy between sexes and in …

Association Between Self-Reported Dyspnea And Depressive Symptoms In New Mexico Uranium Workers, Shiva Sharma

Biomedical Sciences ETDs

BACKGROUND: Uranium workers risk experiencing dyspnea, a lung disease symptom. Previous studies found associations between dyspnea and depressive symptoms in lung disease, but this requires further study in occupational cohorts.

METHODS: This study evaluated the association between dyspnea and depressive symptoms in former uranium workers screened by New Mexico Radiation Exposure Screening & Education Program. Dyspnea and depressive symptoms were evaluated through questionnaires, Modified Medical Research Council Dyspnea Scale and a modified Patient Health Questionnaire-2, respectively. Logistic regression models were applied to cross-sectional analyses. Generalized linear models were applied to longitudinal analyses.

RESULTS: Cross-sectional analyses demonstrated higher dyspnea scores were …

Exosomal Secretion Of A Psychosis-Altered Microrna Regulates Glutamate Receptor Expression; Effects Of Antipsychotics And Maternal Immune Activation, Stephen Amoah

Biomedical Sciences ETDs

MicroRNAs are a subcategory of evolutionarily conserved small non-coding RNAs that are altered in psychiatric conditions. The ability of small secretory microvesicles known as exosomes to influence neuronal and glial function via their microRNA (miRNA) cargo has positioned them as a novel and effective method of cell-to-cell communication. Inflammation, which is evident in schizophrenia (SCZ) and bipolar disorder (BD), and antipsychotics usage may alter miRNA expression in the brain. However, little is known about the role of exosome-secreted miRNAs in the regulation of neuronal gene expression and their relevance for SCZ and BD. Here, we used miRNA profiling and qRT-PCR …

Translating Touches: New Approaches For Modeling And Rescuing Executive Function Deficits After Prenatal Alcohol Exposure, Johnny A. Kenton

Biomedical Sciences ETDs

Background: With a recent meta-analysis indicating that 9.8% of women worldwide consume alcohol during pregnancy, understanding the effects of fetal alcohol spectrum disorder (FASD) is critical for both identifying and treating those afflicted. Traditional preclinical research investigating the behavioral effects of neurodevelopmental disorders, such as FASD, has utilized tasks designed for specific animal models. For this reason, preclinical research into FASD requires a shift into clinically relevant behavioral testing. One such method involves touch-sensitive screens integrated into an operant box to emulate clinical computer-based tasks, such as the Cambridge Neuropsychological Test Automated Battery.

Methods: Here, we utilized a mouse model …

Zinc Finger Dna Repair Protein Targets Of Arsenite In The Nucleotide Excision Repair Pathway, Juliana M. Huestis

Biomedical Sciences ETDs

Environmental arsenic exposure affects around 100 million people worldwide. Recent evidence shows that arsenite, even at low concentrations, may act as a co-carcinogen, by displacing zinc from the zinc finger motifs of certain DNA repair proteins, leaving them vulnerable to redox modification and loss of protein function. This study probes the physical chemistry underlying arsenite and zinc binding to XPA and PARP1, in the hope of illuminating some of the aspects of protein structure that determine susceptibility to arsenite binding. In this study, dissociation constants are determined for zinc and arsenite binding to peptides corresponding to the C4 zinc finger …

Cold-Inducible Rna Binding Protein (Cirp) Impedes Proliferation And Inflammation In The Pymt Mouse Model For Breast Cancer, Daniel Albino Lujan

Biomedical Sciences ETDs

RNA binding proteins (RBPs) regulate gene expression by controlling mRNA export, translation, and stability. When altered, some RBPs allow cancer cells to grow, survive, and metastasize. Cold-inducible RNA binding protein (CIRP) is overexpressed in a subset of breast cancers, induces proliferation in breast cancer cell lines, and inhibits apoptosis. We generated a transgenic mouse model overexpressing human CIRP in the mammary epithelium to ask if it plays a role in mammary gland development. We also assessed the effects of CIRP on breast tumorigenesis using breeding crosses with the PyMT mouse model for breast cancer. CIRP decreased proliferation at the lactational …

Influence Of Metabolic Capacity On The Consequences Of Spreading Depolarization, Katelyn M. Reinhart

Biomedical Sciences ETDs

In the United States, stroke is the fifth leading cause of death and stroke survivors often face long-term disability. After ischemic stroke, the ischemic territory can expand and recruit previously viable tissues into the lesioned area. During this critical period, there is an unsettling lack of interventions that prevent or treat secondary neuronal loss and cognitive decline. Clinical subdural recordings have revealed a close link between infarct progression and waves of spreading depolarizations (SD) in the acutely injured brain. SD is a slowly progressing wave of near-complete neuroglia depolarization that is extremely energetically demanding and this additional metabolic ‘hit’ …

Mechanisms And Consequences Of Myb Gene Activation In Salivary Gland Tumors, Candace Frerich

Biomedical Sciences ETDs

Salivary gland adenoid cystic carcinoma (ACC) is an aggressive tumor with a tendency to infiltrate surrounding nerves and metastasize to distant sites. The standard treatment often fails to control local tumor recurrence and distant metastases and no approved targeted therapeutic options exist for these tumors. The goal of our studies was to reveal the molecular mechanisms driving ACC tumor development and novel drug targets to improve patient morbidity and mortality.

We first analyzed clinical and RNA-sequencing (RNA-seq) data for 68 formalin-fixed paraffin-embedded (FFPE) ACC tumor samples and described previously unappreciated molecular heterogeneity that predicts patient outcome. The poor outcome subgroup …

Neuron Specific Gene 2 (Nsg2): A Novel Regulator Of Excitatory Neurotransmission Via Ampa Receptor Trafficking At A Subset Of Glutamatergic Synapses, Praveen Chander

Biomedical Sciences ETDs

Members of the Neuron-Specific Gene family (NSG1-3) play critical roles in excitatory synaptic transmission via regulation of AMPA receptor surface expression within the post-synaptic density (PSD). While NSG1 and NSG3 regulate AMPAR recycling and endocytosis, respectively, the function of NSG2 has remained elusive. Here we undertook a series of studies to assess the role of NSG2 in excitatory synaptic transmission. We found that a portion of NSG2 punctae localized with HOMER1 and surface AMPARs at excitatory synapses and that NSG2 AMPAR subunits in mouse brain. Knockout of NSG2 selectively impaired the frequency of AMPA mEPSCs, while overexpression caused a significant …

Assessment Of Particulate Matter Toxicity And Physicochemistry At The Claim 28 Uranium Mine Site In Blue Gap, Az, Jessica G. Begay

Biomedical Sciences ETDs

Particulate matter (PM) inhalation is known to have adverse effects on the respiratory and cardiovascular systems. Community concerns in the southwestern US have been expressed in regions of extensive open-pit hard-rock (especially uranium) mining that windblown dusts may pose an excess health risk. To address these concerns, we have undertaken a campaign to detail: (1) the amount and composition of the PM_2.5 in the region, (2) meteorological characteristics, such as wind speed and direction, and (3) and immunological and physiological responses of mice after PM_2.5 exposures.

For toxicity studies, C57BL/6 and apolipoprotein E-null (ApoE^-/-, a vascular …