SURE 2016 Symposium Abstract Booklet

Summer 2016 SURE Research Symposium

SURE Research Symposium

Abstract Booklet

Thursday, August 8th 2016

Summer 2016 SURE Research Symposium

9:30am – 11:30am Presentations

Presenter

Presentation Title Optimization of Cellular Assays for the Study of Zika Virus (ZIKV) Closed-Loop Neural Modulation and Optimization and Consequential Behavioral Effects in Sprague Dawley Rats Pathological Personality as an Evolutionary Adaptation: Implications for Romantic Relationship Strategies Elucidating Novel Cellular Pathways Controlling Biofilms and Motility Identification of cultivable bacteria in monarch eggs, larval guts and milkweed plants WASH and Arp2/3 actin polymerization complexes genetically interact to modulate the structure of Drosophila neuromuscular synapses Optogenetic Inhibition of the Mouse Model Basal Ganglian Snr Effects Motor Behavior in a LickingTask Activity Localization of protein expression in induced pacemaker cardiac spheroids Spatial Distribution of Putative Glutamatergic Neurons in the Neonatal Mouse Spinal Cord Analyzing 2-NBDG Uptake in Lung-Resident Memory CD8 T-Cells Process evaluation assessing a multicomponent dialysis facilty intervention to increase referral for kidney transplantation Structural and functional characteristics of transcallosal pathways in chronic stroke

Olivia Antao Omer Ashmaig Shauna Bowes Laura Briggs Mahal Bugay Guillemette Carrot Jill Cartwright Ruben Casas Tluang Cer Lily Chen Loren Cobb Cassie Fierro

Sarah Fisher

Peripheral Regeneration of Spinal Motoneurons following either Sciatic Nerve Crush or Sciatic Nerve Cut-Repair

Cecilia Garza

Disruption of proline metabolism alters proliferation and survival of colorectal cancer cells

Janay Harris

Administering a low-protein diet rebalances the TGF-β/Smad pathway in diabetic nephropathy

Lindsay Hexter

Keeping it Current: Adding Na+/K+-Pump, Ca2+, and Ca2+-Activated-K+ Currents to a Fly Neuron Model

Summer 2016 SURE Research Symposium

9:30am – 11:30am Presentations

Presenter Austin Howley Sharon Hsieh Lina Jowhar Christy Kang Travise Kinney Maya Lakshman Shoeb Lallani Alexus Landry Fredrick Leon Valerie Linck Josephine Liwang Meagan Naraine Anders Olsen Sahar Panjwani Jonathan Park Sean Perryman Chengcheng Qiu Kaushik Ravipati William Reis

Presentation Title Alterations in the Development of Ventromedial Prefrontal Cortex GABAergic Circuitry Following Neonatal Hippocampus Damage Feasibility of a Sleep Telemedicine Program for Veterans Chemical and Stress Resistances of Clostridium difficile Exaggerated Reductions in Muscle Interstitial pH During Exercise in Chronic Kidney Disease The role of SC35 in Alzheimer's Disease and Tau mRNA processing Eye Fixation to Emotional Faces in Traumatized African American Children Stereological Analysis of CalretininImmunoreactive Interneurons In Transgenic Monkey Models of Huntington’s Disease Cognitive impairment in parkinson disease did not predict benefit from behavioral intervention for urinary symptoms Structural Studies of RSV Fusion Glycoptein Using Cryo-Electron Tomography Inability to Regulate Blood Pressure in Response to Alterations in Dietary Salt in Mice Lacking VDAC3 The Effects of PKC and AMPK on Water Reabsorption in Rat Inner Medullary Collecting Duct Nuclear Factor-kB mediates Zinc DeficiencyInduced NCC Upregulation in Mouse Distal Convoluted Tubular Cells Discovering Phage Antibody Selectivity and Optimization in Bacterial Systems Predicting Real World Functioning Using Neuropsychological Assessments in an Accountability Court Population Role of LCMT-1 in Mouse Fetal Hepatoblast Growth and Division Maternal Depression and Parenting: Testing the Moderating Role of Affect Assessment of Floristic Composition to Elucidate Flora Change at Lullwater Preserve Effects of an M1 Receptor Agonist on Memory Dysfunction in an Alzheimer’s Disease Rat Model

Inhibition of PDE7 Reduces Inflammatory Response Following Renal Ischemia/Reperfusion Injury

Summer 2016 SURE Research Symposium

9:30am – 11:30am Presentations

Presenter

Presentation Title Quantifying Vascular Dynamics in Inferior Frontal Regions using BOLD Signal Derived from Echo Planar Imaging Determine contribution of RSV-CX3CR1 infectivity in Human Epithelial Cells (HEp-2).

C-H Functionalization and Cyclopropanation Using Novel Oxidatively Robust Copper Catalyst Comparative Immunobiology of Mesenchymal Stromal Cells and Hepatic Stellate Cells Regulation of BNDF signaling by the schizophrenia associated microRNA miR-137 2B4 Deficiency Impacts The Expansion And Proliferation But Not Death Of Donor-Reactive CD8+ T Cells Spherical nucleic acid-pMHC multimer for T-cell detection and immunophenotyping Biochemical characterization of the Mycobacterium tuberculosis MazEF-mt6 toxinantitoxin complex Novel, Low Bias Consensus Clustering Method for Analyzing the Dynamics of SICAvar Gene Expression in Plasmodium knowlesi Malaria Infections in silico Effects of Dual Treatment with SDF-1 and AMD3100 with BMP-2 on Osteogenic Differentiation Investigation of the influence of endosymbiont Burkholderia on squash bugs' (Anasa tristis) response towards foreign bacteria ESRD Patients Have Altered Pattern of Heart Rate Variability Responses to Graded Lower Body Negative Pressure Longitudinal Assessment in Means to Evaluate the Efficacy of Neural Progenitor Cell Therapy on Huntington’s Disease Mouse

Abinand Rejimon Nelson Reyes Martin-Louis Riu Jenna Sands Niraj Shah Phoebe Stark Wenzheng Sun Meklit Tilahun

Christopher Tseng

Pooja Venkatesh Junyan Xia Kara Ye Sarah Ye

Summer 2016 SURE Research Symposium

12:30pm-2:30pm Presentations

Presenter

Presentation Title Evaluation of PARP 1 Inhibition to Sensitize Head and Neck Cancer Cells to DNA-Damaging Therapies Defining Early Escape Phenotypes in Kras, Lkb1Mutant Drosophila Wing Epithelia Modeling Fetal Alcohol Syndrome with Cardiomyocytes Derived from Human Pluripotent Stem Cells Effects of optogenetic stimulation of basal ganglia output on locomotive behavior in mice Malaria Alters Neutrophil Response in Streptococcus Pneumoniae Co-Infection Application of C--H activation to the synthesis of disubstituted triazoles Observing the effects of electrostatics on membrane deformation driven by protein crowding The effects of S. marcescens exposure on hostparasitoid interactions Gramicidin Templated Peptide Network Modifying PVC to Create Bacteria Resistant Medical Tubing Slippery when Wet! Tribological Properties of Polyacrylamide Hydrogel Paravalvular Leak Closure: Percutaneous vs. Surgical Intervention Quantifying biofilm formation of Sinorhizobia meliloti by measuring the diffusion coefficients of polystyrene beads in the microfluidic platforms Inter-generational olfactory sensitivity in a mouse model Searching for antagonists of the transcriptional activating function of H2A.Z in Arabidopsis thaliana Utilizing Pollen DNA Metabarcoding for Reconstructing Pollinator Networks in Forests Managed for Biofuel Production

Andrew Alter Luke Arnce Madhumita Baskaran Conrad Bhamani Alexandra Caldwell Yulei Cao Jasmine Carrothers Jae Hoon Cho Jaime Coronado Alexander Crich Nicholas Cuccia Andy Dong Yijun Dong Nandini Doshi Katie Duval Julie Fowler

Summer 2016 SURE Research Symposium

12:30pm-2:30pm Presentations

Presenter

Presentation Title Dysregulated protein synthesis as a biomarker in Fragile X human patient derived cells Pharmacological Blockade of PI3K δ and BcrAbl During ex vivo T cell Expansion Preserves the Naïve Compartment and Improves Yield The Chemical Elucidation and Antibacterial Activity of a Traditional Oil Macerate of Hypericum perforatum

Analyzing the Allylic C-H Functionalization of Certain Aliphatic Alkene Compounds

Sarah Griffith Jasmin Jeffery

Austin Kim

Brittany Lee Yun-Hsuan (Stellina) Lee

Genotypic Variation on Neural Phenotypes Non-homologous end-joining pathway inhibits homologous recombination repair of replicationassociated DNA damage Expression of Cytokines Using Genetically Engineered Probiotic Yeast S. boulardii SRPK2 phosphorylation of AEP in Alzheimer’s Disease Characterization of Physio-Behavioral Changes in Adult Mice with Nerve Injury-Induced Neuropathic Pain Development of a Stem Cell Model of the Pediatric Neuromuscular Disease SMARD1 Women are more at risk for developing PTSD than men: A prospective study

Solah Lee Moon Young Lee Pai Liu Alejandro Lopez Jennifer Min Esperance Mugabekazi

When Oncogene Meets Metabolism: BRAFV600E-specific Regulation of CSGLcAT in Human Melanoma

Zhiyu Qian

Possible Detection of Trans-Methyl Formate Lines in the Millimeter-Submillimeter Regime Changes in Cerebral Blood Flow after Repetitive Concussions in Adolescence

Lindsay Rhoades Wendy Rodriguez

Priya Sathyanarayan

Spherical Aggregation Improves Human Cardiac Progenitor Cells Endothelial Lineage Commitment via Notch-HDAC2 Mechanism

Niraj Shah

Regulation of Dendritic Morphology by BDNF in a non-miR-137 dependent manner

Summer 2016 SURE Research Symposium

12:30pm-2:30pm Presentations

Presenter

Presentation Title A novel genetic screen in Drosophila designed to discover secreted factors that drive glioblastoma initiation and progression.

Effects of SNr inhibition in mice performing a locomotor go task Overground Walking Force Production and Associations with an Observational Gait Assessment in Persons with Chronic Incomplete Spinal Cord Injury A Comparison of Two Disinfectants in the Sanitation of Monkey Automated Feeders Using ATPase-based Microbial Methods ASIC1a/Îą-ENaC Hybrid Channels Contribute to Alveolar Fluid Clearance The Association Between Dietary Sulfur Amino Acid Intake and Vascular Function Human Osteopontin Isoforms Differentially Promote Vascular Smooth Muscle Cell Migration Investigations of Rhodium Pi-Allyl Complexes on Disubstituted Olefins Electrically-Induced Changes in Cardiac Progenitor Cells Assessment of Olfactory-Based Retention of Acquired Fear in Healthy Humans Change in the Number of Cholinergic Interneurons in a Mouse Model of DopamineResponsive Dystonia: Is Age a Factor?

Comparison of Indian Air pollutant Emissions Inventories at National and Regional Levels Pressure Distribution in Models of Arterial Stenosis with Steady Flow Using Blood Flow Simulations Mechanism of tetrameric transmembrane influenza A/M2 proton channel activation Role of Vascular Dysfunction in Alzheimer’s Disease

Nilang Shah Monica Sharma Telissa Spencer

Jazz Stephens Phi Trac Phong Tran Michelle Tsai Alexander Warren Ashley Washington Leah Weingast Lagena Williams Qianru Wu Siqi Xue Jonathan Zawadzki Xiancong Zhang

Summer 2016 SURE Research Symposium

Abstracts

Andrew Alter Evaluation of PARP 1 Inhibition to Sensitize Head and Neck Cancer Cells to DNA-Damaging Therapies Andrew Alter, Erica Werner, PhD Emory University, Paul Doetsch, PhD Emory University

Cisplatin and radiation therapy are two frontline treatments for head and neck cancer (HNC). One mechanism by which both treatments mediate cell death is by the production of reactive oxidative species (ROS), which cause base damage and single-strand breaks in DNA. Poly (ADP-ribose) polymerase 1 (PARP 1), a protein found primarily in the nucleus, is involved in several DNA repair mechanisms, including the base excision repair and single-strand break repair. We hypothesized that inhibiting PARP 1 sensitizes cancer cells to both cisplatin and irradiation treatments. Four HPV- and four HPV+ HNC cell lines were used. Previous studies show that the HPV+ cells contain significantly higher endogenous levels of ROS and are also significantly more sensitive to ROSinduced DNA damage, as a result of overactive NOX2. We predicted that HPV+ cells should therefore have constitutively active PARP 1 and that PARP 1 inhibition should sensitize the HPV+ cell lines to cisplatin and irradiation therapy to a greater extent than it should in the HPV- cell lines.

Results indicated that both HPV+ and HPV- cell lines had constitutively active PARP 1, and diphenyleneiodonium, a flavoenzyme inhibitor that inhibits NOX2, reduced PARP 1 activity all cell lines. PARP 1 inhibition did not sensitize any of the cell lines to cisplatin, and it only sensitized one cell line, which was HPV+, to radiation treatment. Future studies, noting that high PARP 1 activity has shown to be toxic, will explore mechanisms to exploit the constitutive activation of PARP 1 to induce cell death in HNC.

Olivia Antao Optimization of Cellular Assays for the Study of Zika Virus (ZIKV) Olivia Q. Antao, Jacob T. Beaver, Dahnide Taylor, Elena V. Vassilieva PhD, Richard Compans PhD, and Ioanna Skountzou MD PhD

Zika virus has recently spread rapidly in the Western hemisphere, with a widespread outbreak in Brazil beginning in April 2015. Infection of women in the first trimester of pregnancy has resulted in a high incidence of microcephaly in newborn infants. In the U.S., a total of 1404 cases in 46 states were reported as of July 20, 2016 (CDC). These have occurred in persons who had travelled to tropical countries, and were exposed to mosquitoes, although instances of sexually transmitted infection have been reported. In order to develop a vaccine to counteract this virus, we will need to adopt and optimize assays for Zika. The MTT assay is a common practice that uses cell metabolic activity to measure cell proliferation. Yellowish solution of MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide) is added to the live cells where it is reduced in mitochondria to purple water-insoluble MTT-formazan. Since this reaction only occurs in the live cells, the decrease of intensity of the purple color upon addition of study compounds is indicative of cellular toxicity of such compounds. We used this assay to investigate whether three compounds proposed as potential anti-Zika reagents (C, D, U) are cytotoxic. Plaque assays are another practice commonly used to determine viral titer. These assays will be optimized for cell confluency, supplemental media, incubation length, and staining. These techniques are vital for the study of Zika virus pathogenesis and infection, a virus new to our laboratory and the scientific community.

Summer 2016 SURE Research Symposium Luke Arnce Defining Early Escape Phenotypes in Kras, Lkb1-Mutant Drosophila Wing Epithelia Luke Arnce, Emory University , Chang-Soo Seong, PhD Winship Cancer Institute, Melissa GilbertRoss, PhD Winship Cancer Institute

The third most commonly mutated gene in lung adenocarcinoma is the kinase LKB1, where ~25,000patientd have this mutation. Patients co-mutated for LKB1 and KRAS have significantly shorter overall survival compared to either mutation alone and have no targeted therapies. In a genetically engineered Kras mouse model, co-mutation of Lkb1 is sufficient to promote metastasis of lung adenocarcinoma, and primary tumors and metastases show aberrant activation of both EMT and adhesion signaling. Due to its conserved role as a master regulator of cell polarity, we hypothesize that a subset of LKB1-mutant cells use classical EMT to 'lead' cells through the basement membrane, followed by defective adhesion signaling through FAK, which allows LKB1mutant cells to exploit collagen to navigate the micro environment. To test this, we will use the genetically-tractable model organism Drosophila Melanogaster to I) test the mechanism and signaling pathway used by Lkb1-mutant cells to traverse the basement membrane in vivo

Omer Ashmaig Closed-Loop Neural Modulation and Optimization and Consequential Behavioral Effects in Sprague Dawley Rats Mark J. Connolly, Omer Ashmaig, Syed Ali Zaidi, Robert E. Gross M.D./Ph.D. Emory University

Mesial temporal lobe epilepsy (TLE) is the most common form of epilepsy in humans. Deep brain stimulation (DBS) represents a novel therapeutic method and relies on the use of continuous delivery of electrical stimulation through electrodes to regions of the brain. The aim is to reduce both frequency and severity of electrical impulses that cause seizures. The therapeutic effects of DBS are clearly evident in other neurological and movement disorders such as Parkinson ’s disease.

Closed-loop neural modulation is being investigated as a type of neural modulation that may be superior to traditional approaches. However, the main limitation with closed-loop neural modulation is that it determines how to stimulate by reading neural signals, but does not take into account for the behavioral effects of the therapy. Critical to developing novel neural modulation therapies will be rigorous assessment of how they impact behavior. In this project, we aim to assess any resulting behavioral effects due to a closed-loop optimization system using a memory task.

Summer 2016 SURE Research Symposium Madhumita Baskaran Modeling Fetal Alcohol Syndrome with Cardiomyocytes Derived from Human Pluripotent Stem Cells Madhumita Baskaran, Makenna Pierce, Qingling Wu, MS Monalisa Singh MD/PhD Chunhui Xu, PhD

Fetal Alcohol Syndrome affects 6 to 9 out of 1000 children in the United States. It occurs because of excessive alcohol consumption during pregnancy. The exact effects on the developing hearts of the fetus are unknown. So far, the potential solutions for curing the disease has been very limited. In this experiment, human induced pluripotent stem cells were differentiated into cardiomyocytes (hiPSC-CMs) and exposed to various concentrations of alcohol to mimic Fetal Alcohol Syndrome.

The hiPSC-CMs exhibit the same maturity levels as fetal cells in the womb thus making them sufficient models. For data acquisition, Fluorescent Immunocytochemistry (ICC), Microelectrode Array (MEA), and Calcium Imaging were utilized. Results from the ICC showed that the alcoholexposed cells had less intact gap junctions than that of the control cells, smaller cell size, and less defined striations. The results from the MEA showed normal and consistent beating rates among the baseline recordings, and shorter, less frequent, and sometimes irregular beating rates in the alcohol recordings. The calcium recordings indicated similar results as the control group can be characterized by regular, steady beating patterns whereas the alcohol group can be characterized by irregular, varying beating patterns.. Through the analysis of the data in this experiment, it can be concluded that exposing hiPSC-CMs to concentrations of alcohol causes adverse effects on the heart such as less intact gap junctions, smaller size, and less defined striations and may even lead to arrhythmias.

Conrad Bhamani Effects of optogenetic stimulation of basal ganglia output on locomotive behavior in mice Conrad Bhamani, Undergraduate Student Cornell University, Arthur Morrissette, Bachelor's Degree Georgia Institute of Technology, Dieter Jaeger, PhD University of Michigan.

It has been shown that the basal ganglia play some part in behavior and decision-making; however, the exact reason its pathways work is not yet completely understood. Two pathways are described in previous models: the direct and indirect pathway. The direct pathway confirms a certain motor reaction to a sensory stimulus, while the indirect pathway denies competing motor reactions. Most of what is known about the basal ganglia has been derived from diseases that involve the area, such as Parkinson’s disease and Tourette’s syndrome. Recently, the use of optogenetics has made it possible to activate or inhibit specific areas in the brain with a precision of milliseconds.

Optogenetics utilize channel proteins that, when injected into the brain, can change the membrane potential of a neuron when a certain wavelength of light is emitted onto them. Here, we used optogenetics in a female Slc32a (VGAT) mouse by injecting Arch3.0 opsin protein channels unilaterally into the substantia nigra pars reticulata (SNr) of the basal ganglia. This effectively inhibited the activity of the SNr, and change in behavior between optogenetic and non-optogenetic trials was monitored. This was accomplished by head-restraining a mouse on a running wheel and training it to run when given a specific ‘go’ cue. Our results indicate that the use of optogenetics caused a change in the locomotive behavior of the mouse, and thus caused a change in basal ganglia output.

Summer 2016 SURE Research Symposium Shauna Bowes Pathological Personality as an Evolutionary Adaptation: Implications for Romantic Relationship Strategies Shauna M. Bowes, Emory University College of Arts and Sciences, Madeline G. Nagel, Emory University College of Arts and Sciences, Ashley L. Watts, Emory University Department of Psychology, Jessica L. Maples-Keller, University of Georgia Department of Psychology, Robert D. Latzman, Georgia State University Department of Psychology, Joshua D. Miller, University of Georgia Department of Psychology, & Scott O. Lilienfeld, Emory University Department of Psychology

Numerous studies have demonstrated that personality traits relate differentially to both short-term and long-term mating strategies. Two constellations of personality traits, narcissism and psychopathy, may be especially relevant to understanding individual differences in mate-seeking behaviors, as both have been shown to relate to a wide range of sexual and romantic mating behaviors. Although previous studies have demonstrated such relations, there is a paucity of research examining these relations with more specific features of both psychopathy and narcissism.

Thus, in the present study, we examined the relationships among narcissism and psychopathy’s separable subdimensions with different mating and romantic relationship interests and behaviors, namely short- and long-term mating interests, attitudes toward infidelity, and self-reported acts of infidelity, in three large, racially diverse samples of college undergraduates (Ns=308, 357, and 300).

Most aspects of psychopathy and narcissism predicted short-term mating interests, and effects were most pronounced for the subdimensions reflecting impulsivity and exploitativeness (rs ranged from .15 to .21). Following, most aspects of psychopathy predicted the endorsement of more accepting attitudes toward infidelity, with the exception of the bold features of both psychopathy and narcissism. In contrast, only the self-centered and exploitative features of narcissism predicted both emotional and physical infidelity. Our findings raise the possibility that specific aspects of narcissism and psychopathy, namely the disinhibited, entitled, and callous features, have been maintained in the population as evolved alternative romantic mating strategies encapsulating short-term mating interests and looser attitudes toward infidelity.

Laura Briggs Elucidating Novel Cellular Pathways Controlling Biofilms and Motility Laura Briggs, Benjamin Fontaine, Emory University, Parth Jariwala, Emory University, Emily Weinert, PhD Emory University

Intracellular levels of nucleotides such as cyclic diguanylate monophosphate (c-di-GMP) control numerous cellular pathways, including those involved in biofilm formation. Biofilms are microbial communities, consisting of bacteria and an excreted matrix, that protect the bacteria from environmental stress and predation. Using a method to quantify 2’, 3’-cyclic nucleotides (2’, 3’cNMPs) using liquid chromatography-tandem mass spectrometry (LC-MS/MS), an inverse relationship between 2’, 3’-cNMP levels and biofilm formation is observed in Escherichia coli.

Similarly, in Pectobacterium carotovorum, a plant pathogen, our research demonstrates that motility and quorum sensing pathways are influenced by a globin-coupled sensor, an oxygensensing protein that contains a diguanylate cyclase output domain that synthesizes c-di-GMP. This diguanylate cyclase produces c-di-GMP in the cell, allowing the cell to respond to changing oxygen levels by altering its intracellular c-di-GMP-dependent pathways. These findings show strong links between these two types of nucleotides and various cellular phenotypes. Future work is focused on investigating additional proteins involved in both signaling pathways, as well as additional phenotypes controlled by the nucleotides.

Summer 2016 SURE Research Symposium Mahal Bugay Identification of cultivable bacteria in monarch eggs, larval guts and milkweed plants Mahal Bugay, Erica Harris, Nancy Holbrook, Matthew Lipow, Jacobus de Roode, PhD Emory University, & Nicole Gerardo, PhD Emory University

Although there is growing knowledge about the host acquisition of gut microbes and how diet may impact the community structure, our understanding is limited by the lack of cultivable microbe manipulative experiments coupled with culture-independent sequencing. The monarch butterfly makes an ideal, tractable system in which to investigate these questions because of its less diversified, microbial community, larval consumption of host plants with medicinal properties, and the prevalence of a common, protozoan parasite. Bacteria from monarch eggs, larval host guts, and host food plants have been previously cultivated. From these samples, I will extract, amplify, and sequence bacterial DNA to identify the bacterial species present. By identifying cultivable bacterial species from different sample types, we may be able to determine the source and transmission mode of the monarchs’ gut microbes. I will also compare the identified cultivable bacteria to bacteria identified in previous culture-independent sequencing of larval guts fed on alternative host plants. Furthermore, this work will inform future experiments that will manipulate the gut microbial community of monarch butterflies to determine if bacterial species provide parasitic resistance.

Alexandra Caldwell Malaria Alters Neutrophil Response in Streptococcus Pneumoniae Co-Infection Alexandra B. Caldwell, Nathan T. Jacobs, Tracey J. Lamb, PhD University of Edinburgh

Evidence shows that an individual with the malaria parasite is more susceptible to secondary bacterial infections, more specifically, pneumococcal diseases caused by the bacteria Streptococcus pneumoniae. It is known that once a malaria parasite infects its host, it leads to impaired neutrophil function, meaning neutrophils have limited phagocytic activity. This limited phagocytic activity leads to an increase in the production of reactive oxygen species, which in turn, leads to increased susceptibility to secondary bacterial infections. We hypothesize that in the presence of a mixed infection, there will be an increase in neutrophil recruitment and production of reactive oxygen species, implying an impaired immunological response to malaria. Using Plasmodium berghei NK65 and Streptococcus pneumoniae, we found nasal colonization at the beginning of this experiment to marginally increase in the mixed infection in comparison to the single infection of S. pneumoniae.

This lead us to believe that neutrophil recruitment and production of reactive oxygen species would be the same across the nasal cavity, lungs, and spleen. Although the data does show an increase in neutrophil and reactive oxygen species recruitment, there is more neutrophil recruitment to the nasal cavity and the spleen, as well as down regulation of reactive oxygen species production in the spleen, but not the lungs. Therefore, our data suggests that neutrophil recruitment may not be the determining factor that causes pneumococcal disease.

Summer 2016 SURE Research Symposium Yulei Cao Application of C---H activation to the synthesis of disubstituted triazoles Yulei Cao and Annette Neuman, PhD Oxford College of Emory University, Division of Natural Science and Mathematics, Oxford, GA. 30054

Triazoles are present in many biologically active compounds, including antifungal, antitubercular, and antineoplastic agents. Disubstituted triazoles have classically been synthesized by the Huisgen cycloaddition reaction. The drawback of this method is that it often produces mixtures of regioisomers. Further, cycloaddition does not facilitate preparation of compound libraries with varied substituents. Therefore, we seek to develop a method for the preparation of 1,5disubstituted triazoles via a C-H activation strategy. Previous reaction condition of C-H activation of 1,2,3-triazoles produced the expected disubstituted triazoles with extremely low yields. By alternating the solvent and oxidant, the yields increased significantly. A byproduct also formed; initial analysis of the byproduct indicate it to be a dimer of the reactant. Liquid chromatographymass spectrometry (LCMS) was used to identify the composition of the product at each stage of the reaction, and the result indicated the purification method need to be refined.

Guillemette Carrot WASH and Arp2/3 actin polymerization complexes genetically interact to modulate the structure of Drosophila neuromuscular synapses Guillemette Carrot, Victor Faundez, PhD, Emory University, Cortnie Hartwig , Kennesaw State University

The cellular mechanisms underlying schizophrenia related genes are subject to current research.

The Schizophrenia susceptability factor Dysbindin is found in 80% of Schizophrenic human brains.

Dysbindin is a monomer of a larger octameric complex, Bloc-1, which is involved in endosome formation at the synapse. Bloc-1 has homologues in Drosophila melanogaster with subunit mutations resulting in a quantifiable phenotype at the neuromuscular junction (NMJ). Arp 2/3 is a protein involved in actin polymerization that is regulated by the activity of BLOC-1. It has been shown that corresponding mutations of Arp 2/3 and Dysbindin rescue the phenotype observed at the NMJ compared to animals with Dysbindin mutations alone. WASH is a known regulator of the Arp 2/3 complex. We wish to identify whether it is part of this Schizophrenia-related pathway. Our goal is to decode the factors related to this pathway to deepen our understanding of the mechanism. We will observe the effects of WASH and Arp 2/3 mutations on the neuromuscular junction of the female third stage instar larvae and quantify the boutons and muscle area of the muscles 6 and 7 of the second or third junction. WASH overexpression shows a significantly higher bouton count than the wild type. A mutant with both WASH overexpression and Arp 2/3 loss-offunction shows a similar phenotype as WASH overexpression. Therefore we have identified WASH as being upstream of Arp 2/3 but not the only regulatory mechanism.

Summer 2016 SURE Research Symposium Jasmine Carrothers Observing the effects of electrostatics on membrane deformation driven by protein crowding Jasmine Carrothers, Gokul Raghunath, and Brian Dyer, PhD Department of Chemistry, Emory University

Receptor-mediated endocytosis is known to play a significant role in cellular infection. However, there is no clear understanding of the underlying mechanisms that control such processes. To look at this problem from a fundamental perspective, we designed a project to test the role of electrostatic repulsions between peripherally bound proteins, in membrane deformation and reshaping. The methods employed in this study, were isothermal titration calorimetry (ITC) to observe the binding affinity and dynamic light scattering (DLS) to monitor vesicle morphology. To test whether electrostatic repulsions play a role in membrane deformation, we performed experiments where we changed the ionic strength and/or the pH of buffer solutions and observed the changes in binding affinity and vesicle size. We report that, at pH values greater than 6, the binding affinity scaled linearly with decreasing concentrations of protons. We attribute this to the protonation of the Histidine tag of the protein. To further investigate the system, we used different lipid compositions comprising of 80% DPPC (Saturated), POPC (Unsaturated), and DSPC (Saturated with higher number of carbons) and 20% Ni-NTA for our lipid vesicles and His-tagged proteins as interacting partners. Our results indicate that the charged environment of the proteins can potentially decrease the binding strength between Ni-NTA containing lipid vesicles and the respective proteins. Follow up DLS experiments revealed that the propensity for membrane deformation increases at high salt concentrations, especially in the gel phase. We believe that our results further our mechanistic understanding of the membrane bending process, driven by protein binding.

Jill Cartwright Optogenetic Inhibition of the Mouse Model Basal Ganglian Snr Effects Motor Behavior in a Licking-Task Activity Jill Cartwright, Spelman College; Arthur Morissette, Emory University; Dieter Jaeger, PhD, Emory University

There are various structures within the human brain that influence movement coordination.

Among these are the basal ganglia, a subcortical collection of nuclei through which motor activity is largely conducted. Failure or degeneration of the basal ganglia leads to motor disorders, including the development of Parkinson's Disease when dopamine neurons are lesioned. One key structure conveying basal ganglia output is the substantia nigra reticulata (SNr). By inhibiting the SNr using optogenetics in mice, and subsequently examining their behavior while performing a licking task, we are able to make inferences about the innate functionality of the SNr in the basal ganglia when coordinating motor functions.

Summer 2016 SURE Research Symposium Ruben Casas Localization of protein expression in induced pacemaker cardiac spheroids Ruben Casas, San José State University Sandra Gonzalez, BS Emory University and Georgia Institute of Technology Jun Li, PhD Emory University and Georgia Institute of Technology Hee Cheol Cho, PhD Emory University and Georgia Institute of Technology

Abstract withheld.

Tluang Cer Spatial Distribution of Putative Glutamatergic Neurons in the Neonatal Mouse Spinal Cord Tluang Cer, Undergrad Agnes Scott College, Marie-Claude Perreault, PI Emory University

The spinal cord contains local interneurons of different neurotransmitter phenotypes. One important group of interneurons are glutamatergic interneurons. Glutamtergic interneurons release glutamate, the major excitatory neurotransmitter of the CNS. In the spinal cord, the large majority of glutamatergic interneurons express the vesicular glutamatergic transport 2 (Vglut2+).

Vglut2+ interneurons are responsible for activating somatic and autonomic motoneurons which in turn control our muscles and body internal organs. The spatial distribution of Vglut2+ interneurons in the spinal cord remains unknown. In this project, we examine the distribution of Vglut2+ interneurons in the 10th thoracic segment of the spinal cord.

Lily Chen

Analyzing 2-NBDG Uptake in Lung-Resident Memory CD8 T-Cells Lily Chen Sarah Hayward Jacob Kohlmeier, PhD

Tissue-resident memory T-cells (TRM ) play an important role in adaptive immune memory by preventing reinfection at the sites of bacterial and viral entry. By looking at the ability of these cells to take up the fluorescently labeled glucose analogue 2-NBDG, we are able to learn more about Tcell metabolism. Tissue-resident memory T-cells are non-circulating and thus don’t have access to the blood and serum. Due to this localization they aren’t able to access glucose as well as cells in other parts of body. This suggests different pathways that promote cellular metabolism and growth must be used. We looked at the differential glucose uptake in the T-cells located in lung-resident, Bronchoalveolar Lavage (BAL), and Mediastinal Lymph Node (MLN) and found that there’s more 2NBDG uptake in MLN than BAL and lung-resident T-cells.

Summer 2016 SURE Research Symposium

Jae Hoon Cho The effects of S. marcescens exposure on host-parasitoid interactions Jae Hoon Cho, Zachary Lynch, Ph.D. candidate (5th year student), Emory University, Levi Morran, Ph.D., University of Oregon

While it is well established that mutualistic bacteria can protect their hosts from parasitoids, the effects of pathogenic bacteria on host-parasitoid interactions are poorly understood. We explored how parasitoid attack rates and intensities in addition to host development times and survival rates were affected when the bacterial pathogen Serratia marcescens was added to laboratory fruit fly– parasitoid wasp systems. Drosophila melanogaster was exposed to S. marcescens then the wasps Asobara tabida or Leptopilina buolardi to examine the effects on development timing and survival.

We found that S. marcescens exposure followed by wasp exposure significantly lowered fly survival rate compared to either bacteria or wasp exposure alone. A follow-up experiment conducted on S. marcescens, L. boulardi, and D. simulans determined whether this outcome was the result of differential wasp attack rates and intensities on fly larvae. Although we found a clear effect on increased wasp attack rate and intensity in groups exposed to bacteria both before and during wasp exposure, only wasp attack intensity exhibited a significant difference. Finally, we tested the effect of S. marcescens exposure on fly larval mobility as a possible mechanism driving the increased wasp attack intensity experienced by infected fly larvae and found a marginally significant effect of S. marcescens exposure on the mobility of D. melanogaster and D. simulans larvae (p=0.0616).

These results relate the effects of S. marcescens to fly–parasitoid wasp interactions, specifically showing that the decrease in mobility of infected fly larvae may underlie the increase in wasp attack intensities and rates.

Jaime Coronado Gramicidin Templated Peptide Network Anthony Sementilli, Emory University, David G. Lynn, PhD Emory University

The Lynn lab is interested in understanding the interactions that govern peptide self-assembly. The amyloid-like peptides can self assemble into nanotubes that display a bilayer leaflet interface that resemble the membrane structures in cells. Recently, it has be shown that the peptide sequence NFF-CHO can form linear trimers that are stabilized by the formation of N,N-acetal oligomers. When allowed to assemble, these peptides form fibers. However, when seeds from mature nanotube assemblies (KLVFFAL and NFF-CHO) are introduced at the particle stage, the NFF-CHO peptides template and form bilayer nanotube structures. The remarkable plasticity of the NFF-CHO network has inspired the proposal to incorporate functional peptides into the developing assemblies to create an asymmetric structure that extends the functions of these new membranes. Having the ability to insert spanning peptides through the bilayer would extend the range of functions accessible to these membranes. Using cellular organelles as a model of the complex chemical networks in cells, our long-term goal is to create novel mesoscale assemblies that could similarly extend cellular capability. The NFF-CHO network will be templated with gramicidin to create an asymmetric membrane bilayer. Gramicidin is a 15 residue antimicrobial peptide that can form pores in the lipid bilayer of the plasma membrane. If gramicidin is able to incorporate within the bilayer lattice, then it would be possible to incorporate other transmembrane peptides that can facilitate electron transfer and transmembrane transport.

Summer 2016 SURE Research Symposium

Alexander Crich Modifying PVC to Create Bacteria Resistant Medical Tubing Alexander Crich, Allison Geoghan, Jen Beveridge, and Professor M.G. Finn, Ph.D

Bacterial infection of polyvinyl chloride (PVC) medical tubing is a common and often fatal medical problem that lacks an effective preemptive solution applicable to all patients. Lining the tubing with silver is the current leading solution, however it is expensive, not completely effective, and not approved for pediatric patients. This project focuses on addressing these limitations by covalently decorating medical tubing with anti-microbial molecules. This was accomplished by replacing the chlorine atoms on the surface of the PVC with azide groups, and then employing the Cu(I) catalyzed alkyne-azide cycloaddition (CuAAC) “click� reaction to covalently attach various potentially useful molecules to the surface of the PVC. Modifications were characterized by infrared spectroscopy (IR) and X-ray photoelectron spectroscopy (XPS) to determine the effectiveness of the CuAAC reaction.

Promising samples were then cultured with bacterial strains common to hospitals, such as Staphylococcus aureus, in order to quantitatively determine how effectively they prevent bacterial adhesion relative to unmodified medical tubing. To date, several anti-microbial molecules have been successfully attached to PVC tubing. We have shown a tenfold decrease in bacterial adhesion relative to unmodified PVC. In the future, more anti-microbial molecules will be explored, and the reactions will be scaled up in pursuit of creating a commercially available medical tubing product.

Ideally, this product will create a preventive solution to medical tubing-related infections that will highly effective, cost efficient, and applicable to all patients.

Nicholas Cuccia Slippery when Wet! Tribological Properties of Polyacrylamide Hydrogel Nicholas Cuccia Justin C. Burton, PhD Emory University

Spherical, polyacrylamide hydrogel particles have recently become a popular system for modeling low-friction, granular materials. For many years, the various sliding frictions of polymer gels on solid surfaces has been studied. Previously, it has been observed that frictional behaviors of polymer gels do not conform to any of Amontons' laws, but rather depend extensively on the chemical structure of the gel and measurement conditions. Since the behavior of many granular materials depend sensitively on frictional properties of the constituent grains, it is necessary to study the surface frictional properties of polyacrylamide gels. Using a custom-built tribometer with ultra-low force resolution, we have measured and analyzed the sliding friction between two polyacrylamide gels at a variety of shearing velocities. The friction coefficient in all cases is very low (~0.01), yet depends on the sliding conditions and the water saturation in the gel.

Summer 2016 SURE Research Symposium

Andy Dong Paravalvular Leak Closure: Percutaneous vs. Surgical Intervention John A. Wells IV, BS, Jose F. Condado, MD, MS, Andy Dong, Norihiko Kamioka, MD, Andrew Ritter, BS, Stamatios Lerakis, MD, James Stewart, MD, Bradley Leshnower, MD, Kreton Mavromatis, MD, Robert Guyton, MD, Chandan Devireddy, MD, Jessica Forcillo, MD, Ateet Patel, MD, Vinod Thourani, MD, Peter Block, MD, Vasilis Babaliaros, MD

Data comparing the treatment of paravalvular leak (PVL) with percutaneous (PI) and surgical (SI) interventions are limited. We conducted a retrospective cohort study comparing baseline characteristics, procedural details, and 1-year survival in consecutive patients at our center who had PI or SI for ≼moderate PVL from 2007-16. The primary outcome was a composite of death, reintervention for PVL, or re-admission for CHF-related symptoms at 1 year. Of 119 patients, 57 had PI and 62 had SI. Differences in baseline characteristics were limited in that SI patients were younger, had more active endocarditis, and fewer urgent procedures than PI patients. There were no differences in the primary end-point or 1-year survival between groups. The PI group had shorter postoperative stay, shorter ICU stay, and fewer readmissions at 30 days. After adjusting for age, active endocarditis, and procedure urgency, there was still a similar risk of death at 1 year between groups and a trend toward reduced 30-day readmission in the PI group. To conclude, PI for PVL closure results in comparable 1-year survival to SI, but with less resource utilization and 30-day re-hospitalization, even after adjusting for differences in baseline characteristics. Newer percutaneous techniques for PVL closure are resulting in improved outcomes for these patients.

These findings support the increased implementation of PI for PVL closure at US institutions.

Yijun Dong Quantifying biofilm formation of Sinorhizobia meliloti by measuring the diffusion coefficients of polystyrene beads in the microfluidic platforms Yijun Dong and Effrosyni Seitaridou, Ph.D., Department of Physics and Astronomy, Oxford College of Emory University, Oxford, Georgia

A biofilm is a highly structured community of microbes that adheres to a surface and immerses in a self-produced polymeric matrix. Biofilms, such as dental plaques and catheter infections, interact extensively with human beings. In this study, the diffusion coefficient (D) of microspheres in bacterial communities was used as a parameter to quantify bacteria growth and biofilm development of five Sinorhizobia meliloti strains (the biofilm-forming strains Rm8530expR+, Rm8530exoY, Rm9034expG, and the non-biofilm forming strains Rm1021 and Rm9030-2expA1).

The experiments were conducted in microfluidic platforms. As the bacterial colonies grew over a 4day period, the motion of the microspheres was recorded via fluorescence microscopy, and the diffusion coefficients were measured every 24 hours. It was found that each S. meliloti strain displayed a unique pattern of change in diffusion coefficient over time. For a given biofilm-forming strain, the diffusion coefficients depended on the environmental or morphological states of the bacterial community. Thus, the diffusion coefficient can be used to identify different S. meliloti strains, and for the biofilm-forming strains, the diffusion coefficient is also an indicator of the stage of biofilm formation.

Summer 2016 SURE Research Symposium

Nandini Doshi Inter-generational olfactory sensitivity in a mouse model Nandini Doshi, Emory University College of Arts and Sciences Brian Dias, PhD, Emory University School of Medicine & Yerkes National Primate Research Center

Abstract withheld.

Katie Duval

Searching for antagonists of the transcriptional activating function of H2A.Z in Arabidopsis thaliana Katie Duval, E. Shannon Torres, Roger Deal all from Emory University

Chromatin modification is critical for the regulation of gene transcription. In Arabidopsis thaliana, proper transcription of the Flowering Locus C (FLC) gene has been shown to require the histone variant H2A.Z. H2A.Z is conserved in all eukaryotes and has been shown to be critical in stress response and to be misregulated in cancers. Incorporation of H2A.Z into nucleosomes regulates gene expression, sometimes activating and other times repressing it. However, this dual-sided nature of H2A.Z is not well characterized. This project utilized a genetic suppressor screen to identify proteins that make H2A.Z necessary for transcription activation. Mutants lacking the Arp6 protein, a protein required for incorporating H2A.Z into chromatin1, were mutagenized to find suppressor mutations that overcome the need for H2A.Z in FLC transcription. One such mutation, eoa2, has been identified to restore FLC expression and wild-type flowering times. This project aims to discover the identity of this EOA2 gene by introducing candidate genes into arp6/eoa2 mutant plants and determining if each one restores flowering time and FLC levels to that of arp6 plants. The most promising candidate in our screen, BLISTER (BLI), has been found to associate with the polycomb repressive complex, which has also been shown to regulate FLC2. Identification of the EOA2 gene will shed light on how the universally conserved H2A.Z protein promotes transcription.

Summer 2016 SURE Research Symposium

Cassie Fierro Structural and functional characteristics of transcallosal pathways in chronic stroke Cassandra Fierro, Emory University; Kamal Shadi, Georgia Institute of Technology; Michael Borich, DPT, Ph.D Department of Rehabilitation Medicine, Emory University School of Medicine

After stroke, structural and functional changes in the brain contribute to the recovery of motor movement. However, certain changes in brain structure and function are also thought to underlie persistent functional disability after stroke. For example, abnormal communication between the stroke-affected hemisphere and non-affected hemisphere is associated with arm impairment after stroke. The corpus callosum, a large bundle of axons connecting the two hemispheres, is the brain structure mediating interhemispheric communication. Indirect evidence has shown associations between callosal structure and function with level of persistent movement dysfunction in chronic stroke. This project aims to further the understanding of neural pathways in the corpus callosum after stroke using non-invasive brain imaging approaches to directly characterize structural and functional abnormalities that are related to persistent arm dysfunction. Using electroencephalography (EEG) and diffusion imaging we will be able to delineate structural and functional maps of interhemispheric communication. We will evaluate relationships between arm impairment and reorganization of interhemispheric neural pathways after stroke. Results from this project are expected to contribute to better assessment of the degree of impairment and maladaptive restructuring of the brain after stroke in order to better delegate rehabilitation treatments on an individual basis.

Sarah Fisher Peripheral Regeneration of Spinal Motoneurons following either Sciatic Nerve Crush or Sciatic Nerve Cut-Repair Sarah E. Fisher, Dr. Francisco J. Alvarez, and Travis M. Rotterman

Following different types of peripheral nerve injury, nerve regeneration does occur. In this project, the difference in reinnervation rates between sciatic cut/repair and sciatic crush injuries in mice was examined. Using immunostaining, lateral gastrocnemius samples from 2 mice at each post injury time point (3 day, 7 day, 14 day, 21 day, and 2 months) with the above injuries were stained.

The immunostaining fluorescently labeled axons, neuromuscular junctions, and vesicular acetylcholine transporters which are key landmarks of reinnervation. In each animal, fifty neuromuscular junctions were examined for reinnervation. To determine the level of reinnervation at a neuromuscular junction, the alignment of the different stains was examined. If a neuromuscular junction showed full reinnervation, then all three fluorescently labeled structures lined up perfectly. If only partial reinnervation had occurred, then the stains would line up for the most part, but alignment would not be present in certain areas. Finally, if there was no reinnervation, then there would be no alignment at the neuromuscular junction. Full recovery of VAchT following both sciatic crush and sciatic repair injuries is incomplete, with an approximate average of only 75% recovery at the 2 month post-injury time point. Furthermore, at the 2 month post-injury time point in both sciatic crush and sciatic repair models, a nearly complete (~90%) recovery of VAchT is observed. Finally, in all comparisons, the sciatic crush injury model shows a faster rate of reinnervation and axon regeneration as compared to the sciatic repair injury model.

Summer 2016 SURE Research Symposium Julie Fowler Utilizing Pollen DNA Metabarcoding for Reconstructing Pollinator Networks in Forests Managed for Biofuel Production Julie Fowler: Undergraduate Student, Department of Environmental Science, Emory University Karen Bell: Post-Doctoral Researcher, Department of Environmental Science, Emory University Brice Lawley: Undergraduate Alumni, Department of Environmental Science, Emory University Berry Brosi: Primary Investigator, Department of Environmental Science, Emory University

Pollen identification has been shown to have applications in many different fields including those of forensics, airborne allergen monitoring, ecology, and particularly for constructing plant-pollinator networks. Pollinator networks are typically assembled using visitation data, or the recording of visual observations of flower visits, but prior work has confirmed the advantages of pollen identification which integrates information from multiple flower visits by each pollinator and thus can allow for better-resolved networks with less field time. The major shortcoming of this approach, however, is that traditional microscopic identification of pollen is slow, of limited taxonomic resolution, and requires extensive expertise and training. An alternative strategy, DNA barcoding, could potentially enhance the speed and the accuracy of pollen identification using techniques common to almost any basic molecular biology lab. Several recent studies have demonstrated proof-of-concept for pollen DNA metabarcoding; pollen in mixed-species samples can be identified through mixed-amplicon sequencing of standard DNA barcoding markers (e.g. chloroplast rbcL or ribosomal ITS2) on the Illumina MiSeq platform. In this project, these recently fine-tuned DNA metabarcoding methods will be used to construct pollinator networks from beecarried pollen in managed forest ecosystems in the south-eastern USA in order to assess the effects on pollinators of biofuels cultivation in these ecosystems.

Cecilia Garza

Disruption of proline metabolism alters proliferation and survival of colorectal cancer cells Cecilia Garza, Agness Scott College, Yiran Han, M.S. [đ?&#x;?] , đ??šđ??§đ??? Chris Yun, PhD, Division of Digestive Diseases, Department of Medicine, Emory University

The malignant transformation initiated by genetic aberration is enhanced by growth factors and angiogenic factors produced by the tumor cells or surrounding environment. Evidence supports that lysophosphatidic acid (LPA) has growth factor-like effects that stimulates cell proliferation, survival, and migration. Previous studies have shown human CRC patients have an elevated expression of LPA2R, and in mouse models the absence of LPA2R show decreased CRC progress.

Tumors often compensate through alternative energy production to ensure their survival. An unpublished study by the Yun Lab shows that proline synthesis is noticeably decreased in cells lacking LPA2R. The goal of this study is to determine the effects of proline biosynthesis in colon cancer cell proliferation and survival. We used SW620, HCT116 and HT-29 human colon cancer cells with different genetic changes. Cells were transfected to decrease the expression of one of the key enzymes of proline synthesis, pyrroline-5-carboxylate synthase (P5CS). We found that proliferation of cancer cells was decreased in P5CS knockdown cells. SW620 xenograft depicts a similar trend with decreased growth in P5CS deficient cells. The role of P5CS in cell survival was determined by treating SW620 cells with the chemotherapic drug 5-FU. LPA rescued SW620 cells from 5-FU-induced apoptosis only in the presence of P5CS. The current study shows that proline metabolism is important for growth and survival of colon cancer and disrupting proline biosynthesis could have a potential therapeutic use.

Summer 2016 SURE Research Symposium Sarah Griffith Dysregulated protein synthesis as a biomarker in Fragile X human patient derived cells Sarah Griffith, Vanderbilt University, Nisha Raj, Emory School of Medicine Department of Cell Biology, Gary Bassell, PhD, Emory School of Medicine Department of Cell Biology.

Fragile X syndrome (FXS) is an X-linked heritable neurologic disease and the most frequently inherited form of intellectual disability. Caused by the absence of fragile X mental retardation protein (FMRP), FXS is also the most prevalent single-gene cause of autism, the most common autism spectrum disorder (ASD). FMRP is an mRNA binding protein that functions at many synapses to mediate negative regulation of mRNA translation. The loss of this protein often alters synaptic function and morphology, and studies using animal models have shown that in the absence of FMRP, global protein synthesis is elevated. The goal of this project is to use neurons and neural progenitor cells (NPCs) generated from patient fibroblast derived induced pluripotent stem cells (iPSCs) as a more translationally relevant model to study Fragile X syndrome. We predict that FXS patient derived cells will have increased activity of pathways such as PI3K and mGluR, and increased protein synthesis due to the absence of FMRP. We will use western blotting to check for dysregulated protein synthesis and a flow cytometry based metabolic labeling assay to quantify protein synthesis. Ultimately we hope to use the data collected to investigate whether dysregulated protein synthesis can be used as a biomarker for FXS and autism, and whether pharmaceutical interventions targeting this dysregulation is a valid therapeutic strategy.

Janay Harris

Administering a low-protein diet rebalances the TGF-β/Smad pathway in diabetic nephropathy Janay M. Harris, Yirong Wang and Mitsi A. Blount

Diabetes is a chronic disease associated with persistently high glucose levels that lead to renal damage, specifically diabetic nephropathy (DN). The pathological features of DN (glomerular basement membrane thickening, podocyte effacement, renal fibrosis, low-grade renal inflammation) are largely mediated by transforming growth factor (TGF)-β activation of Smaddependent pathways. Patients with fibrotic kidneys, a consequence of DN, demonstrate elevated TGF-β levels, highly active Smad2 and Smad3, and dampened expression of the inhibitory Smad7.

This suggests that restoring the dysregulated TGF-β/Smad pathway may be an effective therapeutic strategy for DN. When caught at an early stage, progression of DN can be slowed with a low-protein diet. Interestingly, a low-protein diet attenuates glomerular expression of TGF-β in rats with nephrotic syndrome. We predict that a low-protein diet protects the diabetic kidney by lowering TGF-β levels and, in turn, dampening Smad2/3 signaling. We will use multiple low-dose injections of streptozotocin to induce DN in mice fed either a low-, normal- or high-protein diet. The extent of renal injury will be measured by alterations in known markers of kidney function and changes in renal pathology. We will also elucidate the role of TGF-β/Smad signaling in response to protein diet by examining protein abundance by Western blotting and protein location by immunohistochemistry.

Summer 2016 SURE Research Symposium Lindsay Hexter Keeping it Current: Adding Na+/K+-Pump, Ca2+, and Ca2+-Activated-K+ Currents to a Fly Neuron Model Cengiz GĂźnay, PhD, Georgia Gwinnett College School of Science and Technology Astrid A. Prinz, PhD, Emory University

Modeling biological systems highlights the increasingly important role of computation in today’s research arena. Specifically, in computational neuroscience, models allow for removal of artifacts and other complications that arise in neuronal recordings. Understanding electrophysiology of the nervous system can lead to insights about brain disease and everyday functioning. Computational models thus provide another method of performing experiments, as when fit to real data, they can simulate experiments that are more difficult to execute in reality. This project focuses on a computational model of a fly neuron, based on data from aCC motoneurons, responsible for larval crawling. The model is written in XPP, while simulations and analysis are done in Matlab and Spike2. RP-2 fly neuron data were used for pump parameter fitting, via frequency-current curves, while aCC data were used for Calcium channel fitting with existing Matlab tools (the PANDORA toolbox); the KCa channels will likely be added with respect to changing Calcium concentration.

Spike dynamics were also considered, such as spike-rate anti-adaptation versus adaptation. Overall, a realistic model will be useful to simulate experiments and predict novel neuron qualities, answering questions about neuron plasticity, firing dynamics, and other important themes.

Austin Howley

Alterations in the Development of Ventromedial Prefrontal Cortex GABAergic Circuitry Following Neonatal Hippocampus Damage Austin Howley, Emory University T.J. Libecap, Emory University Jocelyne Bachevalier, PhD, Yerkes National Primate Research Center and Emory University Hillary Rodman, PhD, Emory University

Effective decision-making, emotional regulation, and aversion have been associated with the ventromedial prefrontal cortex (vmPFC). Schizophrenia is correlated with reduced vmPFC activity, resulting in deficits in predicting outcomes to given actions, appropriate social behavior, and utilizing working memory. The hippocampus, a structure in the medial temporal lobe, is known to be connected with the vmPFC and may be involved in its development. Gamma-Aminobutyric acid (GABA) is crucial to the function and connections of the vmPFC. To further investigate the involvement of the hippocampus in vmPFC development, we plan to use GABA markers, Parvalbumin and Calbindin D-28K, to examine densities of GABAergic neurons in the vmPFC of Rhesus macaque brains with lesioned hippocampi compared to those of control brains. We will use immunohistochemistry to visualize the neurons and stereological procedures to quantify them.

Findings in this project will contribute to the current knowledge on the role of the hippocampus in vmPFC development and their connectivity. Further understanding of the relationship between the hippocampus and the vmPFC can potentially contribute to the development of methods of treating schizophrenia and damage associated with impairments in decision-making and emotional processing.

Summer 2016 SURE Research Symposium Sharon Hsieh Feasibility of a Sleep Telemedicine Program for Veterans Sharon Hsieh, Emory University; Mary Margaret Ciavatta, MPH, PA-C, Division of Sleep Medicine, Atlanta Veterans Affairs Medical Center; Barry Fields MD, MSEd & Octavian Ioachimescu, MD, PhD, Emory University, Department of Medicine, Division of Pulmonary, Allergy, Critical Care and Sleep Medicine

Obstructive sleep apnea (OSA) affects at least 2-4% of the general population and nearly 54% of active duty military personnel (Mysliwiec, 2013). Sleep apnea is associated with heart disease, stroke, and impairment in work and driving performance. This prospective, parallel-group pilot study assessed the feasibility of remote telemedicine-based applications in comparison to a more traditional, in-person care model in the diagnosis and treatment of veteran patients who have OSA. At the Atlanta VA Sleep Medicine Center, one group of patients with OSA (n=70) received in-person evaluation for OSA, while another cohort of patients (n=70) received real-time clinic based video telehealth care. The study quantitatively and qualitatively evaluated the clinical value of the telemedicine health care delivery for the diagnosis and treatment of OSA by comparing the Continuous Positive Airway Pressure (CPAP) usage data, patient symptom and satisfaction scores, and mileage driven to and from clinical appointment sites. There was no significant difference in the treatment efficacy and clinical outcomes between groups after 90 days of treatment.

Furthermore, the telemedicine patients adhered to CPAP treatment significantly more than the inperson treatment patients. Thus, the study illustrated that telemedicine sleep care is feasible for the veteran patients. Further studies should explore both groups' time intervals between visits to evaluate telemedicine's implications on timeliness of care.

Jasmin Jeffery Pharmacological Blockade of PI3K δ and Bcr-Abl During ex vivo T cell Expansion Preserves the Naïve Compartment and Improves Yield Jasmin Jeffery, Hampton University; Christopher Petersen, Emory University; Edmund Waller, MD PhD Emory University

Abstract withheld.

Lina Jowhar Chemical and Stress Resistances of Clostridium difficile Adrianne Edwards, Samiha T. Karim, Ricardo A. Pascual, Lina M. Jowhar, Shonna M

Abstract withheld.

Summer 2016 SURE Research Symposium

Christy Kang Exaggerated Reductions in Muscle Interstitial pH During Exercise in Chronic Kidney Disease Christy Kang1,2, Ryan Downey PhD1,2, Dana DaCosta1,2, Melanie Kankam1,2, Jeanie Park MD1,2 1Renal Division, Department of Medicine, Emory University School of Medicine, Atlanta, GA, 2Research Service Line, Atlanta Veterans Affairs Medical Center, Decatur, GA

Chronic kidney disease (CKD) patients have exercise intolerance and reduced physical capacity that contribute to increased cardiovascular risk. Muscle interstitial pH decreases during exercise in response to H+ ion extrusion to the extracellular space which is largely buffered by bicarbonate.

Reductions in muscle interstitial pH lead to pain, and muscle afferent signaling leading to reflex activation of the sympathetic nervous system during exercise. CKD patients often have overt or occult metabolic acidosis which could lead to decreased capacity to buffer muscle interstitial changes in pH. We hypothesized that CKD patients have exaggerated reductions in muscle interstitial pH during exercise. We measured muscle interstitial pH and oxygenation (SmO2) using near infrared spectroscopy (NIRS) in 9 CKD patients and 10 healthy controls during 3 minutes of low-intensity rhythmic handgrip exercise (RHG 20%), and high-intensity rhythmic handgrip exercise to fatigue (RHG 40%). We observed that CKD patients had an exaggerated reduction in muscle interstitial pH during high-intensity exercise (p<0.05), but not during low-intensity exercise. There was no difference in SmO2 during low-intensity or high-intensity handgrip exercises between the groups. These findings suggest that CKD patients have exaggerated reductions in muscle interstitial pH during high-intensity exercise that is not mediated by reductions in muscle oxygenation. CKD patients may have lower capacity to buffer pH changes in the muscle interstitium. Exaggerated reductions in muscle interstitial pH may contribute to exercise intolerance in CKD patients by increasing muscle fatigue, pain, and reflex activation of sympathetic nerve activity which could contribute to abnormal hemodynamic responses during exercise.

Summer 2016 SURE Research Symposium

Austin Kim The Chemical Elucidation and Antibacterial Activity of a Traditional Oil Macerate of Hypericum perforatum Austin J. Kim, James T. Lyles, PhD, Kate Nelson, Cassandra L. Quave, PhD Emory University

Hypericum perforatum L. (Hypericaceae), also called St. Johnâ&#x20AC;&#x2122;s Wort, is a well known medicinal herb often associated with the treatment of anxiety and depression. Additionally, an oil preparation of the flowers is widely used in traditional medicine across the Balkans as a topical wound salve.

Recent research has shown that this oleolite reduces both wound size and healing time. H. perforatum has been well characterized chemically: multiple compounds have been identified including naphthodianthrones, phloroglucinols, flavonoid glycosides, biflavones and anthocyanidins. The naphthodianthrone hypericin and phloroglucinol hyperforin are effective antibacterial compounds against various Gram-positive bacteria, including B. subtilis, E. faecalis, S.epidermidis, and M. luteus, while ineffective against Gram-negative strains. However, hyperforin is quite unstable with light and heat, and thus should not be present in the aged oil macerate preparation of H. perforatum. Additionally, hypericin can cause phototoxic skin reactions if ingested or absorbed into the skin: a common pharmaceutical warning for fair-skinned individuals taking Hypericum supplements, and evidenced by livestock that develop extreme photosensitivity after grazing on H. perforatum flowers. Therefore, the established chemistry presents a paradox for this H. perforatum oil preparation. The hyperforin responsible for the antibacterial bioactivity should degrade in the sunlight as the traditional oleolite is prepared. Alternately, if hypericin is present in established bioactive levels, then the oleolite should cause photosensitivity, yet none is reported. In this research, various extracts of H. perforatum were compared to traditional oleolites with regards to composition and antibacterial activity to better understand the chemistry of this remedy.

Travise Kinney The role of SC35 in Alzheimer's Disease and Tau mRNA processing Travise Kinney James A. Webster Chadwick Hales

Prior research reveals six isoforms of tau exist, resulting from alternative splicing of exons 2, 3, and 10 of chromosome 17q21 where tau is located. The tau protein results in either a 3R or 4R-Tau isoform. The quantitated value entails the number of microtubule binding domain repeats. When exon 10 is eliminated through the alternative splicing process, the tau is a 3R isoform. During infanthood, the body produces only 3R-Tau. However, as the infants grow older, 4R-Tau is created.

In normal human brains, the 3R-Tau and 4R-Tau levels are equal. An imbalance in the 3R:4R ratio may contribute to the aggregation of Tau in AD and other Tauopathies. SC35 is a protein identified in the alternative splicing of Tau. This protein, regulated by cis-acting proteins, is a serine/arginine rich (SR) type protein that when phosphorylated, splices out exon 10. Research reveals that phosphorylated SC35 suppresses nuclear inclusions inducing 3R-Tau, while SC35 proteins without phosphorylation allows for these inclusions and 4R-Tau formation. Our research seeks to find high levels of SC35 in Alzheimer disease cases as well as replicating an overexpression of SC35 to examine its effects on Tau isoforms.

Summer 2016 SURE Research Symposium

Maya Lakshman Eye Fixation to Emotional Faces in Traumatized African American Children Maya Lakshman, Emory University Maria Briscione, BS Emory University Abigail Powers, PhD Emory University Tanja Jovanovic, PhD Emory University

Background: Given the prevalence of trauma in low-income, inner-city families, risk for posttraumatic stress disorder (PTSD) can begin early in life. Previous research has shown that in traumatized adult populations, those with PTSD demonstrate heightened attention bias towards threatening faces. This study aimed to use eye tracking to investigate age-related changes in hypervigilance or avoidance of threat, and whether attention bias is a biomarker of anxiety in children. Methods: We assessed attention bias using an eye-tracker and a visual dot probe task in 14 trauma-exposed children between the ages of 8 and 14. The task included 160 five-second trials, each with a pair of African American or Caucasian faces (angry/neutral or happy/neutral expressions). While participants completed the task, their eyes were tracked to determine fixation duration towards the expressions. Results: We observed that the fixation duration difference score between the angry and neutral faces was larger than the one between the happy and neutral faces, indicating greater bias towards threatening social stimuli. Age was not correlated with fixation towards angry faces, but was positively correlated with fixation towards happy faces. Conclusion: These results align with our hypothesis, which was that there would be a greater fixation to angry faces. It is likely that threat bias is fully developed by age 8, while happy bias develops with age.

Future studies will look into other variables, such as the sex and race of the faces to understand other associations with threat bias. Longitudinal studies will help determine developmental changes with fixation bias.

Summer 2016 SURE Research Symposium

Shoeb Lallani Stereological Analysis of Calretinin-Immunoreactive Interneurons In Transgenic Monkey Models of Huntington’s Disease Shoeb Lallani - Emory University, Department of Human Genetics Emory School of Medicine, Division of Neuropharmacology and Neurological Diseases Yerkes Rosa M. Villalba - Division of Neuropharmacology and Neurological Diseases Yerkes Yoland Smith - Department of Neurology Emory School of Medicine, Division of Neuropharmacology and Neurological Diseases Yerkes Anthony W.S. Chan - Department of Human Genetics Emory School of Medicine, Division of Neuropharmacology and Neurological Diseases Yerkes

Huntington’s disease (HD) is best characterized by atrophy of projection neurons in the striatum.

Recent works have shown that striatal interneurons could also play a pivotal role in the progression of the disease. Specifically, striatal calretinin (CR) interneurons are of particular interest since they are the most numerous interneurons in the regions of interest (ROI) – caudate nucleus and putamen. Previous work has determined that a nonhuman primate (NHP) model for HD shows similarities in degeneration patterns as seen in HD patients. This study conducts a postmortem, neuropathological analysis of the CR interneuron degeneration as seen in HD and control rhesus macaques, differentiating between the caudal and rostral regions of the striatum. Two transgenic HD NHPs were generated, one given a shorter HTT fragment (MR207L) and one a larger fragment (MR220R), and one wild-type (WT) NHP was used as a control (MR255L). Stereology of the brain sections of the transgenic HD NHPs revealed a considerable decrease in CR neurons in the rostral regions of the striatum when compared to the control, but an increase in density of CR interneurons in MR207L. This data suggests that the CR interneurons may be more resistant to HD degeneration than projecting neurons. These results could aid in future investigations regarding the neuronal targets of HD.

Summer 2016 SURE Research Symposium

Alexus Landry Cognitive impairment in parkinson disease did not predict benefit from behavioral intervention for urinary symptoms Alexus N. Landry(1), Musu Sesay MPH(2), Felicia C. Goldstein PhD(3), Theodore M. Johnson II MD, MPH(4,5) , Kathryn L. Burgio PhD(4,6), Patricia S. Goode MSN, MD(4,6), Jorge L. Juncos MD(3), Camille P. Vaughan MD, MS(2,4,5) (1)Louisiana State Univ, Baton Rouge, LA, USA, (2)Dept of Veterans Affairs Rehabilitation Research & Development Center of Excellence for Visual and Neurocognitive Rehabilitation, Atlanta, GA, USA, (3)Department of Neurology, Emory Univ, Atlanta, GA, USA, (4)Dept of Veterans Affairs Birmingham/Atlanta GRECC, (5)Department of Medicine, Emory Univ, Atlanta, GA, USA, (6)Department of Medicine, Univ of Alabama at Birmingham, Birmingham, AL, USA

Urinary incontinence (UI) is a common non-motor symptom in Parkinson disease (PD). This study sought to determine if baseline cognitive function predicts benefit from behavioral intervention to reduce UI in PD. Participants who completed treatment in an ongoing randomized controlled trial of behavioral intervention for urinary symptoms in PD were grouped based on their response to a validated Benefit assessment. The Benefit assessment was conducted 8 weeks after randomization to either continuous bladder diary alone (self-monitoring) or pelvic floor muscle exercise-based behavioral therapy. A screening test of overall cognitive function and domain-specific neuropsychological assessments were conducted at baseline. A Montreal Cognitive Assessment (MoCA) score <18/30 was an exclusion criterion. UI episodes were recorded in 7-day bladder diaries completed at baseline and 8 weeks and change was calculated as a percent. The No Benefit group (n=8) did not differ from the Benefit group (n=20) with respect to baseline overall cognitive function (MoCA) or PD motor symptom severity (UPDRS motor score). There were no significant differences in gender, age, or years with PD between groups (Table 1). The Benefit group reported greater percent reduction in weekly UI episodes compared to the No Benefit group. (Table 1). The two groups did not differ with respect to any domain-specific cognitive test (Table 2). Behavioral intervention for UI provided a favorable treatment option for persons with PD, even in the setting of mild cognitive dysfunction.

Brittany Lee Analyzing the Allylic C-H Functionalization of Certain Aliphatic Alkene Compounds Brittany Lee, Caitlin Bridges, Simon Blakey, PhD

Abstract withheld.

Summer 2016 SURE Research Symposium Solah Lee Non-homologous end-joining pathway inhibits homologous recombination repair of replication-associated DNA damage Solah Lee, Emory University, Hongyan Wang, Winship Cancer Institute of Emory University, Xiang Wang, Winship Cancer Institute of Emory University, Ya Wang, MD, PhD, Winship Cancer Institute of Emory University.

Abstract withheld.

Moon Young Lee

Expression of Cytokines Using Genetically Engineered Probiotic Yeast S. boulardii Moon Young Lee, Milo B. Fasken, PhD Emory University, Anita H. Corbett, PhD Emory University, Tracey J. Lamb, PhD Emory University

Probiotic yeast Saccharomyces boulardii has recently been proposed as a potential vector for oral vaccine delivery protecting against infectious diseases; such a vector would ease the pain and cost of the traditional needlestick vaccination. S. boulardii is genetically close to the well-characterized yeast S. cerevisiae, but is favorable because it grows well in low pH environments. Vaccination using S. boulardii would rely on the use of auxotrophic mutant yeast for antigen secretion.

Auxotroph yeasts have mutations in genes that produce essential enzymes. Therefore, successful transformations can be easily selected with auxotrophs when plated onto selective media, as only yeasts transformed with a plasmid encoding for the required enzyme would survive. Here, we used the Type II CRISPR-Cas9 gene editing system to generate auxotrophic mutants of S. boulardii.

Spotting assays show these mutants have growth similar to that of wildtype. Finally, we transformed these mutants with plasmids that encode cytokines that can be used to signal secretion of antigens. We have generated effective S. boulardii auxotrophic mutants that can be further tested for oral vaccine delivery.

Yun-Hsuan (Stellina) Lee

Genotypic Variation on Neural Phenotypes Yun-Hsuan(Stellina) Lee, Undergraduate, Emory University Charles Zhao& Kathleen Bates, PhD Candidates, Georgia Institute of Technology Hang Lu, PhD, Massachusetts Institute of Technology

Caenorhabditis elegans serves as an important model system in studying neural circuitry with its optically accessible, compact and well-stereotyped nervous system. This study aims to investigate the relationship between genotype and neural phenotype quantitatively and qualitatively by taking advantage of C. elegansâ&#x20AC;&#x2122; genetic homology to humans and high-throughput functional imaging using microfluidic devices. The advancement of targeted gene editing, now allows us to visualize synaptic densities through whole brain imaging analysis. However, efficient feature phenotyping on tens and hundreds of subjects require high-throughput analysis, motivating this project to utilize image processing and microfluidic techniques to investigate novel and subtle effects of genotype on neural phenotypes. Furthermore, the detailed quantitative categorization of features with the developed algorithm for neural analysis enables accuracy during whole ganglion neuronal analysis and investigation of genetic modulation such as epistasis.

Summer 2016 SURE Research Symposium Fredrick Leon Structural Studies of RSV Fusion Glycoptein Using Cryo-Electron Tomography Fredrick Leon, Emory University, Zunlong Ke, Georgia Tech, Chris C Stobart, PhD Emory University, Hong Yi, MS Emory University, Martin L Moore, PhD Emory University, Elizabeth R Wright, PhD Emory University

Respiratory syncytial virus (RSV) is a prevalent infectious virus that targets infants and the elderly.

RSV causes a mild infection of the upper and/or lower respiratory tract in healthy children and immunocompromised adults, but can be a serious cause of illness in certain demographics. RSV is a major cause of bronchiolitis, or inflammation of the bronchial tubes, which may also lead to pneumonia and death. We use cryo-electron tomography (cryo-ET) to investigate the near-tonative state three-dimensional (3D) structures of RSV surface glycoproteins on released virus particles. In this study, elevated temperature was used to trigger the glycoprotein conformational change in addition to the control group. An understanding of the high resolution structure of glycoproteins would be influential for the development and synthesis of further treatments and controls to counteract the incidence of RSV infection.

Valerie Linck Inability to Regulate Blood Pressure in Response to Alterations in Dietary Salt in Mice Lacking VDAC3 Valerie A. Linck, Li Zou, Laura I Galarza-Paez, Linda Li, Otor Al-Khalili , Douglas C Eaton, Tiffany L. Thai, He-Ping Ma

Mitochondria are not only the primary energy source for cells but also help maintaining intracellular calcium [Ca2+]i homeostasis and signaling in many cell types, especially epithelial cells. In the principal cells of renal cortical collecting duct, [Ca2+]i is polarized in domains that lie beneath the apical and basolateral membranes. Our previous research show that mitochondria in principal cells prevent the movement of [Ca2+]i from one pole to the other by taking up Ca2+ and slowly releasing it. Since the physiological relevance of this mitochondrial Ca2+ uptake is unknown, we obtained a mouse lacking a channel (VDAC) that transports Ca2+ across mitochondria. We predicted that knocking out VDAC would have physiological consequences for principal cell function, specifically affecting sodium regulation and inducing a salt-sensitive phenotype. To test our idea, VDAC3 knock out mice and their wild-type littermates were fed with normal chow, low salt, high salt, or high salt with benzamil, an ENaC inhibitor for 2 weeks. We collected measurements for blood pressure and metabolic cages as well as blood and urine samples to be analyzed for osmolality, electrolytes, and protein. VDAC3 knock out mice had lower blood pressure on low salt diet and significantly higher blood pressure on high salt diet. Furthermore, water intake, urine output, urine sodium increased and urine osmolality, urine potassium decreased in mice fed with high salt diet. In conclusion, disruption of VDAC3 leads to an inability to regulate blood pressure in response to changes in dietary salt levels.

Summer 2016 SURE Research Symposium

Pai Liu SRPK2 phosphorylation of AEP in Alzheimer’s Disease Pai Liu, Emory College of Arts and Science, Zhihao Wang, PhD, Xia Liu, and Keqiang Ye, PhD Emory School of Medicine

SRPK2 (serine-arginine protein kinase 2), a critical kinase regulating alternative splicing, is implicated in Alzheimer’s Disease (AD) pathogenesis. Depletion of SRPK2 leading to reduced Tau phosphorylation can effectively improve learning and synaptic plasticity in APP/PS1 mice. AEP (asparagine endopeptidase), remaining active during the process of aging and in brains of AD patients, is also shown to cleave tau to mediate the neurofibrillary pathology. With inhibition of gene encoding AEP, Tau P301S-transgenic-mice show reduced tau hyperphosphorylation and less synapse loss. We hypothesize that SRPK2 can phosphorylate AEP to increase its enzymatic activity, which elevates APP and Tau cleavage and mediates AD pathogenesis. We have found that SRPK2 directly phosphorylates AEP on S226 in vitro. Primary neuronal cultures (DIV13) were treated with 2 or 20 μM of pre-aggregated Aβ overnight and followed by immunofluorescence. Lysosomes, nuclei, and cytosol were extracted from 15 month-old 5XFAD mice and wild type mice brain samples to conduct subcellular fractionation. Primary antibodies including anti-P-S226, anti-AEP, and anti-LAMP1 were used for Western blot. The result has shown that active AEP is phosphorylated and translocated from neuronal lysosomes to cytoplasm. Also, the level of SRPK2phosphorylated AEP in cytosolic fraction showed an increase in AD models compared to wild type.

SRPK2 can regulate subcellular localization and increase its cleavage and activity of AEP in AD mice.

Summer 2016 SURE Research Symposium Josephine Liwang

The Effects of PKC and AMPK on Water Reabsorption in Rat Inner Medullary Collecting Duct Josephine K. Liwang, Jeff M. Sands MD, Janet D. Klein PhD, Yanhua Wang PhD, Emory University Dept. of Medicine: Renal Division

Nephrogenic Diabetes Insipidus (NDI) is caused by the kidney’s inability to respond to vasopressin.

NDI patients experience severe polyuria and have trouble reabsorbing water back into the body.

The protein responsible for the reabsorption of water is aquaporin-2 (AQP2), which is located on the apical surface of the inner-medullary (IM) collecting duct. Recent studies suggest that protein kinase C (PKC) and adenosine monophosphate-activated protein kinase (AMPK) phosphorylate and activate AQP2; however, it is unknown if they stimulate through parallel signaling pathways or if they are interdependent. Our hypothesis is that PKC and AMPK are able to stimulate AQP2 independently and additively. To prove this, we examined the expression levels of AQP2 and AMPK using Western blot analysis. Rat IM tissues were treated with chelerythrine (PKC inhibitor,10µM), Compound C (AMPK inhibitor, 12.5µM), phorbol dibutyrate (PKC activator, 4µM), Metformin (AMPK activator, 4µM), and H-89 (PKA inhibitor, 5µM) at 370C for 30 minutes. The antibodies used to measure protein abundances: total AQP2, p256-AQP2, p261-AQP2, and p-AMPK. H-89 resulted in maintenance of p-AQP2 and increase in p-AMPK. Inhibition of AMPK did not decrease p256-AQP2.

Inhibition of PKC did not decrease p256-AQP2 or pAMPK levels. Activation of PKC and AMPK together maintained p256-AQP2 and p261-AQP2 levels. Conclusion: PKC and AMPK work independently to stimulate AQP2. However, when activated together, PKC and AMPK do not additively stimulate AQP2 to increase water reabsorption. Understanding the regulation of AQP2 will help us understand how urine-concentrating defects occur in NDI and may suggest novel treatment strategies.

Summer 2016 SURE Research Symposium Alejandro Lopez Characterization of Physio-Behavioral Changes in Adult Mice with Nerve Injury-Induced Neuropathic Pain Alejandro Lopez, San Jose State University, Heidi Kloefkorn, PhD, Mike Sawkchuk, PhD, Mallika Halder, Sandra Garraway, PhD, Bill Goolsby, MS, Shawn Hochman, PhD Emory University

Neuropathic pain syndromes are disabling conditions that include spontaneous periods of pain exacerbation, and are linked to depression and anxiety. There are currently no animal models that assess ongoing neuropathic pain. We tested the hypothesis that peripheral nerve injury-induced neuropathic pain leads to measurable changes in respiratory and motor behaviors, and used affordable non-contact, non-invasive electric field sensors that allow for their continuous detection.

We used chronic constriction nerve injury (CCNI) of the right sciatic nerve in C57Bl/6J mice to model neuropathic pain (n = 6 injured; 2 sham). We undertook weekday recordings of motor and respiratory activity (4-6 hours/day) and compared this with conventional tests of thermal and mechanical hyperalgesia. Recordings were undertaken for 1 week prior and 3 weeks after injury.

Analysis involved comparison of mean changes in thermal and mechanical sensitivity, respiratory rate, and overall motor activity. CCNI led to the expected heightened mechanical sensitivity over time consistent with the development of neuropathic pain. Continuous recordings of activity demonstrated that these mice had measurable reductions in motor activity in the first 2 weeks post-injury, and a trend toward increasing respiratory rate in the first week post-injury. We conclude that continuous recordings identified changes in respiration and motor activity associated with the development of neuropathic pain. We propose that future directions involve the use of electric field sensors to continuously monitor and detect spontaneous changes in cardiorespiratory behavior associated with the development of neuropathic pain.

Jennifer Min Development of a Stem Cell Model of the Pediatric Neuromuscular Disease SMARD1 Jennifer Min, Zachary Mceachin, Gary J. Bassell, Ph.D., Wilfried Rossoll, PhD

Spinal muscular atrophy with respiratory distress type 1 (SMARD1) is a rare but fatal pediatric disorder characterized by distal muscular weakness, diaphragmatic palsy, and respiratory failure.

The etiology of this neuromuscular disease is not yet quite understood, and previous clinical case studies suggest there may be other pathogenic mechanisms underlying the symptomatic respiratory distress. SMARD1 is an autosomal recessive disease with mutations in the geneencoding regions of immunoglobulin helicase Îź-binding protein 2 (IGHMBP2), located on chromosome 11q13. Currently, it is unknown why this mutation selectively causes motor neuron cell death. The aim of this project is to model and investigate SMARD1 by 1) generating an induced pluripotent stem cell (iPSC) line by reprogramming patient-specific SMARD1 fibroblasts and 2) characterize the SMARD1 iPSC line using standard characterization methods. Results from this study will provide the tools and insight to further the study of SMARD1 and provide the identification of potential treatments for the neuromuscular disease.

Summer 2016 SURE Research Symposium Esperance Mugabekazi Women are more at risk for developing PTSD than men: A prospective study Esperance Mugabekazi, Agnes Scott College, Vasiliki Michopoulos, PhD, MS, Emory University School of Medicine, Department of Psychiatry &Tanja Jovanovic, Emory University School of Medicine, Department of Psychiatry and Kerry J. Ressler, MD, PhD, Emory University School of Medicine, Department of Psychiatry Harvard/McLean Hospital, Boston, Massachusetts.

Post-traumatic stress disorder (PTSD) is a mental health condition that's triggered by a traumatic event â&#x20AC;&#x201D; either experiencing or witnessing an event that was life-threatening (Mayo Clinic). While many will experience a life-threatening trauma, only a subset of individuals will go on to develop posttraumatic stress disorder (PTSD). One factor associated with increased risk for PTSD is sex, as cross-sectional, epidemiological studies illustrate that females are more at risk for PTSD than men.

In the current prospective study, we hypothesize that women would be more likely to develop PTSD than men in an Emergency Department (ED) right after the occurrence of a traumatic event.

Meagan Naraine Nuclear Factor-kB mediates Zinc Deficiency-Induced NCC Upregulation in Mouse Distal Convoluted Tubular Cells Meagan K. Naraine, Emory University; Mirandy S. Li, BS Emory University; Jasmine Williams, Emory University; Monisha Mistry, Emory University; Robert S. Hoover, MD Atlanta VA Medical Center & Emory University; and Clintoria R. Williams, PhD Atlanta VA Medical Center & Emory University

Zn2+ deficiency (ZnD), a common disorder in chronic kidney disease, contributes to hypertension.

However, the mechanism is unknown. The Sodium Chloride Cotransporter (NCC) plays a critical role in blood pressure homeostasis. Recently, our lab found that ZnD leads to a hyperactivation of NCC that was reversed with Zn2+ supplementation, suggesting a novel mechanism by which ZnD contributes to hypertension. Other studies have shown that Nuclear Factor-kB (NFkB) is activated in ZnD.1 The purpose of this study was to determine the role NFkB plays in ZnD-induced NCC upregulation. We hypothesize that ZnD activates NFkB, which leads to a hyperactivation of NCC. To investigate this mechanism, mouse distal convoluted tubular (mDCT) cells were treated with the NFkB inhibitor Caffeic acid phenethyl ester (CAPE) for 24 hours, prior to inducing ZnD with the Zn2+ chelator TPEN for an additional 24 hours. NCC expression (via qRT-PCR, Western blot, and immunofluorescence) and NCC localization (via biotinylation) were examined. Compared to Vehicle, TPEN increases NCC mRNA and protein expression. However, CAPE prevents NCC mRNA and protein expression. Based on these results, it is apparent that NFkB plays a role in ZnD-induced NCC upregulation in mDCT cells. Understanding the mechanisms by which ZnD contributes to hypertension may have an important impact on the treatment of hypertensive kidney disease.

Summer 2016 SURE Research Symposium Anders Olsen Discovering Phage Antibody Selectivity and Optimization in Bacterial Systems John Nicosia, Dwight Chambers, Anton Bryksin PhD and Thomas H. Barker PhD

Using therapeutic phage display systems, we can discover unique antibodies that will selectively bind to a desired proteins. Phage display research has many applications including: disease identification, therapeutic antibody production and protein selectivity. H6 proteins is an antibody that was discovered to be highly selective in binding to fibrin over fibrinogen. Fibrinogen is modified into fibrin in order to polymerize. Platelets bind to these polymerized fibrin in order to form blood clots at wound sites. Here, we researched the various growth media that could be used to produce the H6 antibody in bacterial systems. We looked for which media was successful in optimizing the production of the desired H6 protein. Similarly in another project, we used phage display systems to discover an antibody that could differentiate Citrullinated Fibronectin (CitFn) over Fibronectin (Fn). Citrullination is a post translational modification to proteins. This modification is common in cells crippled with cancer, idiopathic pulmonary fibrosis and rheumatoid arthritis. However, citrullination is difficult to to identify and differentiate with antibodies. We used a phage library to discover proteins that could selectivity bind CitFn over Fn. Our findings discovered 22 candidate proteins that could be used as selective antibodies to citrullination.

Sahar Panjwani Predicting Real World Functioning Using Neuropsychological Assessments in an Accountability Court Population Sahar Panjwani, Gershom T. Lazarus M.A., Eugene K. Emory Ph.D ---All Emory University

This study investigated the use of neuropsychological assessments to predict real world functioning in an Accountability Court sample. Research has indicated evidence of cognitive deficits in executive functioning through neuropsychological assessments in chronic substance abusers. Length of substance abuse, type of substance abuse, and performances on the Wisconsin Card Sorting Test, Delis-Kaplan Executive Function System, and the Rey-Osterrieth Complex Figure Test were examined. These were compared to variables that were indicative of normal functioning, such as number of arrests and failed urine analyses. In order to examine these variables, a clinical interview and a neuropsychological battery were administered to each participant in the study. Findings suggest that lengths of use of cocaine and alcohol have a strong positive correlation with number of arrests. The results of the study suggest the need for further research in this area with an extended neuropsychological battery, larger sample size, and control groups for variables, such as gender and education. The mechanisms of action and brain areas implicated in substance abuse reward pathways, as well as the real world applicability of each test examined are briefly discussed.

Summer 2016 SURE Research Symposium Jonathan Park Role of LCMT-1 in Mouse Fetal Hepatoblast Growth and Division37 Jonathan Park, Daniella Gonzalez, Grant D. Schleifer, Chris Cutler and David C. Pallas37, PhD. Emory University School of Medicine37.

Protein Phosphatase 2A (PP2A) is a protein phosphatase involved in many different pathways in cell cycle, morphology and development. LCMT-1 and PME-1 catalyze the methylation and demethylation of the catalytic subunit of PP2A, respectively. Previous studies revealed that knocking out the LCMT-1 gene results in liver and hematopoetic defects in mouse embryos.

Embryos lacking the LCMT-1 gene never survive past 17 days of gestation. In order to examine the liver defects, two primary techniques for data collection were used. First, taking advantage of the strong expression of Dlk-1 antigen on the cell surface of hepatoblasts past embryonic gestation day 10.5 (E10.5), anti-Dlk-1 antibodies were used to compare the number of hepatoblasts in wild type and knockout fetal livers. Second, using Alpha-fetoprotein::mCherry (Afp::mCherry) transgenic LCMT-1 knockout mice present in the Pallas lab, time lapse phase and fluorescent microscopy were performed on isolated hepatoblasts, examining cell proliferation, mitosis, and cell cycle length.

These approaches determined the extent and nature of defects in hepatoblats lacking LCMT-1, providing the first insights into LCMT-1 function in these cells.

Sean Perryman Maternal Depression and Parenting: Testing the Moderating Role of Affect Sean T. Perryman, Katherine A. Cullum, M.A. Emory University, Sherryl H. Goodman, Ph.D. Emory University, & Judy Garber, Ph.D. Vanderbilt University

The relations between past major depressive disorder (MDD), current depressive symptom level, current positive and negative affective state (PA and NA), and parenting behaviors were assessed among 131 mothers of 8-10 year olds. Of these mothers, 50% had a history of MDD. Mothers’ mean age was 40.17 years, 45.3% completed graduate or professional programs, and mean household income was $90,000-$100,000. We hypothesized that both history of MDD and current depression symptom levels would be associated with lower amounts of positive parenting and higher amounts of negative parenting. .We also predicted that mothers’ depression (history and current) and positive parenting would be more strongly related in the presence of mothers’ lower PA, and that mother’s depression (history and current) and negative parenting would be more strongly related in the presence of higher NA. We relied on children’s reports of parenting to avoid problems associated with single-source variance. Our results failed to support the hypotheses. Neither history of depression nor current symptom level predicted positive or negative parenting.

Furthermore, we did not find a moderating role of mothers’ PA in the association of current depressive symptoms and positive parenting (∆R2=.004), nor did we find a moderating role of mothers’ NA in the association of current depressive symptoms and negative parenting (∆R2=.024).

Mothers’ current depression symptom levels and affect scores (PA and NA) were highly intercorrelated, which may have interfered with finding a moderating relationship. In addition, our sample was relatively well resourced, which may have buffered the relation between depression and parenting.

Summer 2016 SURE Research Symposium Zhiyu Qian When Oncogene Meets Metabolism: BRAFV600E-specific Regulation of CSGLcA-T in Human Melanoma Zhiyu (Jason) Qian, Ruiting Lin, and Jing Chen

The importance of metabolic alterations in cancer has been increasingly recognized over the past decade. The metabolic reprogramming and rewiring in cancer cells represent distinct mechanisms underlying changes of metabolic circuits, which are either “hijacked” from normal cells or newly “forged” due to “neo-function” of oncogenic mutants. Although increasing evidence emerges and suggests that different human cancers may share common metabolic properties, one question that remains to be unclear is whether distinct oncogene mutations in different cancer types may require different metabolic properties for tumor development, and thus specifically “rewire” and “reprogram” cancer cell metabolism. To approach to this question, we decided to identify the unique “metabolic vulnerability” required by distinct oncogenic mutations. We designed and constructed a novel shRNA library that targets 1,361 out of 1,417 genes encoding known metabolism-related enzymes and protein factors in the human genome. As our first attempt by using this library, we identified a glucuronyltransferase in chondrogenesis, CSGLcA-T as a “synthetic lethal” partner of oncogenic BRAFV600E in human melanoma cells compare to control cells expressing NRas Q61K/R mutant or wild-type (WT) BRAF and NRas. However, the molecular mechanism of this specific regulation is unclear and need to be further addressed.

Chengcheng Qiu Assessment of Floristic Composition to Elucidate Flora Change at Lullwater Preserve Chengcheng Qiu, Emory University Herbarium, Tharangamala Samarakoon, Emory University Herbarium, and Center for the Study of Human Health of Emory University, Carl Brown & Stefanie T. Pierce, Department of Environmental Studies of Emory University, Cassandra L. Quave, Emory University Herbarium, Department of Dematology of Emory University School of Medicine, and Center for the Study of Human Health of Emory University

Lullwater Preserve is a 132-acre majestic nature preserve located at Emory University in Atlanta, Georgia. It holds an irreplaceable place in supporting many native, rare and endemic Piedmont trees and herbs, as well as a variety of wildlife. This research involves the study of floristic composition in Lullwater Preserve and field observations to investigate interrelated factors that lead to floristic changes over the years. We examined Lullwater specimens collected in 1900s from Emory University Herbarium, made voucher specimens from present plant species, and conducted plant invasion study and field observations to elucidate changes of flora. By the end of our research, we had restored 678 records of Lullwater specimens from 1900s, and had made 99 new specimens collected in 2016. Also, we infer that natural succession, plant invasion, animal selective feeding, and human disturbance should be included in main factors of causing floristic changes in Lullwater Preserve, and treating for invasive plants has a great potential of restoring the natural forest. The collection of Lullwater specimens and inference of factors that are causing floristic changes can be used to understand forest development, raise public awareness of ecological importance of Lullwater Preserve, and help plan future maintenance.

Summer 2016 SURE Research Symposium Kaushik Ravipati Effects of an M1 Receptor Agonist on Memory Dysfunction in an Alzheimerâ&#x20AC;&#x2122;s Disease Rat Model Kaushik Ravipati, Emory University; Claire R. Galloway, Emory University; Joseph R. Manns, PhD Emory University

Alzheimerâ&#x20AC;&#x2122;s disease (AD) is notoriously one of the most destructive neurodegenerative disorders.

The two hallmarks of AD, beta-amyloid plaques and neurofibrillary tangles, are accompanied by cholinergic dysfunction in the brain. As a result, acetylcholine transmission is disproportionately reduced in the hippocampus, producing the recognition memory deficits often seen in AD. Current treatments seek to improve acetylcholine transmission through the use of acetylcholinesterase inhibitors, but are extremely limited in their effectiveness due to a lack of receptor specificity. A more effective solution might be the selective M1 receptor agonist VU0364572 since the M1 receptor, a muscarinic acetylcholine receptor subtype, is present in high concentrations in the areas important for memory, such as the hippocampus, and low concentrations in the periphery. In addition, activation of M1 receptors has been shown to reduce beta-amyloid aggregation and tau pathology. The performance of a transgenic rat model of AD (TgF344) will be compared to wildtype rats on an object-in-location recognition memory task that takes advantage of the innate preference that rats possess for novel objects when compared to repeated objects. In this task, AD rats are expected to have decreased memory performance compared to wild-type rats, followed by an improvement in memory in the former group after administration of VU0364572. AD rats showed worse memory than WT rats on the object-in-location recognition memory task. The memory deficits displayed by AD rats were not alleviated by acute administration of VU0364572.

Future experiments should use different memory tests with a broader range of VU0364572 doses.

Summer 2016 SURE Research Symposium William Reis Inhibition of PDE7 Reduces Inflammatory Response Following Renal Ischemia/Reperfusion Injury William R. Reis and Mitsi A. Blount, Department of Medicine, Emory University School of Medicine

Ischemia/reperfusion (I/R) injury is an unavoidable consequence after kidney transplantation and is clinically associated with kidney graft rejection and/or dysfunction. I/R affects many regulatory systems at the cellular and renal tissue level to promote the pro-inflammatory response through chemokines and cytokines. This may lead to progressive interstitial renal fibrosis and declining renal function that ultimately impacts long-term graft outcome. Because elevated cAMP can offset the this harmful, sustained immune response, identification of a targetable protein to increase kidney cAMP levels may provide a viable option to prevent I/R injury before organ recovery by donor pretreatment. This study aimed to understand the benefits of raising renal cAMP levels through PDE7 inhibition for treating the immune response following I/R insult. Preliminary qRTPCR studies showed that PDE7 inhibition reduced expression of harmful cytokines (IL-2, IL-6, IL12β, IL-18) and chemokines (CCL2, CCL5) to sham levels, when stimulated with TNFι in HK-2 cells.

To model this treatment in vivo, we surgically inhibited bilateral renal circulation of SpragueDawley rats for 30 minutes. Ischemia-reperfusion (I/R) surgery is a well-established model for the transplant injury. After such time, perfusion was restored and the rats recovered in metabolic cages for 24 hours before being sacrificed; blood, urine and kidney tissue were harvested at the time of sacrifice. qRT-PCR analysis of kidney tissue showed that I/R injury increased expression of proinflammatory cytokines and chemokines in sham treated rats (n=3). However, when the rats were treated with PDE7 inhibitor (n=3) TC3.6, cytokine and chemokine expression was reduced to sham surgery (n=6) levels. This was noted specifically with the cytokine IL6 and the chemokine CCL5.

Blood Urea Nitrogen (BUN) levels showed a similar pattern in the samples. Reduced cytokine and chemokine expression, as well as reduced BUN levels in treated versus untreated I/R rats, is evidence that PDE7 inhibition may be an effective pretreatment prior to kidney transplant to prevent the necessary I/R injury associated with the surgical procedure.

Summer 2016 SURE Research Symposium

Abinand Rejimon Quantifying Vascular Dynamics in Inferior Frontal Regions using BOLD Signal Derived from Echo Planar Imaging Abinand Rejimon, Indiana University Bloomington, Keith McGregor and Joe Nocera, Center for Visual and Neurocognitive Rehabilitation Atlanta VA & Emory University Department of Neuroscience

A recent project completed by Drs. Joe Nocera and Keith McGregor, has shown that engagement in aerobic exercise is associated with decreased blood oxygenation level dependent (BOLD) signal in right inferior frontal regions (IFG - right pars triangularis and pars opercularis). These changes are negatively correlated with improved cognitive function measured by language tasks. That is, postaerobic training, participants who show decreased right frontal activity show improved verbal fluency. However, changes in BOLD activity have both a neural and vascular origin. As aerobic exercise is well-known to affect vascular function, dissociating the neural and vascular components is complicated in standard functional MRI analysis. Acknowledging the limitations of the acquired signal, we applied an exploratory statistical analysis using a deconvolution approach to attempt to quantify vascular change in IFG. Two groups of participants engaged in either a 12-week aerobic (spin cycling) exercise intervention or a non-aerobic contact controlled intervention. All participants performed a semantic fluency task during fMRI data acquisition before and after interventions. We performed standard imaging preprocessing using AFNI software. Standard generalized linear model analysis revealed significant differences in right IFG between groups after the interventions. We used these regions (BA 44/45) to extract the voxel timeseries for deconvolution analysis allowing for comparisons of baseline to BOLD signal change using acquired and residual timeseries. Results from this analysis will be used for comparison to future MR imaging acquisitions employing vascular reactivity challenges. This affords us additional insight into the neural component of the BOLD response to an exercise intervention.

Summer 2016 SURE Research Symposium

Nelson Reyes Determine contribution of RSV-CX3CR1 infectivity in Human Epithelial Cells (HEp-2).

Nelson O. Reyes, Universidad del Este, Carolina, PR. Devi Rajan PhD and Tania Chirkova PhD, Department of Pediatrics and Infectious Disease, Emory Children Center, Atlanta GA

The respiratory syncytial virus (RSV) is the major cause of serious respiratory illness in young children. This virus also affects aged persons and the WHO estimates that RSV causes 64million infections annually. Given the failure of the vaccine in 1960’s which led to enhanced disease in children after natural infection, there is no vaccine available to prevent this infection. Despite high titer neutralizing antibody in cord blood and serum prevent the risk for severe disease, infection can still occur. Interestingly, recent studies suggest that many of the questions regarding neutralization antibody and protective immunity may be because the type of cells normally used for neutralization study (continuous cell lines). RSV uses CX3CR1 as a primary receptor in primary human airway epithelial cells (HAECs). Unlike HAECs, in continuous cell lines, RSV uses heparan sulfate proteoglycans (HSPG’s) through heparin binding domains to bind and infect the cells.

Designing continuous cells lines that represent similar infectious processes to HAECs, would be a promising step in neutralizing antibody studies most relevant to human infection and vaccine development. Given that, in this study, we want to determine the infectivity of human epithelial type-2 (HEp-2) cells after transiently transfected with CX3CR1. The cells were pretreated with heparinase to block HSPGs binding and an RT-PCR was run to determine viral replication. Our preliminary results suggest that heparinase treatment partially prevented RSV infection of HEp-2 but, CX3CR1 did not enhance virus replication. Efforts are under way to more efficiently block HPSG’s and better focus on CX3CR1 mediated infection.

Lindsay Rhoades Possible Detection of Trans-Methyl Formate Lines in the Millimeter-Submillimeter Regime Lindsay Rhoades (Emory University), Luyao Zou (Emory University), Susanna Widicus Weaver, PhD (Emory University)

Methyl Formate (MeF) is a complex organic molecule found throughout space, and has two conformers, cis and trans. Trans-MeF is thought to be able to serve as a probe for gas phase chemistry in the Interstellar Medium. The spectrum of trans-MeF is not known beyond 60 GHz. Using a “fast sweep” technique, cis-MeF was supersonically expanded in vacuum and exposed to a discharge in order to excite the molecule over the 2880 K potential barrier between the conformers. The spectrum of the system was taken from 164.6-224.6 GHz. After comparing the discharge spectrum to both a spectrum taken without discharge and a direct absorption flow cell spectrum, two lines were unidentified between 165-166 GHz. These lines may be a detection of trans-MeF in the millimeter-submillimeter regime.

Summer 2016 SURE Research Symposium

Martin-Louis Riu C-H Functionalization and Cyclopropanation Using Novel Oxidatively Robust Copper Catalyst Martin-Louis Riu & Chris Scarborough, PhD Emory University

The selective transformation of C-H bonds could drastically alter the way in which all organic molecules are constructed. Diazo compounds are integral in this field of chemistry. However, regioselective functionalizaion and carbene dimerization of especially reactive acceptor-only diazo compounds, such as ethyl diazoacetate, is especially problematic.1 Furthermore, the field is dominated by 2nd and 3rd row transition metal catalysts, which are expensive and highly toxic.2 There has been some success in copper-based, cyclopropanation and C-H functionalization of alkanes and alkyl aromatics.3 However, these copper catalysts are still susceptible to decomposition and carbene dimerization. Presented, is an array of novel oxidatively robust copper catalysts that address problems associated with catalyst decomposition and dimerization of diazo compounds. Coordinated to the copper center is a synthetically flexible triazacyclononane ligand derivative. These ligands contain oxidatively robust R-groups of adjustable steric bulk that prohibit it from reacting with all but the most active carbenoid catalyst under ambient conditions. Steric bulk associated with these R-groups clutter the environment surrounding the active site and aid in suppressing dimerization. This project integrates synthetically flexible, yet oxidatively robust, triazacyclononane ligand derivatives into the field of copper-based carbene transfer chemistry.

Wendy Rodriguez Changes in Cerebral Blood Flow after Repetitive Concussions in Adolescence Wendy J. Rodriguez, Erin M. Buckley, PhD

This project studiesIncreased risks of long-term effects (up to one year) after repetitive concussions has motivated this study. The purpose of this study is to identify the relationship between CBF and cognitive outcome after repetitive concussions in an adolescence. Using an unpublished adolescent mouse model of human concussion, we looked at the effect of repetitive concussions on cognition and on CBF measured with diffused correlation spectroscopy. This research is important for learning the effects on CBF after repetitive concussions in adolescence and the possibility of using CBF as an objective biomarker to identify a patient's risk of worse cognitive outcome with subsequent concussions.

Summer 2016 SURE Research Symposium

Jenna Sands Comparative Immunobiology of Mesenchymal Stromal Cells and Hepatic Stellate Cells Jenna Sands SURE & PERSE, Raghavan Chinnadurai, Phd & Jaques Galipeau, MD Department of Hematology and Oncology, Emory University School of Medicine, Georgia, USA

Mesenchymal stromal cells (MSCs) are widely being tested in cell therapy clinical trials for auto and allo immune disorders. Investigation of the immunobiology of MSC and â&#x20AC;&#x153;MSC likeâ&#x20AC;? nonhematopoietic cells is important to define additional cell therapeutic mechanisms. Hepatic stellate cells (HSCs) are MSC-like cells found in the liver and are the source of matrix deposition that leads to liver fibrosis. The current understanding of the comparative cellular and immunological characteristics of MSCs and HSCs are not entirely clear, which hampers the clinical translation of MSC based therapy targeting liver injury in auto and allo immune disorders. We aimed to clarify the biological similarities and differences between MSCs and HSCs. MSCs were isolated from the bone marrow of healthy individuals and HSCs were derived from transplant rejected liver grafts. Our data demonstrate that HSCs express identical phenotypic markers of MSCs that are defined by the International Society for Cell Therapy (ISCT). Our results also showed that both MSCs and HSCs upregulate the immunosuppressive enzyme Indoleamine 2,3 deoxygenase (IDO) upon IFNg stimulation, which can also be blocked by JAK-STAT inhibitor (Ruxolitinib). Altogether, our study provide evidence that MSC-like cells respond to inflammation and injury in a similar manner. In addition, identification of IDO pathway consensus between these two cell types provide insights on the central role of IDO mediated immunosuppression by non-hematopoietic immune cells. Further studies of comparative and contrast biology between MSCs and HSCs are therefore urgently needed.

Priya Sathyanarayan Spherical Aggregation Improves Human Cardiac Progenitor Cells Endothelial Lineage Commitment via Notch-HDAC2 Mechanism Priya Sathyanarayan, Johns Hopkins University, David Trac, Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology & Emory University School of Medicine, Michael Davis, Division of Pediatric Cardiology, Department of Pediatrics, Emory University School of Medicine

Congenital heart disease remains a significant issue due to its high mortality rate. For example, hypoplastic left heart syndrome has a 58% survival rate at 5 years after surgical repair and requires cardiac transplantation by 7 years to treat heart failure (1). Surgical repairs are not a long-term solution as patients inevitably suffer from heart failure and require a cardiac transplant. However, stem cell therapy using cardiac progenitor cells (CPCs) is a potential treatment that can be used to avoid transplantations. By elucidating the mechanism behind CPC lineage commitment, we could potentially improve differentiation rates as well as the regenerative potential of CPCs. Aggregating CPCs as spheres robustly increases Notch signaling, as well as CPC lineage commitment. In addition, we have shown that CPC aggregation increases histone deacetylase (HDAC 1 and 2). One way that HDAC could be influencing differentiation is by regulating transcription factors for cardiac development, such as Gata4. Through the characterization of HDAC in aggregated spheres (3D spheres) as well as testing the effects of HDAC inhibitors on lineage commitment markers, we will be able to gain a better understanding of how HDAC is affecting CPC lineage commitment.

Summer 2016 SURE Research Symposium

Niraj Shah Regulation of Dendritic Morphology by BDNF in a non-miR-137 dependent manner Niraj Shah, Emory University, Kristen Thomas, Emory University, Gary Bassell, PhD Emory University

Schizophrenia is a cognitive disorder with many phenotypes, usually being characterized by hallucinations and abnormal social behaviors. Nearly 1% of the world’s population is affected by schizophrenia. The disorder has no known definitive cause making it difficult to develop an effective treatment or cure. A Genome-Wide Association Study (GWAS) identified a microRNA 137 (miR-137) SNP associated with schizophrenia. microRNAs are non-coding RNAs involved in RNA silencing and regulation of gene expression. miR-137 is known to regulate neuronal development, but the mechanism by which miR-137 regulates dendritic morphology remains poorly understood.

The ErbB/neuregulin signaling complex has been associated with schizophrenia and stimulating dendritic outgrowth. miR-137 has several downstream targets in the ErbB signaling pathway, including the mammalian target of rapamycin (mTOR) signaling pathway. The mTOR signaling pathway is important for protein synthesis, more specifically for proteins playing a role in neuronal morphology and synaptic plasticity. We found that inhibiting miR-137 prevents Neuregulin 1α from stimulating dendritic outgrowth. We also found that Neuregulin 1α acts through the mTOR pathway to stimulate dendritic outgrowth and S6 phosphorylation. Our findings led us to the current study which was to determine whether miR-137’s effect is specific to the Nrg1α pathway or if it affects additional pathways that act through the mTOR pathway. Brain derived neurotropic factor (BDNF) stimulates both dendritic outgrowth and the mTOR pathway, similarly to Nrg1α.

Immunofluorescence shows that BDNF stimulates S6 phosphorylation and increases dendritic complexity independent of miR-137. Immunofluorescence also shows that BDNF stimulates S6 phosphorylation independent of the mTOR pathway.

Summer 2016 SURE Research Symposium

Nilang Shah A novel genetic screen in Drosophila designed to discover secreted factors that drive glioblastoma initiation and progression.

Nilang Shah, Clay Coston Rowe, Joanna Wardwell-Ozgo, PhD, Renee D. Read, PhD. Department of Pharmacology, Emory University School of Medicine.

Glioblastomas (GBMs) are highly proliferative, invasive primary brain tumors derived from glia and glial progenitor cells, and are incurable with current treatments. GBMs often possess mutations that activate epidermal growth factor receptor tyrosine kinase (EGFR) and Pi-3 kinase (PI3K) signaling pathways. Studies show that additional factors and mechanisms in the tumor and the tumor microenvironment cooperate with EFGR and PI3K mutations to drive tumorigenesis, although the identity of these factors remains unclear. To address this problem, we sought to identify proteins that, when overexpressed and secreted into the tumor microenvironment, contribute to tumor initiation and progression in glial cells transformed by constitutive co-activation of EGFR and PI3K signaling. To test the activity of secreted factors on GBM initiation and progression, we used a GBM model in the fruit fly Drosophila melanogaster in which co-activation of EGFR and PI3K drives neoplastic transformation of glial progenitor cells. In the Drosophila GBM model, tumorous glia arise in vivo in the central nervous system in live Drosophila larvae, and create large easily visualized tumors that kill their hosts in 5-6 days. Thus, this GBM model system allows us to view the effects of overexpressed secreted factors on tumorigenesis in an intact brain microenvironment. Using this Drosophila GBM model, we performed an enhancer/suppressor screen wherein individual overexpression constructs for nearly every secreted and transmembrane gene in the Drosophila genome were tested using the Gal4-UAS system for their phenotypic effects on tumor growth, tumor progression, over-all brain size, and host survival. Thus, both the cellautonomous and cell-nonautonomous effects of the tested secreted factors were phenotypically assessed in our screen. From this screen, we identified several genes as candidate enhancers. Using immunohistochemistry and fluorescent microscopy to stain and image whole dissected brains from Drosophila larvae, we analyzed the phenotypes caused by overexpression of the candidate enhancer genes on neoplastic glia transformed by co-activation of EGFR and PI3K. The results of our screen and the phenotypes of candidate enhancers will be presented. Our findings could reveal novel genes that, if evolutionarily conserved in humans, may drive GBM initiation and progression in the brain.

Summer 2016 SURE Research Symposium

Monica Sharma Effects of SNr inhibition in mice performing a locomotor go task Monica Sharma, Undergraduate Emory University, Arthur E. Morrissette, Bachelor's Degree Georgia Institute of Technology, Dieter Jaeger, PhD University of Michigan

The substantia nigra pars reticulata (SNr) is the main output structure in the rodent basal ganglia.

The hypokinetic disorder of Parkinsonâ&#x20AC;&#x2122;s is believed to result from a dopamine deficiency that results in increased inhibitory activity in the SNr. Optogenetics might make it possible to better treat this motor disease, but first, one must obtain a more concrete understanding of neural circuitry. In order to find the best strategy to have mice learn making a go decision based on different whisker sensory cues and to learn more about how the basal ganglia affects movement, we engaged a mouse in a sensorimotor discrimination task, in which correct trial performance was measured. It was a go task in which the stimulus was an air puff, and the response was running.

Upon achieving a success rate of 70%, optogenetics was used to inhibit the SNr following the go stimulus. We hypothesized that if the SNr were inhibited following the go signal, then the mouse would continue to run, and reaction time would speed up. As per our statistical analysis, reaction time was higher with optogenetic stimulation. This suggests that optogenetic stimulation influences movement. In the future, we would perform a stop task to further enhance our understanding of basal ganglia output and neural circuity.

Telissa Spencer Overground Walking Force Production and Associations with an Observational Gait Assessment in Persons with Chronic Incomplete Spinal Cord Injury Telissa Spencer, BS University of Texas at Austin Denise M. Peters PT, PhD Emory University Yann Thibaudier, PhD Emory University Randy D. Trumbower, PT, PhD Emory University

Regaining the necessary motor skills to walk efficiently is an important goal for many persons with chronic incomplete spinal cord injury (iSCI). Gait deficits such as reduced speed and increased risk of falling often limit their capacity to ambulate independently in different environments. These impairments suggest that force production could be reduced in this population. However, force profile characterization has never been investigated in persons with iSCI during overground walking. Clinicians often use assessment tools, such as the Spinal Cord Injury Functional Ambulation Inventory (SCI-FAI), as an inexpensive and efficient way to evaluate deficits in persons with iSCI. This assessment examines spatiotemporal parameters of gait and quantifies the quality of walking in persons with iSCI. However, this tool does not provide direct insight on force production during overground walking, and how the SCI-FAI could be a good indicator of force production remains to be examined. Our objectives were to 1) characterize stance-phase force production during overground walking in persons with iSCI compared to able-bodied controls, and 2) examine the association between force production and the SCI-FAI score. Our results showed a reduction in both fore-aft and vertical force production in persons with iSCI, as well as associations between braking force/timing and vertical loading at push off and the SCI-FAI score. The information gained from this study would be most useful in developing targeted therapeutic interventions that focus on improving force production in persons with iSCI, and thus, potentially improving SCI-FAI score and the quality of walking.

Summer 2016 SURE Research Symposium

Phoebe Stark 2B4 Deficiency Impacts The Expansion And Proliferation But Not Death Of Donor-Reactive CD8+ T Cells Phoebe C. Stark, Sonia J. Laurie, B.S, and Mandy L. Ford, Ph.D. Emory Transplant Center, Emory University School of Medicine, Atlanta, GA

Previous studies from our group suggest that expression of 2B4, a cosignaling protein expressed on subsets of CD8+ T cells, may be associated with improved graft survival following transplantation.

In order to elucidate the effect of 2B4 on T cell activation, proliferation, death, and bioenergetics, CD8+ 2B4 knockout TCR transgenic T cells specific for a single, graft-expressed antigen (OT-I) and wild type (WT) OT-I controls were stimulated with cognate peptide in vitro for three days. Flow cytometry was used to assess differences between WT OT-I and 2B4KO OT-I cells at 0, 24, 48, and 72 hrs post stimulation. Interestingly, we observed that fewer 2B4KO OT-I T cells underwent division as measured by dilution of intracellular dye at both 24 and 72 hours post stimulation. We hypothesized this may be due to increased cell death. To test this, we repeated the experiment and analyzed the number of apoptotic T cells. We found no difference in cell death between 2B4KO and WT T cells. However, expression of CD127 and CD62L, two markers that are highly expressed on naïve T cells, was retained on activated 2B4KO T cells relative to WT controls. This indicated that 2B4KO T cells may have a reduced capacity to differentiate into effector cells. In summary, these data are consistent with a costimulatory role for 2B4 in augmenting differentiation and proliferation following T cells activation. These findings may contribute to the development of new 2B4-targeting immunotherapies to inhibit donor-reactive CD8+ T cell responses and prolong graft function following transplantation.

Jazz Stephens A Comparison of Two Disinfectants in the Sanitation of Monkey Automated Feeders Using ATPase-based Microbial Methods Brandon Hughes & Julie Moran & Mark Sharpless & Kelly Ethun, Emory University

The disinfection and sanitation of areas containing possible harmful microbes in animal laboratory settings is very important to maintain the welfare of the animals as well as to provide integrity to possible studies utilizing the animals. The purpose of this experiment is to compare the efficacy and efficiency of two disinfectants (Quaternary and Phenolic) on automated feeders used in a nonhuman primate (NHP) enclosures at the Yerkes National Primate Research Center. I hypothesize the Husky 810 disinfectant will be more effective than the Quatricide PV disinfectant due to various studies that suggest phenolic disinfectants are more effective than quaternary disinfectants in areas containing organic material. The goal is to develop a standard operation procedure that adequately removes potentially harmful biological material to utilize throughout the facility.

Summer 2016 SURE Research Symposium

Wenzheng Sun Spherical nucleic acid-pMHC multimer for T-cell detection and immunophenotyping Wenzheng Sun, Undergraduate Emory University, Victor Pui-Yan Ma, PhD Emory University, Hunter Martinez, Emory University, Brian Evavold & Khalid Salaita, Emory Univeristy.

Interaction between T Cell receptors (TCRs) and peptide-MHC is a key interaction in adaptive immune responses. However, monomeric pMHC has low binding affinities to TCRs, making detection and quantification of antigen specific T cells a challenging process. To address this problem, we are developing multimeric peptide-MHC particles for detecting and phenotyping antigen specific T-cells. The core component of our pMHC particles is a spherical nucleic acid, which consists of densely functionalized DNA on a gold nanoparticle surface, functionalized with antigen specific peptide-MHC. Preliminary flow cytometry results suggest our particles are able to pick up a higher percentage of high affinity CD4+ T cells (SMARTA) when compared to tetramer, the current gold standard used for T-cell detection and phenotyping in the field. Furthermore, the particles are able to detect antigen specific CD4+ and CD8+ T-cells in polyclonal T-cells. Our particles achieved comparable level of staining with a 1000 fold concentration deficit compared to the tetramer staining procedure, making it a robust and effective alternative to the current staining technologies.

Meklit Tilahun Biochemical characterization of the Mycobacterium tuberculosis MazEF-mt6 toxin-antitoxin complex Meklit Tilahun, Eric D. Hoffer, and Christine M. Dunham, Ph.D. Department of Biochemistry, Emory School of Medicine

Antibiotic resistant bacteria are threatening the effectiveness of antibiotics and thus are challenging modern medicine. The activation of toxin-antitoxin gene pairs activated during stress aid bacteria in their transitioning to a persistent state. This persistent state is characterized by the ability of the bacteria to be antibiotic tolerant. This persistent state is moderated by the activation of toxinantitoxin systems such as MazEF toxin-antitoxin complex. E. coli and Bacillus subtilis MazEF toxinantitoxin complex is one such system whereby MazF is an RNase that cleaves free messenger RNA to inhibit growth during stress conditions. During non-stress conditions, MazF is inhibited by a protein antitoxin called MazE. In Mycobacterium tuberculosis, there is a significant expansion of the MazEF family with 6 different paralogs. Here we characterize Mycobacterium tuberculosis MazFEmt6 where the MazF-mt6 toxin was shown to cleave ribosomal RNA to inhibit protein synthesis. We use microbiology, biochemical and structural biology approaches to overexpress, purify and solve the three dimensional structure of the unique Mycobacterium tuberculosis MazEF-mt6 toxin antitoxin co-complex. These results will provide molecular insights into the structure and function of the Mycobacterium tuberculosis MazFE-mt6 system.

Summer 2016 SURE Research Symposium

Phi Trac ASIC1a/α-ENaC Hybrid Channels Contribute to Alveolar Fluid Clearance Phi T. Trac, Emory University, Megan M. Greenlee, PhD Emory University, Tiffany Thai, PhD Emory University, Qiang Yue, Emory University, Otor Al-Khalili, Emory University, Abdel Alli, PhD Emory University, Douglas C. Eaton, PhD Emory University

Pneumonia mortality is caused by abnormal alveolar fluid clearance (AFC) preventing normal gas exchange with consequent hypoxemia. It is reasonable to conclude that α-ENaC is responsible for all AFC because α-ENaC knockout completely prevents newborn mice from clearing lung fluids.

However, complete ENaC inhibition with amiloride decreases AFC by only half. Single channel measurements from alveolar epithelial cells suggest another channel contributes to AFC. This nonselective cation channel (NSC) is present at almost the same frequency as ENaC (highly-selective cation channel, HSC), is non-selective for Na+ over K+, has a larger conductance, and shorter mean open and closed times. HSC are made of ENaC subunits, while the composition of NSC is unknown. It appears that NSC contain α-ENaC because α-ENaC knockdown decreases NSC frequency. We hypothesize that NSC consist of both α-ENaC and ASIC1a protein subunits. ASIC1a, another member of the ENaC/degenerin protein family, is strongly expressed in alveolar cells. We manipulated ASIC1a and α-ENaC presence in heterologous expression systems and alveolar cells and assessed the effect on HSC and NSC. shRNA knockdown of ASIC1 in a lung cell line reduces NSC, but not HSC frequency while α-ENaC silencing reduced HSC and NSC frequency. Mice instilled with ASIC1a shRNA silencing vectors also displayed decreased NSC. ASIC1a/α-ENaC coimmunoprecipitate from lung cell lysates and are closely associated in mammalian two-hybrid assays. We conclude that NSC are ASIC1a/α-ENaC hybrid channels that help maintain AFC. However, the striking differences between NSC and HSC regulation may require a new paradigm to explain AFC regulation.

Summer 2016 SURE Research Symposium Phong Tran The Association Between Dietary Sulfur Amino Acid Intake and Vascular Function Phong H. Tran Erika L. Betterman (BS) Arshed A. Quyyumi (MD) Dean P. Jones (PhD) Thomas R. Ziegler (MD) Jessica A. Alvarez (PhD, RD)

Cysteine and its oxidized form, cystine, along with methionine, are sulfur amino acids (SAAs) that form the major extracellular thiol/disulfide system in human plasma. Imbalances in the thiol/disulfide redox systems are associated with oxidative stress, and has been linked to cardiovascular disease (CVD) and cardiac arterial disease (CAD). Previous research has found positive associations between plasma cystine levels with CVD risk factors. Additionally, studies have found associations between dietary intake of methionine and CVD. Considering that Americans habitually consume diets high in SAAs, and that dietary SAAs are known to influence plasma levels of those amino acids, the purpose of our study was to investigate whether dietary SAA intake was associated with worsened markers of vascular health. Dietary intake of SAAs and measures for pulse wave velocity (PWV), augmentation index (AIX), flow mediated dilation (FMD), lipids, and cholesterols were obtained for 435 subjects from the Emory/Georgia Tech Predictive Health Initiative. Initial Pearson correlations suggested significant (p < 0.05) correlations between dietary reduced GSH and PWV (r = 0.07), dietary cystine and systolic BP (r = 0.11), and dietary cystine and FMD (r = -0.10). However, after using a multiple regression model to control for age, gender, BMI, smoking status, diabetes status, race, and history of heart disease, the correlation between dietary SAAs and markers of vascular health were not significant. Future studies examining the impact of dietary SAAs in individuals with higher CVD risk may be warranted in this cohort.

Michelle Tsai Human Osteopontin Isoforms Differentially Promote Vascular Smooth Muscle Cell Migration Michelle Z. Tsai, Emory University, Sarah Harirforoosh, B.S. Emory University, Grace Sanghee Lee, PhD University of Kentucky, Courtney M. Caroti, B.S. Emory University, Hector Salazar, B.S. University of Georgia, Alicia N. Lyle, PhD Emory University

Obstructive arterial disease is common to most cardiovascular disease (CVD) pathologies and often leads to ischemia. The bodyâ&#x20AC;&#x2122;s normal response to ischemia is to increase the development of functional collaterals to restore blood flow and oxygen to damaged tissues. Functional collaterals are formed through processes such as vasculogenesis, angiogenesis, and arteriogenesis, and involve the proliferation and migration of multiple cell types including vascular smooth muscle cells (VSMCs). Osteopontin (OPN), an extracellular inflammatory protein, is known to be necessary for collateralization and VSMC migration. In humans, OPN exists as three isoforms: OPNa, OPNb, OPNc, generated through the alternative splicing of a single mRNA transcript. Preliminary data from the Lyle lab demonstrate that OPN is necessary for VSMC migration and that isoforms differentially affect VSMC migration; however, exactly how the OPN isoforms influence the migration process remains poorly defined. We hypothesize that the OPN isoforms differentially influence the development of filopodia by differentially associating with their downstream receptors, and thus, have divergent effects on FAK activation. To identify the specific roles of the isoforms in VSMC migration, we utilized modified Boyden Chamber Assays to study VSMC migration and western blots to assess FAK phosphorylation. By understanding how human OPN isoforms exert divergent effects on VSMC migration and collateral formation, we may be able to target specific isoforms to modulate new vessel formation in patients with obstructive arterial disease.

Summer 2016 SURE Research Symposium

Christopher Tseng Novel, Low Bias Consensus Clustering Method for Analyzing the Dynamics of SICAvar Gene Expression in Plasmodium knowlesi Malaria Infections in silico Christopher Tseng, Jung-Ting Chien & Stacey Lapp, Emory Vaccine Center, Mary Galinski, PhD Emory Vaccine Center, Emory University School of Medicine

During the course of the malarial parasitic invasion, distinctive variations of antigens are made by the parasite and are presented on the surface of infected red blood cells (RBCs). These antigens allow infected RBCs to avoid detection by the host immune system, making them an important aspect of malaria’s pathogenesis and a potential target for vaccine development. In that context, this project focuses specifically on schizont-infected cell agglutination (SICA) antigens, variably expressed proteins encoded by the SICAvar gene family in the malaria parasite Plasmodium knowlesi. Using transcriptomic data collected from RBCs infected by P. knowlesi, a normalized time course of gene expression during P. knowlesi’s 24 hour life cycle was constructed utilizing a pipeline of RNA sequencing and bioinformatics approaches. Clustering methods were then applied to group together P. knowlesi genes that share similar expression dynamics, with a consensus of the results revealing 1714 genes classified together with SICAvar, indicating likely co-expression. By studying the cellular mechanisms and pathways of these genes that may play a role in regulating SICAvar, we can better understand the factors that contribute to antigen variability and malaria’s virulence.

Pooja Venkatesh Effects of Dual Treatment with SDF-1 and AMD3100 with BMP-2 on Osteogenic Differentiation Pooja Venkatesh, Jean Ndong, Ph.D Emory University, Manvinder Kumar, Yvonne Stephenson, Ivory Williams, Steven Goudy, M.D Emory University

Increasing studies suggest stromal derived factor-1 (SDF-1), a chemokine signaling molecule, plays important roles in bone formation via differentiation of progenitor cells, recruitment, and proliferation. The only current therapy for craniofacial bone defects is surgical correction, a highly invasive therapy with a relatively high complication rate. The use of signaling molecules to potentially allow patient bone to repair its own defects can avoid the risks involved with surgery.

Previously conducted studies suggest SDF-1 has great potential to induce bone regeneration and is the focus of this lab. A secondary focus is AMD3100, a drug shown previously to inhibit osteoblast differentiation. This study provides evidence that treatment with SDF-1 alone, induces a slight increase in osteoblast differentiation while dual treatment with BMP-2 inhibits differentiation.

Treatment with AMD3100 inhibits new bone formation and the drug's inhibitory effects cannot be rescued by growth factor BMP-2.

Summer 2016 SURE Research Symposium Alexander Warren Investigations of Rhodium Pi-Allyl Complexes on Disubstituted Olefins Alexander Warren, Jacob Burman, Simon Blakey

The synthetic community is still in need of many methodologies that can bring together cheap feedstock chemicals to construct C-C and C-N bonds in a controlled manner for drug discovery and pharmaceutical process synthesis purposes. My work with the Blakey group focused on assisting my mentor Jacob Burman with C-C and C-N bond formation using C-H Functionalization as the core methodology. The synthesis of complex molecules usually requires several oxidation and reduction reactions, decreasing the yield of the final product. With C-H Functionalization methods, however, chemist can shorten this process by activating C-H bonds to produce new C-C or C-N bonds. Our research focuses on cationic rhodium(III) catalysts that form Ď&#x20AC;-allyl complexes via activation of inert sp3 C-H bonds to produce new C-C and C-N bonds. Successful isolation and characterization of a novel Ď&#x20AC;-allyl complex on 1,2-disubstituted olefins shows that electron deficient derivatives of RhCp* can be used to form an isolatable metal-complex. Because of this ability to be isolated, these Ď&#x20AC;-allyl complexes can then be used stoichiometrically with the desired nucleophile whether carbon or nitrogen based, to produce new C-C or C-N bonds. Running competition experiments for the complexation using RhCp* and [RhCpECl2]2 and monitoring the kinetics of the reactions has also determined RhCp* is not sufficiently cationic to sustain as an isolatable intermediate. Along with determining a more cationic Rh catalyst than RhCp* such as [RhCpECl2]2 is required to stabilize complex enough for isolation, the kinetics have been determined to identify the reaction requires ~24hrs to reach equilibrium.

Ashley Washington Electrically-Induced Changes in Cardiac Progenitor Cells Ashley Washington, Brown University, Eric Santana, Druid Hills High School, Michael Davis, PhD Emory University, Joshua Maxwell, PhD Emory University

The emergence of cardiovascular regenerative medicine as a potential therapeutic strategy for pediatric heart failure has provided new directions for treatment with a focus on replacing/regenerating the damaged myocardium in order to restore normal cardiac function. Stem cell based therapies have shown beneficial effects on several cardiovascular diseases in adults, and cardiac progenitor cells (CPCs), a progenitor cell found in the myocardium, have been met with early success in clinical trial. Although it has primarily been tried in adults, stem cell therapy is relatively untested in the pediatric population. Because of this, it is crucial to develop new methods of exploring the regenerative potential of these cells in order to improve therapeutic interventions in the pediatric population. It has been shown that regenerative potential of adult CPCs can be enhanced by ex vivo manipulation by genetic engineering or exposure to environmental, chemical, and biological treatments prior to delivery. Previous studies and reports have shown that electrical stimulation of adult cardiac adipose tissue-derived progenitor cells can cause changes to the phenotype and genetic machinery of cells. This allows them to become more suitable for cardiac regenerative approaches. This treatment works via activation of calcium (Ca2+) signaling in the adult cells. The central hypothesis of this proposal is that ex vivo activation of Ca2+ signaling via electrical stimulation will enhance the expression of cardio-specific genes in cardiac progenitor cells.

Summer 2016 SURE Research Symposium Leah Weingast Assessment of Olfactory-Based Retention of Acquired Fear in Healthy Humans Leah Weingast,Atlanta Veterans Affairs Medical Center,Emory University School of Medicine, Maria Briscione, Atlanta Veterans Affairs Medical Center,Emory University School of Medicine, Kristie Garza, Emory University School of Medicine, Yerkes National Primate Research Center, Vicki Wing,Emory University School of Medicine, Filomene Morrison, Emory University School of Medicine, Yerkes National Primate Research Center, Brian Dias,PhD, Emory University School of Medicine,Yerkes National Primate Research Center, PhD, Tanja Jovanovic, PhD, Emory University School of Medicine, Seth Norrholm, PhD, Atlanta Veterans Affairs Medical Center, Emory University School of Medicine

Abstract withheld.

Lagena Williams Change in the Number of Cholinergic Interneurons in a Mouse Model of Dopamine-Responsive Dystonia: Is Age a Factor?

Lagena Williams: Yerkes National Primate Research Center, Udall Ctr of Excellence for PD. Gul Yalcin-Cakmakli: Yerkes National Primate Research Center, Udall Ctr of Excellence for PD Sam J. Rose: Duke Univ Durham, NC Ellen J. Hess: Department of Neurology, Department of Pharmacology, Emory University Atlanta, GA Yoland Smith: Yerkes National Primate Research Center, Udall Ctr of Excellence for PD, Department of Neurology.

Abstract withheld.

Qianru Wu Comparison of Indian Air pollutant Emissions Inventories at National and Regional Levels Eri Saikawa, Assistant Professor, in the Department of Environmental Sciences , Emory University Zhong Min, postdoctoral fellow in the Department of Environmental Sciences , Emory University

Air pollution in India has negative impact on local residents' health. Being able to obtain accurate and precise estimation and simulation is essential in order to fully understand the situation of Indian air pollution. There are many Indian air emission inventories, and the discrepancies exist among them. Some studies have already compared the Indian air emission inventories, but most of them are at national level, not the regional level. The main objective of this project is to compare different Indian air emission inventories at both national and regional levels.

Summer 2016 SURE Research Symposium

Junyan Xia Investigation of the influence of endosymbiont Burkholderia on squash bugs' (Anasa tristis) response towards foreign bacteria Junyan Xia, Tarik Acevedo, Nicole Gerardo

Symbiotic microbes can have crucial influences on host survival and development, such as facilitating growth and providing nutrients. Symbiotic microbes might also affect a host’s resistance towards foreign bacteria and a host’s ability to vector pathogen if the host is a vector. Squash bug (Anasa tristis) is a vector of Serratia marcescens, the plant pathogen that causes cucurbit yellow vine disease (CYVD) . It is of particular interest to study symbiosis in A. tristis given its potential agricultural application. In this project, we will investigate how the presence or absence of the Burkholderia endosymbiont influences A. tristis’ ability to clear out foreign symbiotic Burkholderia and pathogenic S. marcescens after being infected. The result shows that symbiont positive A. tristis has significantly stronger bacteria clearance than symbiont negative A. tristis after being injected S. marcescens ZO1. The presence of symbiont Burkholderia SQ4A might cause A. tristis to exhibit stronger immune response towards foreign bacteria. No significant difference of bacteria clearance between symbiont positive and symbiont negative A. tristis was observed when A. tristis were injected symbiotic bacteria Burkholderia SQ4A. It is possible that A. tristis’ immune system has adapted to the presence of symbiont Burkholderia SQ4A, and thus a subsequent introduction of the symbiont doesn’t elicit a strong response. More replicates, however, are needed to confirm the results.

Siqi Xue Pressure Distribution in Models of Arterial Stenosis with Steady Flow Using Blood Flow Simulations Siqi Xue; Boyi Yang, PhD; Alessandro Veneziani, PhD Department of Mathematics and Computer Science, Emory University, Atlanta, GA

Partial occlusion of vessels - due to lipid deposit or any other reasons - is one of the main causes of many arterial diseases.In this research, arterial stenosis was studied using mathematical models and computational fluid dynamics (CFD) simulations. Six geometries of vessel segments was built with different stenosis symmetry (axisymmetric and non-symmetric), percentages stenosis, and lengths of stenosis, each was applied to CFD simulations with boundary conditions of steady inflows. The blood pressure distribution along the walls and the pressure drop data were obtained for Reynolds numbers varying from 200 to 3000, and compared. Results showed that the geometry of stenosis affects the pattern of blood pressure drop.

Summer 2016 SURE Research Symposium

Sarah Ye Longitudinal Assessment in Means to Evaluate the Efficacy of Neural Progenitor Cell Therapy on Huntingtonâ&#x20AC;&#x2122;s Disease Mouse Sarah Ye, Emory University, In Ki Cho, PhD Emory University, Emory School of Medicine, Yerkes National Primate Research Center, Carissa Hunter, Emory University, Yerkes National Primate Research Center, Jasmine Hope, Yerkes National Primate Research Center, Anthony Chan, PhD Emory University, Emory School of Medicine, Yerkes National Primate Research Center

Huntingtonâ&#x20AC;&#x2122;s disease (HD) is an autosomal dominant neurodegenerative disease characterized by neuronal dysfunction and cell loss in the striatum. HD is caused by expanded CAG trinucleotide repeats in exon 1 of the Huntingtin (HTT) gene1. Recent developments in stem cell research and regenerative medicine has made cell replacement therapy a viable long-term treatment of HD2.

Induced pluripotent stem cells (iPSCs) can be differentiated to lineage specific progenitor cells.

Progenitor cells have a lower chance of developing teratoma after transplantation and are easier to maintain and expand in cell culture2. Neuroprogenitor cells (NPCs) have the capability to differentiate into neuronal and glial cells and have a potential to replace the lost cell population due to the disease progression. However, one caveat is that iPSC and NPC established from the patient sample carries the same genetic mutation. One alternative is to use gene therapy to either alter the mutation or suppress the gene expression of the mutated gene2. Cell replacement therapy coupled with gene therapy using small hairpin RNA (shRNA) to suppress the expression of mHTT is a promising treatment option for HD 2. Our lab has established NPCs derived from iPSCs from wildtype (WT) (WT-NPCs) and transgenic HD monkeys (HD-NPCs)3. Both cells express GFP (green fluorescent protein) and HD-NPCs express mHTT3. In this study, we investigated if the grafted NPCs in WT and HD mice are able to survive and expand in vivo and if shRNA targeting the HTT (shHTT) can promote the cell survival.

Summer 2016 SURE Research Symposium

Kara Ye ESRD Patients Have Altered Pattern of Heart Rate Variability Responses to Graded Lower Body Negative Pressure Kara Ye (1,2), Ryan M. Downey Ph.D (1,2), Peizhou Liao(3), Dana R. DaCosta B.S.(1,2), and Jeanie Park M.D.(1,2) 1. Renal Division, Department of Medicine, Emory University School of Medicine, Atlanta GA 2. Research Service Line, Department of Veterans Affairs Medical Center, Decatur GA 3.

Department of Biostatistics and Bioinformatics, Rollins School of Public Health, Emory University, Atlanta, GA

End stage renal disease (ESRD) patients have autonomic neuropathy that contributes to increased cardiovascular (CV) risk. Normal autonomic response to orthostatic stress is to increase sympathetic nervous system (SNS) and decrease parasympathetic nervous system (PNS) activty in order to maintain arterial blood pressure. Prior studies have shown that healthy humans have decreases in heart rate variability (HRV) in response to increased orthostatic stress induced by graded lower body negative pressure (LBNP) reflective of increased SNS and decreased PNS activation to the heart. Given that ESRD patients have autonomic neuropathy, we hypothesized that ESRD patients have lower HRV at baseline, and an inability to adjust SNS and PNS activity in response to orthostatic stress. We measured continuous beat-to-beat blood pressure and electrocardiogram (ECG) at baseline and during increasing doses of LBNP in ESRD and healthy subjects. HRV was quantified as NN interval and SDNN, reflective of total autonomic activity, and RMSSD and pNN50, reflective of PNS activity. We observed baseline SDNN (p=0.001), RMSSD (p=0.004), and pNN50 (p=0.016) were significantly lower in ESRD patients compared to controls, suggestive of chronically high SNS and low PNS activity. The slopes of change in RMSSD and pNN50 with increasing LBNP were significantly different between ESRD and controls (p=0.004 and p<0.001, respectively). Whereas controls exhibited gradual decreases in RMSSD and pNN50 with increasing LBNP, these values remained constant in ESRD patients. This suggests ESRD patients are unable to adjust PNS activity in response to orthostatic stress which may contribute to orthostatic intolerance and increased CV risk.

Summer 2016 SURE Research Symposium

Jonathan Zawadzki Mechanism of tetrameric transmembrane influenza A/M2 proton channel activation Jonathan Zawadzki, Ban-Seok Jeong, R. Brian Dyer

The Influenza virus is ubiquitous and infection is often fatal. In order to prevent a flu pandemic, four FDA approved drugs have been developed. Of those four, two target the viral M2 proton channel. Unfortunately, influenza has developed resistance to the drugs. Understanding the M2 proton channel will pave way for the next generation of effective drugs. As a result, M2 proton channel’s structure and function have been intensively studied. In this study, to confirm the suggested model of M2 proton channel activation, Forster Resonance Energy Transfer (FRET) is used to measure the change in distance of the C-termini of the M2 proton channel in different pHs. In FRET there is a non-radiative energy transfer between a donor and an acceptor, which is highly sensitive to the distance between them. As the M2 proton channel is activated, the distance between the C-termini should increase and therefore the energy transfer should increase as well.

The primary objective of this experiment is to measure the distance between the C-termini of the M2 proton channel as pH changes (pH 6,7,8). As a secondary aim, the distance will be measured after the M2 proton channels are saturated by amantadine. Amantadine is a drug used to inhibit influenza by the blockage of the M2 proton channel. Understanding the mechanism by which the structure of M2 proton channel changes under various conditions may help develop new treatments.

Xiancong Zhang Role of Vascular Dysfunction in Alzheimer’s Disease Xiancong Zhang, Emory University, Roy L. Sutliff, Ph.D Emory University

Alzheimer’s disease is characterized by a state of reduced cerebral blood flow, which may partly be secondary to cerebral artery vascular dysfunction. Recent studies show that vascular dysfunction may precede Alzheimer’s disease. However, the role of vascular dysfunction in the cognitive decline has not been well studied. Two models were evaluated to examine the effects of AD on vascular function. The first model characterizes the effects of brain homogenates from patients with or without tauopathies on vascular function using isolated rat aorta segments incubated with homogenates following maximal relaxation. The second model determines if vascular dysfunction precedes cognitive defects using a rat Alzheimer’s disease model (Tg F344AD) by studying vascular function at 12 months and 24 months of age. Aortas were isometrically mounted and vascular function was assessed by examining contractile responses to KCl and phenylephrine and relaxation responses to methacholine and sodium nitroprusside. Preliminary data from the wild type rats suggest that the tauopathies negatively affect vascular relaxation. Preliminary data from 1-year-old and 2-year-old rats suggests that the aortas from older AD rats generate less force in response to phenylephrine, compared to wild type rats and that the older AD rats develop endothelial dysfunction. These results help to define the relationship between AD and vascular dysfunction and provide unique models to further understand AD-associated vascular disease.