• The Inhibition of NADPH Oxidase 1/Protein Disulfide-Isomerase Increases BIM in Pancreatic Cancer

      Knox, Henry John Douglas; Biomedical Sciences (Augusta University, 2022-05)
      Pancreatic cancer requires elevated protein synthesis. To maintain endoplasmic reticulum (ER) homeostasis, chaperones are overexpressed. Persistent ER stress activates the unfolded protein response (UPR), which can become overwhelmed and lead to cell death. Reactive oxygen species (ROS) from NADPH oxidases (Noxs) can regulate ER homeostasis. In particular, the chaperone protein disulfide-isomerase (PDI) colocalizes with Nox1 in the ER to regulate its activity. Because Nox1 is expressed in the stroma of pancreatic cancer, I studied the extent to which Nox1-derived ROS help establish an adaptive ER homeostasis via PDI in pancreatic cancer cells. I generated a pancreatic cancer mouse model lacking the Nox1 gene in the whole body (Nox1-null KPC mice). Nox1-competent KPC mice were used as a control. The induction of Cre recombination was done by administration of tamoxifen for five consecutive days. After 2 months of tamoxifen induction, the pancreas was harvested. Western blotting was carried out to evaluate PDI, UPR activation, ER stress and cell death. The Nox1-null KPC mice were viable. The lack of Nox1 reduced PDI, C/EBP homologous protein (CHOP) in KPC mice. The UPR sensor IRE1α and the growth arrest and DNA damage-inducible 34 (GADD34) were absent in Nox1-competent KPC mice, but they were recovered in Nox1- null KPC mice. In Nox1-null KPC mice, the apoptotic protein BIM was increased. I concluded that in pancreatic cancer, the lack of Nox1 decreased PDI, which led to an accumulation of unfolded proteins and activation of UPR system. GADD34 recovery increased protein synthesis, worsening the scenario. Persistent ER stress overwhelmed the UPR system, which caused BIM-induced cell death.
    • Identification and Characterization of Force Sensitive Domains using an In Vivo Dorosphila Synthetic Biology Platform

      Harman, Jacob Henry; Biomedical Sciences
      The Notch protein is a family of highly conserved transmembrane signaling proteins that are found in all metazoan life. This protein family plays many roles in developmental and regulatory pathways in these organisms such as the development of the human vertebral column and the Drosophila adult wings. The Drosophila Notch protein has been the focus of recent research as this protein has shown to be dependent on a unique signaling mechanism that relies on the force being applied to the receptor by the endocytosis of a bound ligand for a cleavage event to occur. This protein has been shown to only signal in the presence of this endocytosis force which allows for cleavage at the S2 and S3 sites on the protein which allows for a transcription factor to be released from the membrane and enter the nucleus. In synthetic biology, this protein has found a role in creating customizable cell-cell signaling platforms known as Synthetic Notch or Syn-Notch. As demonstrated in previous research, Notch proteins contain highly modifiable ligand binding and transcription factor domains that can be interchanged with other ligand binding and transcription factor domains from non-Notch proteins without affecting the protein signaling. In addition to the ligand-binding and transcription factor, the Notch protein contains a Negative Regulatory Region (NRR) which functions as a force sensor domain and allows for the force signaling activation seen in the native Notch proteins. This region of the Notch protein is often conserved in Syn-Notch systems as a means of controlling the signaling of Syn-Notch proteins used in precision medicine. The unique force-sensing function of this domain has raised questions regarding the characteristics of this domain that allow for it to act in a force-sensing manner and if this domain is as interchangeable as its other components. In this study, domains from other non-Drosophila Notch proteins and non-Notch protein domains were evaluated in an in vivo Drosophila platform for the ability to function as a force sensor domain. In this screen, several unique force sensor domains were identified as having the ability to recapitulate the force sensing characteristic of the canonical Notch NRR. While a common thread between these domains in terms of defining characteristics remains elusive, this study was able to demonstrate the feasibility of this screening method to identify force-sensitive domains and identify 4 domains that exhibit this characteristic. The identification of these domains which are force sensitive not only allows for the characteristics to function as force-sensitive domains but also provides alternative force sensors with differing force sensitivities when compared to the canonical Notch NRR for use in Syn-Notch systems. The development of these receptors will not only contribute to the field of synthetic biology as means to create synthetic platforms for use in future research, but these receptors will allow for the development of even more precise treatments using these synthetic receptor systems in clinical therapies.
    • Prevalence of Chronic Medical Conditions Among the 1990-1991 Gulf War Female Veterans

      Ansa, Benjamin E; Public Health (Augusta University, 2022-05)
      Background: The care of the large number of women joining the U.S. military is receiving increased attention. Currently, 56% of all women veterans alive served during the 1990–1991 Gulf War (GW). Veterans of the GW have reported poorer health outcomes than their counterparts who did not deploy because of the potential health risks associated with military service during the GW, and the possible use of chemical and biologic warfare agents. Military service and deployment affect women differently than men, however gaps exist in the research of common chronic conditions among veterans due to the lack of gender-specific results from published studies. Purpose: The specific aims of this study were 1) to examine and compare the prevalence of chronic medical conditions (CMCs) among deployed GW female veterans and GW era female veterans that did not deploy; 2) to examine the prevalence of exposure to environmental toxicants among deployed GW female veterans and compare the prevalence of CMCs between exposed and unexposed females; and 3) to examine and compare the prevalence of CMCs among deployed male and deployed female veterans of the GW. Methods: Data from three longitudinal epidemiologic studies of the health of the 1990-1991 U.S. GW veterans were analyzed for this study. Cross tabs and logistic regression were done to calculate the prevalence and prevalence ratios (PRs) of seventeen self-reported CMCs among the study population. PRs were also used for assessing the relationship between self-reported exposure to environmental toxicants and the prevalence of CMCs among deployed female veterans. Results: The most prevalent CMCs reported by both deployed and non-deployed female veterans were headaches and high blood cholesterol. Compared to non-deployed females, significantly higher proportions of deployed females reported migraine headaches, osteoarthritis, and skin problems. In addition, the prevalence of headaches was almost three times higher for deployed female veterans, compared to deployed male veterans. Anthrax vaccine, oil well fires, pyridostigmine bromide, petroleum combustion products and chemical weapons were the toxicants with the highest proportions of reported exposure among deployed female veterans. Compared to unexposed females, significantly higher proportions of exposed females reported migraine headaches, headaches (other than migraines), high blood cholesterol, hypertension, acid reflux/GERD, irritable bowel syndrome, colon polyps, thyroid disease, osteoarthritis, tinnitus, asthma, chronic lung disease, and skin problems. Conclusions: Compared to their counterparts, higher prevalence of some CMCs were observed among deployed female veterans of the 1990–1991 GW, especially among those that reported toxicant exposures. This information about CMCs and exposures is useful for guiding medical assessments and policy development affecting this group of female veterans.
    • The Perioperative Synergy Index

      Elliott-Dawe, Cheryl Ann; Nursing (Augusta University, 2022-05)
      The presence of synergy within perioperative services has a significant bearing on a hospital’s viability and positive patient outcomes. Synergy is a positive emergence that arises as a value-added return on investment when all contributing factors and personnel work toward common goals. The purposes of this research were to develop a perioperative synergy model and to create an index to evaluate perioperative synergy. Research questions were: (a) What are the antecedents of synergy in the context of perioperative services? (b) What research is currently being conducted on high functioning perioperative services across perioperative disciplines? (c) What are the items necessary to ensure face and content validity of a perioperative synergy index? (d) How should the items be worded to ensure clarity, readability, and usability of a perioperative synergy index? (e) What are the underlying constructs that constitute perioperative services synergy? and (f) What are the psychometric properties of the perioperative synergy index? A three-phase survey methodology was adopted. For Phase 1, a directed content analysis of 20 years of research literature on high performing ORs was conducted using nine domains of a theoretical perioperative model. The analysis yielded 221 items that were presented to a panel of perioperative experts who rated each item’s relevance to perioperative success on a 4-point scale. Expert agreement on content was evaluated using a content validity index. In Phase 2, experts judged the wording and clarity of each item and of the instructions. Inter-rater agreement on clarity of items and instructions was evaluated using Gwet’s AC1 coefficient. For Phase 3, perioperative managers across the nation were surveyed and asked to rate levels of agreement with the items on a 6-point Likert scale as each item pertained to their hospital. Confirmatory factor analyses were completed on the responses, resulting in a model consisting of nine constructs and 53 indicators. Construct, convergent, and discriminant validity were assessed. Reliability was evaluated using composite reliability, inter-item correlations, and average inter-item correlations. Factor scores were employed to calculate a perioperative synergy index coefficient that ranged from 0 to 1, with higher values indicative of higher functioning perioperative services at the hospitals involved in the study. Managers can use the index to identify areas in need of improvement and can use the model to guide process improvements. Following further refinement, the index may serve as a new way to evaluate performance and standardize protocols across healthcare systems.
    • Twist1 Evokes Matrix Metalloproteinase 9 and Collagen IV Synthesis in Activated Pancreatic Stellate Cells

      Geister, Emma; Biomedical Sciences (Augusta University, 2022-05)
      Background: Pancreatic cells are embedded in an extracellular matrix (ECM) and are also surrounded by a thin sheet of specialized extracellular matrix known as basal lamina. In healthy pancreas, pancreatic stellate cells (PaSCs) are responsible for the synthesis of basal lamina, which is mainly composed of collagen IV and laminin. In chronic pancreatitis (CP), PaSCs are responsible for the synthesis of fibrotic tissue, which is mainly composed of fibronectin and collagen I. Reactive oxygen species (ROS), which are predominant inflammatory mediators in CP, evoke the formation of fibrotic tissue by PaSCs. One of the sources of ROS is NADPH oxidase (Nox) enzymes. In particular, Nox1 has been associated with both fibrogenesis in a CP mouse model and an up-regulation of the transcription factor Twist1 and matrix metalloproteinase (MMP) 9 in CP-activated PaSCs. I sought to determine the functional relationship between Twist1 and MMP-9, as well as other PaSC-produced proteins. Therefore, my aim was to assess the extent to which Twist1 up-regulates MMP-9 and other PaSC-produced proteins. Methods: To overexpress Twist1, I infected activated PaSCs from Nox1-null mice with retroviruses expressing either mouse Twist1 or the empty backbone. I compared the relative expression of MMP-9 and other PaSC-produced proteins using quantitative PCR. To examine whether MMP-9 expression is regulated by Twist1 at the transcriptional level, I carried out a dual-luciferase reporter assay using a pGL2 Luciferase Reporter Vector carrying the human MMP-9 (pGL2-hMMP-9) promoter. I co-transfected HEK293 cells with human Twist1 in pCMV6 vector, or the empty backbone (negative control) along with human pGL2-hMMP-9 promoter vector or pGL3-control vector using Lipofectamine 3000. As a positive control, I co-transfected HEK293 cells with human NF-ĸB1 in pFUW-tetO vector along with the pGL2-hMMP-9 promoter. After 48 h, I performed the dual-luciferase reporter gene assay. Results: I found that the up-regulation of Twist1 in culture-activated PaSCs from Nox1-null mice increased MMP-9 mRNA level, but it did not modify the expression of PaSC-produced proteins linked to fibrosis [e.g., collagen I, fibronectin, α-smooth muscle actin, transforming growth factor-β (TGF- β), and interleukin-6 (IL-6)]. Therefore, I studied the expression of collagen IV, a component of basal lamina and found that the expression of Twist1 in activated PaSCs from Nox1-null mice increased collagen IV at the mRNA level. I also found that Twist1 increased the expression of MMP-9 at the transcriptional level in a NF-ĸB dependent manner using a dual-luciferase assay. Conclusion: Twist1 in PaSCs induced the expression of MMP-9 and collagen IV, at the transcriptional level. NF-ĸB was required for the transcriptional up-regulation of MMP-9 by Twist1. The expression of other PaSC-produced proteins, including collagen I, fibronectin, IL-6, α-smooth muscle actin, and TGF-β, were not affected. Since Twist1 in activated PaSCs was responsible for the synthesis and remodeling of the basal lamina in healthy pancreas, I concluded that the overexpression of Twist1 using a retrovirus approach was not sufficient to change the phenotype of PaSC from a basal lamina “maker” to a fibrotic tissue “maker.”
    • T cells and NLRP3 Mediate High Fat Diet-Induced Increases in Blood Pressure and Adiposity in Female and Male Dahl Rats

      Ramirez, Lindsey; Biomedical Sciences (Augusta University, 2022-05)
      Cardiovascular disease (CVD) is the leading cause of death worldwide in both sexes, with hypertension being an important modifiable factor. To increase the number of people with adequate blood pressure control, a better understanding of the mechanisms controlling the development of hypertension is necessary. Chronic consumption of a high fat diet (HFD) increases blood pressure and adiposity. Furthermore, clinical data suggest that women may be predisposed to worse cardiovascular health in instances of excess adiposity. Thus, these experiments were designed to test the central hypothesis that a HFD will increase blood pressure and adiposity more in females vs males. Sprague Dawley rats were resistant to HFD-induced increases in blood pressure or fat mass. Conversely, 10 weeks of HFD treatment increased blood pressure, fat mass, and adipose tissue weight in female and male Dahl Wild Type (WT) rats. This was accompanied by a pro-hypertensive T cell profile in both sexes. This finding pointed to a role for T cells in mediating the morbidity observed, T cell-deficient rats were utilized. T cell knock out (KO) blunted the HFD-induced hypertension in both sexes, but only the HFD-induced increase in adiposity in males. Due to the observed protective role of T cell KO, it was crucial to determine what could be activating T cells. NLRP3 is a pattern recognition receptor which activates T cells and adipogenesis, NLRP3 deficient rats were randomized to a Control (Ctrl) or HFD treatment. NLRP3 KO blunted the HFD-induced adiposity in both sexes. It is well known that free fatty acids (FFAs) can activate NLRP3, so a FFA panel was performed in Dahl WT animals. Females had a more pro-hypertensive FFA profile. The Dahl WT rat is a good model to use to study the loss of protection of females that are on a HFD. Additionally, these data demonstrate that T cells and NLRP3 are playing sex-specific roles to promote the HFD-induced increases in adiposity and blood pressure. Finding sex-specific mechanisms that mediate these morbidities will reduce the number of people that develop hypertension and subsequently, CVD, the number one killer of both men and women in the US.
    • High Dimensional Bayesian Multinomial Logistic Regression

      Dow, James Francisco; Biostatistics (Augusta University, 2022-05)
      Classification is an important area in statistics and machine learning where one is interested in determining class membership based on a set of covariates or predictors, which are possibly high-dimensional. A common application is in genomic studies, where finding genes (predictors) associated with a categorical phenotype such as disease status or tissue type can be useful for prevention or determining prognosis. Regression-based modeling allows for linking these predictors, or combinations of them, probabilistically to the different categories of the phenotype of interest. Multinomial logistic regression is a natural statistical framework that provides a way to model the effects of predictors on the probability of a multi-category outcome variable with two or more classes. We propose a Bayesian hierarchical model that uses multinomial logisitic regression with point mass mixture priors to select best subsets of predictors for classification. We develop a Markov chain Monte-Carlo algorithm based on Gibbs sampling to compute the corresponding posterior distribution for the model. We do so by exploiting the P\'{o}lya-Gamma mixture of normal representation that has been derived in the logistic regression setting. Further, we prove that the proposed hierarchical model is model selection consistent in the strong sense. Finally, we compare the proposed methodology, in terms of model selection and classification, to several popular statistical and machine learning procedures in simulation and on a real data set.
    • Activation of a molecular chaperone (sigma 1 receptor) in a murine model of autosomal dominant retinitis pigmentosa.

      Barwick, Shannon; Biomedical Sciences (Augusta University, 2022-05)
      Retinitis pigmentosa (RP) is a devastating group of inherited retinal diseases that leads to visual impairment and eventually complete blindness. Currently no cure or treatment exists for RP patients, thus research into prolonging the vision in these patients is imperative. Sigma 1 receptor (Sig1R) is a promising small molecule target that appears to have neuroprotective benefits in the retina of fast degenerating mouse models. However, it is not clear whether Sig1R activation can provide similar neuroprotective benefits in more slowly progressing RP models, which are more similar to human patients. In this study, we examined whether Sig1R activation can be neuroprotective (i.e. prolong vision) in a more slowly progressing mouse model of autosomal dominant retinitis pigmentosa, RhoP23H/+. Current studies in the field give a brief overview of the RhoP23H/+ degeneration, but do not give a complete characterization of disease progression. Aim 1 of this study sought to further characterize the degeneration of the RhoP23H/+ mouse using 3 in vivo methods, Optomotor Response (OMR), Electrophysiology (ERG), and Spectral Domain-Optical Coherence Tomography (SD-OCT). A slow retinal degeneration was observed in both male and female RhoP23H/+ mice when compared to WT. Visual acuity showed a gradual decline through 10 months. Interestingly, visual acuity was still detectible, albeit significantly reduced through 10 months in both male and female mutant mice. Females appeared to have significantly lower visual acuity than males. These RhoP23H/+ mice showed a gradual decline in scotopic and photopic responses. Aims 2 and 3 sought to investigate the neuroprotective benefits of Sig1R activation in the RhoP23H/+ mouse model. Mutant mice were treated with a high specificity Sig1R ligand (+)-pentazocine ((+)-PTZ) 3x/week and examined using OMR, ERG, SD-OCT. A significant retention of visual function was observed in both males and females at 10 months of age, with treated females retaining ~50% greater visual acuity than non-treated mutant females. Using ERG, significant retention of scotopic and photopic b-wave amplitudes were observed at 6 months in both male and female mice treated with (+)-PTZ. Further, in vivo analysis of ONL thickness revealed a significant retention in both male and female treated mice. Histological studies using retinal cryosections showed significant retention of IS/OS length (~50%), ONL thickness, and number of rows of PRC nuclei at 6 months in both male and female (+)-PTZ-treated mice. Interestingly, electron microscopy revealed preservation of OS discs in (+)-PTZ treated mutant mice. Taken collectively, the in vivo and in vitro data represent the first report of Sig1R activation rescuing visual function and structure in the RhoP23H/+ mouse model. These results are promising and lay the framework for future studies to investigate Sig1R as a potential therapeutic target in retinal degenerative disease.
    • CU TRANSPORTER ATP7A AND EXTRACELLULAR SOD FUNCTION IN ADAPTIVE ANGIOGENESIS AND REGENERATION OF SKELETAL MUSCLES IN RESPONSE TO EXERCISE OR INJURY: ROLE OF EXOSOMES

      Abdelsaid, Kareem; Biomedical Sciences (Augusta University, 2022-05)
      The overall goal of this dissertation is to elucidate novel mechanisms by which communication of skeletal muscle (SKM) and endothelial cells (ECs) through “exosomes” regulates adaptive angiogenesis and regeneration of SKM in response to physical exercise in type 2 diabetes mellitus (T2DM) (project 1) or injury (project 2) by focusing on the Copper (Cu) transporter ATP7A-extracellular SOD (ecSOD, SOD3) pathway. In project 1, we investigated the role of exercise-induced angiogenic effects on ECs in T2DM. We isolated plasma exosomes from control, T2DM and SOD3-/- mice with two weeks of voluntary wheel exercise using differential ultracentrifugation. Isolated exosomes were characterized by ZetaView, TEM and exosome markers (CD63 and Tsg101). Both SOD3 and ATP7A proteins were significantly induced in plasma exosomes by exercise in both mice and humans. Plasma exosomes from T2DM and SOD3-/-/T2DM sedentary mice impaired ECs angiogenic function compared to control mice, which were restored by exercise in only T2DM but not SOD3-/-/T2DM mice. Furthermore, exosomes overexpressing SOD3 significantly enhanced angiogenesis in ECs by increasing local H2O2 levels in a heparin binding domain-dependent manner and restored wound healing and angiogenesis in T2DM or SOD3-/- mice. In project 2, we investigated the role of Cu transporter ATP7A in myogenic differentiation and skeletal muscle regeneration upon injury. C2C12 myoblasts differentiation to myotubes was associated with an increase in ATP7A and SOD3 expression in cell lysates and exosomes isolated from conditioned media. ATP7A depletion with shRNA in myoblasts significantly inhibited myogenic differentiation and angiogenic effects of conditioned media-derived exosomes. Mechanistically, silencing SOD3, chelating Cu by TTM or inhibiting lysyl oxidase (LOX) activity by BAPN significantly inhibited myogenic differentiation, suggesting that ATP7A-SOD3/LOX axis is required for Cu-dependent myogenesis. Cardiotoxin-induced SKM injury model reveals that ATP7A expression was increased in activated, but not quiescent Pax7-positive satellite cells in injured SKM. Functionally, both Cu transporter defective ATP7A mutant and SOD3-/- mice showed impaired SKM regeneration, diminished myofiber sizes, decreased number of centralized myonuclei and satellite cells. In conclusion, our study reveals a novel role of Cu transporter ATP7A and exosomal SOD3 in promoting adaptive angiogenesis and SKM regeneration serving as a novel promising therapeutic tool for resolving impaired angiogenesis.
    • LEADERSHIP PROCESSES DURING THE COVID-19 PANDEMIC: IMPLICATIONS FOR LEADERSHIP PREPARATION AND TRAINING

      Glover, Michell; Advanced Studies Innovation (Augusta University, 2022-05)
      The roles of school leaders have transformed significantly over time, making necessary shifts to place student learning at the core of what principals do. School leadership preparation has also evolved to keep up with the changing responsibilities and challenges that principals encounter. Although effective school leaders are recognized for their character and exemplary practices that contribute to and build collaborative school communities, principals have multiple responsibilities and often experience challenges while leading their schools. The role of the school leader has changed over the course of history. In response to these role shifts, the standards and practices providing the structure for post-secondary preparation and training programs, state certification, and formal evaluation programs have also had to adjust to keep up with societal changes and responsibilities of school leaders. While principals perform their regular responsibilities, with accompanying challenges, the COVID-19 pandemic has created a myriad of new challenges for school leaders around the globe. Although challenges are not new to the principalship, this study seeks to investigate the challenges K-12 school leaders experienced during the COVID-19 pandemic, how their leadership preparation and training did or did not prepare them to manage these challenges, and investigate the solutions principals implemented to counter the challenges experienced. Keywords: , , , professional development
    • LEADERSHIP PROCESSES DURING THE COVID-19 PANDEMIC: IMPLICATIONS FOR LEADERSHIP PREPARATION AND TRAINING

      Workman, Joseph Barnett; Advanced Studies Innovation
      The roles of school leaders have transformed significantly over time, making necessary shifts to place student learning at the core of what principals do. School leadership preparation has also evolved to keep up with the changing responsibilities and challenges that principals encounter. Although effective school leaders are recognized for their character and exemplary practices that contribute to and build collaborative school communities, principals have multiple responsibilities and often experience challenges while leading their schools. The role of the school leader has changed over the course of history. In response to these role shifts, the standards and practices providing the structure for post-secondary preparation and training programs, state certification, and formal evaluation programs have also had to adjust to keep up with societal changes and responsibilities of school leaders. While principals perform their regular responsibilities, with accompanying challenges, the COVID-19 pandemic has created a myriad of new challenges for school leaders around the globe. Although challenges are not new to the principalship, this study seeks to investigate the challenges K-12 school leaders experienced during the COVID-19 pandemic, how their leadership preparation and training did or did not prepare them to manage these challenges, and investigate the solutions principals implemented to counter the challenges experienced.
    • Leadership Processes During the COVID-19 Pandemic: Implications for Leadership Preparation and Training

      Bogans, Adrianne Melva; Educational Leadership
      The roles of school leaders have transformed significantly over time, making necessary shifts to place student learning at the core of what principals do. School leadership preparation has also evolved to keep up with the changing responsibilities and challenges that principals encounter. Although effective school leaders are recognized for their character and exemplary practices that contribute to and build collaborative school communities, principals have multiple responsibilities and often experience challenges while leading their schools. The role of the school leader has changed over the course of history. In response to these role shifts, the standards and practices providing the structure for post-secondary preparation and training programs, state certification, and formal evaluation programs have also had to adjust to keep up with societal changes and responsibilities of school leaders. While principals perform their regular responsibilities, with accompanying challenges, the COVID-19 pandemic has created a myriad of new challenges for school leaders around the globe. Although challenges are not new to the principalship, this study seeks to investigate the challenges K-12 school leaders experienced during the COVID-19 pandemic, how their leadership preparation and training did or did not prepare them to manage these challenges, and investigate the solutions principals implemented to counter the challenges experienced.
    • Role of ADAM17 and JAM-A/F11R in aging-related arterial wall shear stress mechanosensing and abnormal vascular remodeling

      Tian, Yanna; Biomedical Sciences
      Physiological and pathological vascular remodeling is uniquely driven by mechanical forces from blood flow in which wall shear stress (WSS) mechanosensing by the vascular endothelium plays a pivotal role. This study aimed to determine the novel role for a disintegrin and metalloproteinase 17 (ADAM17) in impaired WSS mechanosensing, which was hypothesized to contribute to aging-associated abnormal vascular remodeling. Without changes in arterial blood pressure and blood flow rate, skeletal muscle resistance arteries of aged mice (30-month old vs. 12-week old) exhibited impaired WSS mechanosensing and displayed inward hypertrophic arterial remodeling. These vascular changes were recapitulated by in vivo confined, AAV9-mediated overexpression of ADAM17 in the resistance arteries of young mice. An aging-related increase in ADAM17 expression reduced the endothelial junction level of its cleavage substrate, junctional adhesion molecule-A/F11 receptor (JAM-A/F11R). In cultured endothelial cells subjected to steady WSS, ADAM17 activation or JAM-A/F11R knockdown inhibited WSS mechanosensing. The ADAM17-activation induced impaired WSS mechanosensing was normalized by overexpression of ADAM17 cleavage resistant, mutated JAM-A/F11RV232Y both in cultured endothelial cells and in resistance arteries of aged mice, in vivo. These data demonstrate a novel role for ADAM17 in JAM-A/F11R cleavage-mediated impaired endothelial WSS mechanosensing and subsequent development of abnormal arterial remodeling in aging. ADAM17 could prove to be a key regulator of WSS mechanosensing, whereby it can also play a role in pathological vascular remodeling in diseases.
    • DEVELOPMENT OF POTENTIAL DRUG CANDIDATES FOR SARS-CoV-2 USING MOLECULAR HYBRIDIZATION APPROACH

      Wyman, Kailey; Biomedical Sciences
      ABSTRACT In December 2019, an unknown viral infection originated from a local fish and wild animal market in Wuhan city, China. Since then, the virus has rapidly spread across mainland China followed by the rest of the world. On February 11, 2020, the World Health Organization (WHO), identified the virus as the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). In March 2020, WHO declared the coronavirus outbreak a global pandemic. As of March 2022, SARS-CoV-2 has affected about 500 million people with over 6 million deaths worldwide. Although the world has survived numerous pandemics in the past, this one is an unprecedented global health challenge that has greatly impacted our lives and the socioeconomic of the world. At present therapeutic options for SARS-CoV-2 are very limited. Researchers adopting various approaches to develop new drug candidates for SARS-CoV-2 and the drug repurposing approach is one of them. We adopted the molecular hybridization approach to overcome the limitations of repurposing drugs. We designed and synthesized a new set of small molecules using a quinoline scaffold (from hydroxychloroquine and chloroquine), indole nucleus (from umifenovir), and rhodamine (a very well-investigated medicinally important moiety) via an optimized multistep synthetic route. All the synthesized molecules were characterized by analytical spectral studies. All the synthesized conjugates were screened against the SAR-CoV-2 virus and their cytotoxicity was determined. Computational studies were carried out to validate the biological data. Some synthesized compounds showed promising anti-viral properties against SAR-CoV-2.
    • CHARACTERIZATION OF AN INDUCIBLE p65 KNOCKOUT MOUSE MODEL FOR STUDYING GLIOBLASTOMA

      Trang, Amy; Biomedical Sciences (Augusta University, 2022-05)
      Glioblastoma (GBM) is a malignant primary brain tumor that results in patient death within two years following diagnosis. The GBM tumor microenvironment has an important impact on the formation, progression, and drug resistance of this lethal disease. The GBM tumor microenvironment is composed of a variety of cell types that can support tumor growth like microglia. Microglia are immune cells that can switch from a classical, tumoricidal M1 phenotype to an alternative, tumor-promoting M2 phenotype. However, there are limited studies about the specific effect(s) that microglia have on GBM tumors. Given the importance of the tumor microenvironment to GBM tumor progression, targeting key signaling pathways in tumor-associated cells like microglia could be an effective way to reduce tumor progression. One signaling pathway involved in causing different cancers is the nuclear factor-kappa B (NF-κB) pathway, and it has been implicated in M1 to M2 phenotype polarization. Therefore, this study focuses on the characterization of the p65fl/fl/CX3CR1creER/+ mouse model, which is an inducible p65 knockout mouse model for studying how inhibition of canonical NF-κB signaling in microglia affects GBM tumors. After tamoxifen is administered to the mouse, p65, a transcription factor of the canonical NF-κB pathway, should be deleted in microglia due to the activation of a Cre recombinase (CreER) under the control of the CX3CR1 promoter. Characterization of this mouse model is necessary to determine if the p65 gene is effectively deleted in microglia from these mice. Liquid chromatography-mass spectrometry analysis has shown that the orally administered tamoxifen was present and quantifiable in the brains of tamoxifen-treated mice to induce recombination. Based on data from PCR and western blots, tamoxifen given to the p65 knockout mice induced partial deletion of the p65 gene in isolated microglia when compared to control groups. Preliminary flow cytometry data suggested that p65 deletion in microglia occurred in p65fl/fl/CX3CR1creER/+ mice that received GBM implantations and tamoxifen treatment compared to a vehicle control group. Overall, the data from this study suggests that the p65 gene is partially deleted in microglia from the tamoxifen-treated p65 knockout mice compared to the vehicle control mice.
    • THE ROLE OF c-JUN IN ENHANCING TCR-T FUNCTION IN TREATMENT OF HEPATOCELLULAR CARCINOMA

      Hussein, Mohamed; Biomedical Sciences (Augusta University, 2022-05)
      Liver cancer is the sixth most common cancer and the third leading cause of cancer death in the world, indicating the treatment gap and the urgent need for novel effective therapies. The majority (90%) of liver cancers are hepatocellular carcinoma (HCC), with an overall 5-year survival rate of ~20%. The efficacy of adoptive cell therapy (ACT) targeting malignant tumors has been limited due to lack of T cell activation in vivo with poor expansion and short persistence of tumor-infiltrating T cells. We proposed that engineering T cells to overexpress c-Jun, a transcription factor required for T cell development and activation, would activate T cells and enhance their persistence, function, and anti-tumor efficacy. In this study, using T-cell receptor- engineered T-cells (TCR-Ts) against the HCC-associated antigen Alfa-fetoprotein (AFP), we demonstrated that human TCR-Ts engineered to overexpress c-Jun (TCR-JUN) have better expansion potential in culture with enhanced functional capacity against HepG2 tumor cells. Additionally, TCR-JUN cells were less apoptotic and more resistant to exhaustion after HepG2 tumor stimulation. In HCC xenograft tumor model, c-Jun overexpression enhanced TCR+ T cell expansion and extended overall survival of the tumor bearing mice. Importantly, the TCR-JUN T cells were less exhausted and demonstrated enhanced tumor infiltration, persistence, and functionality. We conclude that engineering T cells to overexpress c-Jun could be a potential approach to enhance the antitumor efficacy of TCR-Ts.
    • *BARRIERS TO DEVELOPING EFFECTIVE MUSIC PROGRAMS IN THE RIVER CITY SCHOOL SYSTEM: AN ANALYSIS OF STAFF PERCEPTIONS

      Baxley, Pamela Marie; Advanced Studies Innovation
      Abstract Participation in healthy music programs has demonstrated positive associations with academic achievement, cognitive abilities, and other socio/affective outcomes. Despite these benefits, music programs often struggle against funding, policy, and high stakes testing and accountability practices - particularly in urban or otherwise diverse populations. The purpose of this case study was to evaluate the health of music programs in the River City School System (RCSS) by studying teachers’ perceptions of limits to their own agency in developing those programs and teachers’ perceptions of barriers to student involvement in music programs. Based on a broad review of literature, researchers studied four hypothesized factors that might act as barriers to teacher agency or student involvement in music programs – culturally responsive pedagogy, scheduling, recruiting, and professional learning & collaboration. This study used a mixed-methods approach employing a survey of all music teachers within the district followed by semistructured interviews for volunteers. Results of the study indicated logistical challenges, relational issues, and philosophical perspectives could act as barriers. From these results and in alignment with extant research, a conceptual map of findings was developed on which to provide recommendations for RCSS and other districts hoping to analyze the health of their own music programs.
    • SUCCEEDING IN INTRODUCTORY STEM COURSES AT COMMUNITY COLLEGES: STEM INSTRUCTORS’ PERCEPTIONS OF ESSENTIAL SKILLS AND BARRIERS TO SUCCESS

      Perkins, Ashlei; Advanced Studies Innovation (Augusta University, 2022-05)
      Community colleges play a key role in providing non-traditional and underrepresented minority students with access to careers in Science, Technology, Education and Math (STEM) fields, which are critical for the economic success of the United States. However, national studies reveal that student persistence in STEM majors is alarmingly low, particularly at community colleges. Therefore, it is important to conduct more studies that contribute to the understanding of STEM course success in college. To that end, this study gathered the following data: 1) instructors’ perceptions of essential skills required to succeed in introductory STEM courses at community colleges; 2) instructors’ perceptions of existing skills students have prior to STEM course instruction at community colleges; and 3) instructors’ perceptions of barriers that may prevent community college students from being successful in introductory STEM courses. This pragmatic, qualitative inquiry included interviews with STEM course instructors at a community college and employed a constant, comparative analysis approach. The study also juxtaposed findings against the literature to determine if there was congruence, in terms of critical STEM skills and knowledge for STEM occupations. Findings showed a significant overlap between essential skills required to succeed in STEM courses and important skills for STEM occupations and that essential skill development is critical but lacking. Keywords: STEM skills, essential STEM course skills, STEM persistence in community colleges, barriers to STEM course success
    • SUCCEEDING IN INTRODUCTORY STEM COURSES AT COMMUNITY COLLEGES: STEM INSTRUCTORS’ PERCEPTIONS OF ESSENTIAL SKILLS AND BARRIERS TO SUCCESS

      Smith, William Pierce; Advanced Studies Innovation (Augusta University, 2022-05)
      Community colleges play a key role in providing non-traditional and underrepresented minority students with access to careers in Science, Technology, Education and Math (STEM) fields, which are critical for the economic success of the United States. However, national studies reveal that student persistence in STEM majors is alarmingly low, particularly at community colleges. Therefore, it is important to conduct more studies that contribute to the understanding of STEM course success in college. To that end, this study gathered the following data: 1) instructors’ perceptions of essential skills required to succeed in introductory STEM courses at community colleges; 2) instructors’ perceptions of existing skills students have prior to STEM course instruction at community colleges; and 3) instructors’ perceptions of barriers that may prevent community college students from being successful in introductory STEM courses. This pragmatic, qualitative inquiry included interviews with STEM course instructors at a community college and employed a constant, comparative analysis approach. The study also juxtaposed findings against the literature to determine if there was congruence, in terms of critical STEM skills and knowledge for STEM occupations. Findings showed a significant overlap between essential skills required to succeed in STEM courses and important skills for STEM occupations and that essential skill development is critical but lacking. Keywords: STEM skills, essential STEM course skills, STEM persistence in community colleges, barriers to STEM course success
    • Succeeding in Introductory STEM Courses at Community Colleges: STEM Instructors' Perceptions of Essential Skills and Barriers to Success

      Payne, Daniela; Advanced Studies Innovation (Augusta University, 2022-05)
      Community colleges play a key role in providing non-traditional and underrepresented minority students with access to careers in Science, Technology, Education and Math (STEM) fields, which are critical for the economic success of the United States. However, national studies reveal that student persistence in STEM majors is alarmingly low, particularly at community colleges. Therefore, it is important to conduct more studies that contribute to the understanding of STEM course success in college. To that end, this study gathered the following data: 1) instructors’ perceptions of essential skills required to succeed in introductory STEM courses at community colleges; 2) instructors’ perceptions of existing skills students have prior to STEM course instruction at community colleges; and 3) instructors’ perceptions of barriers that may prevent community college students from being successful in introductory STEM courses. This pragmatic, qualitative inquiry included interviews with STEM course instructors at a community college and employed a constant, comparative analysis approach. The study also juxtaposed findings against the literature to determine if there was congruence, in terms of critical STEM skills and knowledge for STEM occupations. Findings showed a significant overlap between essential skills required to succeed in STEM courses and important skills for STEM occupations and that essential skill development is critical but lacking. Keywords: STEM skills, essential STEM course skills, STEM persistence in community colleges, barriers to STEM course success