Life Sciences Commons^™

Symbiosis Under Stress: Unraveling The Interplay Of Ocean Acidification And Rising Temperatures On Acropora Samoensis (Staghorn Coral), Jerald William Smolkin

Theses and Dissertations

This experiment explores the response of marine organisms to the combined challenges of ocean acidification and rising temperatures, with a focus on coral reef ecosystems. As global climate change threatens profound declines in coral reefs, understanding the multifaceted impacts of these stressors becomes crucial. The study observes cellular changes in A. samoensis tissues in four different groups, including control, Temp, pH, and Temp and pH. Results indicate significant breakdown of membrane compartmentalization and cell junctions, with notable degradation and calcium carbonate crystallization in pH-stressed samples. Additionally, chlorophyll extraction data support coral bleaching due to the expulsion of zooxanthellae. These findings …

Pharmacokinetic Modeling Of Metonitazene And Isotonitazene Using In Vitro And In Vivo Models And Comparison To Human Postmortem Casework, Justine P. Sorrentino

Theses and Dissertations

Metonitazene and isotonitazene are two benzimidazole opioids that are contributing to the opioid crisis nationwide. Limited pharmacokinetic data has been published on these nitazene compounds, therefore this study sought to determine if these two structurally similar opioids would have comparable half-lives and clearance utilizing in vitro and in vivo models. Using the data determined in the pharmacokinetic study, and by testing human postmortem cases, this study aimed to inform interpretation of these drugs detected in human casework. Sample analysis for all aspects of the study was performed utilizing LC/MS/MS. In vitro pharmacokinetics of both drugs were examined using a human …

Streptozotocin-Induced Diabetes Increases Kidney Toxicity In Rats Exposed To Ochratoxin A, Belkys Gonzalez

Theses and Dissertations

Due to the rising occurrence of diabetes in the population and the prevalence of OTA exposure via ingestion of contaminated foodstuffs, there is a need to assess the possible renal interaction of a diabetic disease state in concurrence with OTA exposure. Sprague Dawley rats were treated with streptozotocin to induce experimental diabetes and then orally exposed to 0.5 mg/kg body weight OTA once a day for 21 days. Renal biomarkers and light microscopy were evaluated to assess the interaction between diabetic nephropathy and OTA exposure. LLC-PK1 proximal tubule cells were used as an in vitro model to assess the potential …

Exposure To Dithiocarbamate Fungicide Ziram Results In Hepatic And Renal Toxicity In Long-Evan Rats, Jeffrey Chen

Theses and Dissertations

Ziram is a broad spectrum fungicide that is used on a variety of crops. It has been shown to display a wide array of toxicological effects, however, there is still a scarcity of mechanistic data regarding how it facilitates those effects. One of its presumed mechanisms of toxicity is due to the production of oxidative stress. However, whether the production of oxidative stress is due to its dimethyldithiocarbamate backbone, its metal moiety, or a combination of both, is still unknown. Contributing to the production of oxidative stress is ziram’s ability to alter essential metal homeostasis through transchelation of other essential …

Determination Of Genetic Variation In Opioid-Resistant Hospice Patients Using A Pharmacogenomics Approach, Daniel Bianculli

Theses and Dissertations

Managing pain requires a delicate balance between the advantages of opioid analgesia and the possible adverse effects. This balance becomes even more difficult in hospice and palliative care environments where dosage regimens are frequently ambiguous. Some hospice patients experience opioid resistance, in which opioids no longer produce an analgesic effect on a patient. Evidence-based analyses frequently omit this vulnerable population, resulting in a need for concrete procedures. The emergence of pharmacogenomic analysis and personalized treatment presents a solution to this problem by using each patient's genomic data to interpret opiate dosing. This pharmacogenomic study aimed to single nucleotide polymorphisms (SNPs) …

Production Of Vp3-Only Aav2 Vlp In E. Coli, Chengyu Fu

Theses and Dissertations

Adeno-associated virus (AAV) is a widely used vector for gene therapy. However, few studies have focused on producing AAV-based virus-like particles (VLP) in E. coli. This project aimed to express VP3 of AAV2 or co-express VP3 of AAV2 with other proteins, such as E. coli chaperones and/or assembly-activating protein (AAP) in E. coli under different expression conditions and explore the potential to produce VP3-only VLP of AAV2 in situ in E. coli. The constructed plasmid pET15ΔHis-VP3 was used to express VP3 under different conditions. The results demonstrated that most expressed VP3 was in inclusion bodies in all tested conditions. Although …

Evaluation Of Microglia’S Structural Variations Within An Organotypic Hippocampal Slice Model After Regionalized Toxic Injury, Jesus Antonio Trejos

Theses and Dissertations

The dendritic cells of the CNS, the microglia (MG), are an initiation point of the immunological response within the post blood-brain barrier (BBB) compartment. Microglia morphology drastically changes in response to cell stress to a much different non-dendritic morphology. This investigation postulates that if the first MG responses to toxic injury are isolated and studied in greater morphological detail there is much to be learned about MG metamorphosis from a baseline (M2) to an activated (M1) state. The organotypic hippocampal slice was the experimental setting used to investigate microglial response to toxic injury. This isolates dendritic cellular responses and associated …

Interplay Between Rna Editing And Alternative Splicing Modulates Biophysical And Pharmacological Properties Of The Ion Channel Cav1.3, Willy Munyao

Theses and Dissertations

Alternative splicing and adenosine-to-inosine (A-to-I) RNA editing are post-transcriptional mechanisms that promote proteomic diversity in the brain. The impact of RNA editing on brain physiology is underscored by its association with the etiology and pathogenesis of various neurological and neurodegenerative diseases. Alternative splicing of exon 43 within the CaV1.3 C-terminus produces long (CaV1.3L) and short (CaV1.343S) variants exhibiting distinct gating and pharmacological properties. While the physiological role of CaV1.3 A-to-I RNA editing in brain function is still unfolding, recent findings indicate that unedited CaV1.3 channels display larger calcium influx and enhanced neuronal excitability, suggesting that A-to-I RNA editing may act …

Mobocertinib Antagonizes Multidrug Resistance In Abcb1- And Abcg2-Overexpressing Cancer Cells, Xingduo Dong

Theses and Dissertations

The resistance of cancer cells to multiple drugs, known as multidrug resistance (MDR), remains a significant obstacle that leads to the ineffectiveness of cancer chemotherapy. The overexpression of ATP-binding cassette (ABC) transporters, particularly ABCB1 and ABCG2, has been strongly correlated with MDR. There is an urgent need to explore and identify novel inhibitors targeting ABCB1 and ABCG2 to overcome the related MDR. Mobocertinib (TAK-788) is an orally administered, irreversible EGFR/HER2 inhibitor approved for the treatment of adult patients with locally advanced or metastatic non-small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) exon 20 insertion mutations. This study …

Combined Cytotoxicity Of The Herbicide Glyphosate And The Fungicide Mancozeb In Mouse Neuroblastoma Cells, Heidi Ebid

Theses and Dissertations

The herbicide glyphosate and the fungicide mancozeb are globally applied on a variety of agricultural crops. Residues of both pesticides are frequently detected in food and water. Combined exposure to both compounds is therefore possible. Glyphosate and mancozeb have been separately investigated in previous studies and were shown to induce neurotoxicity. The current study investigates the cytotoxicity of different glyphosate and mancozeb combinations including mixture of mancozeb + glyphosate, combination of mancozeb followed by glyphosate and combination of glyphosate followed by mancozeb and compares neurotoxicity seen with the combinations to that of glyphosate or mancozeb using mouse neuroblastoma Neuro-2a cells. …

The Splice Index As A Prognostic Biomarker Of Strength And Function In Myotonic Dystrophy Type 1, Marina Provenzano

Theses and Dissertations

Myotonic dystrophy type 1 (DM1) is a slowly progressive, multisystem disorder caused by a CTG repeat expansion in the DMPK 3’UTR that leads to global dysregulation of alternative splicing. The resulting decline in physical function is slow, and no reliable biomarkers exist for predicting disease progression; however, an RNA mis-splicing biomarker associated with weakness may have utility in predicting functional outcomes. Here we validate the Splice Index (SI) as a potential biomarker of DM1-associated strength and function. Muscle biopsies of the tibialis anterior were collected from DM1-affected individuals at baseline (n = 46) and 3-months (n = 34), along with …

Persistence Of Wastewater-Associated Antibiotic Resistant Bacteria In River Microcosms, Aoife P. Mahaney

Theses and Dissertations

The spread of antibiotic-resistant bacteria (ARB) associated with wastewater is a significant environmental concern, but little is known about the persistence and proliferation of these organisms in receiving water bodies after discharge. To address this knowledge gap, we performed a series of microcosm experiments in which river water was amended with either untreated or treated wastewater, and the abundance of viable ciprofloxacin-, Bactrim-, and erythromycin-resistant bacteria was monitored for 72 hours.

Both types of wastewater amendments increased the initial abundance of ARB compared to microcosms containing only river water. The increase was greatest with untreated wastewater, but that effect decreased …

An Investigation Into Splicing Variation And Expression Patterns Of Insulin-Like Peptide 4 (Ilp4) Within The Drosophila Genus, Zachary C. Griffith

Theses and Dissertations

The insulin-signaling pathway plays a crucial role in regulating metabolic homeostasis in both humans and insects. Dysregulation or mutation within this pathway can lead to various diseases, such as diabetes mellitus and insulin resistance. The insulin-like peptide 4 (ILP4) in Drosophila is an interesting subject for study due to its similarity to mammalian insulin and its role in regulating sugar metabolism. In this study, we investigate a novel splicing variation within the Drosophila takahashii Insulin-Like Peptide 4 (ILP4) gene. Our investigation identified a splicing variation found in four Drosophila species that introduces an additional codon in one isoform, resulting in …

Optimizing Z/R Relationship For Radar-Based Precipitation Estimates, Jalon Gary

Theses and Dissertations

Z/R relationship is a power law relationship between radar reflectivity (Z) and rainfall rate (R). The relationship is expressed as Z = aRb where a and b are correlated with drop size distribution (DSD) and vary across locations. Using a Z/R relationship to translate radar reflectivity to rainfall rate can allow for real-time precipitation monitoring with a high temporal and spatial resolution. A dataset of 12 cases in Georgia from December 2021 to November 2022 was examined using the Georgia Weather Network rain gauge system and the KFFC radar to optimize the best Z/R relationship for each case. Radar-based precipitation …

Advancing Explainability In Multi-Label Classification For Tomato Disease Detection Using Machine Learning Interpretability Techniques, Md. Nurullah

Theses and Dissertations

Plant diseases pose a significant threat to global food security, affecting crop yield, quality, and overall agricultural productivity. Traditionally, diagnosing plant diseases has relied on timeconsuming visual inspections by experts, which can often lead to errors. With the rapid growth of technology, machine learning (ML) and artificial intelligence (AI) have opened new possibilities for automating this process. One of the most promising technologies for plant disease diagnosis is Convolutional Neural Networks (CNNs), which have proven effective in image classification tasks. Plant leaves, often exhibiting symptoms such as discoloration and irregular textures, serve as key indicators for disease detection. By processing …

Investigating Ammonium Toxicity In The Performance Of Recirculating Periphytic Algal Wastewater Treatment Systems, Oluchuku Richie Ochonogor

Theses and Dissertations

Freshwater nutrient enrichment from wastewater facilities and other sources can lead to freshwater eutrophication, a threat to global aquatic ecosystems. Mechanical and chemical ways to curb this threat are either too expensive or not sustainable, and thus, not feasible. Compared to mechanical and other methods, sustainable, inexpensive biological methods (for example, algal treatment systems) have therefore been developed for the removal of excess nutrients from wastewater. By design, secondary waste treatment facilities (WWTF) remove organics and solids and lower oxygen demanding substances; however, they may not remove enough nutrients to protect freshwater ecosystems in all cases. While algae-based biological methods …

A Phytochemical Analysis Of Secondary Metabolites In Eupatorium Serotinum, Elise Snow

Theses and Dissertations

Phytochemistry, a subset of Natural Products Chemistry, studies chemically derived products from natural substances. Advances in this field have important applications in medicinal research due to the distinctive and unique properties of natural compounds. All metabolites are produced from the metabolic cycle, which is a pathway that provides energy for the organism. These metabolic compounds vary greatly and are classified based on their function within the organism. Primary metabolites are abundant and contribute to organism growth, development, and maintenance of energy. Secondary metabolites are found in lesser quantity and correlate to natural defense mechanisms. For example, salicylic acid is a …

Range-Wide Analysis Of The Genetic Diversity And Genetic Structure Of The Spotted Turtle (Clemmys Guttata), Madison S. Whitehurst

Theses and Dissertations

Understanding the spatial distribution of genetic diversity and structure in endangered species is vital for effective conservation management. This research discusses the results of an initial analysis of the genetic structure and variation of the spotted turtle (Clemmys guttata), a species of conservation interest. The study sampled 913 individuals from 78 unique locales across 16 states in the eastern portion of the species' range. Utilizing ddRADSeq, a de novo genome assembly was developed to identify nearly 20,000 potential loci. A total of 926 were selected for this analysis. Results revealed no spatial clustering of genetic diversity or structure, …

Calcium Permeability Of Rat Trpv1 In A Close State When Expressed In Xenopus Laevis Oocytes, Zinnia Adhikary

Theses and Dissertations

Transient Receptor Potential V1 (TRPV1) is a nonselective cation channel, which is expressed in nociceptors/ pain detecting receptors. Activated by a wide variety of exogenous and endogenous physical and chemical stimuli such as heat, TRPV1 is involved in temperature detection, body temperature regulation, and pain perception. It is generally accepted that TRPV1's conductance to cations, including Ca2+, depends on its conformational change from a closed to an open state following gating. In the current study, I observed that the plasma membrane of Xenopus laevis oocytes, upon exogenous expression of rat TRPV1, exhibited weak permeability to Ca2+, as evidenced by the …

Searching For Novel Mutations That Regulates The Mouse Polycystin-2 (Trpp2) Ion Channel, Akhilraj Rajan Pillai

Theses and Dissertations

Polycystin-2 (PC2) is an ion channel belonging to the polycystin subfamily of the transient receptor potential (TRP) cation channel superfamily. Mutations in PC2 are associated with autosomal dominant polycystic kidney disease (ADPKD), one of the most common genetic diseases in humans. Despite extensive studies over the past decades, the role of PC2 in normal and ADPKD kidneys remains largely unknown, and the activation and regulation of this ion channel are elusive. To gain a better understanding of the structure and function of the mouse PC2 channel, this study focused on generating five pore mutations in a gain-of-function (GOF) mutant of …

Exploring The Dual Role Of Vegf165 In Bleomycin Induced Lung Injury And Recovery In A Aav Mice Model, Igor Ban

Theses and Dissertations

Pulmonary fibrosis (PF) poses a significant challenge in respiratory medicine due to its complex etiology and progression characterized by excessive lung scarring, leading to compromised pulmonary function. The multifaceted roles of Vascular Endothelial Growth Factor A (VEGF-A) and specifically its isoform VEGF-A165 are critical in this context, given their involvement in angiogenesis, lymphangiogenesis, and tissue repair. The dual nature of VEGF-A165, promoting angiogenesis and potentially exacerbating or mitigating fibrosis, underscores its significance in both the progression and recovery phases of PF. Our work, explores the nuanced functions of VEGF-A165 using AAV1 as a gene delivery vector to overexpress this growth …

Therapeutic Intervention Of Neuroinflammation In Alzheimer Disease By Inhibition Of Classical Complement Pathway Using Anti-C1r Loaded Exosomes, Terjahna Richards

Theses and Dissertations

Alzheimer’s disease (AD) is a neurodegenerative disease associated with memory decline and cognitive impairment. It is a growing global health and socioeconomic concern, affecting more than 50 million people worldwide. Current therapeutics focus solely on symptomatic management instead of preventative or curative strategies. This is mainly a result of the complexity of the pathophysiological mechanism and the poor understanding of the disease progression. Recently, studies have focused on bridging knowledge gaps giving rise to substantial evidence on the vital role of neuroinflammation in the progression of AD. Neuroinflammation has been shown to be a precursor to apparent amyloid plaque accumulation …

Hjp 272, An Endothelin Receptor Antagonist, And Its Role In Cancer Cell Migration And Invasion, Nabeela Fatima Baig

Theses and Dissertations

Endothelins are a family of versatile peptides composed of 21 amino acids with three isoforms: ET-1, ET-2, and ET-3. As the most abundant of the three isoforms, ET-1 is involved in various biological processes, such as regulation of vascular tone, humoral homeostasis, and neural crest development. However, focus is now being directed towards investigating the functions of the Endothelin axis in the progression of different tumor types including ovarian, prostate, breast, lungs etc. HJP 272 is a novel ETAR antagonist and while our group has previously researched its effects on lung inflammation and preterm birth, this study marks the first …

Characterizing The Role Of Epigenetic Regulator Menin In Pediatric Neuroblastoma Growth Using Pre-Clinical Models, Rameswari Chilamakuri

Theses and Dissertations

Epigenetic regulators such as Menin and MLL1 drive cancer progression, metastasis, and relapse. MLL1 forms a COMPASS complex with its binding partners, including Menin, to function as an epigenetic enzyme H3K4 methyltransferase. H3K4 methyltransferases are known to promote cancer stem cells' (CSCs) self-renewal capacity and metastasis. CSCs are one of the major causes of tumor relapse and metastasis conditions, ultimately leading to poor overall patient survival. Neuroblastoma (NB) is an extracranial solid tumor that accounts for 8-10% of all pediatric cancers, and approximately 50% of patients display metastasis at the time of diagnosis. In this study, we hypothesized and used …

Opioid System's Potential Involvement In Ketamine's Cognitive And Antidepressant-Like Effects In Rats, Fan Zhang

Theses and Dissertations

Depression stands as one of the most pervasive mental health challenges worldwide, including the United States. Conventional antidepressants typically display a delayed onset of therapeutic action (4-8 weeks) and lack efficacy in a substantial portion of patients (over 40% of depressed patients are treatment-resistant). The dissociative anesthetic ketamine, which is a glutamatergic N-methyl-D-aspartate (NMDA) antagonist, has emerged as a promising novel treatment due to its rapid and prolonged antidepressant effects, and it is especially beneficial for treatment-resistant depression. Nevertheless, the precise neurochemical mechanisms underlying ketamine’s antidepressant properties remain elusive, hampering the development of safer and more efficacious treatments. The present …

Mechanistic Analysis Of Four-Way Dna Junctions And Cytokine-Binding Aptamers For Therapeutic Interventions, Roaa S. Mahmoud

Theses and Dissertations

DNA is inherently dynamic and topologically diverse and can fold into many different structures. Besides the canonical Watson-Crick structure, other higher-order structures such as G-quadruplexes (G4), i-motifs (iM), and four-way DNA junctions are possible. Although these high-order DNA structures are known to form transiently, they are important due to the crucial roles they play in many cellular processes including DNA replication, recombination, and repair. Among these DNA structures, 4-way junctions (also known as Holliday junctions, HJ) which are formed during the repair of double-strand DNA breaks (DSBs) and interact with proteins have garnered significant attention due to their central role …

Redesign, Evaluation, And Validation Of A Commercially Viable High-Resolution Melt Based Mixture Screening Tool, Chastyn Smith

Theses and Dissertations

Analysis of evidentiary samples containing DNA from multiple contributors (“mixtures”) is a time intensive process for a forensic analyst and one where the contributor nature of a sample is not revealed until the end of the traditional forensic workflow. Often, at this stage, retesting or additional testing of mixture samples may not be possible, particularly if the DNA collection device did not preserve the DNA well enough; consequently leaving only trace amounts of a contributor’s DNA present. Thus, a new collection device that would allow for the increased preservation/integrity of evidentiary samples as well as a method that would allow …

Exploring The Relationship Of Intact Limb Knee Joint Loading With Step Lengths And Step Length Symmetry In Individuals With Lower Limb Amputation, Edward D. Syrett

Theses and Dissertations

Individuals with unilateral lower limb amputation (LLA) frequently experience intact limb knee pain, dysfunction, and osteoarthritis. These problems can worsen pre-existing reductions in mobility and quality of life that this population regularly experiences. It is widely accepted that knee joint loading is implicated in the development of knee joint issues, and intact limb knee joint load alterations are thought to be widespread in individuals with LLA. Prior work suggests that gait deviations frequently present in this population may be a contributory cause of these altered forces. These deviations can take many forms, but individuals with LLA commonly demonstrate alterations in …

Investigating The Regulation Of Epithelial-Mesenchymal Transition Through Fibronectin-Mediated Tgf-Β1 Mechanotransduction Using In Vitro Models Of Triple-Negative Breast Cancer, Michael M. Sofroniou

Theses and Dissertations

TGF-β1 is a potent regulator of epithelial-mesenchymal transtion (EMT), a cellular
transdifferentation process that promotes cellular motility and matrix remodeling, critical to tumorigenesis and metastasis. TGF-β1-induced EMT promotes the assembly of FN, which synergistically binds latent TGF-β1 to aid in its activation. The purpose of of this work is to further probe the effects of autocrine TGF-β1 signaling during EMT progression by utilizing CRISPR-Cas9 deletion of the TGFB1 gene in in vitro models of triple-negative breast cancer. The studies outlined in this work demonstrate that autocrine TGF-β1 maintains a baseline level of mesenchymal marker expression. It also functions to desensitize …

Illuminating Function And Enabling Inhibition At The Protein-Membrane Interface Of Glutathione Peroxidase 4., Courtney L. Labrecque

Theses and Dissertations

Membrane proteins act as gateways to cells and are responsible not only for signaling, lipid transport, and cellular homeostasis, but they are also integral in disease pathologies. One class of membrane proteins, peripheral membrane proteins (PMPs), reside in aqueous compartments of the cell until they are recruited to the membrane to perform their function. Glutathione peroxidase 4 (GPx4) is a PMP enzyme responsible for engaging the membrane and reducing lipid hydroperoxides, thereby preventing cellular death via ferroptosis. Due to its central function surrounding ferroptosis, GPx4 is an interesting drug target that has recently gained interest for inflammatory diseases as well …