Doctor of Philosophy, The Ohio State University, 2004, Molecular, Cellular, and Developmental Biology

Ets transcription factors have been implicated in the development, regulation, and survival of numerous hematopoietic derived cells, including B and T-lymphocytes. E26 avian leukemia oncogene-2 (Ets-2) is an Ets family member transcription factor. Strong expression of Ets-2 is observed in developing thymocytes from the DN1 stage and in B-cells from the pro-B stage. A conserved sequence within the Ets-2 pointed domain amino acids 69 to 73 PLLTP is able to interact with the Map kinase ERK. Threonine seventy-two is phosphorylated by ERK1, and permits transactivation by Ets-2. Mutation of this residue from a threonine to an alanine, Ets-2 T72A, abrogates Ras mediated transactivation of Ets-2. Based on this evidence, we hypothesized that Ets-2 was important in T- and B-cell development, activation, and function. To test this hypothesis we analyzed Ets-2 activation in T-cell lines, and created two transgenic mice lines that express Ets-2 T72A in either developing T-cells or B-cells. Transactivation of Ets-2 can be induced by both Ras activation and by phorbol 12-myristate 13-acetate and ionomycin stimulation in the Jurkat T-cell line. In-vitro analysis of Ets-2 activation by ionomycin results in a rapid, but transient Ets-2 band shift as evidenced by western blot analysis of protein extracts from thymocytes. Transgenic mice that over-express Ets-2 T72A in the thymus displayed a dramatic reduction in thymus size associated with hypocellularity. This reduction was associated with a partial developmental block at the double negative 2 and double negative 3 stage of development, a twenty-fold increase in c-Kit+ expression, and a five-fold increase the CD5low population. Further, thymocytes from the transgenic mice have increased apoptosis both in-vitro and in-vivo compared to non-transgenic littermates. Transgenic mice that over-express Ets-2 T72A in B-lymphocytes revealed a loss of the B220Hi population of B-cells in the bone marrow. This population consists of mature B-cell (open full item for complete abstract)

Committee: Natarajan Muthusamy (Advisor) Subjects: Biology, Cell

Doctor of Philosophy, The Ohio State University, 2023, EDU Teaching and Learning

Abstract This research attempted to begin to examine the problem of why youth and consequently, adults with complex disabilities, are not being employed in competitive, integrated environments. As policy, WIOA mandates this status with few exceptions. The study asked, “who and/or what state, and local agencies provide services that mean to support employment and community access?” The study also questioned what services are successful in providing employment supports to consumers with complex disabilities, as well as the barriers to providing supports. The study used a theoretical framework that combined a Stages-Heuristic policy model and Organizational Niche Theory in which to view the problem. A qualitative research design was used along with descriptive statistics of an electronic survey sent to 123 VR, DD agency and Blind/VI services personnel. A Focus Group was facilitated to expand on survey responses and help discover or add to emerging themes from open- ended survey questions. The results of the survey were limited, with only 17 participants responding to questions. The Focus Group with three members, was also limited in geographical regions, as well as representative agencies. Therefore, it was impossible to answer the main questions regarding what state agencies provide specialized services to individuals with complex disabilities for the purpose of competitive, integrated employment. However, the open-ended questions/responses gave good insight into what services are provided, which are successful, and what barriers the responding agencies have in providing specialized services. The Focus Group discussion added to the survey responses, and several themes were apparent. The responses also reinforced much of the current literature (of which there is little) around employment attainment for individuals with significant disabilities.

Committee: Tiffany Wild PhD (Advisor) Subjects: Education; Public Policy; Special Education

Master of Arts, Miami University, 2022, Economics

International efforts to mitigate climate change constitute a global coordination game between world actors regarding their contribution to and prevention of a warming global environment. While national climate policy decisions are likely focused on domestic political pressures, a second driving force exists in this international coordination: climate policy adopted by each individual nation impacts the inherent payoff of policy decisions available to other nations. We study this coordination specifically between the United States (US) and the European Union (EU). Implementing an event study approach, our results indicate that US policies which loosen climate protections are associated with decreased market pricing for EU emissions allowance futures, while the opposite is true for US policies which tighten climate protections. We interpret these results as suggestive evidence that markets perceive the US and the EU to engage in a common interest game with respect to climate policy in which their policy decisions are strategic complements.

Committee: David Lindequist (Advisor); Nam Vu (Committee Member); Jing Li (Committee Member) Subjects: Climate Change; Economics; Environmental Economics; Finance

Doctor of Philosophy, The Ohio State University, 2021, Biomedical Sciences

Ewing sarcoma is an aggressive bone and soft tissue-associated cancer affecting pediatric, adolescent, and young adult patients. Despite general improvement in pediatric cancer outcomes due to novel therapeutic options, Ewing sarcoma treatment, which consists of high-dose chemotherapy, radiation, and/or local surgical control, has remained largely unchanged for several decades and patients with metastatic disease continue to see poor outcomes. Although pediatric cancers often have far fewer mutational events than adult cancers, Ewing sarcoma is particularly interesting as the disease is often characterized by a sole chromosomal translocation event: These chromosomal translocations fuse one of the FET protein family members, a group of putative RNA-binding proteins, to a member of the ETS transcription factor family. As these FET/ETS fusion proteins have been determined to function as oncogenic transcription factors responsible for driving Ewing sarcomagenesis, it is critical that the biological mechanisms these fusions utilize to facilitate this process are elucidated. Despite discovery of several FET/ETS translocations, the majority of studies in the field focus on EWS/FLI, as it is the most common fusion observed in patients. Although these studies have provided a breadth of knowledge surrounding oncogenic function of the protein, there is a great deal of uncertainty how alternative FET/ETS fusions should be diagnosed and treated in the clinic. Herein, we characterize a novel FET/ETS fusion and perform the first comparative analysis on EWS/FLI and alternative, rarer FET/ETS fusion proteins. Our results reveal general similarities in DNA-binding and transcriptional regulation properties between the broad FET/ETS fusion group and provide the first tangible body of evidence to support that these fusions should indeed be classified as bona fide Ewing sarcoma tumors. Furthermore, we sought to characterize contributions of the FLI protein to overall EWS/FLI funct (open full item for complete abstract)

Committee: Stephen Lessnick MD/PhD (Advisor); Timothy Cripe MD/PhD (Committee Member); Lawrence Kirschner MD/PhD (Committee Member); Mark Parthun PhD (Committee Member) Subjects: Biomedical Research; Cellular Biology; Molecular Biology; Oncology

MS, University of Cincinnati, 2021, Medicine: Immunology

Background and Aims: Atopic diseases are associated with increased proliferation of T cells and an increase in levels of Th2 cytokines. Memory T cells are capable of producing cytokines much faster than naive T cells upon exposure to antigen. This ability of memory T cells correlates to the presence of positive histone marks at the cytokine gene loci. Additionally, data has shown that constitutively bound transcription factors, such as ETS-1, may play a role in keeping regulatory elements important for cytokine production open in resting memory cells. This suggests that histone modifications and/or TF constitutive binding play a major role in the poising of genes for transcriptional activity and hence, they can influence the expression levels of genes. We hypothesize that ETS-1 is one of several TFs integral to keeping chromatin sites open. We also hypothesize that by targeting the histone modifications at the Th2 cytokine locus, we can regulate the expression of the cytokines that play a role in atopic disease progression. Methods: CUT&Tag using an ETS-1 antibody was performed on Jurkat cells and two subpopulations of human CD4+ T cells, Naive and T Central Memory, which were either activated for 2.5 or 5 hours or left resting. The resulting fragments were then sequenced onto the human genome to determine if ETS-1 is able to constitutively bind important activation sites in memory cells when compared to naive cells. The histone modifiers G9a, LSD1, and SUV39h1 were added to a dCas9 backbone and assembled into a lentiviral vector. D10.G4.1 cells were then transduced by these vectors. Cells expressing these vectors were to be selected through single-cell cloning, followed by addition of gRNA to locate the fusion protein to sites of interest. Cell expression was to be analyzed using RT-qPCR. Results: The CUT&RUN approach has allowed us to identify ETS-1 binding sites in the Jurkat cell line. The ETS-1 binding was associated with the deposition of positive h (open full item for complete abstract)

Committee: Artem Barski Ph.D. (Committee Chair); Jonathan Katz Ph.D. (Committee Member); Marat Khodoun Ph.D. (Committee Member) Subjects: Immunology

PhD, University of Cincinnati, 2020, Medicine: Molecular and Developmental Biology

The formation of a functional vascular system is required for proper embryonic development in all vertebrates. Being one of the first organ systems to form during embryogenesis, ¬the cardiovascular system, comprising the heart, arteries, veins and capillaries, transports nutrients, oxygen, metabolites, waste, immune cells and hormones throughout the body. Thus, the formation of organ systems during development and subsequent maintenance throughout life is reliant upon a normal functional vascular system. This dissertation focuses on the molecular mechanisms that govern vascular development in zebrafish embryos. We utilize zebrafish embryos due to their rapid development, optical transparency and high amenability to genetic manipulation.

Committee: Saulius Sumanas Ph.D. (Committee Chair); Elisa Boscolo Ph.D. (Committee Member); Brian Gebelein Ph.D. (Committee Member); Joshua Waxman Ph.D. (Committee Member); Katherine Yutzey Ph.D. (Committee Member) Subjects: Cognitive Psychology; Developmental Biology

Doctor of Philosophy, The Ohio State University, 2015, Molecular, Cellular and Developmental Biology

In this thesis I examined the regulation of two Drosophila genes: RNase P RNA (RPR), which codes for the ribozyme component of an essential pre-tRNA processing enzyme, and vein (vn), which encodes a secreted ligand for the Epidermal growth factor receptor (Egfr). These two genes represent two different modes of gene regulation—while RPR is ubiquitously expressed, vn has a complex pattern of expression in specific tissues. Further, RPR is present in the intron of the Drosophila ATPsynC gene and transcriptionally co-regulated with the recipient gene. In contrast, vn is an independent gene with a complex promoter. The transcriptional regulation of Drosophila RPR is intriguing because it lacks signals for Pol III transcription and is in the intron of a Pol II transcribed gene. This is in contrast to other eukaryotic RPR genes studied thus far, which are all typical Pol III regulated genes. Using biochemical analyses, I have demonstrated that the annotated gene, the only copy of RPR in the genome, codes for the bona fide Drosophila RPR. My reporter gene study demonstrated that RPR is produced in a splicing independent fashion and its biogenesis is dependent on the Pol II promoter of the recipient gene. Pol II dependent transcription of RPR seems to be a hallmark of two major groups in Arthropods - Hexapods and Vericrustaceans. Current data supports a genetic event that caused the switch from an independent Pol III transcribed RPR to a Pol II dependent RPR that occurred approximately 500 million years ago. After the initial change the Pol II transcribed RPR moved again, as evident in the different in RPR recipient genes in different orders of Hexapoda. The orthologs of RPR recipient genes in D. melanogaster are expressed throughout development in all tissues, suggesting that ubiquitous expression may be one of the characteristics of RPR recipient genes. While the transcriptional coupling leads to the ubiquitous expression of RPR, vn is expressed in a dynamic pattern in t (open full item for complete abstract)

Committee: Amanda Simcox (Advisor) Subjects: Biochemistry; Bioinformatics; Biology; Genetics; Molecular Biology

Doctor of Philosophy, The Ohio State University, 2014, Molecular, Cellular and Developmental Biology

Despite recently declining rates of incidence and mortality, breast cancer remains the second leading cause of cancer death in American women. Much of the difficulty in treating this disease stems from the extreme diversity of cell types that compose the mammalian breast and the complexity of communication among them. Recently, macrophages and cells of the myeloid lineage have been discovered as key mediators of tumor progression, yet the precise mechanism through which this occurs remains relatively unsolved. The answer may lie in studying the ETS family of transcription factors, known to regulate many genes in cancer-relevant cell processes such as growth, proliferation, and motility. Ets2 in particular has been an attractive target as previous research suggests it has stroma-specific oncogenic potential. Through transgenic mouse tumor models, here we show that disabling Ets2 specifically within tumor-associated macrophages results in stunted tumor growth at both the primary and metastatic sites. Gene expression profiling and chromatin immunoprecipitation experiments reveal Ets2 directly binds and represses key anti-angiogenic genes, resulting in reduced tumor vasculature, thus limiting breast tumor growth and spread. Furthermore, PU.1, an ETS family member with wide influence over the cells of the hematopoietic lineage, was found to collaborate with Ets2 to regulate expression of key oncogenes Hif1a and IL-6. These results reveal that ETS factors work together as master regulators to influence angiogenesis and inflammation from within the macrophage compartment. These findings have implications for novel breast cancer therapies and prognostic tools. Macrophage-specific Ets2 deletion resulted in dysregulation of hundreds of genes. Comparing these genes with human breast cancer patient datasets resulted in a novel gene signature capable of retroactively predicting patient survival in multiple breast cancer subtypes including the highly aggressive ER-negat (open full item for complete abstract)

Committee: Michael Ostrowski PhD (Advisor); Gustavo Leone PhD (Committee Member); Thomas Ludwig PhD (Committee Member); Qianben Wang PhD (Committee Member) Subjects: Biology; Biomedical Research; Cellular Biology; Genetics; Health; Molecular Biology; Oncology

PhD, University of Cincinnati, 2008, Medicine : Epidemiology (Environmental Health)

Exposure to environmental tobacco smoke (ETS) is associated with onset of wheeze in children. There may be genetically at risk subgroups with an inability to metabolize components of ETS exposure components or have increased susceptibility to lung injury. Cotinine, the predominate metabolite of nicotine, is metabolized by the cytochrome p450 2A6 gene (CYP2A6). Polymorphisms in this gene have been shown to cause problems in nicotine metabolism resulting in decreased cotinine formation. Children with variant CYP2A6 alleles may have decreased cotinine values even though they have similar exposure as wild-type children. The GSTP1 gene produces the predominant detoxifying enzyme of reactive oxygen species (ROS) in the lung. A variant in this gene causes a decrease in ROS detoxification, which can lead to oxidative stress and lung injury. Children born to atopic parents were genotyped for four variants of the CYP2A6 gene and one GSTP1 variant. Hair samples were analyzed for nicotine and cotinine. Parents reported the number of cigarettes smoked by each resident in the home, number of hours around smoke per day, and ETS exposure in the car. Parents were queried yearly about their child's wheezing history. Recurrent wheeze was defined as two or more episodes in the past 12 months, and persistent wheeze was defined as recurrent wheeze at ages two and three. In contrast to other studies, African-American parents reported significantly higher ETS exposure, 50% versus 30% for Caucasians (p<0.01). There was a 15% decrease in mean cotinine level among African-Americans with the CYP2A6*9 variant (p=0.1). When cotinine level was dichotomized into high (upper 33%) and low, children with high cotinine and any CYP2A6 variant had increased prevalence of recurrent and persistent wheeze. Among subjects with high cotinine levels, the GSTP1 variant allele was associated with a significantly decreased prevalence of recurrent wheeze (p=0.03). These findings may indicate that African American (open full item for complete abstract)

Committee: Grace LeMasters PhD (Committee Chair); Ranjan Deka PhD (Committee Member); Gurjit K. Khurana Hershey MD, PhD (Committee Member); Linda Levin PhD (Committee Member) Subjects: Epidemiology

PhD, University of Cincinnati, 2007, Medicine : Molecular and Developmental Biology

Epithelial FGF signaling during lung organogenesis has been shown to be essential for branching morphogenesis, specification of surfactant producing type II cells and the maintenance of epithelial progenitors. In order to determine epithelial targets of FGF signaling, E11.5 mouse lungs were cultured for 24 hours in the presence of the FGF receptor antagonist SU5402, which inhibited branching morphogenesis. Affymetrix gene chip analysis identified a number of epithelial genes regulated by FGF signaling, including Elf5, a member of the Epithelial Specific Ets family of transcription factors. In situ hybridization revealed that Elf5 had a dynamic pattern of expression during lung development, transitioning from the distal epithelium at E11.5 to the proximal airway epithelium in late lung development. It was also determined that expression of Elf5 was induced by FGF7 and FGF10, ligands of epithelial specific FGFR2b. In order to further define the pathways by which FGFs activate Elf5 expression, E11.5 lung tips were also cultured in the presence of inhibitors of PI3-Kinase/Akt-mediated signaling (LY294002) and MAP Kinase/Erk-mediated signaling (U0126). It was found that LY294002 significantly reduced Elf5 expression, whereas U0126 had no effect. The observation that proximal airway epithelium (a tissue that lacks expression of distal epithelial markers) highly expressed Elf5 in late gestation suggested that Elf5 may play a role in negatively regulating distal epithelial specification/differentiation. To test this hypothesis, a transgenic mouse model was generated containing a doxycycline inducible HA-tagged Elf5 transgene under the control of the lung epithelium specific Sftpc promoter to overexpress Elf5 in the distal lung epithelium. The results indicated that high Elf5 expression disrupted branching morphogenesis and negatively regulated distal epithelial differentiation of the lung epithelium while repressing type II cell enriched genes Erm, Napsin and Sftpc. Further (open full item for complete abstract)

Committee: Dr. John Shannon (Advisor) Subjects:

MS, University of Cincinnati, 2004, Medicine : Epidemiology (Environmental Health)

This study examined infants' risk for developing upper respiratory symptoms (URS) in relation to their exposure to environmental tobacco smoke (ETS) and mold. Eligible infants (n=633) were identified by birth records and had one parent that was atopic by positive skin prick test (SPT). Exposure information was collected at the time of parent SPT, and the average age of the infants was seven months. Parents were asked to complete monthly diaries related to their infants' URS. Our analysis included two models, one using parental report of mold/mildew and a sub-analysis (n=342) using in-home assessor report of none, low or high mold exposure. When controlling for race, gender and socioeconomic status, multivariate logistic regression showed an increased risk of sinus infection with parental report of mold/mildew (OR 1.91; 95%CI 1.14-3.23) and exposure to >20 cigarettes/day (2.38; 1.26-4.49); in the in-home assessor report, high mold was even more significant (6.7; 2.27-19.79). Ear infection was also associated with assessor report of mold, in both the low (1.77; 1.17-2.67) and high (3.07; 1.32-7.12) categories. Rhinitis was significantly associated with parental report of mold/mildew (1.32; 1.31-1.47) and exposure to >20 cigarettes/day (1.74; 1.14-2.65). Although significant associations were not observed with allergic rhinitis, elevated odds ratios were observed with both parent and assessor report of mold. This analysis suggests that mold is a risk factor of sinus infection, ear infection and rhinitis, while ETS is a risk factor of sinus infections and rhinitis. This analysis suggests that mold exposure and not ETS is associated with ear infections in infants. The sub-analysis findings also may suggest that mold exposure is a stronger risk factor than ETS for sinus infection, while ETS may be more closely associated with rhinitis.

Committee: Dr. Grace LeMasters (Advisor) Subjects: Environmental Sciences

Doctor of Philosophy, The Ohio State University, 2004, Molecular Genetics

Ets family transcription factors are regulated by signal transduction pathways. Phosphorylation of the Ets-1 and Ets-2 at a single conserved threonine residue (T38 and T72, respectively) by ras-MAPK pathways leads to their activation and persistent target genes expression. The phosphorylation of Ets-2 affected its activity, as well as its protein partnership. Ets-2 interacted with Brg-1 or BS69 co-repressors in a phosphorylation dependent manner, and repressed target gene expression. Ets-2 knockout mice are embryonic lethal due to an extra-embryonic defect, whereas ets-2T72A/T72A mice are viable and fertile, with no obvious abnormality. Ets-2 is constitutively phosphorylated in macrophages derived from motheaten-viable (mev) mice, while its phosphorylation is tightly regulated in wt cells. The aberrant Ets-2 phosphorylation correlated with increased target gene expression and cell survival. To directly test the role of Ets-2 phosphorylation in inflammation, the ets-2T72A allele was introduced into mev mice. In contrast to ets2+/+, mev/mev mice, ets-2T72A/T72A, mev/mev mice were fertile, had increased life span and body weight, elevated macrophage apoptosis in the absence of CSF-1, but reduced inflammation and expression of inflammatory genes, including cytokines (TNFa), chemokines (MIP1a, b), extracellular matrix proteases (MMP9), cell adhesion molecule (integrin aM) in lungs and macrophages. Both ets-1-/- mice and Ets-2 T72A/T72A mice are viable and fertile. To reveal Ets-2 function possibly masked by gene redundancy, we mated ets-2T72A/T72A mice with Ets-1 knockout mice. Ets-1-/-, Ets-2T72A/T72A mice died between embryonic day 11.5 to 14.5, with dramatic angiogenesis and cardiovascular defects. Compared to control embryos, the double mutant embryos expressed lower levels of Ets target genes, such as Ang1, Tie2, MMP3, MMP9, and Fli-1, but elevated levels of VEGF. Therefore, Ets-2 phosphorylation is important in immune response, angiogenesis and cancer. To further e (open full item for complete abstract)

Committee: Michael Ostrowski (Advisor) Subjects: Biology, Genetics

Master of Science (MS), Ohio University, 1991, Chemical Engineering (Engineering)

A mathematical simulation of ETS' limestone emission control (LEC) process using a moving bed configuration

Committee: Michael Prudich (Advisor) Subjects: Engineering, Chemical

Master of Science (MS), Ohio University, 1989, Chemical Engineering (Engineering)

A mathematical simulation of ETS' limestone emission control process using the method of characteristics: Fixed-bed configuration/gas-phase mass transport control

Committee: M. Prudich (Advisor) Subjects: Engineering, Chemical

Master of Sciences, Case Western Reserve University, 2008, Physics

Online Distance Learning has made it possible to use a large set of training resources customized to the users and the environment. The study identifies an effective approach for the development and marketing of a GRE physics online guided study program based on the available literature and the experience and materials developed by Professor Robert Brown at Case Western Reserve University. The findings conclude that the approach is cost‐effective for the student, will serve approximately 30% of the 12,500 GRE physics candidates every year and will generate an annual income of $80,000 if continued as a university publishing project or more if transformed into an online training platform with multiple revenue streams. The program also serves as a proof of concept towards structuring future online instructional methodologies to position Case Western Reserve University and the Department of Physics at the forefront of learning technologies.

Committee: Dr. Robert Brown (Advisor); Cyrus Taylor (Committee Member); Edward Caner (Committee Member) Subjects: Business Community; Business Costs; Education; Educational Software; Marketing; Mass Media; Physics; Science Education; Teaching