Now showing items 21-40 of 1070

    • 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.
    • Statistical Methods for reaction Networks

      Odubote, Oluseyi Samuel; Department of Biostatistics and Epidemiology
      Stochastic reaction networks are important tools for modeling many biological phenomena, and understanding these networks is important in a wide variety of applied research, such as in disease treatment and in drug development. Statistical inference about the structure and parameters of reaction networks, sometimes referred to in this setting as model calibration, is often challenging due to intractable likelihoods. Here we utilize an idea similar to that of generalized estimating equations (GEE), which in this context are the so-called martingale estimating equations, for estimation of reaction rates of the network. The variance component is estimated using the approximate variance under the linear noise approximation, which is based on partial dierential equation, or Fokker-Planck equations, which provides an approximation to the exact chemical master equation. The method is applied to data from the plague outbreak at Eyam, England from 1665-1666 and the COVID-19 pandemic data. We show empirically that the proposed method gives good estimates of the parameters in a large volume setting and works well in small volume settings.

      Goldman, Chandler; Biomedical Sciences
      The establishment of cell polarity is critical for performing complex functions including division and migration. As such, the loss of polarity is implicated in many diseases including cancers. To establish polarity, many cell types rely on the asymmetric sorting of messenger RNAs or mRNAs. These mRNAs are held in a translationally-repressed state until reaching their destinations. Upon arrival, translation is allowed to commence, giving rise to spatially restricted proteins. Often, mRNAs are transported to their destinations along microtubules via linkage to one of the sub families of microtubule motors, Dynein or Kinesins. The mechanism by which mRNAs are linked to these motors is unknown for the vast majority of localizing mRNAs. Drosophila oocytes and embryos display a great number of mRNAs that are localized to specific regions. The protein Egalitarian (Egl) has been shown to directly bind several mRNAs and participates in their microtubule-based transport by promoting linkage to cytoplasmic Dynein. In Aim 1 of this thesis, we aim to determine the mechanism by which Egl and its interacting partners, Dynein light chain (Dlc/LC8), and Bicaudal D (BicD), tether mRNAs to the Dyenin motor for transport. Dlc is required for Egl homodimerization, which promotes binding to mRNAs. BicD then preferentially associates with mRNA-bound Egl and links the complex to the Dynein motor. In Aim 2, we further investigate Egl’s role in mRNA transport by determining the critical amino acid residues within its RNA binding domain required for mRNA association.
    • An Essential Oil Intervention for Chemotherapy-Induced Peripheral Neuropathy in Breast Cancer Survivors: A Mixed Methods Study

      Langley-Brady, Dawn Louise; Nursing (Augusta University, 2020-12)
      Chemotherapy-induced peripheral neuropathy (CIPN) is a painful, debilitating consequence of cancer treatment and is considered the most adverse of non-hematologic events. Pharmacological approaches to CIPN are often ineffective and cause adverse effects. A problem faced by many breast cancer survivors is poor CIPN treatment coupled with practitioners’ lack of understanding about their subsequent quality-of-life (QOL). Essential oils (EOs) are an underutilized non-pharmacological approach to pain reduction. EO mechanisms of action include non-competing inhibition of 5-HT, AchE, and Substance P and antagonism of TRPA1 and TRPV1. The study aims were to ascertain the effect of an EO intervention (EOI) on CIPN and quality-of-life (QOL) in breast cancer survivors and develop a deeper understanding of CIPN QOL using photovoice methodology. This mixed methods research design employed a randomized, single-blind, placebo-controlled trial. Twenty-six breast cancer survivors with chronic lower extremity CIPN were enrolled in the quantitative strand using purposive sampling. Participants were stratified by baseline pain score and randomized to intervention (n = 13) and placebo (n = 13) groups. Participants topically applied an EOI (containing Curcuma longa, Piper nigrum, Pelargonium asperum, Zingiber officinale, Mentha x piperita, and Rosmarinus officinalis ct. cineole) or placebo three times a day for six weeks. Pain was assessed weekly using the Short-Form McGill Pain Questionnaire-2 (SF-MPQ-2) and daily using the Visual Analogue Scale (VAS). QOL was assessed using the QOL:CIPN20 and QOL Adult Cancer Survivor questionnaires(QLACS) at baseline, midpoint, and endpoint. Data were analyzed in SPSS using generalized estimating equations. Test of model main effects were significant for visit (SF-MPQ-2, p = .000; VAS, p = .008; QLACS Pain subdomain, p = .026), but not for visit*group interaction effects. SF-MPQ-2 and VAS positively correlated with QOL:CIPN20 scores (r = .843, r = .671); however, QOL:CIPN20 model main effects were not significant. The VAS %Δ for intervention and placebo groups was -14.67 and -7.57 respectively. This was not statistically significant, but is clinically important. Regardless of group assignment, pain and QOL improved. The EOI was well-tolerated and demonstrated 50% more pain reduction than placebo. A subset of participants(n = 9) were enrolled in the qualitative strand, received photovoice-related training, and spent four weeks photographing their life with CIPN. Participants participated in photo-interviewing, a photo discussion focus group, a photovoice exhibition at a local art gallery, and an event de-briefing focus group. Data were analyzed in NVivo using thematic, visual content, and iconographical analyses. Six primary themes emerged from the data: (a) advice for clinicians, (b) positive photovoice experience, and CIPN (c) causes pain, (d) affects relationships, (e) causes disruptions, and (f) alters self-image. Further research is needed to enhance EO pain-reducing efficacy as a natural nursing intervention. CIPN greatly impacts breast cancer survivors’ QOL and is multifaceted. Nursing care for breast cancer survivors should include education regarding the potential severity and lifelong effects of CIPN and benefits of study participation and group support.
    • The Effect of Instructor Mindset on Student Motivation and Self-Efficacy

      Restrepo, Leigha; Department of Psychological Sciences (Augusta University, 2020-07)
      Dweck’s theory of mindset proposes two different mindsets a person may have: fixed or growth (Dweck, 2007). A person with a fixed mindset believes intelligence is fixed and a person with a growth mindset believes that they can improve their intelligence with effort (Dweck, 2007; Murphy & Dweck, 2016). The present study was designed to examine the effect of an instructors’ apparent mindset on the expectations of success and persistence in STEM disciplines among students. Students were presented with sample syllabi that portrayed an instructor with either a fixed or growth mindset and completed questionnaires and a short, written reflection to measure their perception of mindset, self-efficacy, and motivation. Results of this study revealed that students expected a higher grade, reported more academic self-efficacy, and had a positive perception of the instructor after reading the growth syllabus. Overall, Black students reported more academic self-efficacy than White students and reported more academic self-handicapping after reading the growth syllabus. Students reported that the attributions (gender, minority, status, effort/ hard work, luck, difficulty of the course, intelligence/ ability) contributed more to their grade in the class after reading the growth mindset syllabus than the fixed syllabus, with difficulty of the course and intelligence/ ability significantly contributing to their perceived grade in the class after reading the fixed syllabus. The mindset portrayed by an instructor can have an impact on the student through a decrease in their overall academic performance. Examining the different ways in which a change in the mindset that is portrayed can help to increase student motivation and expectations.

      Packer, Jonathan; Department of Psychological Sciences (Augusta University, 2020-07)
      This study set out to determine the effectiveness of using tianeptine as a treatment for traumatic brain injury (TBI). A controlled cortical impact model was utilized to induce a bilateral moderate TBI in the frontal cortex of the rat. Sham surgeries were performed to ensure an accurate control group. Rats received 30mg/kg tianeptine, or an equal volume of saline one hour following injury and once a day for nineteen days following surgery. Rats were tested for behavioral, motor, and cognitive deficits using the following tasks: Morris water maze (reference and working memory), foot fault task, forelimb use asymmetry task, open field task, and the passive avoidance task. As well, the brains were analyzed for differences in remaining cortical tissue following injury. Significant improvement was found in the Morris water maze reference memory task, the foot fault task, and the open field task for injured rats receiving tianeptine. Similarly, significant improvement was found in the remaining cortical tissue following injury in rats receiving tianeptine. Taken together, these results indicate tianeptine may be a viable treatment for improving recovery following TBI in rats.

      Littlejohn, Rodney; Department of Cellular Biology and Anatomy (Augusta University, 2020-07)
      Background. Early cardiac development is a tightly regulated process, involving spatiotemporal coordination of multiple signaling pathways and heterogenous cell populations, both generated de novo and externally sourced. While the roles of transcription, environmental, and epigenetic factors have all been studied extensively in the context of heart development, the roles of post-translational protein modification in regulating this process remain to be elucidated. NEDD8 (neural precursor cell expressed developmentally downregulated 8) is a novel ubiquitin-like protein modifier. Conjugation of NEDD8 to protein targets, a process termed neddylation, has been shown to regulate cell proliferation, cell signaling, and protein homeostasis, and play important roles in multiple physiological and pathological events. We have previously shown that neddylation is developmentally downregulated in the developing heart and is essential for mid-to-late gestational ventricular chamber maturation. However, whether and how neddylation regulates early cardiogenic events remains unknown. Methods and results. Mice with constitutive, cardiac progenitor cell-specific, cardiomyocyte- and vascular smooth muscle cell-specific deletion of NAE1, a regulatory subunit of the NEDD8-specific E1 activating enzyme, were created. Constitutive deletion of NAE1 led to early embryonic lethality before E9.5. Nkx2.5Cre-mediated deletion of NAE1 decreased neddylated proteins in the heart, disrupted normal cardiogenesis and resulted in embryonic lethality by embryonic day (E) 12.5 due to heart failure. Similarly, SM22αCre-driven deletion of NAE1 also caused cardiac failure and embryonic lethality by E13.5. The striking cardiac phenotypes were associated with myocardial hypoplasia, ventricular hypo-trabeculation, and pronounced endocardial and/or epicardial defects in both models. Unbiased transcriptomic analysis revealed dysregulated expression of genes associated with cardiomyocyte differentiation, proliferation, and maturation in NAE1-deficient hearts. Indeed, inhibition of neddylation disturbed cardiomyocyte proliferation, and myofibril assembly in vitro and in vivo. Moreover, defects in cardiomyocyte differentiation and maturation were linked to downregulation of Nkx2.5 and Mef2c, two key transcription factors regulating early cardiogenesis. Conclusion. Collectively, our findings demonstrate that neddylation in cardiac progenitor cells and cardiomyocytes is essential in the regulation of cardiogenesis in transgenic mouse models. Our results uncover a previously unknown role of post-translational modification in the regulation of cardiac development via potential roles in mediating cardiomyocyte proliferation, differentiation, and maturation.
    • Characterization of a Cyclic Peptide (ADO5) as a Novel Inhibitor of the Hsp90 Chaperoning Machine

      Fang, Wayne; Department of Biological Sciences (Augusta University, 2020-05)
      Protection of oncogenic proteins is the foundation of many hallmarks of cancer. Based on this, hsp90 inhibitors have emerged as a potentially potent strategy for cancer treatment. The clinical efficacy of the earlier Hsp90 inhibitors remains unsatisfactory, in part due to their induction of heat shock response and anti-apoptotic mechanisms in cancer cells. To identify alternative therapeutic agents without these effects, we have developed a cell-free high-throughput screen (HTS) platform based on the folding of progesterone receptor (PR) by the core components of the Hsp90 chaperoning machine. During our initial screening of 175 natural products from North African medicinal plants, we discovered the cyclic peptide AD05 as a novel Hsp90 inhibitor. AD05 has shown a powerful antitumor activity against various cancer cell lines including HeLa, Hs578T, MDA-MB231, MDA-MB453, E0771, THP1, and U937. Western blot analysis revealed that AD05 destabilizes Hsp90 client proteins without inducing heat shock response as indicated by lack of upregulation of Hsp70, Hsp40 and Hsp27. Remarkably, AD05 does not induce apoptosis but rather triggers autophagy in various cell lines.
    • The Effect of PFOA on ERα+ and ERα- Human Breast Cancer Cell Lines

      Gaw, Victoria; Department of Biological Sciences (Augusta University, 2020-05)
      Perfluorooctanoic acid (PFOA) is a man-made chemical that belongs to a large group of fluorotelomers. PFOA is used to manufacture both industrial and consumer products and individuals can be exposed to PFOA through ingesting PFOA-contaminated water or food. While the long-term effects of perfluorooctanoic acid are largely unknown, there is increased evidence suggesting it to be an endocrine disruptor. Studies have shown that PFOA binds to and activates the peroxisomeproliferator-activated receptor α (PPARα), which can regulate the expression of other genes and receptors such as the other PPAR isoforms as well as estrogen receptor α (ERα). Previous experiments in our lab demonstrated that PFOA treatment of MCF-7 breast cancer cells (an ERα positive cell line) decreased ERα mRNA and protein levels, and decreased cell viability by ~20% within 48h of treatment. However, these cells were treated in the absence of fetal bovine serum (FBS), a cell culture additive that contains important growth factors. When we repeated these experiments without serum withdrawal, we initially noted a tendency towards increased proliferation in MCF-7 cells treated with 50µM and 100µM PFOA at both 24h and 48h compared to control. To further examine the role of ERα in PFOA-induced proliferation, we carried out additional experiments in another ERα positive cell line, T47-D, as well as an ERα negative cell line, MDA-MB-231. All three cell lines showed a tendency for increased viability. These data suggest that the PFOA-induced increase in cell viability in these cell lines is not dependent on ERα expression. In addition, the opposing effects of PFOA on proliferation in MCF-7 cells in the presence and absence of FBS demonstrates the importance of accurately and completely reporting cell culture and treatment conditions.
    • Development of Chemically Defined Culture Conditions for in vitro Expansion of Human Wharton’s Jelly Stem Cells

      Shaikh, Arika; Department of Biological Sciences (Augusta University, 2020-05)
      Mesenchymal stem cells (MSCs) are multi-potent and capable of differentiating into a variety of cell lineages. While MSCs have commonly been isolated from bone marrow for treatment of various diseases, Wharton’s Jelly (WJ), an extra-embryonic umbilical cord tissue rich from hyaluronic acid (HA), represents an alternative source for a safer and less invasive isolation of MSCs. Typically, WJ-MSCs are isolated and cultured in undefined media containing fetal bovine serum (FBS), of which use has been associated with different complications, including reproducibility of studies, transmission of infectious agents, and induction of immunologic reactions. To overcome these complications, and thus to facilitate clinical applications of WJ-MSCs, this project aimed to develop chemically defined and safe culture conditions for human WJ-MSCs. We hypothesize that undifferentiated growth of WJ-MSCs will be supported by an HA-based extracellular matrix and fortified DMEM/F12 supplemented with defined macromolecules, antioxidants, lipids and growth factors found in platelet lysate. This hypothesis was tested by comparing the growth kinetics and morphology of WJ-MSCs cultured in defined and undefined media. WJ-MSCs were isolated via enzymatic digestion from discarded human umbilical cords. Following phenotyping and sorting by evaluating expression of relevant markers (i.e., CD105, CD73, and CD90) using flow cytometer, WJ-MSCs were randomly distributed and cultured in five different defined media plus an undefined control medium. The best alternative in terms of cell morphology and proliferation was the medium 3 consisting of DMEM/F12 supplemented with glutamine, ITS (define), antioxidant mixture, lipid mixture, and growth factor mixture. Medium 3 was further improved by adding increasing concentrations of ethanolamine. These results are of significance for therapeutic applications of MSCs. Further research is needed to optimize compositions of extracellular matrix and growth factors while examining the plasticity of MSCs.
    • Assessing Local Parks For Their Infrastructure Availability And Use Along With Physical Activity Levels Of The Local Children

      Shabu, Elizabath; Department of Kinesiology (Augusta University, 2020-05)
      This research project assessed public park physical activity infrastructure use in Richmond and Columbia Counties. Prior research has shown that children not only enjoy outdoor time, but also consider parks as a place for socializing. Furthermore, research has shown that playground time positively impacts children’s imagination. The playground also aids in the physical fitness of children by offering interactive experiences that can add to the recommended 60 minutes of physical activity a day. Park assessments were conducted utilizing the Physical Activity Resource Assessment (PARA) Tool and found that overall, parks in both counties were well taken care of. Additionally, there was a wide variety of amenities available to utilize. There were some areas of concern in the parks, however, including cigarette buds, alcohol containers, trash, and cracked sidewalks. In both counties, parks were observed to see how much children utilized the playground equipment. Observations concluded that the majority of children utilized the different variety of equipment, with the swings and slides being the most common. In conclusion, this presentation will describe the diversity of amenities, challenges in maintenance, and the overall use of public parks in both Richmond and Columbia Counties.
    • Profiling the HCA Receptor Family through BRET Analysis of GPCR-G-Protein and GPCR-Arrestin Interactions

      Saj, Dalia; Department of Biological Sciences; Department of Pharmacology & Toxicology (Augusta University, 2020-05)
      Increasing obesity rates have put the American population at higher risk for developing obesity-related medical conditions such as hypertension, heart disease, and diabetes. The hydroxycarboxylic acid (HCA) receptor family is a family of G protein-coupled receptors (GPCRs) that are expressed in adipose tissue and function as metabolic sensors, making them potential pharmaceutical targets in the treatment of obesity and other metabolic disorders. The HCA receptor family consists of the HCA1, HCA2, and HCA3 receptors, which are activated by hydroxycarboxylic acids such as lactate and 3-hydroxybutyric acid. We utilized bioluminescence resonance energy transfer (BRET) to study agonist-induced coupling of luciferase-tagged HCA receptors to Venus fluorescent protein-tagged G protein heterotrimers or arrestins. Our results indicate that the three HCA receptors couple to the Gαi/o subfamily of G proteins. The data additionally confirms a lack of coupling to the other G protein subfamilies (Gαs, Gαq, and Gα12/13), and lacks evidence of arrestin recruitment to HCA receptors. Overall, our study highlights the use of BRET as a powerful tool for analysis of GPCR signaling and demonstrates its possible use for future studies to determine the potency of potential drugs targeting HCA receptors as a therapy for health-related problems such as obesity.
    • Fast- Track Approach Following Heart Surgery in Infancy and Early Childhood: Outcome Analysis and Predictors of Failure

      Esquivel, Raquel; Department of Biological Sciences (Augusta University, 2020-05)
      Early extubation (EE) has become a critical determinant in perioperative management following congenital heart surgery (CHS) during early childhood. Fast track (FT) strategies and EE, when feasible, can have beneficial effects on clinical outcomes. The authors sought to determine the impact of EE on clinical outcomes, total hospital costs, identify predictors of failure and suggested criteria for new patients. A retrospective chart review of children ≤6 years old (n=64) who underwent CHS between January-December 2017 was performed. EE was defined as successful removal of the endotracheal tube in the operating room or upon arrival in intensive care unit (ICU). Groups were identified as (A):EE/Fast track and (B):no EE. Determinants for EE failure were assessed, and cost analysis pursued. The authors found 39 patients with EE compared to 25 that were not. Children who were EE (mean=6.795 days, sd= 4.250) spend significantly less (p < 0.0001) overall time in the ICU compared to non-EE patients (mean=19.960 days, sd=13.081). The authors also found that the total hospital stay for patients who were EE (mean=6.976 days, sd=4.090) was significantly reduced compared to those who were not (mean=21.783 days, sd=13.450) (p <0.0001). Furthermore, the authors found that children who were EE had a significant reduction (p <0.0001, sd= 23,196.203) in total hospital cost than patients who were not EE. Based on our analysis, we concluded that EE is feasible following CHS during early childhood but requires team approach and thoughtful use of FT protocols.
    • How Do Culturally Constructed Identities Influence Musical Preference?

      Ramos, Lindsay; Department of Communication (Augusta University, 2020-05)
      Through surveys and interviews, this study takes an interdisciplinary approach from both the fields of anthropology and communication to analyze music preference in relation to aspects of identity that are culturally constructed, specifically sex, race, and sexuality. Initially, my research was solely focused on the anthropological aspects of this topic; however, having studied both disciplines, once the data collection began it became clear that the theories and practices can be intertwined, and both are needed in order to fully explain this study. While my research is based on theories and concepts in these fields that will be discussed below, my study sheds light on aspects of this topic that have not been widely studied before,specifically the roles that intersectional, personal, and collective identities play in music choice as well as stereotypes.
    • The Historical Emergence of Art Therapy, its Modern Day Usage, and Possible Alternative Application

      Owen, Connor; Department of Communication (Augusta University, 2020-05)
      This paper focuses on the evolution of art therapy from the founding of the field of Psychology under Freud to the current day’s use in a variety of populations. Art therapy is defined by the American Art Therapy Association as “an integrative mental health and human services profession that enriches the lives of individuals, families, and communities through active art-making, creative process, applied psychological theory, and human experience within a psychotherapeutic relationship.” (AATA, 2020) Art therapy falls within the field of client or patient based approaches with the addition of creative work. The paper also addresses a need in research surrounding effectiveness for college-aged students. It is important to note that in the research there appears to be two broad thought currents that descend from Jung’s research and reconvene as modern art therapy, leading to differing opinions on its path from conception to modern use. The paths are one of theoretical philosophy based interventions and practical handson approaches ultimately both utilizing art making as therapeutic practice.
    • Diabetes Attenuation of the Estrogen-Mediated Increase in Endothelial Function is Associated with Circulating SIRT1

      O’Bryant, Sinéad; Department of Biological Sciences (Augusta University, 2020-05)
      Cardiovascular disease (CVD) is the most prominent killer within Type 1 Diabetes (T1D) with endothelial dysfunction as a major player in the development of CVD. Women with T1D experience an accelerated CVD risk despite the apparent sex-specific cardioprotection from circulating endogenous estrogen experienced by heathy pre-menopausal women. Animal models have shown the modulation of SIRT1, a NAD+ histone deacetylase, by estrogen as a CVD protector. This study sought to test the hypothesis that lower circulating SIRT1 is associated with reduced endothelial function in T1D women. Change in flow mediated dilation (FMD), a clinical measure of endothelial function, and SIRT1 over the menstrual cycle exhibited contrasting trends between T1D women and healthy women: increases of FMD and SIRT1 as estrogen increases in healthy women and decreases of FMD and SIRT1 as estrogen increases in T1D women. This provides evidence that signaling roles by circulating estrogen may be attenuated in T1D and that the effects of decreased SIRT1 contributes to endothelial dysfunction, resulting in determinant effects on vascular health in T1D women.
    • LGBTQ+ College Student’s Well-being and Physical Activity

      Nix, Dalanie; Department of Kinesiology (Augusta University, 2020-05)
      Many college students experience a downswing in mental well-being once beginning college. Studies have shown that the mental well-being of many college students are negatively impacted by alcohol consumption, cigarette use, and lower grades. Along with those factors, poor sleep habits are also linked to poor performance and overall well-being of students. LGBTQ+ college students experience discriminatory stressors, such as bullying, compounded with the stressors of college life which can lead to increased rates of anxiety, depression, and suicidal thoughts. Physical activity has been shown to improve well-being and depression symptoms. It has also been proven to be as effective as psychological and drug therapies. Many LGBTQ+ college students are turned away from sports due to LGBTQ+ cultural norms, as well as bullying from peers. This project employed a case study narrative approach of LGBTQ+ college students. 5 participants, ranging from 18-21 years of age and various sexual orientations, were interviewed about how physical activity has affected their well-being. We predict that LGBTQ+ college students who participate in regular physical activity will express lower levels of anxiety and depression along with greater levels of well-being.
    • Identification of the CAP1-Binding Domain of Adenylyl Cyclase 3 in Humans

      Gunby, Kimberly; Department of Biological Sciences (Augusta University, 2020-05)
      My research is aimed at finding the cyclase-associated protein 1 (CAP1) binding domain on human adenylyl cyclase 3 (AC3). Previous studies in our lab show that the interaction between CAP1 and AC3 inhibits migration and invasion of pancreatic cancer cells. The inhibitory mechanism is thought to involve the binding of AC3 and CAP1, causing the inhibition of globular-actin polymerization needed for filopodia formation and cell motility. A better understanding of this interaction will help facilitate the discovery for drugs that inhibit the migration and invasion of pancreatic cancer cells. To locate the binding region, we constructed mutants of WT AC3 plasmid using a Site-Directed Mutagenesis kit. We substituted a highly conserved proline residue at position 307 for an arginine residue (P307R) and a glutamate residue at position 308 for an alanine residue (E308A). The mutations were confirmed by sequencing. We then transfected pancreatic cancer cell line PANC-1 with WT and mutant AC3 plasmids and confirmed the expression using Western-blotting. To test whether the mutated AC3 could still interact with CAP1, we performed co-immunoprecipitation. We found that the residues proline and arginine in AC3 are not required for the interaction with CAP1. Further substitutions of other conserved residues are underway.
    • CBD Analysis in Oils and Foods

      Foley, Joanna; Department of Chemistry and Physics (Augusta University, 2020-05)
      Cannabidiol (CBD) has become a very prominent topic in the medical community and popular marketplace because of its widespread consumer use. Tetrahydrocannabinol (THC) and other similar molecules can be present in commercial CBD products, so testing is necessary to determine the presence of the CBD. Existing methods of analysis for CBD oils are only known on GC-FID (gas chromatography – flame ionization detector) and these methods are not optimal for the wide variety of commercial CBD products available. Thus, a GC-MS (mass spectroscopy) method, based on a published GC-FID method, was created to optimize the detection of CBD because not only separation but also identification can be obtained. This method can be applied to a wide variety of foods, gummies, and other items that may contain CBD and similar molecules. The method has been optimized by varying GC column temperature, and sample preparation, to find a balance between analysis time, analyte detection, and resolution for the various types of cannabinoid molecules present in commercial CBD oil samples. The optimized method was able to determine that a 1:3 ratio of oil to solvent gave optimal signal of all CBD oils tested. The optimized method was then tested on a variety of commercial and self-prepared CBD edibles to determine that CBD was still present and was not degraded into THC.