• A Study of Human Skin Color, a Natural Sunscreen: Physiology, Molecular Evolution, Public Health and Student Learning

      Ayala, Juan; Mukhopadhyay, Soma; Biological Sciences; Mukhopadhayay, Soma; Augusta University (1/30/2020)
      Human skin coloration is a combination of pigmentation, ultraviolet (UV) exposure, gene expression and natural selection. Skin tone is also associated with several physiological processes, such as vitamin D synthesization, calcium homeostasis, maintaining proper blood folate concentration, and the production of serotonin. In recent years, the study of molecular evolution has become very significant not only to understand the human body but also becoming an integral part for understanding public health and other fields of medical science. Our goal of this project was to create an interactive course module for Anatomy and Physiology students to show how skin physiology was driven by evolutionary pressures. Also, the module was intended to show how exposure to some UV radiation is important for certain biological processes and to offer protection against cancer and on the other hand how overexposure might cause damage and lead to cancer. Students were introduced to molecular evolution of skin color and the production of different pigments, eumelanin and pheomelanin to shield DNA from harmful UV light. Additionally, UVAB and UVC irradiance were measured and compared to the UV index which indicates the strength of UV radiation for the day to make people aware of the environmental factors arou
    • ATAD3A: a critical driver for head and neck cancer

      Caleb Jensen; Yong Teng; Liwei Lang; Biological Sciences; Yong Teng; Augusta University (1/15/2020)
      For patients with head and neck cancer whose tumors are HPV negative HPV(-), current therapy does not lead to significant longevity and most succumb to loco-regional recurrence of the primary tumor. We discovered that HPV(-) head and neck squamous cell carcinoma (HNSCC) highly expressed ATPase family AAA-domain containing protein 3A (ATAD3A). ATAD3A is the mitochondrial protein, which has been demonstrated as an oncogene in breast and lung cancer. However, nothing has been reported regarding its role in HNSCC. Using the HPV(-) HNSCC cell line HN12 as a cell model, we show here that knockout of ATAD3A expression by CRISPR-CAS9 in HNSCC cells, leading to reduced cell proliferation and decreased the ability of colony formation and anchorage-independent growth in soft agar. Importantly, ATAD3A loss also significantly suppressed HNSCC cells to grow in 3D culture. Together, these findings suggest the potential oncogenic role of ATAD3A in HNSCC cells, and implicate that ATAD3A represents a promising target for better treatment of patients with HPV(-) HNSCC.
    • Characterization of a Cyclic Peptide AD05 as a Novel Inhibitor of the Hsp90 Chaperoning Machine

      Fang, Wayne; Lu, Sumin; Jilani, Yasmeen; Debbab, Abdssamad; Chadli, Ahmed; Debbab, Abdssamad; Biological Sciences; Chadli, Ahmed; Augusta University; Heinrich Heine Universität Düsseldorf (1/31/2020)
      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.
    • Characterization of Proton Sensitive G protein-Coupled Receptors

      Nam, Alisha; Okasha, Najeah; Spencer, Angela; Lambert, Nevin; Biological Sciences; Department of Pharmacology and Toxicology; Chemistry and Physics; Spencer, Angela; Augusta University (1/31/2020)
      G protein-coupled receptors (GPCRs) are membrane-bound receptors that can stimulate an intracellular signaling pathway following activation by a ligand. According to the International Union of Basic and Clinical Pharmacology (IUPHAR) database, GPR4, GPR65, and GPR132 are Class A orphan GPCRs with protons reported as their putative endogenous ligand; however, these receptors are currently understudied. After confirming whether these receptors are pH-sensitive, the purpose of our study was to investigate the interactions between GPR4, GPR65 and GPR132 and G protein subtypes (G?s, G?i, G?q, and G?12) upon stimulation with an acidic solution. Using bioluminescence resonance energy transfer (BRET), we studied the coupling between luciferase-tagged GPR receptors and fluorescent protein (Venus)-tagged G proteins in response to pH changes. Data indicated that all three receptors responded to pH changes. Upon extracellular response to pH changes, the receptors activate different G protein subtypes and thus, different signaling pathways: GPR4 activates G?i, G?q, and G?12; GPR65 activates all four subtypes; and GPR132 activates G?i�and weakly activates G?q, and G?12. Identifying these receptors as true proton sensors leads the way in understanding the role they play in maintaining acid-base homeostasis and will be critical for the development of novel drugs combatting acid-base related disorders.

      Duncan, Leslie; Jensen, Caleb; He, Leilei; Lang, Liwei; Teng, Yong; Biological Sciences; Oral Biology and Diagnostic Sciences; Georgia Cancer Center; Teng, Yong; Lang, Liwei; et al. (1/16/2020)
      A promising arsenal of histone deacetylase (HDAC)-targeted treatment has emerged in the past decade, as the abnormal targeting or retention of HDACs to DNA regulatory regions often occurs in many cancers, including head and neck squamous cell carcinoma (HNSCC). However, few has been studied regarding the beneficial role of HDAC inhibition in anti-HNSCC therapy and the underlying molecular mechanisms remain elusive. Epidermal growth factor receptor (EGFR) is commonly expressed at high levels in HNSCC (more than 90%) and serves as a prime target for new anti-HNSCC therapy. Interestingly, Trichostatin A (TSA), one of HDAC inhibitors, not only inhibits EGFR phosphorylation, but also induces repression of EGFR total protein amount in HNSCC cells. We further show that TSA induces EGFR degradation through the ubiquitin-proteasome pathway in HNSCC cells, which is associated with downregulated AKT and ERK1/2 signaling pathways. The study uncovers that EGFR is one of targets of HDAC-based treatment, providing mechanistic insight into the action of HDAC inhibitors. As there is an increasing interest in using HDAC inhibitors for cancer treatment in the clinic, the outcomes from the present study would be significantly beneficial for the development of new rational HDAC-targeted anticancer modalities.
    • Diagnosis of Mental Illness in the Narrator of Charlotte Gilman's "The Yellow Wallpaper" Using the DSM-5

      Fang, Wayne; Sadenwasser, Tim; Biological Sciences; English and Foreign Languages; Sadenwasser, Tim; Augusta University (12/10/2019)
      Charlotte Gilman's "The Yellow Wallpaper" explores mental illness, freedom, and the faults of the rest cure by exploring the life of a wife who has been diagnosed with neurasthenia. With this story Gilman describes an increasingly common practice during her time, and how problematic it was for the individuals who were diagnosed. Through the wife's narration, Gilman shows how many women felt trapped since they were forced to undertake the rest cure due to one-sided relationship dynamics. Using this narrative of the wife's deteriorating mental health, Gilman argues for equality in relationships as well as better treatments for mental health. In this presentation, I will use the Diagnostic and Statistics Manual of Mental Disorders (DSM-5) as well as other scholarly sources to diagnosis the wife's mental illness. To do this, I will take the wife's narration and compare it to diagnostic criteria as presented in the DSM-5. By examining the narrator's thoughts and actions I will be able to examine the progression of her mental illness. Examining the wife's mental health can show how many women of her may have felt trapped. This in turn can explain how many women faced unequal power dynamics in their marriages.
    • Effect of NF-κB Deletion on Bone Marrow Macrophage Respiratory Burst Ability

      Soni, Karan; Bradford, Jennifer; Biological Sciences; Bradford, Jennifer; Augusta University (1/31/2020)
      The nuclear factor-kappaB (NF-κB) signaling pathway is very important in normal immune system function and is also often aberrantly regulated in many different types of cancers. As many cancers are characterized by elevated numbers of infiltrating monocytes/macrophages, we have developed an animal model that lacks canonical NF-κB signaling in bone-marrow derived macrophages (BMDMs). As BMDMs can infiltrate solid cancers, the aim of this particular study was to assess the functionality of phagocyte oxidase ability in NF-κB deficient BMDMs. A respiratory burst assay involves stimulating the phagocyte oxidase enzyme in macrophages to release reactive oxygen species (ROS) so that they can degrade and combat invading pathogens as well as cancer cells. Based on our recent experiments that showed BMDMs lacking p65 had poor phagocytosis ability and low nitrite production, we hypothesize that BMDMs lacking NF-κB signaling will have a decreased respiratory burst response compared to control BMDMs.
    • Establishing a GFP Marker in Zebrafish to Study the Localization of Tinagl1

      Blackburn, Helena; LeMosy, Ellen; Biological Sciences; Department of Cellular Biology and Anatomy; LeMosy, Ellen; Augusta University (1/27/2020)
      Tinagl1 is a secreted protein found in the basement membrane under epithelial cells. The LeMosy lab previously showed that tinagl1 knockdowns resulted in abnormal spinal development and heart orientation during zebrafish development. These data, together with changes in length of motile cilia, suggested that tinagl1 is involved in cilia function during development. The mechanism of this interaction is unknown, and it is unclear whether Tinagl1 is only in basement membranes at the basal side of cells, or if it also localizes to the apical side of cells where most cilia project. A deeper understanding of the localization of Tinagl1 during development is a logical next step in understanding how this protein functions. Zebrafish provide an excellent model for studying this localization as they display strong phenotypic effects that can be easily imaged. The localization of Tinagl1 will be tracked using a Tinagl1-GFP fusion construct developed through PCR and insertion into a Tol2 transposon vector. This construct will be injected into early embryos together with transposase mRNA to create mosaic fish showing Tinagl1-GFP in selected tissues. Successful germline integration of the tinagl1-GFP DNA will lead to the development of a transgenic line of zebrafish allowing imaging of Tinagl1 localization during development.

      Hitchens, Samantha; Saul, Bruce; Biological Sciences; Saul, Bruce; Augusta University (1/31/2020)
      Invasive wild pigs (Sus scrofa) have a destructive impact across the world. The variety of cultures affected make the development of more effective and diverse management methods vital. Although wild pigs are often hunted with dogs, this method is not suitable or legal in all areas. Considering this, and pigs highly developed sense of smell, the following hypothesis was developed: Can a natural scent function as a satisfactory pig repellant? To test our hypothesis, we attracted wild pigs into areas baited with corn, and performed separate trials by adding hair from four different mammal species (dog, cat, horse and coyote). Our experimental design forced pigs to interact with the hair before consuming the bait. Trail cameras monitored each location over a five month period and wild pig behaviors were recorded. The presence and absence of pigs throughout the study trials was analyzed and compared with images captured during the control trials (corn only). Image totals were evaluated to determine if the hair prevented pigs from entering any areas, and the duration of any absences was noted. The results supported our hypothesis that a natural scent (dog hair) can decrease wild pig activity and potentially serve as a repellant.

      Gunby, Kimberly; Sabbatini, Maria E.; College of Science and Mathematics; Biological Sciences; Sabbatini, Maria; Augusta University (1/30/2020)
      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.
    • Investigating Signaling Pathways Involving the HCA Receptor Family

      Saj, Dalia; Spencer, Angela; Okashah, Najeah; Lambert, Nevin; Biological Sciences; Chemistry and Physics; Department of Pharmacology and Toxicology; Spencer, Angela; Augusta University (1/31/2020)
      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 Gi/o�subfamily of G proteins. The data additionally confirms a lack of coupling to the other G protein subfamilies (Gs,�Gq,�and G12), 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.

      Gaw, Victoria; Glenn, Manderrious; Cannon, Jennifer; Biological Sciences; Cannon, Jennifer; Augusta University (1/31/2020)
      Perfluorooctanoic acid (PFOA) is a synthetic chemical belonging to a larger group of fluorotelomers. These compounds have been used in the production of both industrial and consumer products as surfactants and are environmentally persistent pollutants. While the long-term effects of PFOA are largely unknown, there is increasing evidence suggesting it to be an endocrine disruptor. Studies have shown that PFOA binds to and activates peroxisome-proliferator-activated receptor ? (PPAR?), which can regulate the expression of other genes and receptors. Previous experiments in our lab demonstrated that PFOA treatment of MCF-7 breast cancer cells (an ER?-positive cell line) decreased expression of ER? mRNA and protein, and decreased cell viability by ~20% within 48h of treatment compared to DMSO controls. However, these cells were treated in the absence of fetal bovine serum (FBS).� When we repeated these experiments without serum withdrawal, we initially noted a tendency towards increased proliferation in MCF-7 cells treated with 50�M and 100�M PFOA at both 24h and 48h compared to control. To further examine the role of ER? in this PFOA-induced proliferation, we carried out additional experiments in MCF-7 cells along with experiments in another ER?-positive cell line, T47D, as well as an ER?-negative cell line, MDA-MB-23.
    • Salty or Slightly Salty: Is Fish Species Richness affected by an obsolete navigational cut

      Patterson, Rebecca; Matthews, Loren; Reichmuth, Jessica; Saul, Bruce; Mathews, Loren; Biological Sciences; Biology; Reichmuth, Jessica; Augusta University; Georgia Southern University (1/30/2020)
      As rivers flow toward the coast, freshwater mixes with saltwater in estuaries. The mixing here creates a wide range of environments for many organisms. The Satilla River Estuary has been cut eight times, which has altered the salinity gradients that are a result of natural tidal flow. Altered salinity gradients pose a threat to migratory fish species because they are no longer able to pick up on directional cues these gradients provide, ultimately affecting species richness in the estuary. The purpose of this study is to determine if Noyes Cut has affected salinity gradients at five collection sites in Umbrella and Dover Creeks. Experimental gill nets were set one hour before max flood tide and soaked for two hours.� All fish were identified to species with total and fork lengths measured to the nearest centimeter. Noyes Cut and Parsons Creek had the most species richness while River Marsh Landing and Todd Creek experienced the lowest diversity.� We believe these large salinity fluctuations are due to a sediment deposit that blocks water flow as a result of Noyes Cut. When Noyes Cut is closed, we expect fish to redistribute into Umbrella and Dover Creeks as a result of restored salinity gradients.�

      Miranda Henderson; Canela, Jenelly; Fischer, Jeff; Reichmuth, Jessica; Biological Sciences; Fischer, Jeff; Reichmuth, Jessica; Augusta University (1/30/2020)
      Microsporidia are spore-forming obligate intracellular parasitic fungi that infect eukaryotic organisms. They are ubiquitous in nature and infections occur worldwide in terrestrial and aquatic hosts. Some species of Microsporidia have been shown to infect the hepatopancreas of shrimp, which may affect their ability to obtain nutrients, stunt their growth, and increase their susceptibility to additional diseases. Microsporidiosis in shrimp has been shown to negatively impact the commercial shrimp industry, resulting in great economic loss specifically to the state of Georgia since this fishery is the largest and most lucrative. This study was conducted to evaluate the prevalence of microsporidia in shrimp from the Satilla River Estuary in Georgia because of man-made cuts that have altered water quality conditions that could affect shrimp health specifically. Shrimp were caught at four collection sites using 6.1m (20ft) otter trawls and cast nets and were transported on ice back to the lab where they were frozen until dissection. Using bright-field light microscopy and a previously established staining technique, microsporidian spores were detected in hepatopancreas� extracts in greater than 30% of the shrimp analyzed.�

      Thomas, Eyana; Lian, Eric; Roberts, Kimberly; Young, Lufei; Biological Sciences; Department of Medicine; College of Nursing; Georgia Highlands College; Young, Lufei; Augusta University (1/31/2020)
      Patients with diabetes are two to four times more likely to die from cardiac diseases than other without this diagnosis. Cardiac rehabilitation (CR) is known to improve the physical functioning, reduce risk factors in cardiac patients with diabetes. The aim of this study is to evaluate if cardiac patients with comorbidity of diabetes had improved physical functioning (measured by six-minute walk test [6MWT]) and reduced risk factors (measured reduced fasting blood sugar, lipid profile, overweight, fat composition) after the cardiac rehabilitation program. A retrospective observational cohort longitudinal study using secondary data from electronic medical records was conducted. Clinical data were collected from the individual cardiac treatment plan form used by a cardiac rehabilitation center. Among 93 patients, improvement in physical functioning (i.e.: six-minute walk distance, and METs) was significant (p-value = <.01). This was done by comparing pre and post 6MWT scores.