• ADIPOSE HDAC9 DELETION PROTECT AGAINST DIET INDUCED OBESITY IN MICE THROUGH REGULATING ENERGY EXPENDITURE

      Hassan, Nazeera; Zarzour, Abdalrahman; Department of Biological Sciences; Department of Medicine; College of Allied Health Sciences; Kim, Ha Won; Weintraub, Neal; Augusta University (2019-02-13)
      Our group has previously identified histone deacetylase 9 (HDAC9) as a regulator of adipocyte differentiation, and its expression levels were elevated in diet induced obese (DIO) mice.� We also reported that global HDAC9 deletion protected mice against DIO through promoting beige adipogenesis. Here, we hypothesized that adipose HDAC9 correlate with human obesity similar to murine models, and its deletion is sufficient to protect against DIO. To test this hypothesis we crossed HDAC9 floxed mice with adiponectin-cre mice to generate adipose-specific HDAC9 knockout mice (AdipCre-HDAC9), which exhibited 30% less weight gain when fed high fat diet compared to control despite increased food intake, in association with increased energy combustion & O2 consumption, improved insulin sensitivity and glucose tolerance. However, unlike global HDAC9 deletion, this was not associated with increased beige adipogenesis nor increase in brown adipose tissue function. Interestingly, AdipoCre-HDAC9 mice fed normal chow diet didn�t exhibit altered energy expenditure nor weight differences when compared to littermate controls. These finding suggest that adipose HDAC9 regulate energy expenditure in response to high fat diet and can be a promising therapeutic target to combat obesity.
    • APOROSA OCTANDRA: STUDY THE PROTECTIVE EFFECTS OF ITS BARK EXTRACT AGAINST D-GALACTOSE INDUCED COGNITIVE IMPAIRMENT AND OXIDATIVE STRESS IN MICE AND ITS PHYTOCHEMICAL INVESTIGATION

      Schinder, Sonya; Department of Chemistry and Physics; Panda, Silva; Augusta University (2019-02-13)
      Aging is a multifarious natural process, linked with several biochemical and morphological variations in the biological system. Aging not only challenges the increased vulnerability as well as homeostasis network to the cognition and locomotion but also to physical, mental or social activities. Medicinal plants have been used since ancient time to cure and prevent various diseases. Several natural compounds such as isoflavones, anthocyanins, and catechins isolated from plant sources act as a potent antioxidant against ROS (Reactive Oxygen Species).�Antioxidants, especially natural antioxidants are recommended for the prevention of aging. In this study, we utilized an unexplored traditional medicinal plant�Aporosa octandra�(Buch.-Ham. ex D. Don) that�belongs to the family�Euphorbiaceae,�sub-family Phyllanthaceae that is shrub to tree, up to 15 m high and comprises of 50 species, which are distributed throughout Asian regions. This plant is enlisted as a medicinal plant and is used for centuries in the Ayurvedic system. We investigated phytochemical contents of the plant and evaluated the biological activity.
    • CHARACTERIZING THE ROLE OF PANCREATIC STELLATE CELLS IN THE TRANSITION OF CHRONIC PANCREATITIS TO PANCREATIC CANCER

      Godoy, Catalina; Department of Biological Sciences; Department of Pharmacology & Toxicology; Csanyi, Gabor; Sabbatini, Maria; Augusta University (2019-02-13)
      Background- Chronic pancreatitis (CP) and pancreatic cancer are two diseases that share a mutual histological feature known as fibrosis produced by pancreatic stellate cells (PaSCs). In response to pancreatic inflammation, PaSCs are activated from quiescent phenotype into myofibroblast-like cells, which express extracellular matrix components. PaSCs are also known to facilitate the migration and invasion of pancreatic cancer cells, which are accompanied by increased matrix metalloprotease (MMP) production and epithelial-to mesenchymal transition (EMT). NADPH oxidase (Nox) is a family of enzymes that catalyze the transfer of an electron from NAD(P)H to oxygen to generate superoxide or hydrogen peroxide. Because Nox1 is expressed in PaSCs, the objective of this study was to assess the extent to which Nox1 in PaSCs facilitates the migration and invasion of pancreatic cancer cells by regulating the expression of MMP and genes involved in EMT. Results/Discussion-We found that the lack of Nox1 lowers the expression of MMP-9 mRNA and the EMT-induced gene Snail in PaSCs. Further studies need to be done in PaSCs from mice with CP and CP-associated oncogenic KRas-driven pancreatic cancer.
    • DESIGN AND SYNTHESIS OF HYBRID CONJUGATES AS POTENTIAL ANTI-INFECTIVE AGENTS

      Littlefield, William E; Wade, Margaret; Makkanal, Tina; College of Science and Mathematics; Department of Chemistry and Physics; Panda, Silva; Augusta University (2019-02-13)
      Infectious diseases caused by pathogenic microorganisms are major challenges despite all the steps taken to control or cure. New drug development with high efficacy/selectivity for infectious diseases is a point of interest for many researchers. It has reported that tuberculosis is one of the ten major causes of death in the world. Multi-drug resistance (MDR) is another major concern in bacterial and fungal infections. The present study deals with the development of new conjugates of pyrazinoic acid and isoniazid linked via an amino acid. The synthesized conjugates show promising and interesting results against a variety of microbial strains, tuberculous and non-tuberculous mycobacteria. Molecular modeling studies were used for understanding and validation of the experimental data.
    • DESIGN AND SYNTHESIS OF QUINOLONE-TRIAZOLE CONJUGATES AS POTENTIAL ANTIBACTERIAL AGENTS

      Honkanadavar, Hitesh H; Tran, Queen; Department of Chemistry and Physics; Panda, Silva; Augusta University (2019-02-13)
      Quinolones are one of the most important synthetic antibacterial agents have been widely used in the treatment of diverse infections including urinary tract, respiratory and bone joint as well as sexually transmitted diseases, prostatitis, pneumonia, and acute bronchitis. However, quinolone resistance increases towards many Gram-negative and Gram-positive species. Molecular conjugation has been known for the rational design of new biologically active entities by fusion of compounds and/or pharmacophores recognized and derived from known bio-active molecules. The present study directs towards the construction of novel quinolone-triazole conjugates and investigation of their antimicrobial properties. The detail results will be discussed at the conference.
    • DEVELOPMENT OF TRANSGENIC ZEBRAFISH MODEL FOR INVESTIGATION OF THE FUNCTION OF MICROGLIA

      Sura, Survasha; Department of Biological Sciences; Department of Biochemical and Molecular Biology; Georgia Cancer Center; Rajpurohit, Surendra K; Augusta University (2019-02-13)
      Zebrafish have emerged as a powerful model organism for elucidating the development and function of microglia. Generation of new transgenic reporter lines and imaging tools strengthen the zebrafish model in microglia study�in-vivo. The aim is to develop a novel compound transgenic line to study the inflammatory process mediated by NF-kB in microglia cells. This novel compound transgenic line will establish a new model for microglia study. To generate the novel compound zebrafish transgenic model for microglia, we are crossbreeding microglia transgenic line zebrafish (Tg(mpeg1:mCherry) with the NF-kB Tg(6xNFkB:EGFP) transgenic progeny. We first generate a heterozygous F1 progeny which will be bred to generate an F2 homozygous progeny. Once the F1 progeny of the Microglia-NfkB transgenic line is developed, they will be crossbred to develop the Homozygous compound transgenic line. Fluorescent Microscopy will be used to screen the larvae generated from the breeding events. By developing the compound transgenic line, we are optimizing microglia isolation and sorting methodology by using the related antibodies as the marker. The NF-kB microglia transgenic line will provide a unique platform for drug screening to address microglial based ailments, thus furthering the understanding and treatment of human disease.
    • ECOTOXICOLOGY OF YELLOW-BELLIED SLIDERS (TRACHEMYS SCRIPTA) AND MUSK TURTLES (STERNOTHERUS ODORATUS) IN NATURAL WETLANDS

      Hammesfahr, Rachel; Department of Biological Sciences; Cromer, Robert; Augusta University (2019-02-13)
      Glyphosate is one of the active ingredients in many different herbicidal products such as Roundup. Preliminary research has suggested that glyphosate is a possible endocrine disruptor, can cause developmental defects, and is a potential carcinogen. Due to its potential harmful effects on different organisms, we seek to monitor the levels of glyphosate in wetland areas. This will be done by analyzing samples from two commonly found indicator species, the yellow-bellied slider turtle,�Trachemys scripta, and the musk turtle,�Sternotherus odoratus. Research will be done on turtles caught at Reed Creek Nature Center and Brick Pond Park. Physical measurements will be taken, and blood will be drawn from each turtle. Analysis of the glyphosate levels in the blood samples will be completed using a glyphosate ELISA kit. While this research will not prove that glyphosate has harmful effects on the turtles, it will quantify the amount of the chemical present. If there are high concentrations, this will indicate a need for more research on how glyphosate affects different organisms so long-term effects on the environment can be estimated.
    • EFFECT OF GABAERGIC NEURONS IN SUSCEPTIBLE VERSUS RESILIENT MALE RATS TO PTSD

      Dixon, Rachel; Mandavilli, Rohan; Department of Phychological Sciences; Department of Biological Sciences; Departmetn of Pharmacology & Toxicology; Bunting, Kristopher M; Alexander, Khadijah; Vazdarjanova, Almira; Augusta University (2019-02-13)
      Post-traumatic stress disorder (PTSD) is a psychological disorder that can occur after a traumatic event. Individuals with PTSD exhibit extreme anxiety and learning and memory difficulties. Once exposed, 12-35% develop PTSD with women twice as likely to be affected than men. Our goal is to discover underlying mechanisms to prevent PTSD, as we investigate the relevance of glutamic acid decarboxylase positive (GAD+) neurons on susceptible (SUS) and resilient (RES) male rats. SUS and RES phenotypes were assessed using the highly advanced RISP protocol to reveal susceptibility to a PTSD-like phenotype. The increase of GAD+ cells in the medial prefrontal cortex (mPFC) informs us that more GABAergic neurons are present, which can cause inappropriate recall. We will be examining if there is a difference in the number of GAD+ cells in the RES versus SUS male rats. To investigate, we used cryosectioned brains from SUS or RES rats. The brains were stained using immunohistochemistry to isolate the GAD+ neurons in the mPFC and were counted. The results of this experiment will be determined and examined at a later date closer to our presentation. We expect to see a SUS male rats to have a higher number of GAD+ neurons.
    • THE EFFECTS OF ACUTE AND CHRONIC KETOSIS ON GOLF ACCURACY

      Blume, Grant; Department of Kinesiology and Health Science; Holland, Maleah; Augusta University (2019-02-13)
      Ketones, molecules produced as a byproduct of fat breakdown by the body, can be used as an alternate source of energy when glucose supplies are low. Several studies have shown that ketones may help with cognitive recovery and motor learning. This study examined the effects of both acute and chronic ketone supplementation on golf putting accuracy in middle-aged adults to determine if elevated circulating ketone levels improved accuracy. The results demonstrate no significant differences between the putting accuracy of the ketone group or placebo group. Data collection for this study will be continued to determine if a measurable difference can be seen with more data.
    • EFFECTS OF CHRONIC ALOHOL AND GLUCOSE EXPOSURE ON VIABILITY OF ALVEOLAR MACROPHAGES

      Keller, Elizabeth; College of Science and Mathematics; College of Allied Health; Curry-McCoy, Tiana; Thomas, Amanda; Augusta University (2019-02-13)
      The adverse health risks associated with alcohol abuse and obesity are widely known by the general population. Although lesser known, studies have presented the lungs as secondary organs affected by such lifestyle factors. Healthy lungs are protected against infection and harmful airborne particles by macrophages, the working entities of the immune system which fight potential sources of infection. When these immune-responsive cells are compromised and unable to perform their functions, lung health may deteriorate. Therefore, a healthy pulmonary alveolar macrophage population is vital for adequate lung function. Chronic alcohol abuse and obesity have been shown to suppress alveolar macrophage function, thus lowering the lungs� first line of defense. The objective of this study is to determine the effects of exogenous ethanol and increased glucose concentration on macrophage size and viability via an�in vitro�study on NR8383 rat alveolar macrophages. The study measures macrophage viability under treatment conditions.
    • THE EFFECTS OF CIRCULATING ESTROGEN ON SIRT1 LEVELS IN PREMENOPAUSAL WOMEN

      O'Bryant, Sinead; Department of Biological Sciences; Harris, Ryan; Augusta University (2019-02-13)
      According to epidemiological data, healthy premenopausal women exhibit greater protection from cardiovascular disease (CVD) when compared to men of a similar age. �It has been hypothesized that estrogen, one of the primary female sex hormones, is responsible for this protection. Previous studies have shown that the nicotinamide-adenine-dinucleotide-(NAD+)-dependent histone-deacetylase Sirtuin 1 (SIRT1) provides protection from hyperglycemia, metabolic and endothelial dysfunction, all which contribute to CVD development. �The aim of this analysis was to show the potential relationship between circulating endogenous estrogen and SIRT1 in premenopausal women and its association with HbA1c, an indicator of glycemic state.� By utilizing the menstrual cycle, when estrogen is at its highest range during the follicular phase and at its lowest during meneses, the effects based on circulating estrogen on SIRT1 concentrations were evaluated in women by ELISA of participant blood plasma. �The results show SIRT1 and circulating estrogen have a significant positive correlation, and SIRT1 and HbA1c have a trending negative correlation. It was also seen that SIRT1 concentrations were found to be protected against high HbAlc levels when endogenous estrogen was high, providing evidence that circulating estrogen may act as a mediator of SIRT1, functioning as a protective mechanism from CVD in premenopausal women.
    • EFFECTS OF VITAMIN D3 DEFICIENCY, VITAMIN D RECEPTOR KNOCKOUT, AND DIABETES ON CORNEAL EPITHELIAL NERVE DENSITY

      Vick, Sarah; Department of Biological Sciences; Department of Cellular Biology and Anatomy; Lu, Xiaowen; Watsky, Mitchell; Augusta University (2019-02-13)
      It is estimated that 41.6% of the US population suffers from vitamin D deficiency, with Blacks (82.1%) and Hispanics (69.2%) at even greater risk�Vitamin D deficiency can be caused by a variety of sources and given the wide range of causes, it is important to understand what measures this population might take to proactively prevent greater harm, or to reverse harm that might have already occurred. This project is designed to test the general hypothesis that Vitamin D deficiency exacerbates preexisting primary corneal pathologies. Previous research has established that the corneal epithelium in diabetic mice heals at a faster rate than the epithelium in diabetic vitamin D receptor (VDR) knockout (KO) mice. It is known that within diabetic mice, the corneal nerve density is decreased. However, it is unknown how VDR KO mice or vitamin D deficient with diabetes will affect corneal nerve density. In order to identify variabilities within the nerves that indicate slow wound healing, the mouse corneas will be collected, stained for confocal microscope observation, and analyzed through image processing to determine nerve density.
    • EVALUATION OF CANIS HAIR AS A POTENTIAL WILD PIG REPELLENT ON COWDEN PLANTATION, JACKSON, SC

      Hitchens, Samantha Rae; Department of Biological Sciences; Hull College of Business; Saul, Bruce; Augusta University (2019-02-13)
      Invasive wild pigs (Sus scrofa) continue to have a destructive impact across the world. The uncontrolled spread of these intrusive animals has affected many cultures, making it important to develop management methods across many locales. Wild pigs are often hunted with dogs; however, this method may not be suitable or legal in all areas impacted by this animal. In consideration of this fact, along with the knowledge that pigs have a highly developed sense of smell, led us to the hypothesis: Can a natural scent function as a satisfactory pig repellant? Based upon our past testing trials of potential scents in the Savannah River swamp near Jackson, SC, dog hair appeared to have a potential effect. We designed this experiment to attract wild pigs into an area baited with corn, and subsequently applied dog hair to the same area. Trail cameras were used at study locations to observe the normal patterns of wild pigs before and after dog hair applications. We analyzed our data by noting the presence and absence of pigs throughout our study trials. Image totals were also examined to determine if the dog hair dissuaded the pigs from entering the area. Our results supported our hypothesis.
    • AN EXAMINATION OF MORAL PANICS: HOW THE FEAR OF SATANISM AFFECTED TABLETOP ROLE PLAYING

      Williams, Travis; Department of Biological Sciences; Department of Communication; Department of Anthropology & Philosophy; Johnson, Edgar; McClelland-Nugent, Ruth; Augusta University (2019-02-13)
      Moral panics around youth entertainment have been an occurrence as long as culture has been established. As long as youth entertainment has values that can be seen as going against the established values of the preceding generation, a moral panic could take place. The purpose of this research was to analyze how moral panics centered on youth entertainment begin and gain traction. To do this, the research was focused on the 1980s moral panic around tabletop roleplaying games, specifically�Dungeons & Dragons. By tracing the origin of the moral panic to the fear of cults and occult from the 1970s, we can find more context as to why some individuals believed that role playing games could cause adolescents to use the games as a style of dangerous escapism or as a gateway to the occult. To further understand this moral panic, an analysis of some of the major detractors of role playing games was done, as well as researching the role the media played in cultivating the moral panic. With a greater understanding on how moral panics begin and gain traction, this research can be used to compare and contrast other moral panics around youth entertainment.
    • EXAMINING THE EFFECTS OF AETOKTHONOS HYDRILLICOLA EXTRACT ON OXIDATIVE STRESS IN C6 CELLS

      Ward, Kayla; Department of Biological Sciences; Wiley, Faith; Augusta University (2019-02-13)
      Aetokthonos hydrillicola, a species of cyanobacteria, has colonized an invasive species of hydrilla in the lakes of the Southeastern United States. This cyanobacterium is suspected to cause Avian Vacuolar Myclinopathy (AVM). AVM is a neurological disease that affect birds. Bald eagles and American coots have been primarily studied and known to be affected by AVM. Symptoms of AVM consist of brain lesions, loss of basic motor skills, and the disease often leads to death. Extracts of A. hydrillicola are toxic to C6 cells, and this cell line is used as a model to examine the mechanism of toxicity. The aim of this research project is to understand the role of oxidative stress in A. hydrillicola cytotoxicity and determine if antioxidant compounds may protect the cells. Common oxidative stress inhibitors, Gingko biloba extract and selenium, have been tested in different concentrations in order to determine if oxidative stress is present and preventable. These compounds did not prevent toxicity in the C6 cells exposed to the cyanobacterial extracts. The presence of oxidative is currently being further investigated using a 2-7 dichlorofluorescin diacetate (DCFH-DA) assay, which indicates the presence of reactive oxygen species.
    • FLATFISH ASSEMBLAGE AND ABUNDANCE DIFFERENCES BETWEEN THE SAINT CATHERINES ISLAND AND SATILLA RIVER ESTUARIES

      Bickle, Abigail; Coleman, Alex; Brown, Jason; Thiruvaiyaru, Dharma; Sethuraman, Sankar; Department of Biological Sciences; Department of Mathematics; Reichmuth, Jessica; Saul, Bruce; Augusta University (2019-02-13)
      St. Catherines Island is a pristine uninhabited barrier island off of the Georgia coast, and is believed to have experienced less anthropogenic alteration when compared to mainland estuarine habitats. In contrast, the Satilla River estuary has been historically affected by human activity, especially during the construction of navigational "cuts" through the marsh in the early 1900s. These cuts were used to support economic gain no longer in the area. Because of the differences in human influence, we hypothesize the fish assemblages will be different. We compared abundance of various resident flatfish species captured when trawling and using gill nets in these two systems. As benthic species, flatfish may be especially affected by anthropogenic disturbances of the estuarine substrates. This study provides insight into the effects of human disturbances on benthic fish species populations and assemblages. We compared catch-per-unit effort for six flatfish species between data among several sites, seasonally, between 2015 and 2018. We also looked at environmental variables when comparing abundance. The data are reflective of differences that exist in resident flatfish populations, and this condition could be explained by anthropogenic activities.
    • INVESTIGATING THE REQUIREMENT OF HOB1 ON THE SENSITIVITY OF�SCHIZOSACCHAROMYCES POMBE�AFTER EXPOSURE TO VARIOUS DNA DAMAGING AGENTS

      Qureshi, Arman; Department of Biological Sciences; Abdulovic-Cui, Amy L; Augusta University (2019-02-13)
      It is inherently important that when damaged, DNA is repaired efficiently and with high accuracy. BIN1 encodes a protein that plays a role in genomic stability, specifically in cell cycle regulation, chromatin remodeling, and DNA repair. Previous research has shown that the protein Bin1 exhibits an inhibitory role in the double strand break repair pathway of non-homologous end joining (NHEJ). The homolog of�BIN1,�HOB1, is found in the fission yeast,�Schizosaccharomyces pombe. To understand the role�HOB1�has on yeast survival after damage, two strains of�S. pombe, a wild type strain (WT) and a strain without�HOB1�(hob1?), were exposed to various DNA damaging agents. Each treatment introduced different types of DNA damage that require repair by different DNA repair pathways. These treatments included UV radiation, hydrogen peroxide treatment, Bleomycin treatment, and Cisplatin Treatment. After treatment with each respective agent, the death response of each strain was calculated and the % of surviving cells at multiple doses was graphed logarithmically. The data collected overwhelming support the idea that the presence of�HOB1�has a positive role on the survival of yeast after DNA damage. The WT strains tested survived better than the�hob1?�counterparts.
    • IS EXERCISE THE BIGGEST INFLUENCER OF HAPPINESS? RESEARCHING HOW INFLUENTIAL EXERCISE IS IN COMPARISON TO OTHER VARIABLES IN DAILY LIFE

      Collins, Megan; Department of Sociology, Criminal Justice & Social Work; Davies, Kim; Augusta University (2019-02-13)
      There have been multiple studies that indicate that there is a relationship between exercise and happiness. In this research, I test whether exercise is the strongest factor when compared to other common variables in a person's life for predicting happiness. Using the 2012 General Social Survey (GSS) data, I test several different variables in order to determine which of them has the strongest correlation with a person's general happiness. The 2012 GSS consisted of 4,820 respondents that ranged in age from 18 to 89. Using logistic regression, I compare the variables of sex, marriage, age, frequency of exercise, employment, and income and found that exercise has a positive relationship with a person's general happiness and that is the most strongly correlated variable. Other variables were also found to be significant and will be discussed in the poster.
    • LAVANDULA ANGUSTIFOLIA USE FOR DYSMENORRHEA IN YOUNG WOMEN: A REVIEW OF THE LITERATURE

      Nelson, Brenda; College of Science and Mathematics; College of Nursing; Langley-Brady, Dawn; Augusta University (2019-02-13)
      Dysmenorrhea effects 20% of women causing missed school and work days and interferes with daily life. Dysmenorrhea is caused by menstrual uterine contractions which may result in pain, nausea, vomiting and headaches. Many women utilize pharmacological symptom management, but experience side effects such as edema, libido reduction and increased symptom severity. Aromatherapy is a holistic non-pharmacological approach to symptom reduction. Aromatherapy is the use of essential oils via inhalation or topical application to relieve pain, stress and more.� The purpose of this project is to review the literature surrounding�Lavandula angustifolia�(lavender) and dysmenorrhea to give a foundation for future research. PubMed, TRIP, and Cochrane Library databases were searched for peer-reviewed journals articles in English and published within the last 10 years with the following keywords: dysmenorrhea, lavender, aromatherapy and human. The literature review resulted in six articles meeting inclusion criteria. These articles established the effectiveness of lavender in reducing dysmenorrhea pain in the first three days of menstruation, through inhalation and abdominal application. Lavender essential oil is also effective in reducing nausea and headaches resulting in an alternative for women experiencing dysmenorrhea. Aromatherapy has fewer risks than pharmacological and surgical approaches to dysmenorrhea management and should be studied further.
    • THE MECHANISM OF INVERSE AGONISTS ON HISTAMINE RECEPTORS, HISTAMINE RECEPTOR H1, AND HISTAMINE RECEPTOR H2

      Patel, Shrey P; Department of Phychological Sciences; Department of Pharmacology & Toxicology; Lambert, Nevin; Augusta University (2019-02-13)
      The experiment discusses the role of inverse agonist binding to receptors and how its effect cell signaling. The specific receptors that was focused on in the project was histamine receptor H1 (HRH1) and histamine receptor H2 (HRH2) which are types of G-protein coupled receptors (GPCR). Both receptors are activated when a ligand, specifically a histamine molecule, which binds to the receptor and activates the signaling pathway within the cell. The main protein within the signaling pathway is the G-protein which helps the cascade effect of the signal to other molecules. G-proteins are activated through GTP. An inverse agonist works like an agonist but will have an opposite end effect within the cell. It was originally thought that inverse agonist works the same way as an agonist to recruit a GTP and activate a G-protein for signaling. The experiment being tests tries to explain the opposite that the inverse agonist could activate the protein without GTP and continue to have its effect on the cell. Human embryonic cells were transfected with plasmids that contain sequences for the receptors and the G-protein, which were also tagged with a fluorophore to measure any bioluminescence with interaction of G-protein and the receptor when the ligands binds. From collecting data from the bioluminescence effect, it shows that there is an interaction a receptor and G-protein complex when the inverse agonist is bound.