Bachelor of Sciences, Ohio University, 2024, Biological Sciences

Fat-specific protein 27 (FSP27) plays a key role in regulating lipid metabolism in both adipose tissue and skeletal muscle (SkM) function. Emerging evidence suggests a link between SkM health and cognitive function, leading us to hypothesize that FSP27 might significantly impact neurocognition. To test this hypothesis, we utilized SkM-specific FSP27-knockout (Fsp27-/-) and SkM-humanized FSP27-transgenic (M-hFSP27tg) mouse models, representing loss- and gain-of- function of FSP27 in muscle, respectively. Behavioral assessments, including the Morris Water Maze and the Classic Labyrinth maze, were conducted to evaluate the effects of FSP27 manipulation on spatial memory, executive function, and motor coordination. Our results revealed that MhFSP27tg mice demonstrated superior cognitive performance, with enhanced memory retention and accelerated learning curves, while M-Fsp27-/- mice exhibited impaired spatial memory and problem-solving abilities. These findings indicate a significant role for FSP27 in neurocognitive function, likely mediated by its influence on SkM metabolism and interaction with brain processes. This research underscores FSP27's potential in addressing the reciprocal relationship between SkM and cognition, with future studies planned to elucidate its molecular mechanisms in neurocognition.

PHD, Kent State University, 2025, College of Arts and Sciences / School of Biomedical Sciences

Part I: The protein phosphatase isoforms PP1γ1 and PP1γ2, alternate spliced products of the Ppp1cc, are identical except for their 8 or 22 amino acid C-termini. The PP1γ2 isoform is enriched in testis and is present in sperm from all placental mammals. Targeted disruption of Ppp1cc results in male infertility. Testis specific expression of PP1γ2 restores fertility in mice lacking Ppp1cc. However, replacing PP1γ2 with PP1γ1 in testis and sperm through CRISPR /Cas9 gene editing impairs sperm motility, ATP generation, and fertility. The interactors of protein phosphatase 1 (RIPPO) - PPP1R2/I2, PPP1R7/SDS22, and PPP1R3/I3 - are present in sperm as regulators of PP1γ2 during sperm motility initiation and maturation in the epididymis. Reversible phosphorylation of these RIPPOs results in PP1γ2 activation or inhibition. The spatiotemporal expression of I2, I3, and sds22 matches the expression of PP1γ2 in developing testicular spermatogenic cells. The enzyme PP1γ2 and the inhibitors colocalize within sperm. In PP1γ1 knock-in mice, I2, I3, and SDS22 localization within sperm are dramatically altered. While PP1γ2 and the RIPPOs are present in the functional regions of the sperm head, PP1γ1 along with I2, I3, and SDS22 are absent. Protein levels, of the inhibitors determined by mass spectroscopy and western blot are similar to that found in testis and sperm of WT mice. The mRNA levels of SDS22, I2, I3 are also roughly similar in testis KI mice. We have determined levels of phosphorylation of I2, I3, and SDS22 in sperm from PP1γ1 KI PP1γ2 mice compared to WT mice in caudal sperm. Serine phosphorylation of PPP1R7 and PPP1R2 were significantly altered in sperm from PP1γ1 knock-in mice while phosphorylation of PPP1R11 was unchanged, Phosphoproteome analysis also showed altered phosphorylation of hexokinase and mitochondrial pyruvate dehydrogenase. The phosphorylation of mono carboxylate transporter along with its associated protein embigin and spetin present in sperm midpiece and (open full item for complete abstract)

PHD, Kent State University, 2025, College of Arts and Sciences / School of Biomedical Sciences

Cardiovascular disorders remain the leading cause of morbidity and mortality worldwide, with atherosclerosis as the predominant underlying pathology. In individuals with Metabolic Syndrome (MetS), which encompasses conditions such as obesity, diabetes, dyslipidemia, and hypertension, the development of atherosclerosis is accelerated. Vascular smooth muscle cells (VSMCs) play a pivotal role in atherosclerotic plaque formation through phenotypic switching—a process influenced by various extracellular factors, including TSP1. TSP1, a matricellular glycoprotein, has emerged as a key mediator of vascular inflammation, remodeling, and cell behavior. However, its precise role in VSMC differentiation and atherosclerosis in the context of MetS remains unclear. The significance of my dissertation lies in addressing the critical knowledge gap surrounding TSP1's role in VSMC phenotypic modulation within atherosclerotic lesions, particularly under the compounded metabolic stresses of MetS. The goal of this study was to elucidate the effect of TSP1 on VSMC behavior and plaque development. We specifically investigated the link between TSP1 expression, atherosclerosis, and SMC differentiation in MetS and the regulatory role of smooth muscle-specific TSP1 in SMC phenotypic changes in diabetes and the molecular pathways involved. We found that agouti KKAy+/-ApoE-/- mice (MetS) exhibited pronounced metabolic disturbances, including significant increases in body weight, non-fasted blood glucose levels (>250 mg/dL), and plasma lipid levels (cholesterol and triglycerides) compared to non-agouti KKAy-/-ApoE-/- controls (non-MetS). Male MetS mice demonstrated severe glucose intolerance, although insulin sensitivity was unaffected across groups. Male MetS mice exhibited significantly greater lipid burden and plaque area in the aortic root associated with upregulation of TSP1 and lesser testosterone levels compared to non-MetS males. However, female MetS mice did not display the same les (open full item for complete abstract)

Psy. D., Antioch University, 2025, Antioch Seattle: Clinical Psychology

This dissertation presents a comprehensive analysis of the current research on scrupulosity, a subtype of obsessive–compulsive disorder (OCD) characterized by intrusive thoughts and compulsive behaviors related to religious and moral concerns. The dissertation identifies key similarities and differences from OCD, and directs focus to thematically related yet unsubstantiated theoretical work in psychology that helps elucidate the core features and etiological factors of scrupulosity as differentiated from other OCD subtypes. The study addresses the critical dearth of research on scrupulosity, aiming to fill significant gaps in the literature regarding its historical context, varied presentation and prevalence in different cultural contexts, and potentially effective treatment approaches to address better the needs of a significant number of people worldwide. Beginning with an exploration of historical conceptualizations from the 2nd through the early 21st centuries, the dissertation traces the recognition of scrupulosity and recommendations for treatment across various cultural traditions and major world religions including Islam, Judaism, Hinduism, Buddhism, and Christianity, from both Protestant and Catholic sources, as well as non-religious belief systems. It highlights notable historical figures who exhibited scrupulous behaviors contextualizing them with a modern psychological lens. As the leading theologians of their faiths, they often ironically v advised its treatment from their own experience as the most influential theologians of each of their faiths. These historical writings still have wisdom to impart today. The history of scrupulosity is, in many ways, a history of religion across time and culture, as well as of the birth and first 150 years of psychology itself. Key schools of psychological thought are explored for relevance to developing contemporary evidence-based treatments. Due to few qualitative or quantitative studies on scrupulosity compared t (open full item for complete abstract)

PHD, Kent State University, 2025, College of Arts and Sciences / School of Biomedical Sciences

New insights into cellular interactions and key biomolecules involved in LC bone metastasis will have remarkable therapeutic benefits. Using a panel of four LC cells, we investigated the interaction between LC and bone (LC-OC interaction) by exposing differentiating OCs to LC cells directly in a co-incubation setting or indirectly via treatment with LC secretomes (CM or exosomes). LC-OC interaction facilitated the production of large-sized OCs (nuclei >10) coupled with extensive bone resorption pits on bovine bone slices. Proteomics and western blotting analysis identified Gal3bp as a potential biomarker which was primarily released by LC exosomes. The enhancement of OC differentiation and function by LC-exosomal Gal3bp was supported by studies where recombinant Gal3bp and anti-Gal3bp were applied. Our results indicated that dysregulation of crucial OC markers, Gal3bp and Gal3 during LC-OC interaction possibly contributed to the stimulation of osteoclastogenesis. Overall, this work implicated LC-exosomal Gal3bp in osteolytic metastasis of LC which warrants further studies to assess its potential prognostic and therapeutic relevance. BM is a significant complication of solid malignancies such as LC, frequently manifesting in advanced stages and affecting 30-40% of patients with LC, resulting in poor prognoses. Research on the LC-exosomal marker Gal3bp indicates its potential as a predictive biomarker due to its role in promoting osteoclastogenesis, although its in vivo implications remain to be elucidated. To investigate Gal3bp in vivo, we utilized murine BM models with intratibial and intracardiac administration of A549-Luc2 cells. Skeletal samples from athymic (nu/nu) mice were analyzed for structural deformities using 3D reconstruction and quantitative parameters (cortical and trabecular). The results demonstrated that intratibial rGal3bp supplementation of LC cells significantly increased bone resorption and compromised bone structure due to enhanced osteoclast (open full item for complete abstract)

Doctor of Philosophy (PhD), Wright State University, 2024, Biomedical Sciences PhD

DNA replication can be perturbed by various agents that slow or stall the replication forks, causing replication stress. If undetected, stressed forks may collapse, causing mutagenic DNA damage or cell death. In response to replication stress and DNA damage, the eukaryotic cell activates the DNA replication checkpoint (DRC) and DNA damage checkpoint (DDC) pathways to promote DNA synthesis, repair, and cell survival. The two cell cycle checkpoint pathways are controlled by the protein sensor kinases Rad3 (hATR/scMec1) and Tel1 (hATM/scTel1) in fission yeast, although Tel1 plays a minimal role in checkpoint functions. Rad3 and Tel1 belong to a family of phosphatidylinositol-3-kinase-related kinases (PIKKs), whose stability is regulated by the heterotrimeric TTT (Tel2-Tti1-Tti2) complex. The current model suggests that the TTT complex works with Hsp90 and R2TP complex in the co-translational maturation of all PIKKs for their proper folding and stability. We have previously reported a tel2-C307Y mutant with a moderately reduced Rad3 protein level (~60% of wild-type cells). This mutation eliminates Rad3 mediated signaling in the DRC pathway but moderately reduces signaling in the DDC pathway. This result suggests that Tel2 of the TTT complex may specifically regulate the DRC pathway. In this study, we investigated this possibility by taking a genetic approach to analyze the functions of Tti1, the largest subunit of the TTT complex. We randomly mutated the tti1 gene and integrated the mutations at the genomic locus by pop-in and pop-out recombination strategy. As a result, 100 primary tti1 mutants were successfully screened, based on their increased sensitivities to hydroxyurea (HU) which depletes cellular dNTPs and/or the DNA damaging agent methyl methanesulfonate (MMS). Preliminary characterization of the primary Tti1 mutants, based on their relative sensitivities to HU, MMS or both agents, led us to focus on a collection of 24 mutants. Among the 24 mutants, DNA seq (open full item for complete abstract)

Doctor of Philosophy (PhD), Wright State University, 2024, Environmental Sciences PhD

Gold has long been recognized for its medicinal applications, dating back centuries. The use of N-heterocyclic carbenes (NHCs), stable, tunable ligands, has revolutionized the development of metal complexes, particularly those involving gold. Among these, Au-NHC complexes have shown significant promise in treating various diseases, especially arthritis and cancer. More recently, our research has focused on synthesizing dual-targeting naphthoquinone-annulated NHC-based gold(I) complexes, which have demonstrated impressive therapeutic potential in cancer treatment. This emphasizes the growing importance of gold-based complexes in developing more selective and effective cancer therapies. By strategically tuning the electronic and steric properties of these complexes, we have explored a variety of functionalized naphthoquinone-annulated NHCs. In Chapter 1, we detail the synthesis of two isomers of nitro-functionalized naphthoquinone-annulated NHC precursors (imidazolium salts). The successful synthesis of these compounds was confirmed through various characterization techniques, including 1H and 13C NMR and FT-IR spectroscopy. We also investigated the electrochemical properties of these NHC precursors (imidazolium salts), to better understand their redox chemistry. In Chapter 2, we expanded our study by complexing the newly generated NHCs with iridium metal centers to examine their electronic and electrochemical properties. NHC-iridium cyclooctadiene (Ir-COD) and NHC-iridium carbonyl complexes were synthesized for this purpose. The introduction of nitro groups at two distinct positions on the ligands led to noticeable variations in their electrochemical behavior. Additionally, FT-IR spectroscopy was used to examine the carbonyl stretching frequencies of the Ir(CO)₂Cl complexes. The values obtained from IR provided significant insights into the donating abilities of newly generated NHCs. This was further quantified through Tolman Electronic Parameter (TEP) value cal (open full item for complete abstract)

Doctor of Philosophy in Clinical-Bioanalytical Chemistry, Cleveland State University, 2024, College of Arts and Sciences

Ulcerative colitis disease can progress into cancer over many stages of dysplasia if not treated at early stages. Inflammatory responses are the main factors in the progression of ulcerative colitis. They are triggered by epithelial lesions as well as microbiota, which infiltrate into the interstitium layer and augment inflammatory responses. The inflammatory responses are mediated by cytokines and chemokines, which are induced by colon fibroblasts and epithelial cells, immune cells, as well as the microbiome. This study focuses on one potent, pro-inflammatory chemokine, CXCL8, which attracts immune cell to the lesion areas and augments disease. The first part of the dissertation focuses on the signaling pathways that regulate upregulation of CXCL8 levels in colitis-associated and colitis-associated cancer (CAC) fibroblasts. Using unbiased mRNA sequencing analysis, we identified that the progression of the fibroblasts is associated with an inflammatory status and involves the injury-induced senescence pathway. Testing three immune-derived cytokines (TNF, IL-1 and IFN) as well as two bacterial signals (LPS and Flagellin/FliC) demonstrated that the NFB signaling pathway is the major signaling pathway associated with upregulation of CXCL8 expression levels in colon fibroblasts in vitro and in vivo. In addition, investigating mycoplasma as a representative in vivo stimulus also resulted in upregulated CXCL8 expression. Moreover, normal colon fibroblasts, colitis-associated fibroblasts and colon epithelial cell demonstrate overlapping, but also cell type-specific ways to activate CXCL8 expression. Finally, reprogramming colon fibroblasts demonstrates an epigenetic memory in colon fibroblasts from ulcerative colitis that is associated with elevation of CXCL8 expression. The second part of the dissertation focuses on the regulation of microRNAs (miRNAs) in downregulating CXCL8 expression levels. Investigating E2F7/E2F8, cMyc, and miRNAs of the miR-17 family demonstra (open full item for complete abstract)

Master of Science (M.S.), University of Dayton, 2024, Bioengineering

Electrospun nanofiber (ESNF) membranes have attracted widespread interest in many applications due to their advantages in high specific surface area, high porosity, and structural controllability. This study combines gap electrospinning and electrolyte-assisted electrospinning techniques to develop a novel electrospinning approach for producing nanofiber mats of arbitrary geometry. A 3D printed conductive gelatin-based polymer electrolyte (GPE) solution is used as a geometric collector to focus the deposition of electrospun mats. The method utilizes syringe extrusion 3D printing of the GPE solution to produce a shape upon which ESNF are focused. The printable GPE ink is formulated to ensure it possesses the necessary conductivity, shear-thinning, and thixotropic properties. We have developed a gelatin-based GPE ink, enhanced with Laponite to improve shear-thinning properties and salts to increase conductivity. The 3D printing equipment then extrudes the GPE solution on the surface of the target device according to the pre-designed pattern. The optimized GPE solution formulation contained 8% w/v gelatin, 0.2% w/v Laponite, 2 mol/L sodium chloride, and 14.3% v/v glycerol, which was shown to meet the dual requirements of 3D printing and assisting electrospinning. The ink's conductivity was 8.02 S/m measured using a custom developed four-point probe system for gels. Rheological analysis demonstrated that the ink exhibits shear thinning (fluid behavior index n=0.223), which allows GPE ink to maintain a balance between easy extrusion and structural stability. We tested the electrospinning solution used during the experiment and investigated and characterized electrospinning operating parameters to explore several relationships between unrestricted mat diameter (UMD) and electrospinning operating parameters and GPE patterning threshold. The performance of the GPE ink was thoroughly examined experimentally: under the conditions of 25°C and 26% relative humidity, the (open full item for complete abstract)

Doctor of Philosophy, Case Western Reserve University, 2025, Pathology

The COVID-19 pandemic claimed the lives of millions of people and affected communities worldwide. SARS-CoV-2, the virus that causes COVID-19, continues to be a global health concern, as does the inevitable threat of new viral outbreaks. We must therefore learn from this virus in the hope of better preparing for future pandemics. We investigated the SARS-CoV-2 accessory protein ORF3a and its roles in modulating host antiviral strategies, namely inflammatory signaling and autophagy regulation. ORF3a activates NF-κB signaling, which induces an inflammatory response in infected cells and can also prime certain cells for inflammasome assembly and subsequent cell death. We found that, unlike the homologous protein SARS-CoV ORF3a, SARS-CoV-2 ORF3a does not depend on its N-terminal TRAF-binding sequence to activate NF-κB. The ORF3a homologs thus affect NF-κB signaling through different mechanisms. Second, SARS-CoV- 2 ORF3a blocks autophagy by binding to the human protein VPS39, a member of the complex that facilitates membrane fusion between autophagic compartments. We discovered that the predicted β-propeller domain of VPS39 is critical to its interaction with ORF3a. Regulating autophagy is important for productive SARS-CoV-2 infection; disrupting the ORF3a:VPS39 interaction could therefore be a future strategy to hinder SARS-CoV-2 propagation.

PhD, University of Cincinnati, 2024, Medicine: Pathobiology and Molecular Medicine

Myocardial infarction (MI) is currently responsible for over 150,000 deaths annually in the United States. Even when successfully treated, MI imparts lasting cardiovascular defects that lead to increased morbidity and mortality. One of the contributors to these observed outcomes is cardiac reperfusion injury (CRI), which manifests as cardiomyocyte death after ischemia is treated. CRI is caused by several events triggered by the ischemic microenvironment during the MI, including metabolic changes, perturbation in Ca2+ signaling, pro-thrombotic inflammation, endothelial dysfunction, and reactive oxygen species (ROS) formation. ROS scavenging in particular has drawn the attention of many investigators as a means of preventing the CRI. However, the rapid nature of free radical formation during reperfusion is a major pharmacokinetic obstacle in using antioxidants for this indication. Controlled hypoxic reperfusion (CHR) is the transient introduction of hypoxic reperfusate into the ischemic heart and subsequent reperfusion with a fully oxygenated solution. CHR was shown to be effective in reducing the infarct size and improving the heart function compared to normal reperfusion. It has been hypothesized that the cardioprotective effects of CHR are achieved by reducing the amount of ROS formed in the heart after reperfusion. Radhakrishnan et al. first showed in 2016 that during acoustic droplet vaporization (ADV), the ultrasound-mediated conversion of liquid perfluorocarbon droplets into gas microbubbles, oxygen (O2) diffused from the surrounding fluid into the microbubbles. This process has since been extensively described in buffers. In this dissertation, O2 scavenging from physiologically prepared whole blood is evaluated and a novel method for quantifying phase transition efficiency via flow cytometry is proposed. First, whole blood was prepared to mimic arterial whole blood in vivo. Next, oxygen scavenging and was assessed in an in vitro flow phantom via blood gas a (open full item for complete abstract)

MS, University of Cincinnati, 2024, Engineering and Applied Science: Chemical Engineering

This work presents crucial advances towards developing a novel, electricity-free PCR thermocycler utilizing the Curie pendulum mechanism. Designed specifically for resource-limited and remote environments, this thermocycler aims to overcome the challenges posed by the high cost, large size, and significant power requirements of conventional PCR systems. By leveraging the Curie temperature effect in nickel strips, the system enables passive heating and cooling without external power or temperature sensors, providing a portable solution for molecular diagnostics. The Curie pendulum setup comprises a nickel strip mounted on a lightweight pendulum arm those swings freely in response to changes in temperature. A stationary magnet and a flame-drive heat source are strategically positioned to induce oscillatory thermal cycling as the nickel strip repeatedly loses and regains magnetic properties at its Curie point. This design enables consistent thermal cycling. Key parameters, including magnetic strength, heat source selection, strip orientation, thickness, and length, were optimized to ensure consistent and effective thermocycling. Additionally, participation in the NSF I-Corps program provided valuable insights from clinicians and researchers that informed the design, emphasizing the practical needs of healthcare workers in both developed and developing regions. The Curie pendulum-based thermocycler offers a cost-effective and portable alternative for nucleic acid amplification in resource-limited settings, with potential applications extending beyond PCR to other biochemical assays and educational use. This system represents a significant advance in point-of-care diagnostics, bringing us closer to a fully electricity-free PCR testing platform.

Master of Computer Science (M.C.S.), University of Dayton, 2024, Computer Science

The mouse cerebellar cortex, with its layered structure of molecular, Purkinje, and granular layers, refines motor control, coordination, and balance. Purkinje cells, the primary output neurons, process signals from parallel and climbing fibers and send inhibitory outputs to cerebellar nuclei, enabling motor learning and cognitive processing. Nowadays we have a better understanding of the cerebellar cortex's crucial roles in brain function and its implications in neurological disorders thanks to recent developments in neuroimaging, optogenetics, and electrophysiology that have clarified mechanisms underlying synaptic plasticity and signal processing within the region. Segmenting the cerebellar cortex is crucial for analyzing its cellular structure and connectivity, helping us understand motor control and learning processes. It enables detailed study of neural circuits and is key for investigating structural changes related to neurological disorders, aiding in targeted research and potential treatments. In this thesis, we apply various segmentation models to identify mouse brain cells through instance segmentation, focusing on precise classification of cell types and structures within the cerebellar cortex. We collect and annotate a dataset of 1000 images of mouse brain cells. We further evaluate different instance segmentation methods in two families, namely, transformer and non-transformer-based methods. Results indicate that non-transformer models outperformed transformer models in the small sized cells whereas transformer-based methods achieve better performance on large-sized cells. This thesis paves way to further research in using computer vision to understand mouse brain cells.

Doctor of Philosophy, Case Western Reserve University, 2025, Pathology

Plasmodium vivax induces blood stage infections by invading red blood cells (RBCs) using the Duffy (Fy) blood group antigen system. Individuals with the Fy blood group-positive alleles (FY*A and/or FY*B) are susceptible to infection, while Fy-negative (null) individuals with the erythrocyte silent, ES, allele (FY*AES and FY*BES) show resistance. Recent studies challenge this paradigm by reporting that Fy null populations across sub-Saharan Africa show incidences of P. vivax infection. In addition, our recent breakthrough discovery in Cell Host Microbe demonstrates Fy “null” cells may in fact express the Fy protein. Studies suggest that sites of hematopoiesis, such as the bone marrow or spleen, may act as a biological niche for P. vivax invasion into reticulocytes (the parasite's preferred target cells), but this has not been substantiated in vivo. To investigate this, a reliable culturing protocol and method of assessment is required. Research shows that culturing P. vivax in vitro is not only challenging, but impossible on a long-term scale, necessitating more innovative methods. We demonstrate that human bone marrow can be utilized to culture P. vivax and propagate the parasite on a long-term basis (over 1 year) for a series of experiments. To address our limitations, we have introduced the more easily cultured P. knowlesi (the closest related Plasmodium species to P. vivax) that has been genetically modified to replace its endogenous Fy binding protein (DBP) orthologue with PvDBP to create a transgenic parasite, PkPvDBPOR. Using this model, we evaluated the culture under different perturbations (e.g. Fy-specific antibodies) and discovered low level in vitro invasion of Fy null recipient cells.

Doctor of Philosophy, Case Western Reserve University, 2024, Genetics

Background: Cystic Fibrosis (CF) is a progressive, systemic disease caused by mutations in the CFTR gene. Despite the availability of highly effective CFTR modulators, there is continued need for the development of CF therapeutics because there is a portion of people with CF not eligible for modulator therapy due to their mutations. CF mouse models are powerful tools to study mutation-specific CF biology and to screen novel therapeutics. This dissertation describes the creation of several novel mouse models using different genetic approaches. Methods & Results: Human-exon (hEx) replacement mouse strains in which a mouse (m)Cftr exon-of-interest was replaced with the corresponding human exon's sequence were designed. hEx replacement strains carrying either human exon 3, 11, 12, or 26 expressed chimeric Cftr from the endogenous mCftr locus. WT hEx3, hEx11, hEx12, and hEx26 strains displayed phenotypes similar to non-chimeric WT controls and highly dissimilar to CF mice. Additionally, a mouse model containing the W1282X mutation in the mCftr gene was created. W1282X mice recapitulated common CF manifestations including low Cftr mRNA expression, poor survival and growth, and altered electrophysiology in various tissues. Forskolin induced swelling (FIS) was performed on intestinal organoids derived from the W1282X mouse and an existing G542X mouse treated with various pharmacologics. Both organoids demonstrated FIS upon treatment with readthrough agents, nonsense-mediated decay inhibitors, and CFTR modulators; however, the specific drug combinations that elucidated the most robust rescue differed in W1282X and G542X organoids. Discussion: The hEx replacement mice provide a novel genetic approach for creating humanized mouse models. hEx 3, 11, 12, and 26 strains displayed WT phenotypes with no evidence of CFTR dysfunction therefore validating the hEx approach for modeling CF mutations. The W1282X mouse is the first W1282X-specific in vivo CF model. It (open full item for complete abstract)

Doctor of Philosophy, The Ohio State University, 2024, Molecular, Cellular and Developmental Biology

Cardiovascular disease in the leading cause of death worldwide. While the etiology of heart disease is diverse, a well-documented unifying theme in the progression of heart disease is the development of cardiac fibrosis. Cardiac fibrosis, defined as excess collagen and extracellular matrix deposition in the heart, is mediated by the activation and differentiation of resident quiescent fibroblasts into myofibroblasts. Pathologic fibrotic remodeling in the heart occurs in response to persistent stress stimuli, as well as injury. Fibrotic remodeling leads to stiffening of the ventricles, impaired relaxation, and progressive impairment in cardiac function. While current standard-of-care medications have been vital in the treatment of heart diseases over the last several decades, there are currently there are currently no effective therapies in clinical use specifically targeting fibrosis in the heart. This highlights a significant need to identify novel therapeutics that target fibroblasts and fibrotic remodeling. Our group recently identified Sertad4 (Serta Domain Containing Protein 4) as a potential regulator of fibroblast activation in the heart. However, the function of Sertad4 is unknown. In this dissertation, I sought to investigate the role of Sertad4 in pathologic cardiac remodeling through a combination of in vitro and in vivo studies. To study the role of Sertad4 in vivo we utilized a reporter mouse and a global loss of function mouse. We also utilized three models of cardiac dysfunction: 1) myocardial infarction through coronary LAD ligation; 2) chronic angiotensin II/phenylephrine infusion through osmotic minipumps and 3) chronological aging as a model of diastolic dysfunction. We observed that Sertad4 is predominantly expressed in activated fibroblasts and its expression is significantly elevated in failing human and mouse hearts. Loss of Sertad4 in mice led to preserved cardiac function post-MI and decrease in cardiac hypertrophy and fibrosis. Using is (open full item for complete abstract)

Doctor of Philosophy, The Ohio State University, 2024, Biomedical Sciences

Viral pandemics are caused by viruses spilling over from animal reservoirs and subsequently adapting to efficiently infect, replicate, and spread in human hosts. Given that coronavirus and avian influenza virus outbreaks have occurred in recent years in geographic regions in which human deficiencies in the Interferon-Induced Transmembrane Protein 3 (IFITM3) antiviral protein are common, we investigated whether IFITM3 may play a role in interspecies virus infection and adaptation. We found that both IFITM3-deficient mice and human cells could be infected with low doses of avian influenza viruses that failed to infect WT counterparts, identifying a new role for IFITM3 in controlling the minimum infectious viral dose threshold. Additionally, we used a panel of 11 diverse avian influenza viruses and 3 swine viruses, 2 of which are known to have transmitted to humans, to infect human cells with or without IFITM3. Each animal virus showed increased infection of human cells lacking IFITM3 as compared to controls, even upon interferon treatment. Remarkably, influenza viruses passaged through Ifitm3-/- mice exhibited enhanced host adaptation, a result that was distinct from passaging in mice deficient for interferon signaling, which caused virus attenuation. Passaging of a SARS-CoV-2 beta strain and an Omicron BA.4 strain through WT or Ifitm3-/- mice both resulted in elimination of tissue culture adaptations prevalent in the parent stocks of these viruses. Further, virus passaged 20 times through Ifitm3-/- animals gained >1-log replicative advantage and ability to induce weight loss in WT mice while WT-passaged virus did not significantly change in its ability to replicate or induce weight loss. Mouse adaptation of both influenza virus and SARS-CoV-2 was associated with discrete changes in the viral genomes resulting in amino acid substitutions, suggesting that enhanced virus replication in the absence of IFITM3 may facilitate adaptive mutations. Our data demonstra (open full item for complete abstract)

Doctor of Philosophy, Case Western Reserve University, 2024, Pathology

The steroid hormone progesterone (P4), acting via the nuclear P4 receptors (PRs) in decidual stromal cells (DSCs) is essential for pregnancy maintenance, and loss of P4/PR signaling induces parturition. Inflammation at the chorion-decidua interface (CDI) triggers term and preterm parturition; however, the mechanism is uncertain. This study tested the hypothesis that pro-labor inflammatory stimuli induce parturition by increasing expression of the P4-metabolizing enzyme, aldo-keto reductase 1C1 (AKR1C1), which converts P4 to an inactive form, thus causing localized intracellular loss of P4/PR signaling. Here, we utilized human CDI tissue, a decidual stromal cell line, and Rhesus macaque and mouse models of preterm birth to determine that 1) AKR1C1 is induced by inflammation and metabolizes P4 thereby starving the receptor to induce a local P4 withdrawal, which can be prevented with an AKR1C1 inhibitor BPSA and 2) Inflammation-induced AKR1C1 transcription is dependent on MAPK and NF-kB signaling. Taken together, our data support the hypothesis that inflammatory stress promotes labor by inducing AKR1C1-mediated P4 withdrawal in DSCs. Importantly, our data suggest that risk for inflammation-induced preterm birth could be reduced by targeting AKR1C1 activity in uterine P4 target cells. This can be achieved via inhibition of AKR1C1 activity (e.g., with BPSA), by preservation of PR activity by treatment with a P4 analog that is not metabolized by AKR1C1 (e.g., R5020), and/or by inhibition of IL-1β-induced AKR1C1 expression in DSCs (e.g., by treatment with an IL1 receptor antagonist such as Anakinra). Furthermore, P38 inhibition may be a viable therapeutic strategy to prevent induction of AKR1C1 transcription.

Doctor of Philosophy, The Ohio State University, 2024, Biophysics

Fragment-based quantum chemistry methods provide a way to circumvent the steep nonlinear scaling of electronic structure calculations, enabling the investigation of large molecular systems using high-level methods. First, we present calculations on enzyme models containing 500-600 atoms using the many-body expansion (MBE) and compare them to benchmarks where the entire enzyme-substrate complex is described at the same level of density functional theory (DFT). When amino acid fragments contain ionic side chains, the MBE exhibits oscillatory behavior under vacuum boundary conditions, but rapid convergence is restored using low-dielectric boundary conditions. This suggests that full-system gas-phase calculations are unsuitable as benchmarks for assessing errors in fragment-based approximations. A three-body protocol maintains sub-kcal/mol accuracy compared to supersystem calculations, as does a two-body approach combined with a low-cost full-system correction. In the next section, we use fragmentation to compute protein–ligand interaction energies in systems with several thousand atoms. Convergence tests using a minimal-basis semi-empirical method (HF-3c) indicate that two-body calculations, with single-residue fragments and simple hydrogen caps, are sufficient to reproduce interaction energies obtained using conventional supramolecular electronic structure calculations, to with 1 kcal/mol at about 1% of the cost. Additionally, we show that semi-empirical methods can be used as an alternative to DFT, to assess convergence of sequences of quantum mechanics (QM) models (of increasing size) generated by different automated protocols. Two-body calculations afford a low-cost way to construct a “QM-informed” enzyme model. This streamlined, user-friendly approach to building ligand binding-site models requires no prior information or manual adjustments, making it accessible and practical for a wide range of applications. For the latter parts of this work, we will be focusi (open full item for complete abstract)

PHD, Kent State University, 2024, College of Arts and Sciences / School of Biomedical Sciences

Wound healing is a very complex process requiring a well-orchestrated integration of multiple cellular and molecular events involving many players for antimicrobial activity and the promotion of new vascular formation. Acute wounds are healed by following the normal process of repair including inflammation, proliferation, and remodeling phases. If acute wounds fail to progress through the normal healing phases, it can develop into delayed healing or non-healing chronic wounds, particularly associated with the presence of bacterial biofilm (a community of bacteria encased in a protective matrix) and impaired angiogenesis. Conventional antibiotics frequently develop resistance and have limited efficacy against biofilm-associated wound infections. Approaches for promoting pro-angiogenic activity for wound healing have been relied on the use of bioactive molecules or growth factors, which have limitations in developing cost-effective treatment options. Additionally, thus far, each of the above aspects for antimicrobial and proangiogenic activities have been separately investigated to a great extent and an integrated approach to simultaneously addressing these three issues in a single drug delivery platform has yet to emerge. Wound scaffolds are biomaterial platforms designed to support tissue regeneration and enhance wound healing. In particular, nanoparticle-based scaffolds hold promise for treating chronic wounds due to their characteristics to exhibit higher reactivity due to high surface to volume ratio for improved cellular interactions, and easiness in surface functionalization, and controlled release of bioactive molecules. Additionally, their small size enables deep penetration into the wound bed. In this study, we propose to harness the unique characteristic of copper ion that can exhibit antibacterial and pro-angiogenic properties towards developing a cost-effective 33 scaffold for treating chronic wounds. Copper has emerged as an essenti (open full item for complete abstract)