• OPTIMIZED ISOLATION AND QUANTIFICATION OF IN VIVO DISTRIBUTION OF EXOSOMES FOR POTENTIAL TARGETED THERANOSTIC APPLICATION

      Rashid, Mohammad Harun; Department of Biochemistry and Molecular Biology (Augusta University, 2019-07)
      Exosomes are critical mediators of intercellular crosstalk and regulators of the cellular/tumor microenvironment. Exosomes have great prospects for clinical application as a theranostic and prognostic probe. Nevertheless, the advancement of exosome research has been thwarted by our limited knowledge of the most efficient isolation method and the in vivo trafficking. Here we have shown that a combination of two size-based methods using a 0.20 μm syringe filter and 100k centrifuge membrane filter followed by ultracentrifugation yields a greater number of uniform exosomes compared to other available methods. We demonstrated the visual representation and quantification of the differential in vivo distribution of radioisotope 131I-labeled exosomes from diverse cellular origins, e.g., tumor cells with or without treatments, myeloid-derived suppressor cells and endothelial progenitor cells. We also determined that the distribution was dependent on the exosomal protein/cytokine contents. Further, we also generated engineered exosomes expressing precision peptide for targeting CD206 positive M2-macrophages. M2-macrophages participate in immune suppression, epithelial to mesenchymal transition, invasion, angiogenesis, tumor progression and subsequent metastasis foci formation. Given their pro-tumorigenic function and prevalence in most malignant tumors with lower survival, early in vivo detection and intervention of M2-macrophages may boost the clinical outcome. To determine in vivo distribution of M2-macrophages, we adopted 111In-oxine based radiolabeling of the targeted exosomes and SPECT. When injected these radiolabeled targeted exosomes into 4T1 breast tumor-bearing mice, exosomes accumulated at the periphery of the primary tumor, metastatic foci in the lungs, in the spleen, and liver. Ex vivo quantification of radioactivity also showed similar distribution. Injected DiI dye-labeled exosomes into the same mice showed the adherence of exosomes to the CD206 positive macrophages on ex vivo fluorescent microscopy imaging, confirming the targeting efficacy of the exosomes. In addition, we utilized these engineered exosomes to carry the Fc portion of mouse IgG2b with the intention of augmenting antibody-dependent cell-mediated cytotoxicity. We have auspiciously demonstrated that M2-macrophage targeting therapeutic exosomes deplete M2-macrophages both in vitro and in vivo, and reduce tumor burden in a metastatic breast cancer model. The applied in vivo imaging modalities can be utilized to monitor disease progression, metastasis, and exosome-based targeted therapy.
    • Development and Characterization of a Closed-Head Mild Traumatic Brain Injury Model

      Alverson, Katelyn; Clinical Laboratory Sciences (Augusta University, 2020-12)
      Traumatic brain injury (TBI) is a leading cause of death and disability worldwide and places an enormous economic burden on both families and health care systems that provide support for survivors. The majority of TBI cases are deemed mild (mTBI) and go undetected due to the less discernable signs and symptoms. However, there is increasing evidence that mTBI can lead to detrimental chronic consequences. Unfortunately, the mTBI research field is still in its infancy. We set out to develop a model of closed-head mTBI that recapitulated mTBI in the clinic. Using a murine controlled cortical impact model, we show no structural damage, increased edema, behavioral deficits, cell death and decreased synapses in the acute time after the mTBI. We also evaluated the chronic behavioral changes from two weeks to three months post TBI. All in all, our mTBI model showed significant cellular changes, but did not give robust chronic behavioral results. A secondary outcome of our study was the evaluation of a potential therapeutic: remote ischemic conditioning (RIC). Acutely, RIC improved edema, behavioral outcomes, cell death, and synapse loss. Overall, our study does identify key areas that should be recapitulated in further development of the model: no structural damage, little to no edema, cell death and decreased synapses, and behavioral changes. This model also requires further investigation into the chronic consequences of mTBI as well as the use of RIC.
    • THE ROLE OF GPR109A IN REGULATION OF RETINAL ANGIOGENESIS AND BLOOD-RETINAL BARRIER AS A POTENTIAL THERAPEUTIC TARGET IN DIABETIC RETINOPATHY

      Abdelrahman, Ammar; Department of Biochemistry and Molecular Biology (Augusta University, 2020-12)
      Currently, treatments of diabetic retinopathy (DR) have limited therapeutic benefits and limited accessibility to the growing diabetic population at risk because of the high expenses and complicated procedures. Inflammation, endothelial dysfunction, and microvascular damage are common features of diabetic complications including DR. GPR109A is the metabolite sensing receptor of beta-hydroxybutyrate (BHB) the principal ketone body in humans. Our previous studies have shown the role of GPR109A expression in promoting anti-inflammatory response in retinal pigmented epithelial (RPE) cells and the relevance of the receptor in DR. Expression of the GPR109A in microvascular endothelial cells (ECs) has been reported recently. However, the relevance of GPR109A expression and activation to retinal EC functions are yet to be studied. Our goal in this study was to identify the role of GPR109A expression and activation in barrier and angiogenic functions of retinal ECs in context of diabetic retinopathy. We used electrical cell impedance sensing (ECIS) technology to evaluate barrier functions in primary human retinal endothelial cells (HRECs) which constitute the inner BRB. Knocking down GPR109A in HRECs with siRNA decreased the transendothelial electrical resistance (TEER) compared to scrambled siRNA. Treating HRECs with BHB increased their TEER and counteracted VEGF-induced barrier disruption through activation of GPR109A and increasing zonula occludens-1 (ZO-1) expression. Treatment of STZ-diabetic mice with exogenous BHB for one month protected against the pathologic albumin leakage induced by diabetes and improved the visual acuity of this animal model of diabetes. Using the mouse model of oxygen induced retinopathy (OIR), we showed that Gpr109a-/- mice had slower vascular recovery from pathologic angiogenesis compared to age matched wild type mice. Moreover, physiologic revascularization of vaso-oblitrated retinas was impaired by loss of GPR109a and associated with dysregulated inflammatory and angiogenic signaling. Collectively, these data point to a role for GPR109A in the regulation of barrier and angiogenic mechanisms in retinal ECs and, promote the receptor as a potential druggable target for impacting these mechanisms in microvascular retinal diseases such as DR.
    • Cassandra Radical Feminist Nurses Network: Feminism, Nursing, and a History for the Present

      Dillard-Wright, Jessica Susan; Nursing (Augusta University, 2020-12)
      As the last light of the Equal Rights Amendment (ERA) faded in 1982, a group of radical feminist nurses coalesced around their shared outrage at nursing’s disciplinary failure to engage deeply with feminist causes. The 1982 American Nurses Convention coincided with this last gasp of the ERA, held in a hotel in Washington, D.C. where thousands of nurses, overwhelmingly women, converged for professional development and camaraderie. And although the city outside the hotel roiled in protest, the Convention unfurled with nary a mention of the constitutional amendment that would secure legal equality irrespective of gender. Incensed by this omission, and with nursing’s general resistance to political engagement, these radical nurses descended on the hotel bar and began organizing what would become Cassandra Radical Feminist Nurses Network. Cassandra Radical Feminist Nurses Network (“CASSANDRA” hereafter, in the convention established by the organization in their Newsjournal) was an activist network active from 1982 until 1991. This study used historical research methods to document CASSANDRA’s legacy while unpacking the complex interrelationship between nursing and feminism. This includes examining the influences of race, gender, and sexuality, influences that shapes normative understandings of nursing from its Victorian origins to the present. CASSANDRA was unusual in its overt affiliation as a nurses’ organization with a radical feminist allegiance during an era when feminism and nursing were frequently at odds. As a decentralized, radical feminist “web,” the aim of CASSANDRA was to “create and develop a group that would truly provide an open forum for feminist nurses from all walks of life and how to avoid the usual male-oriented hierarchy and rigidity of most national organizations” (LaGodna, 1982, p. 1). In unfurling the nuances of gender and sexuality that CASSANDRA navigated, it is clear that the work of CASSANDRA envisioned a radical space for collective resistance and connection, reflecting the normative expectations in nursing that stemmed from nursing’s Victorian imaginary. Even while CASSANDRA’s work around gender and sexuality was bold and transgressive, their engagement with race was poorly articulated. Because of this, the organization’s work reinforced white normativity. Ultimately, like mythological Cassandra, CASSNADRA would eventually quiet to a whisper. What understanding the thrums of CASSANDRA, of nursing’s rich and complex history can do is provide a clear view of nursing’s disciplinary history. This is a fundamental prerequisite for a more just, equitable nursing future.
    • Impact of general health on the outcomes of center-based cardiac rehabilitation

      Roberts, Kimberly A; Nursing (Augusta University, 2020-12)
      Background: Cardiovascular disease is the number one cause of death, and cardiac rehabilitation (CR) is effective in reducing the risks of disease progression and mortality by improving physical functioning and quality of life. Despite the significant impact of CR, the completion of CR is low. Factors influencing CR completion have been widely studied, but little is known about the impact of general health on CR completion self-care behaviors, and physical functioning associated with CR completion. Therefore, the purpose of the study was to explore relationships between health beliefs, general health, CR completion, physical function, and self-care behavior. The conceptual framework shows how health belief perceptions, support systems, sociodemographic factors, and general health influence CR completion, physical function, and self-care behavior. Methods: This was a retrospective cohort study using a sample of participants completing the outpatient CR center at a large medical center in the southeastern United States. Multiple linear regression was used to determine the predictors of CR completion, the improvement of physical function, and self-care behavior. Results: HbA1C predicted CR completion, and there was no significant relationship between CR completion and general health. Gender and general health (sitting time, fatigue, anxiety, and depression) predicted the improvement of physical functioning. Age and general health (sitting time, self-care complexity, and disease burden) predicted self-care behaviors. There was no relationship between health beliefs and CR completion. Conclusion: CR completion was predicted by glycemic control, while physical functioning improvement and self-care behavior were predicted by general health indicators. Additional research is needed to validate the findings and develop a sensitive screening tool to identify high-risk patients who are likely to drop out from a CR program. Further research to develop strategies to prevent CR incompletion and poor outcomes prior to patients’ participation is warranted.
    • A Molecular Basis of Chemoresistance in Bladder Cancer

      Lahorewala, Sarrah; Biochemistry and Cancer Biology (Augusta University, 2020-12)
      Background: In advanced bladder cancer (BC), development of resistance to the frontline chemotherapeutic drugs Gemcitabine and Cisplatin contributes to the poor prognosis of patients. Newly discovered chondroitinase, HYAL-4 V1 (V1), drives malignant transformation in BC. We evaluated V1’s role and the downstream molecules involved in the mechanistic regulation of chemoresistance in BC. Experimental Design: HYAL-4 expression was evaluated by RT-qPCR and IHC in metastatic muscle-invasive BC patients who received Gemcitabine plus Cisplatin chemotherapy. HYAL-4 wild-type and V1 were stably expressed or silenced in three BC and one normal urothelial cell line. Transfectants were analyzed for Gemcitabine and Cisplatin sensitivity, and for Gemcitabine influx and efflux to determine the mechanism of Gemcitabine resistance. The effect of cytidine deaminase (CDA) inhibition on Gemcitabine sensitivity was evaluated in vitro and in xenograft models. Results: HYAL-4 expression was an independent predictor of disease-specific mortality and treatment failure in our clinical cohort, and stratified patients into higher risk for both those outcomes. V1-expressing BC and normal urothelial cells were resistant to Gemcitabine due to the upregulation of cytidine deaminase (CDA) expression and activity, resulting in increased Gemcitabine metabolism and efflux; treating cells with tetrahydrouridine (THU), a CDA inhibitor, abrogated the chemotherapeutic resistance. Gemcitabine-resistant V1 cells demonstrated increased expression of V1’s substrate CD44 and phosphorylated STAT3. Si-RNA mediated CD44 knockdown and STAT3 inhibition both sensitized cells to Gemcitabine in vitro. In xenograft models, treatment with a combination of Gemcitabine and THU completely inhibited tumor growth. Conclusions: This project discovered V1 as a novel determinant of Gemcitabine resistance and potential predictor of treatment response in BC. V1 drives resistance to Gemcitabine through CD44-STAT3 mediated upregulation of CDA, and inhibiting this pathway sensitizes tumor cells to the therapy in preclinical models of BC.
    • Improving Heart Failure Outcomes Using Guideline

      Crew, Deborah; Flores, Yolanda; Augusta University Health; College of Nursing (2019-05-01)
      This poster presents a project plan and intervention to improve heart failure.
    • A Hospital Pastoral Care Program to Improve Advance Care Planning Discussion in Patients with Heart Failure

      Crew, Deborah; Young, Lufei; Flowers, William; Dunlap, Stephanie; Schafer, Pascha; Weintraub, Neil; Augusta University Medical Center; College of Nursing (2020-10-08)
      Heart failure (HF) is a chronic disease associated with poor quality of life (QoL), high rates of readmission and high inpatient cost (Sadeghi et al., 2016). Advance care planning (ACP) improves quality of life, and reduces readmission and its associated cost (Schichtel et al., 2020). The uncertain trajectory of HF makes it difficult to find the ‘right time’ to initiate ACP communication, hindering patients, caregivers and health care providers to plan and prepare for the future care (Ahluwalia, S. C., & Enguidanos, S., 2015). Hospitalization experience makes HF patients and caregivers more receptive and desire to discuss end of life care. Therefor, hospitalization may provide a timely opportunity to initiate ACP discussion and palliative care before discharge (Schichte et al., 2020). The goal of this project is to assess the completion rates of ACP discussion and documentation among hospitalized HF patients and identify barriers to initiate ACP communication.
    • The effects of altered salivary viscosity on the incidence and extent of dental caries in rats

      Biesbrock, Aaron Reed; Department of Oral Pathology (Augusta University, 1989-05-02)
      The research was intended to determine whether alterations in salivary viscosity brought about by altering dietary components affected the incidence and extent of dental caries in rats.
    • The stability of an increased vertical dimension of occlusion in growing and non-growing golden hamsters

      Berman, Scott C.; Department of Oral Biology (Augusta University, 1993-03)
    • Psychosocial Factors as Predictors for Patient Outcomes in Rehabilitation of Upper Extremity Injury Caused by Trauma:

      Holley, Ashlyn; Saren, Madison; Wygle, Sarah; Deese, Abigail; Payne, Regan; Department of Occupational Therapy (Augusta University, 2020-08-26)
      At the conclusion of this presentation, attendees will be able to: 1) Explain three ways in which psychosocial factors have the ability to alter rehabilitation outcomes in individuals who have sustained a traumatic upper extremity injury, 2) Discuss two gaps in the literature regarding the impact of psychosocial factors on rehabilitation outcomes in individuals recovering from traumatic upper extremity injuries, 3) Identify two methods, strategies, or assessments to be implemented in practice in order to evaluate and address psychosocial factors as a component affecting functional outcomes of clients with traumatic UE injuries.
    • Telehealth Interventions to Address Chronic Disease Self-Management Interventions within the Scope of Occupational Therapy: A Scoping Review

      Albritton, Liz; Fish, joJo; Henkel, Jeff; Lee, Shelby; Luttrel, Rachel; Rackleff, Layne; Department of Occupational Therapy (Augusta University, 2020-09-02)
      At the conclusion of this presentation, attendees will: 1) Distinguish between the types of telehealth interventions for chronic disease self-management within the scope of occupational therapy and 2) Describe the outcomes of using telehealth for chronic disease self-management based on the presented results of a scoping review of the literature.
    • The Effects of Interprofessional Education on Occupational Therapy Student Practitioner Outcomes: A Systematic Review

      Adams, Emma; Carlto, Meg; Kali, Todd; Department of Occupational Therapy (Augusta University, 2020-09-01)
      Objectives of Presentation: 1) Understand and communicate the effects that various evidence based interprofessional education interventions have on occupational therapy student outcomes. and 2) Discuss the gaps in the literature, indicated by this systematic review, as they relate to interprofessional education and its effects.
    • Benefits of International Fieldwork for Occupational Therapy Students

      Vickman, Hannah; Carter, Krissy; Dittmer, Chandler; Nettles, Taylor; Wang, Caroline; McCarley, Trinity; Department of Occupational Therapy (Augusta University, 2020-09-01)
      At the conclusion of this presentation, attendees will: 1) Identify the perceived benefits for international fieldwork ascertained from the research and how that relates to professional and personal development, and 2) Identify the key clinical experience differences between OT students participating in international and domestic fieldwork, as established from the presented research.
    • Evidence supporting interventions within the scope of occupational therapy for addressing

      Arnold, Jackie; Cyr, Emily; Garner, Kathryn; Long, Tori; Robles, Alexis; Department of Occupational Therapy (Augusta University, 2020-08-26)
      At the conclusion of this presentation, attendees will: a) List three effective interventions to address cognitive and psychological factors in chronic pain based on the presented scoping review of the literature, and b) Articulate the importance of cognitive/psychosocial factors in relation to chronic pain.
    • Outcomes of Music, Dance, & Movement Interventions for Children with Autism Spectrum Disorder Within the Scope of Occupational Therapy: Scoping Review

      Bankson, Baylee; Cox, Ashlyn; Fulmer, Haley; Hausman, Lydia; Longfellow, Danielle; Department of Occupational Therapy (Augusta University, 2020-08-31)
      At the conclusion of this, attendees will: 1) Identify frequently measured outcomes of using music, dance, and movement-based interventions with individuals with autism spectrum disorder, as identified through a scoping review of the literature and 2) List specific music, dance, or movement-based interventions that are available to OT practitioners working with individuals with autism spectrum disorder (ASD), as identified through a scoping review of the literature.
    • Biomechanical behavior related to structure in normal and congenitally disordered elastic arteries

      Beall, Arthur C.; Department of Pharmacology and Toxicology (Augusta University, 1992-12)
    • The effects of retinoic acid-induced differentiation on neurotransmitter receptor content and signal transduction in a human neuroblastoma cell line

      Baumgartner, Melissa K.; Department of Pharmacology & Toxicology (Augusta University, 01/23/1993)
      The purpose of the present study was to establish the effects of retinoic acidindttced differentiation on muscarinic receptor populations and signal transduction pathways in the human neurroblastoma Sk-N-SH cells. The human neuroblastoma cell line Sk-N-SH was induced to differentiate by treatment with 1 uM retinoic acid for 7 days. Differentiation was characterized by profuse neurite outgrowth, a decrease in cell growth, and a 2~3 fold increase in the protein content of each cell. Muscarinic receptors were labelled-using [3H]N-methyl scopolamine. Muscarinic receptor density increased by approximately 36% after treatment for 7 days with retinoic acid (Bmax, control = 126 ± 13 fmol/mgprotein; Bmax, retinoic acid-treated= 170 ± 17 fmol/mg protein; p<0.05), corresponding to a 170% increase in receptor content per cell. The affinity of [3H]NMS for the receptors was somewhat lower in the differentiated cells (KD, control = 0.14 ± 0.04 nM; KD, retinoic acid-treated = 0.25 ± 0.0.4 nM; p<0.05). The guanine nucleotide sensitivity of agonist (carbamylcholine) binding to Sk-N-SH muscarinic receptors Was slightly decreased by differentiation. Reverse transcriptase/polymerase chain reaction (PCR) analysis using muscarinic receptor subtype specific primers revealed that the undifferentiatied Sk-N-SH cells transcribed mRNA for all 5 receptor subtypes; this pattern was not affected by differentiation. [3H]NMS displacement curves with subtype- selective receptor ligands (pirenzepine, m1; AFDX-116, m2; 4-DAMP, m3) indicated the predominant expression of m1 and m3 receptor subtypes, and differentiation did not affect the pharmacological profile of the expressed muscarinic receptor populations. Differentiation did not affect basal G protein GTPase activity. However, acetylcholine (100 uM) stimulation of G protein GTPase activity was decreased in differentiated cells (18 ± 1.8 pmol/min/mgprotein) compared to the undifferentiatied cells (23 ± 1 .0 pmol/ min/ mg protein) (p<0.05). Inhibition of acetylcholine--stimulated GTPase activity with selective muscarinic receptor antagonists indicated that the m3 antagonist (4-DAMP) was as effective as atropine in inhibiting activity by 80-100%. Selective m1 and m2 antagonists were less effective (30-40%) at inhibiting stimulated GTPase activity. There were no differences in inhibition of stimulated GTPase activity after differentiation. Immunoblots of control and retinoic acid-treated cells revealed no change in Goa, Gsa or Gp content after differentiation; however, 0.1% ethanol and retinoic acid-treated cells displayed a 30% decrease in expression of Gia3, and Gqa. Muscarine (0.1-100 uM) stimulated 45Ca influx into Sk-N-SH cells, and this uptake was inhibited by preincubation with atropine. The magnitude of the muscarinic receptor-mediated uptake was 50-60% lower in the differentiatied cells. Basal adenylate cyclase activity was depressed in the differentiated cells (2.5 pmol / min / mg protein) compared to the undifferentiated cells (8.4 pmol / min / mg protein) (p< 0.05). Forskolin (5 - 50 uM)-stimulated adenylate cyclase activity was not altered, however fractional stimulation was significantly (p<0.0001) increased in the differentiated cells. Differentiated cells displayed a slightly greater receptor-mediated inhibition of the adenylate cyclase activity by carbamylcholine (1 uM- 1 mM). It is demonstrated that in Sk-N-SH cells, retinoic acid-induced differentiation: 1) increases the size of the muscarinic receptor population (Bmax) while decreasing [3H]NMS binding affinity, 2) does not alter muscarinic receptor pharmacology, or the expression of. muscarinic receptor subtypes, 3) decreases muscarinic receptor-stimulated 45Ca flux 50-60% compared to undifferentiated cells, 4) depresses basal adenylate cyclase activity, increases fractional stimulation of forskolin-stimulated activity of adenylate cyclase, and may increase muscarinic receptor-mediated inhibition of adenylate cyclase activity, 5) does not alter basal G protein GTPase activity but depresses muscarinic receptor-stimulated high affinity GTPase activity suggesting muscarinic receptor-G protein coupling is altered, and 6) does not alter expression of Goa, Gsa and Gp content while Gia3 and Gqa are depressed in differentiated as well as in 0.1% e.thanol treated cells.
    • Toxicity of visible light-cured denture resins

      Barron, Dara Jewell; Department of Oral Biology (1992-04)
      In this study three commercial formulations of visible light-cured (VLC) denture resins have been analyzed. The products used are those suggested for the reline, repair and fabrication of dentures to improve their fit. The biocompatibility of these resins was investigated by measuring RNA and DNA synthesis of oral epithelial cells in vitro. The extent to which oral cells recover from toxic resin exposure, the conversion of monomer into polymer, the presence of inorganic filler, and resin leacha~ility have also been studied. It was shown that VLC denture resins inhibit the synthesis of RNA and· DNA relative to a heat-cured resin control (p~0.05). Although epithelial cells appeared to recover from toxic resin exposure, this recovery was inconsistent among experiments. Infrared spectroscopy illustrated chemical group differences that occurred before and after photo-polymerization. Using these differences, the conversion of monomer into polymer ranged from 77% to 97%. This conversion was significantly affected (p< .003) by the type of curing unit, duration of photo-polymerization, and surface exposed to visible light. Soluble substances in cured and uncured resin products were analogous using HPLC. The range of inorganic filler present was 0-15%. These investigations suggest that visible light-cured denture resins may impair the replication of oral epithelial cells. This effect may be related to the leachability of unpolymerized resin constituents, the presence or absence of filler particles, or polymerization by-products.